Book of Abstracts

BOOK OF
ABSTRACTS
PRESENTATION
The Brazilian Meeting on Analytical Chemistry (20th ENQA) returns to Rio Grande do
Sul state, where a previous edition (10 th ENQA) was organized in the Federal University of
Santa Maria (UFSM), in 1999. This is the first edition of ENQA after the beginning of Covid-19
pandemic. In addition to the inherent difficulties of organizing any presential meeting, we still
face challenges related to this pandemic. Nobody imagined in 1999 that in this century we
would be facing anti-vaccine and anti-science movements. However, despite all the losses and
adversities, we have much to celebrate because good science proved again that it is the only
safe way to manage such problems. In this sense, analytical chemistry has provided many
contributions to help in the management of this pandemic as well as to the science
development.
The 20th edition of ENQA will be hosted together with the 8th Ibero-American
Congress of Analytical Chemistry (CIAQA). It is worth noting that the historical and cultural
affinity among members of the analytical chemistry communities of Ibero-America is important
to promote the integration among researchers from these regions.
Therefore, the 20th ENQA/8th CIAQA aims to provide a discussion forum on the
advances in analytical chemistry through conferences, workshops, oral sessions, technical
lectures, short courses, presentations of posters and company exhibitions. From
September 25 to 28th, 2022, the scientific advances in analytical chemistry will be presented
by national and international conference attendees and speakers, contributing to the update of
knowledge in this area. 20th ENQA/8th CIAQA will also be fostering direct contact among
participants and the leading companies active in the field of analytical chemistry.
In addition to the scientific program, the city of Bento Gonçalves offers many tourist
attractions and excellent hospitality and gastronomical infrastructure.
Welcome to 20th ENQA/8th CIAQA.
Érico Marlon de Moraes Flores

Chair of the 20th ENQA/8th CIAQA
3
–––––––––––––––––––– 20th ENQA and 8th CIAQA September 25 to 28th, 2022 ––––––––––––––––––––
TABLE OF CONTENTS
PROGRAM………………………………………………………………………………………… 5
ELECTROANALYTICAL ........................................................................................................ 6
ATOMIC SPECTROMETRY .............................................................................................. 96
MASS SPECTROMETRY .................................................................................................. 127
INSTRUMENTATION AND AUTOMATION ..................................................................... 161
SEPARATION METHODS ............................................................................................... 193
SAMPLE PREPARATION .................................................................................................. 214
ENVIRONMENTAL CHEMISTRY ................................................................................ 31212
ANALYTICAL CHEMISTRY & ENERGY ......................................................................... 389
CHEMOMETRICS ........................................................................................................... 414
MISCELLANEOUS .......................................................................................................... 473
4
PROGRAM
5
ELETROANALYTICAL
6
RAPID AND SELECTIVE ELECTROCHEMICAL SCREENING METHOD FOR
MEPHEDRONE DETECTION IN FORENSIC SAMPLES USING A GRAPHENE
SCREEN-PRINTED ELECTRODE
Larissa M. A. Melo (PG)a*, Lívia M. S. Aranha (UG)b, Luciano C. Arantes (R)c, Pablo A.
Marinho (R)d, Camila D. Lima (R)a, Izabela F. Schaffel (PG)e, Nathália S. Conceição (PG)e,
Rafael Q. Ferreira (R)e, Wallans T. P. dos Santos (R)a,b
a Universidade Federal dos Vales do Jequitinhonha e Mucuri, Departamento de Química, Diamantina, Minas Gerais,
Brazil, 39100-000
b Universidade Federal dos Vales do Jequitinhonha e Mucuri, Departamento de Farmácia, Diamantina, Minas Gerais,
Brazil, 39100-000,
c Instituto de Criminalística, Polícia Civil do Distrito Federal, Seção de Perícias e Análises Laboratoriais, Brasília, Distrito
Federal, Brazil, 70610-200

d Instituto de Criminalística, Polícia Civil de Minas Gerais, Belo Horizonte, MG, Brazil, 30180-060,
e Universidade Federal do Espírito Santo, Departamento de Química, Vitória - ES, Brazil, 29075910
*e-mail: larissamamelo@gmail.com
Mephedrone is an illicit stimulant drug used for recreational purposes, also known as 4-
methylmethcathinone (4-MMC) of the synthetic cathinones (SC) class, which have widely been
reported in forensic analysis. The preliminary identification of 4-MMC and other SC in seized
samples is of great interest for forensic investigation in order to perform a fast and simple screening
test of these drugs. In this context, we present the electrochemical detection of 4-MMC in forensic
samples using, for the first time, two redox processes of this SC on screen-printed electrodes (SPE)
based on working electrodes of graphite (SPE-Gr) and graphene (SPE-GP). The electrochemical
behavior of 4-MMC at the SPEs presented one oxidation process and one reduction process, which
are independent and irreversible, both with mass transport controlled by adsorption and diffusion.
The 4-MMC detection at SPEs was optimized in Britton-Robinson buffer solution (0.1 mol L−1) pH
10.0 with adsorptive stripping differential pulse voltammetry (AdSDPV). The optimal conditions for
4-MMC determination were obtained with amplitude of 90 mV, modulation time of 50ms, step
potential of 10 mV, time interval of 0.5s, and accumulation time of 1min. In order to evaluate the
sensitivity of the electrochemical detection of 4-MMC using AdSDPV, SPE-Gr and SPE-GP were
compared. The SPE-GP presented anodic (jpa) and cathodic (jpc) peak current densities
approximately 4-fold and 8-fold higher than the jpa and jpc of the SPE-Gr, respectively. The higher
sensitivity obtained for 4-MMC detection with SPE-GP is due to the larger real surface area available
for adsorption of this analyte on graphene working electrode than graphite. Thereby, the SPE-GP
was chosen to perform the 4-MMC electrochemical screening in forensic samples. The proposed
method presented a wide linear range for oxidation (2.6 to 56 µmol L−1) and reduction (6.9 to 112
µmol L−1) processes, which provided limits of detection (LODs) of 1.9 µmol L−1 and 5.5 µmol L−1,
respectively. The obtained LODs by the proposed method are low enough for application in seized
forensic samples containing the 4-MMC. Furthermore, a good stability of the 4-MMC electrochemical
responses at the SPE-GP was obtained using the same or different electrodes (N = 3), with RSD <
5.0% for both redox processes. Interference studies for a common adulterant (caffeine) and thirteen
other illicit drugs (phenylethylamines, amphetamines, and other SC) demonstrated that the proposed
method is highly selective for electrochemical screening of 4-MMC in forensic analysis. In addition,
seized samples were identified with 4-MMC by the proposed method (preliminary test) and confirmed
by LC-MS and CG-MS (definitive methods). Addition-recovery studies of 4-MMC in these samples
were also performed by the proposed method, with recoveries around 100% using both
electrochemical processes for detection of this drug. Therefore, the analytical performance of SPE-
GP with AdSDPV technique for 4-MMC detection demonstrates a selective and sensitive screening
method for application in forensic analysis, providing an on-site analysis with a fast and simple
preliminary identification of this SC in seized samples.
[UFVJM, CNPq, CAPES, FAPEMIG]
––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

7
DEVELOPMENT OF A IMPEDIMETRIC SENSOR BASED ON Pi-CONJUGATED
POLYMER AND GOLD NANOPARTICLE FOR DETECTION OF DISSOLVED
OXYGEN
Miquéias L Portugal (UG)a, André O. Oliveira (R)a, Marcos F.S. Teixeira (R)a*
a São Paulo State University, Department of Chemistry and Biochemistry, School of Science and Technology Presidente
Prudente, São Paulo, Brazil, 19060-900
*e-mail: marcos.fs.teixeira@unesp.br
The π-conjugated polymers are used as transducer support in the development of chemical sensors
due to their low cost, flexibility, conductivity and presence of active redox sites in their structure. An
interesting strategy in device preparation is the formation of metal and polymer nanocomposites.
This strategy can aid in the electrochemical signal due to the increase in active surface, thus
improving detection sensitivity. In the present study a new sensory platform using a nanocomposite
with poly(1,2-phenylenediamine) (poly(o-PD)) and gold nanoparticles (AuNP) was studied for
impedimetric detection of dissolved oxygen. The poly(o-PD)-AuNP nanomaterial on the FTO surface
was prepared by a single-step technique1 by potential cycling in a deaerated solution containing 2.00
mmol L-1 1,2-phenyldiamine and 1.00 mmol L-1 HAuCl4 in 0.10 mol L-1 H2SO4 medium. By analyzing
the voltammetric response of the sensor in the presence of molecular oxygen in electrolytic solution,
it was possible to observe a decrease in the reduction overpotential of oxygen, followed by a
significant increase in the cathodic current of the system. The performance of the poly(o-PD)-AuNP
film as a chemiresistor material for the sensing of molecular oxygen was evaluated by impedance
measurements at the applied potential of -0.30 V in PBS (7.4). The charge transfer resistance values
change substantially in the presence of oxygen in solution. The effect of oxygen on the resistivity of
the device is attributed to changes in the oxidation state of the polymer. The impedance
measurements in real time (chrono-impedance) with the poly(o-PD)-AuNP sensor with different
oxygen concentrations revealed good linearity behavior between the charge transfer resistance and
the molecular oxygen concentration (0.02-5.56 mmol L-1). The new O2 detection platform will be used
to monitor gradients in static cultures of adherent cells exposed to graded oxygen both at rest and
after metabolic stimulation.
poly(o-PD)-AuNP poly(o-PD@AuNP
2,0 4,2
1,8 (a) (b) FIT

r = - 0,99
1,6 2
r = 0,98
1,4 -1
0.02 mmol L 4,0
1,2
-2
log Rredox ()
1,0
Z'' /k cm
0,8
[O2]
3,8
0,6
0,4
0,2
-1
5.56 mmol L
3,6
0,0 [O2]
-1 -1
0.02 mmol L 5.56 mmol L
0,0 0,2 0,4 0,6 0,8 1,0 1,2 1,4 1,6 1,8 2,0
-2
Z' /k cm -4,5 -4,0 -3,5 -3,0 -2,5 -2,0
log [O2]
1 Olean-Oliveira A et al., ACS Sensors, 4, 2019, 118.

8
A NOVEL ELECTROCHEMICAL SENSOR BASED MOLECULARLY IMPRINTED
POLYMER FOR THE SENSITIVE DETECTION OF CAFFEINE
Natalia M. Silva (UG)a, Heitor F. Trevizan (PG) a, André Olean-Oliveira (PG)a,
Marcos F. S. Teixeira (R)a*
a São Paulo State University, Department of Chemistry and Biochemistry, School of Science and Technology Presidente
Prudente, São Paulo, Brazil, 19060-900
Cocaine is the most consumed drug in the world. Commonly trafficked in powder form, cocaine
receives adulterants such as caffeine, xylocaine, and benzocaine. The cocaine dilution carried out
by the trafficker has as main objective to obtain an increase in the volume of the drug and
consequently a greater profit on it. To control this serious global problem, an accurate and easy-to-
use sensory platform is needed to identify and quantify, helping to map the origin of the drug. In this
work, a new electrochemical sensor was developed based on molecular imprinting technology for
caffeine detection. The formation of the molecularly imprinted polymer (MIP) [1] was carried out by
electropolymerization of Bismarck Brown Y monomer and caffeine as molecular template in HCl
solution.
Figure 1. A) DPV response of the MIP sensor in aqueous solution of KCl 0.5 mol L-1 (pH=2.0) for different caffeine
concentrations. Pulse amplitude = 25mV. B) Analytical curve of the caffeine MIP sensor.
The electrochemical behavior of the sensor at different concentrations of caffeine was verified by
differential pulse voltammetry (DPV) applied a potential range of -0.3 to 0.8V vs. SCE. From Figure
A, we can observe the increase in the anodic current of the sensor with the variation of the caffeine
concentration in electrolyte solution. The current increment and the displacement of the oxidation
potential are indicative of the caffeine interaction with the specific cavities generated in the imprinted
polymer. The MIP sensor showed a linear response relationship (Fig. B) between 0.05 to 0.5 µmol
L-1 of caffeine (Ip (µA cm-2) = 0.631 + 11.4 [Caffeine] (µmol L-1), r = 0.9969) with detection limit of
0.02 mol L-1. The MIP electrochemical sensor showed promising analytical performance for caffeine
detection.
1 Trevizan HF, Olean-Oliveira A, Cardoso CX, Teixeira MF, Sensors and Actuators B: Chemical, 2021, 130141.

9
A HIGHLY SENSITIVE CAPACITIVE ALBUMIN SENSOR BASED ON
MOLECULARLY IMPRINTED POLYMER
Heitor F. Trevizan (PG)a, André Olean-Oliveira (PG)a, Patrícia Monteiro Seraphim (R)a,
Marcos F. S Texeira (R)a*
a São Paulo State University (UNESP), Faculty of Science and Technology, Pres. Prudente, SP, Brazil, 19060-900
Albumin is an important protein in the body and plays a critical role in the maintenance of nutrition
balance, plasma pressure, and transportation of different substances. Moreover, the change of
albumin concentration in the blood or tissue fluids can be used as a biomarker in human health and
disease1. For example, the low concentrations of albumin in blood may suggest changes in the
immune system, caused by viral infections such as human immunodeficiency (HIV) and Coronavirus
(COVID-19)2,3. In this study, a novel molecularly imprinted polymer (MIP) sensor based on poly(azo-
BBY) for the electrochemical detection of albumin is reported. The poly(azo-BBY) MIP film on
electrode fluorine tin oxide (FTO) was prepared by electropolymerization of Bismarck Brown Y in the
presence of albumin as target molecule. The electrochemical sensing properties were investigated
by electrochemical impedance/capacitance spectroscopy to confirm the binding sites and verify the
electroanalytical behavior of the sensor.
(A) (B)
-1 -1
3.0 ng mL 0 ng mL
1.35
1.38
10
C'' / F cm-2
1.30
1.25
8 1.32
C'' / F cm-2
1.20
C''/ F cm-2
6
11.00 11.05 11.10 11.15 1.26
-2
C' / F cm
4
1.20
2
1.14
2 4 6 8 10 0.0 0.5 1.0 1.5 2.0 2.5 3.0

-2
C' / F cm -1
Albumin (ng mL )
Figure 1. (A) Complex capacitance spectra (Nyquist capacitive) of the poly(azo-BBY)-MIP sensor in different
concentrations of albumin at +0.05 V vs SCE in 0.05 mol L-1 PBS (pH=7.3) over the N2 atm. (B) Analytical curves of the
imaginary capacitance values for different albumin concentrations obtained at 0.10 Hz. N = 3.
Analyzing the imaginary capacitance as a function of the applied frequency, the MIP film capacitance
system decreased to the low frequency region with increasing albumin concentration (Fig. 1A). Due
to the specific cavities for albumin on the large surface of the MIP sensor, the target molecule can
easily be rebinding on these cavities. The device has a linear range of response in the concentration
range of 0.25–1.0 ng mL-1 (Fig. 1B) with good correlation between the imaginary capacitance and
the albumin concentration (r = 0.9979). The prepared sensor shows a sensitivity of 0.156 µF cm2 mL
ng-1 and a limit of detection (LOD) of 0.215 ng mL-1 (S/N = 3).
1 Wang L, Zhao L, Colloids and Surfaces A: Physicochemical and Engineering Aspects, 632, 2022, 127843.
2 Gandhi M, Indiramma J, Jayaprakash NS, Kumar AS, Journal of Electroanalytical Chemistry, 2022, 116018.
3 Meng XM, Li ZR, Zheng XY, Liu YX, Niu WJ, Qiu XY, Lu HZ, European Journal of Pharmaceutical Sciences, 167, 2021,
105986.
[CAPES]

10
STUDY OF PHOTOELECTROCATALITICAL BEHAVIOR OF PALLADIUM-
SALEN/GRAPHENE NANOSTRUCTURES APPLYING IN WATER OXIDATION
Gabriel M. Silva (UG)a, Yuri A. Oliveira (PG)a, Andre Olean-Oliveira (PG)a, Marcos F. S.
Teixeira (R)a*
aSão Paulo State University (UNESP), Faculty of Science and Technology, Pres. Prudente, SP, Brazil, 19060-900
The catalytic oxidation of water is the basis of hydrogen-based fuel cells and one of the challenges
faced in its use is the high energy demand, requiring the use of alternative routes to reduce the
overpotential of activation of the water oxidation reaction (OER). Solar energy can be converted into
other forms of energy, such as electrical, chemical and thermal, using it as an additional energy
source can reduce the overpotential energy for the oxygen evolution reaction, facilitating the process
at the beginning of catalysis1,2.With this objective in mind, electrocatalytic materials have been
studied. The Poly[Pd(Salen)]/GO electrocatalyst has shown promising results for the reduction of
OER activation energy barriers, this is due to the π-conjugated bonds present in the salen ligand
and in graphene oxide (GO), facilitating the passage charge, decreasing the energy needed for the
reaction to take place.2
40
400 A) 0,90 B) No light
6W
Potential Onset (V vs ECS)
0,88 35
350 6.5 W
0,86
9W
0,84
30 16 W
Density current (A cm-2)
300 0,82
30 W
0,80
250 0,78
25
0,76
-Z''/k
200 0,74 20
5 10 15 20 25 30
Potential lamp (W)
150
15
100
10
50
5
0
0
0 5 10 15 20 25 30 35 40
-0.2 0.0 0.2 0.4 0.6 0.8 1.0
Potential (V) vs ECS Z'/k
Figure 1. (A) Linear sweep voltammetry in a 0.1 mol L-1 H2SO4 solution in a deaerated N2 solution, v= 25 mV s-1. with
diferent lamp powers(30w, 16w, 12w, 9w, 6.5w and 6w respectively). Inset: Onset potential varying according to the power
of the lamp. (B) Spectrum in the Nyquist complex plane of the electrode coated with poly[Pd(Salen)] in 0.1 mol L -1 H2SO4
solution and pH 3 in deaerated N2 solution, applied potential of 0.5 V vs ECS.
The study of the photoresponse of the Poly[Pd(Salen)]-GO film and its photoelectrocatalytic behavior
was investigated with the absence and presence of light (dichroic lamps of different powers), by
voltammetry linear sweep technique was possible to verify a displacement of the potential for the left
as the lamp power increases (6w, 6.5w, 9w, 12w, 16w and 30w), thus indicating a reduction in the
starting potential for the OER. Studies using electrochemical impedance spectroscopy (EIE) and
electric circuit model was possible to verify that the film resistance decrease as the lamp power
increases, thus indicating greater ease for the load to pass through the system, this reduction in the
overpotential for the OER, this reduction can be attributed to the structure of the Poly[Pd(Salen)]-
GO film in an alternating system.3
1 Blakemore JD, Crabtree RH, Brudvig GW, Chem. See. 115, 2015, 12974.
2 Oliveira YA, Olean-Oliveira A, Teixeira MFS, Journal of Electroanalytical Chemistry, 880, 2021, 114928.
3 Tang R, Zhou S, Zhang Z, Zheng R, Huang J. Advanced Materials. John Wiley and Sons Inc, 33, 2021, 2005389.
[CNPq]

11
OXYGEN PLASMA-TREATED GRAPHITE SHEET ELECTRODES FOR MDMA
SENSING
Jian Felipe da Silva Pereira (PG)a*, Larissa Magalhães de Almeida Melo (PG)b, Rogério
Valentim Gelamo (R)c, Wallans Torres Pio dos Santos (R)b, Eduardo Mathias Richter (R)a,
Rodrigo Alejandro Abarza Muñoz (R)a
a Universidade Federal de Uberlandia, Instituto de Química, Uberlândia, Minas Gerais, Brazil, 38405140.
b Universidade Federal dos Vales do Jequitinhonha e Mucuri, Departamento de Química, Diamantina, Minas Gerais,
Brazil, 39100000.
c Universidade Federal do Triângulo Mineiro, Instituto de Ciências Tecnológicas e Exatas, Uberaba, Minas Gerais, Brazil,
38064200.
*e-mail: jianfelipe555@gmail.com
Several different synthetic drugs are reported in apprehensions, and among them, 3,4–methylene-
dioxymethamphetamine (MDMA) has gained attention since 2000 due to its psychoactive
characteristics1,2. Therefore, its detection has great importance in the forensic scenario. Amid the
techniques used to determine MDMA, electrochemical methods have emerged because of the
capacity to miniaturize the system, low cost and effective approach to identify this drug, and several
working electrodes (WE) have been proposed.3,4 Graphite sheets (GS) rise among the other
electrodes due to its characteristics, such as flexibility, high conductivity, low cost and low
background currents, and those attributes are enhanced when coupled with plasma surface
treatment.5,6 This work presents the application of GS with O2 plasma treatment to determine MDMA.
SEM images show higher presence of edge planes in the treated surface (GS-O2) when compared
with the untreated, which corroborates with cyclic voltammograms in the presence of 1 mmol L -1 of
[Fe(CN)6]3-/Fe(CN)6]4-, who presents a higher peak current (Ipa = 50.6 µA and Ipc = 50.6 µA for GS-
O2, and Ipa = 25.1 µA and Ipc = 29.3 µA for GS) and a lower ∆Epeak for the GS-O2 surface when
compared with GS (180 mV and 840 mV respectively). In cyclic voltammograms containing 0.5 mmol
L-1 of MDMA it was possible to identify two well defined oxidation peaks and a higher peak current
for GS-O2, demonstrating that this electrode presents a better platform to electroanalytical purposes.
The influence of pH was evaluated using 0.12 mol L-1 Britton-Robinson buffer solutions, and pH 10
was selected to be used for next experiments. Analytical curves were performed using differential
pulse voltammetry (DPV) with the concentration of MDMA varying from 0.5 to 15.0 µmol L -1 using
both GS and GS-O2 as WE (step potential: 9 mV, modulation amplitude: 100 mV, modulation time:
10 mV). It was observed that GS-O2 indeed presented a better performance, and the details of the
comparison is presented in Table 1. The GS-O2 surface presented lower quantification (LQ) and
detection (LD) limits and a wider linear range, which confirms an improved performance for the
treated surface.
Table 1. Performance comparison between GS and GS-O2 electrodes for MDMA detection.
Electrode R2 Slope LD (µmol L-1) LQ (µmol L-1)
Linear range
(µmol L-1)
GS 0.99 3.64 10-8
0.8 2.6 3.0 – 15.0
GS-O2 0.99 1.14 10-7 0.09 0.3 0.5 – 15.0
It was concluded that the O2 plasma surface treatment was able to enhance the graphite sheet
performance to detect MDMA, providing a higher slope, lower LD and LQ and wider linear range.
1 Ministério da Justiça. Polícia Federal. Drogas Sintéticas. Brasília, 2018. Relatório. https://www.gov.br/pf/pt-br/acesso-
a-informacao/acoes-e-programas/relatorio-de-drogas-sinteticas-2018/drogas_sinteticas_2018_eng.pdf
(Accessed July 22, 2022).
2 Christophersen AS, Toxicology Letters, 112-113, 2000, 127.
3 Krotulski AJ, Mohr ALA, Fogarty MF, Logan BK, Journal of Analytical Toxicology, 42, 2018, 544.
4 LR, Smith, JP, Zuway KY, Sutcliffe OB, do Carmo DR, Banks CE, Analytical Methods, 8, 2016, 142.
5 Oliveira AG, Munoz RAA, Angnes L, Electroanalysis, 22, 2010, 1290.
6 Pereira JFS, Borges PHS, Moura GM, Gelamo RV, Nossol E, Canobre SC, Richter EM, Munoz RAA, Electrochemistry
Communications, 105, 2019, 106497.
[FAPEMIG, CAPES, CNPq]

12
DEVELOPMENT OF AN ELECTROCHEMICAL SENSOR BASED ON IMPRINTED
MOLECULARLY AZO-POLYMER FOR CAPACTIVE DETECTION OF CYTOSINE
Tainá B. Rodrigues, Heitor F. Trevizan (PG), André Olean Oliveira (PG), Marcos F. S. Teixeira
(R)*
São Paulo State University (UNESP), School of Science and Technology, Pres. Prudente, São Paulo, Brazil, 19060-900
Cancer is a genetic disease that arises from an alteration in the cell's DNA, causing the disorganized
multiplication of abnormal tissue cells. DNA plays crucial roles in the living body, which contains four
bases (guanine, adenine, thymine and cytosine) that carry the genetic information of disease. Thus,
the detection of qualitative or quantitative alterations of the four nucleobases is of great importance
both for clinical diagnosis and for obtaining genetic information from cells. In the present abstract,
we report the use of an electrochemical sensor for cytosine using a molecularly imprinted polymer
(MIP) as a molecular recognition platform. The MIP film was deposited on a fluorine tin oxide (FTO)
electrode from the electropolymerization of Bismarck Brown Y in the presence of cytosine as a target
molecule. To compare, a non-imprinted polymer (NIP) electrode was constructed identically without
cytosine template. The MIP and NIP were characterized electrochemically by cyclic voltammetry in
KCl 0.50 mol L-1 in the presence and absence of cytosine (Fig. 1).
(A) Molecularly Imprinted Polymer (B) Non-Imprinted Polymer
10
)
-2
Current density ( cm )
-2
Current Density ( cm
10
0
0
-10 -10
Absence of cytosine
Presence of cytosine Absence of cytosine
-20 Presence of cytosine
-20
-0.3 0.0 0.3 0.6 0.9 -0.3 0.0 0.3 0.6 0.9
Potential (E vs ECS) Potential (E vs ECS)
Figure 1. Electrochemical response of MIP (A) and NIP (B) electrode in the absence and presence of 1.0 mmol L -1 cytosine
(pH 6.7) at a scan rate of 50 mV s−1.
By analyzing Fig.1A, a significant increase in the anodic capacitive currents of the MIP electrode
was observed in an electrolyte solution containing cytosine. Comparatively, the electrochemical
response of the NIP electrode (Fig.1B) to cytosine in solution did not show a significant increase in
capacitive current. This result is related to the increase in the porosity of the film and the intrinsic
permeability of cytosine to the redox site of the polymer through the cavity generated by the
molecular imprint. This idea will be reinforced when we apply electrochemical impedance study to
obtain real impedance versus real capacitance graphs1. Analytical curves based on capacitance
measurements are being analyzed in order to verify the sensor performance.
1 Trevizan HF, Oliveira OA, Teixeira MFS. Sensors and Actuators B: Chemical, 343, 2021, 130141.

13
IMPEDIMETRIC PLATFORM FOR FORENSIC IDENTIFICATION OF
ALCOHOLIC BEVERAGE CONTAMINANTS USING PRINCIPAL COMPONENT
ANALYSIS
Vinicius Vessane (UG), Heitor F. Trevizan (PG), André Olean-Oliveira (PG),
Celso X. Cardoso (R), Marcos F. S. Teixeira (R)*
São Paulo State University (UNESP), School of Science and Technology, Pres. Prudente, São Paulo, Brazil, 19060-900
Electrochemical sensors based on electrochemical impedance spectroscopy have gained

prominence in several areas due to their sensitivity for verifying changes in interfacial phenomena.
The present abstract shows a systematic electrochemical impedance study on the electrooxidation
of methanol, ethanol, 1-propanol, ethylene glycol and glycerol by nickel-salen metallopolymer. The
study led to the elaboration of alcohol analysis diagrams based on multivariate analysis statistics
using the physicochemical parameters composed by the electrochemical impedance spectroscopy
(EIS). The EIS measurements were carry out in a solution of 0.10 mol L-1 NaOH solution at an applied
potential of and +0.47 V (vs. SCE). The chosen potential is found immediately after the peak of
oxidation, related to the oxidation process of the metallic center from Ni2+ to Ni3+, responsible for the
oxidation of alcohol molecules in solution. Furthermore, in this potential, the phenomenon of mass
transport will be minimized and that the variations of impedimetric parameters will be directly related
to the phenomenon of alcohol oxidation. The impedimetric parameters (real Z' and imaginary Z''
impedance; real C' and C'' imaginary capacitance, absolute impedance |Z| at 5.4 Hz) obtained with
the sensor were used as variables in the multivariate analysis. From the PCA graph (Fig. 1) can be
clearly seen that the system distinguishes the types of alcohol with impedimetric variables for
different alcohol concentrations.
1-Propanol
2
PC 2 (38.70%)
1 Ethanol
Ethylene Glycol
0
-1
Methanol
-2
Glycerol
-2 -1 0 1 2 3
PC 1 (61.05%)
Figure 1. PCA using five impedance values parameters obtained from fixed frequency of 5.4 Hz for three different alcohol
concentrations (0.123 to 0.476 mmol L-1).
The next classification study was carried out to identify the clustering pattern in alcoholic mixture.
The simulations were performed in solution of ethanol with different ratios of methanol and ethylene
glycol in the proportions of 0%; 0.2%; 1%; 2%; 5%; 10%; 30% and 60%. The PCA results show an
identification distance between the groups allowed the differentiation of pure and mixed samples
from the groups containing ethanol-methanol and ethanol-ethylene glycol.

14
NANODIAMOND BASED ELECTROCHEMICAL SENSOR DEVELOPMENT FOR
PARACETAMOL DETERMINATION
Deborah O. Lopes (UG)a*, Patrícia B. Deroco (R)b,c, Orlando Fatibello Filho(R)d, Geiser G.
Oliveira (R)a
a FederalUniversity of Tocantins, Collegiate of Chemistry and Engineering of Bioprocesses and Biotechnology, Gurupi,
Tocantins, Brazil, 77402-970
b University of Campinas, Institute of Chemistry, Campinas, Sao Paulo, Brazil,13083-970
c Autonomous University of Barcelona, Catalan Institute of Nanoscience and Nanotechnology, Bellaterra, Barcelona,
Spain, 08193.
d Federal University of São Carlos, Chemistry Departament, São Carlos, São Paulo, 13565-905
*e-mail: oliveira.lopes@mail.uft.edu.br
Paracetamol (PAR) is the most used drug in the world with analgesic and antipyretic properties,
which is used to reduce and relieve pain and fever. It is easily found in pharmacies and drugstores,
and self-medication along with ease of purchase contributes to the excessive and abusive use of
this type of medicine, which can cause acute liver failure or even be fatal in some cases1. Thus, the
development of methods for PAR determination and monitoring are important. The use of
electrochemical methods has attracted attention because they provide accurate, sensitive, and low-
cost analyses2. In this work, we report the development and application of a glassy carbon electrode
(GCE) modified with a nanodiamond (ND) film for PAR determination in pharmaceutical formulations.
Initially, the electrochemical behavior PAR was investigated in 0.10 mol L−1 H2SO4 by cyclic
voltammetry (CV) technique at a GCE and ND/GCE at a potential scan rate of 50 mV s −1, in the
presence of 5.0 × 10−4 mol L−1 PAR (Fig. 1), showing that the incorporation of ND on the GCE surface
caused a remarkable increase of peak current and a decrease of the peak potential (from 0.804 V
for GCE to 0.750 V for ND/GCE). Reduction peak was not observed in the reverse potential scan for
both electrodes, characteristic of an irreversible electrochemical process.
16 GCE
ND/GCE
I/ µA
0.2 0.4 0.6 0.8 1.0

(E vs Ag/AgCl)/ V
Figure 1. Cyclic voltammograms obtained for presence of 5.0 × 10 −3 mol L−1 PAR in 0.10 mol L−1 H2SO4 with sweep rate
50 mV s−1.
The effect of the supporting electrolyte on the PAR oxidation peak was investigated, and a 0.10 mol
L−1 sulfuric acid solution provided the best voltametric profile. Finally, the optimization of square-
wave voltammetry (SWV) parameters was studied, selecting a frequency of 80 Hz, a pulse amplitude
of 40 mV and a sweep increment of 6 mV. The analytical curve was linear in the PAR concentration
range from 9.9 x 10−7 to 1.3 x 10−4 mol L−1, represented by the equation Ipa(A) = 0.07 + 0.11 [PAR]
(mol L−1), with a correlation coefficient equal to 0.999 and a detection limit of 1.63 × 10−7 mol L−1.
1 Espinosa BM et al., J. Pharm Biomed Anal, 42, 2006, 291

2 Simioni NB et al., Diamond and Related Materials, 74, 2017, 191
[PIBIC/UFT, FAPT, CNPq, CAPES, FAPESP]

15
RESORCINOL ELECTROCHEMICAL DETERMINATION USING A FULLY 3D
PRINTED APPARATUS
Jocieli Fabri (UG)a, Daniel R. Cocco (PG)a, Rodrigo A. A. Muñoz (R)b,*, Diego P. Rocha (R)a
a Federal Institute of Parana, Pitanga, Paraná, Brazil, 85200-000
b Federal University of Uberlandia, Institute of Chemistry, Uberlândia, Minas Gerais, Brazil, 38400-902
*e-mail: munoz@ufu.br
Resorcinol (RS) is a phenolic compound which is considered an emerging contaminant of soil and
natural waters. RS can cause acute and chronic diseases involving the central nervous,
reproductive, immune, and respiratory systems1. In this regard, RS monitoring is of relevant
environmental and scientific interest. Traditional methods for RS determination are based on liquid
and gas chromatography techniques, which are expensive, require highly specialized operators, and
do not allow for on-site analysis (something very desired nowadays in analytical chemistry)1. In this
sense, electrochemical techniques combined with the use of 3D printing can be a low-cost alternative
to chromatographic techniques. This work aims to use a fully 3D printed manufactured apparatus
(cell and electrodes) for the electrochemical determination of RS. A previous step is required in the
use of 3D printed electrodes composed of polylactic acid and carbon black since in such electrodes
there is a large amount of non-conductive material, making the transfer of electrons difficult with a
consequent poor electrochemical performance. Therefore, a chemical/electrochemical treatment in
an alkaline medium (0.5 mol L-1 NaOH) was used to remove excess of non-conductive thermoplastic
and expose the electroactive sites present on the electrode surface2. By the cyclic voltammetry
technique, it was observed that after treatment the performance of the electrode for the oxidation of
the RS improves significantly, obtaining an anticipation in the oxidation potential as well as a great
increase in the current (Fig. 1). Employing Square-Wave Voltammetry, the proposed method
achieved a linear range from 5 to 400 µmol L-1 with a detection limit of 3.4 µmol L-1. In addition, an
RSD value of 4.0% was obtained for five consecutive analyses of a solution containing 100 µmol L-
1
RS. To show the applicability of the developed method, four different real water samples were
analyzed (tap, river, well, and mine waters) and satisfactory recovery values between 90% and 108%
were acquired. Finally, the method developed allows field analysis, is simple and inexpensive, does
not produce toxic residues, and provides better or comparable results to traditional methods.
Figure 1. Cyclic voltammetric recordings before (black lines) and after (red lines) chemical/electrochemical treatment in
the presence (solid lines) and absence (dashed lines) of 1 mmol L-1 of RS in 0.1 mol L-1 PBS (pH 7.2) (background
electrolyte). Experimental conditions: scan rate of 50 mV s -1 and step potential of 5 mV.
1 Iftikhar T, Asif M, Aziz A, Ashraf G, Jun S, Li G, Liu H, Trends in Environmental Analytical Chemistry, 31, 2021, e00138.
2 Rocha DP, Squissato AL, Silva SM, Richter EM, Munoz RAA, Electrochimica Acta, 335, 2020, 135688.

16
BIOCOMPATIBLE, REPRODUCTIBLE, AND SCALABLE WEARABLE LEAF
SENSORS FOR SENSITIVE AND REMOTE MONITORING OF THE WATER LOSS
FROM PLANTS
Júlia A. Barbosa (PG)a,b*, Vitória M. S. Freitas (R)a, Lourenço H. B. Vidotto (R)c, Gabriel R.
Schleder (R)d, Ricardo A. G. Oliveira (R)a, Jaqueline F. Rocha (R)a,e, Lauro T. Kubota (R)c,
Luis C. V. Silveira (R)a, Hélio C. N. Tolentino (R)f, Itamar T. Neckel (R)f, Angelo L. Gobbi (R)a,
Murilo Santhiago (R)a,e, Renato S. Lima (R)a-c,e
a Brazilian Center for Research in Energy and Materials, Brazilian Nanotechnology Laboratory, Campinas, São Paulo,
Brazil, 13083-970
b University of São Paulo, São Carlos Institute of Chemistry, São Carlos, São Paulo, Brazil, 09210-580
c University of Campinas, Institute of Chemistry, Campinas, São Paulo, Brazil, 13083-970
d Harvard University, John A. Paulson School of Engineering and Applied Sciences, Cambrige, Massachusetts, United
States, 02138
e University of ABC, Santo André, State, Brazil, 09210-580
f Brazilian Center for Research in Energy and Materials, Brazilian Synchrotron Light Laboratory, Campinas, São Paulo,
Brazil, 13083-970
*e-mail: julia.barbosa@lnnano.cnpem.br
Impedimetric wearable sensors are a promising strategy for determining the loss of water contents
(LWC) from leaves as they can afford on-site and nondestructive quantification of cellular water from
a single measurement.1,2 Since the water content is a key marker of leaf health, the monitoring of
LWC can lend key insights into the daily practice in precision agriculture, toxicity studies, and
development of agricultural inputs.1,2 Ongoing challenges with this monitoring are the on-leaf
adhesion, compatibility, scalability, and reproducibility of the electrodes, especially when subjected
to long-term measurements. This paper introduces a set of sensing material, technological, and data
processing solutions that overwhelm such obstacles. Mass production-suitable electrodes consisting
of stand-alone Ni films obtained by well-established microfabrication methods 3 and eco-friendly
pyrolyzed paper4 enabled the reproducible determination of LWC from soy leaves with optimized
sensibilities of 27.0 (Ni) and 17.5 kΩ %–1 (paper). Moreover, the freestanding design of the Ni
electrodes was key to deliver high on-leaf adhesion and long-term compatibility. Their impedances
remained unchanged under the action of wind at velocities of up to 2.00 m s–1, whereas X-ray
nanoprobe fluorescence assays5 allowed us to confirm the Ni sensor compatibility by closely
monitoring the soy leaf health in an electrode exposed area. Both electrodes operated through direct
transfer of the conductive materials on hairy soy leaves using an ordinary adhesive tape. We used
a handheld and low power potentiostat with wireless connection to a smartphone to determine LWC
over 24 h. Impressively, a machine learning model was able to convert the sensing responses into
a simple mathematical equation that gauged the impairments on water contents at two temperatures
(30 and 20 °C) with reduced root mean square errors (0.1% up to 0.3%). This data suggests a broad
applicability of the platform by enabling the direct determination of LWC from leaves even at variable
temperatures. Overall, our findings6 may help to pave the way to translate ‘sense–act’ technologies
into widespread use toward on-site and remote investigation of plant drought stress. These platforms
can provide relevant information for aiding efficient data-driven management and guiding decision-
making steps.
1 Kim JJ, Allison LK, Andrew TL, Sci. Adv., 5, 2019, eaaw0463.
2 Kim JJ, Fan R, Allison LK, Andrew TL, Sci. Adv., 6, 2020, eabc3296.
3 Giordano GF, Freitas VM, Schleder GR, Santhiago M, Gobbi AL, Lima RS, Appl. Mater. Interfaces,13, 2021, 35914.
4 Damasceno S, Corrêa CC, Gouveia RF, Strauss M, Bufon CCB, Santhiago M, Adv. Electron. Mater., 6, 2020, 900826.
5 Kopittke PM, Punshon T, Paterson DJ, Tappero RV, Wang P, Blamey FPC,Van der Ent A, Lombi E, Plant Physiol., 2,
2018, 507.
6 Barbosa JA, Freitas VMS, Vidotto LHB, Schleder GR, de Oliveira RAG, da Rocha JF, Kubota LT, Vieira LCS, Tolentino,
HCN, Neckel IT, Gobbi AL, Santhiago M, Lima RS. Appl. Mater. Interfaces, 2022, DOI: 10.1021/acsami.42c02943.
[FAPESP, Brazilian Synchrotron Light Laboratory]

17
EFFECT OF FDM PRINTING PARAMETERS ON THE ELECTROCHEMICAL
PROPERTIES OF 3D-PRINTED CARBON ELECTRODES
Raquel G. Rocha (PG)a*, Lucas V. Faria (R)a, David L. O. Ramos a, Rafael L. Germscheidtb,
Diego P. dos Santos (R)b, Juliano A. Bonacinb, Rodrigo A. A. Muñoz (R)a, Eduardo M. Richter
(R)a
aInstitute of Chemistry, Federal University of Uberlandia, Uberlândia, Minas Gerais, Brazil, 38400-902
b Institute of Chemistry, University of Campinas, Campinas, São Paulo, Brazil, 13083-859
*e-mail: raquel_rocha1793@hotmail.com
Thermoplastic filaments containing conductive carbon materials have contributed tremendously to

innovations in the scientific scenario, however, the high resistivity of available materials sets a
challenge for electrochemical sensing proposals. In order to avoid this problem, several research
groups have proposed chemical and physical post-treatments that are time-consuming and affect
the structural integrity of materials.1,2 The selection of suitable printing parameters can be a feasible
alternative to improve the response of electrodes produced by FDM 3D printing 3 Herein, we
systematically investigated the influence of printing parameters (orientation, layer thickness, number
of perimeters and print speed) on the electrochemical performance of 3D-printed sensors. For these
studies, an affordable filament of carbon black integrated polylactic acid (CB/PLA) was selected, and
measurements by cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS)
using [Ru(NH3)6]2+/3+ as redox probe were performed. The results showed that electrodes printed
under vertical orientation, with lower layer thickness (0.05 mm) and speed (30 mm s-1) using two
perimeter numbers provided the best electrochemical performance. Furthermore, it was highlighted
that even using chemical/electrochemical surface treatment, the printing parameters control is
relevant, as observed by cyclic voltametric experiments (Figure 1). To understand the improvement
of electrochemical responses, experiments by Raman spectroscopy and multivariate curve
resolution by alternating least squares (MCR-ALS) were carried out which showed greater availability
and homogeneity of conducting sites under the selected conditions. Thus, it can be inferred that 3D-
printing parameters are important tools to allow the manufacture of improved carbon electrochemical
platforms.
Figure 1. CV profiles obtained for electrodes printed in vertical (red line) and horizontal (black line) orientation after
chemical/electrochemical activation in basic medium [15]. Redox probe: 1 mmol L -1 [Fe(CN)6]3-/4-; supporting electrolyte:
0.1 mol L-1 KCl solution; Printing conditions: layer thickness: 0.05 mm; perimeter number: 2 shell; infill density: 100%;
printing speed: 30 mm s-1
1 Omar MH, Razak MN, Wahab MN, Hamzah HH, RSC Adv, 11, 2021, 16557.
2 Stefano JS, Kalinke C, Rocha RG., Rocha, DP, Silva VAOP, Bonacin JA, Angens L, Richter EM, Janegitz BC, Munoz,
RAA, Analytical Chemistry,17, 2022, 6417.
3 Abdalla A., Hamzah HH., Keattch O., Covill D., Patel BA. Electrochimica Acta, 354, 2020, 136618.
[CAPES, CNPq, FAPEMIG, FAPESP, INCTBio]

18
SCREEN-PRINTED ELECTRODE ELETROCHEMICALLY MODIFIED BY
MOLECULARLY IMPRINTED POLYPYRROLE FOR SARS-CoV-2 DETECTION
Milena P. Ferreira (PG)a*, Felipe A. Gorla (PG)a,b, César R. T. Tarley (R)a,c
a State University of Londrina, Department of Chemistry, Londrina, Parana, Brazil, 86051-990
b Federal Institute of Paraná, Campus Assis Chateaubriand, Assis Chateaubriand, Parana, Brazil, 85935-000
c National Institute of Science and Technology in Bioanalysis (INCTBio), State University of Campinas (UNICAMP),
Department of Chemistry, Institute of Chemistry, Campinas, Sao Paulo, Brazil, 13083-970

*e-mail: milenapf41@hotmail.com
Given the challenges inherent to the methods usually used in the diagnosis of COVID-19, such as
RT-PCR (gold standard), the use of electrochemical diagnosis is an opportune way to overcome the
inconveniences related to long response time, the need for qualified technicians, and expensive
equipment. Electrochemical sensors based on Molecularly Imprinted Polymer (MIP) have benefits
such as operational simplicity, on-the-spot detection, cost-effectiveness, fewer sample amounts
required, faster response times, and pronounced selectivity.1 In this work, a screen-printed electrode
(SPE) electrochemically modified by MIP using pyrrole as monomer and the protein spike S1 as
template was developed, named MIP-Ppy. The number of cycles used in the electropolymerization
and the monomer and template concentrations were optimized. All optimization steps were
monitored by CV and EIS using a [Fe(CN)6]3-/4- redox probe. The thickness of the optimized sensors
was estimated by SEM analysis. Initially, the surface of SPE was activated with 0.1 mol L -1 H2SO4
using 15 voltammetric cycles to increase the conductivity and for removing possible contaminants.2
As expected, the activation was confirmed by the reduction in the resistance to electronic transfer
and by the increase in the current signal in the CV. Upon activation, the variables that play an
important role on the electropolymerization and the selectivity of MIP were investigated, including
pyrrole concentrations ranged from 0.01 to 0.3 mol L-1, protein S1 from 10 to 60 µg mL-1, and the
number of cycles from 2 to 20. The best conditions were defined as: pyrrole at 0.3 mol L-1, protein
S1 at 40 µg mL-1, and 4 cycles. SEM images confirmed the formation of the polymeric film, since
MIP-Ppy showed a thickness of 25-30 nm, NIP-Ppy (non-imprinted sensor) 20-23 nm, and bare
SPCE 15-19 nm. It was observed low current for the probe [Fe(CN)6]3-/4- using the MIP-Ppy sensor.
However, upon template protein S1 removal using a 10% (w/v) HAc solution and 0.02% (w/v) sodium
dodecyl sulphate for 2 h, a significant increase on the current was observed, which confirms the
removal of template from the cavities of MIP-Ppy and, thus, making the transfer of electrons easier.
Under optimized conditions, rebinding assays using 0.01 µg mL-1 of protein S1 in phosphate saline
buffer 0.05 mol L-1 (pH 7.4), showed a decrease on the current of probe when using the MIP-Ppy
regarding the NIP-Ppy ones. Such behaviour reveals the selective interaction of protein S1 with the
imprinted cavities. In order to reduce the concentration of protein S1 in the rebinding assays aiming
at attaining the required sensitivity in the diagnosis of SARS-CoV-2, a preconcentration step by
applying + 0.5 V has been evaluated, whose results demonstrates the feasibility of sensor to detect
fg mL-1. Further studies will be performed to direct detection of SARS-Cov-2 in saliva samples.
1 Mahshid SS, Flynn SE, Mahshid S, Biosensors and Bioelectronics, 176, 2021, 1129052021.
2 Martin C, Grgicak C, ECS Transactions, 61, 2014, 26.
[CAPES, CNPq, INCTBio, UEL, IFPR]

19
IN SITU NANOCOATING ON PYROLYZED PAPER: A PIONEERING STRATEGY
TO PROVIDE SENSITIVE ELECTROCHEMICAL ASSAYS IN BIOLOGICAL
FLUIDS
Caroline Y. N. Nicoliche (PG)a,b, Aline M. Pascon (R)a,c, Ítalo R. S. Bezerra (PG)a,c*, Ana C. H.
Castro (R)b, Gabriel R. Martos (UG)d, Jefferson Bettini (R)a, Wendel A. Alves (R)b, Murilo
Santhiago (R)a,b, Renato S. Lima (R)a,b,c,e
a BrazilianCenter for Research in Energy and Materials, Brazilian Nanotechnology National Laboratory, Campinas, São
Paulo, Brazil, 13083-100
b State University of Campinas, Institute of Chemistry, Campinas, São Paulo, Brazil, 13083-970
c Federal University of ABC, Center for Natural and Human Sciences, Santo André, São Paulo, Brazil, 09210-580
d Pontifical Catholic University of Campinas, Faculty of Chemistry, Campinas, São Paulo, Brazil, 13087-571
e University of São Paulo, São Carlos Institute of Chemistry, São Carlos, São Paulo, Brazil, 13566-590
*e-mail: italo.bezerra@lnnano.cnpem.br
The analytical performance of electrochemical biosensors is significantly reduced when subjected to

complex biological media (such as plasma and serum), which is due to the non-specific adsorption
of biomolecules, especially proteins. Therefore, numerous studies in the literature report the use of
semipermeable layers (e.g., polyethylene glycol)1 to avoid this biopassivation on the surface of the
electrodes. Although these strategies have been promising as anti-biofouling (a-BF) strategies,
these semipermeable layers are mostly detrimental to the sensitivity of the devices as they act as
high impedance layers that inhibit redox reactions on the electrode, decreasing the electrochemical
signal. In addition, these layers may have low durability, reproducibility, and scalability. 2,3 To solve
this problem, our research group studied the use of pyrolyzed graphitic paper (PGP) in combination
with the addition of a neutral surfactant, polysorbate 20, commercially known as Tween® 20 (T20).
Such an off-the-shelf polysorbate provides the in-situ formation of a hydrophilic nanocoating on the
electrode (2 nm according to transmission electron microscopy assays) that prevents biopassivation
while providing the gradual capillarity-assisted penetration of the sample through the paper pores.
These cooperative effects lead to a gradual increase in the electrochemical responses. While state-
of-the-art a-BP strategies compromise sensitivity, our approach leads to peak currents that are up
to 12.5 times greater than the original faradaic currents after 1-h exposure to unprocessed human
plasma. The nanointerface was strictly characterized using a set of microscopic and spectroscopic
techniques. Further, the clinical performance of the device was successfully challenged in clinically
important samples, i.e., sera from healthy individuals and patients infected with the Coronavirus
disease 2019 (COVID-19), for screening purposes. For these assays, we adapted the approach to
develop an electrochemical impedance spectroscopy-based affinity immunosensor by anchoring the
Spike (S) protein on gold nanoparticles (AuNPs). S protein was used as bioreceptor for selectively
targeting SARS-CoV-2 virus antibodies. The serum samples could be distinguished from each other
with 100% accuracy. Such results were recently published in the literature.4 The scalability and
simplicity of the platform combined with its unique ability to operate in biofluids with an enhanced
sensibility may help to pave the way for a new generation of electrochemical biosensors capable of
translating field-deployable analyses and mass screening into the real world. This multidisciplinary
work originated from a collaboration among research groups with expertise in distinct science areas,
including electrochemistry, nanomaterials, microfabrication, and microscopy.
1 Pelaz B, del Pino P, Maffre P, Hartmann R, Gallego M, Rivera-Fernández S, de la Fuente JM, Nienhaus GU, Parak
WJ. ACS Nano 2015, 9, 6996.
2 Dhanapala L, Jones AL, Czarnecki P, Rusling JF. Analytical Chemistry 2020, 92, 8021.
3 Lin S, Yu W, Wang B, Zhao Y, En K, Zhu J, Cheng X, Zhou C, Lin H, Wang Z, Hojaiji H, Yeung C, Milla C, Davis RW,
Emaminejad S. PNAS 2020, 117, 19017.
4 Nicoliche CYN, Pascon AM, Bezerra ÍRS, Castro ACH, Martos GR, Bettini J, Alves WA, Santhiago M, Lima RS. ACS
Applied Materials & Interfaces 2022, 14, 2522.
[CNPEM, UNICAMP, UFABC, CAPES, FAPESP, CNPq]

20
GRAPHITE-BASED 3D PRINTED ELECTRODE FOR THE SEQUENTIAL
DETERMINATION OF PARAQUAT AND CARBENDAZIM IN REAL SAMPLES
Luiz Ricardo G. Silva (PG)a, Jéssica S. Stefano (R)a*, Roberta C. F. Nocelli (R)a, Bruno C.
Janegitz (R)a
a Department of Nature Sciences, Mathematics and Education, Federal University of São Carlos, Araras, SP, Brazil,
13600-970.
*e-mail: jessica.s.stefano@gmail.com
One of the biggest problems facing the world today is the contamination of waters, the environment,
and food by toxic chemicals such as pesticides1. Pesticides are heavily employed compounds
capable of protecting crops from insects, weeds, and diseases, however, these compounds can be
extremely harmful to human health, when used in wrong quantities. The possibility of performing fast
analysis, with high sensitivity, in a miniaturized system with no need of sample treatment makes the
electrochemical methods a powerful tool for the analysis of pesticides in food and environmental
samples2, and the use of 3D printing technology enables the manufacturing of simple, versatile and
low-cost analytical systems for on-site analysis3. In this work, we have presented the use of a new
electrochemical 3D printed sensor, based on a graphite conductive polylactic acid (PLA) filament
(Gpt-PLA) manufactured in laboratory for the sequential voltammetric determination of paraquat
(PQ) and carbendazim (CBZ) in water, honey, juice, and milk samples. A ready-to-use labmade
conductive filament with a high graphite loading (40% wt.) on a PLA matrix, was used for 3D printing
the counter, reference and working electrodes. Also, the base for coupling of the electrodes was 3D
printed using raw PLA filament. For the analysis, a pH study was performed using Britton-Robinson
(BR) solution, and pH = 4.0 enabled satisfactory electrochemical response for both PQ and CBZ.
The electrochemical detection of CBZ and PQ was performed by sequential analysis using square
wave voltammetry under optimized conditions for each compound. Initially, a cathodic voltammetric
scan was recorded from 0 to -1.5 V, followed by the obtention of the anodic scans, from 0 to +1.5 V,
after a waiting time of 10 s, enabling the reduction of PQ at around -0.7 V, and the subsequent
oxidation of CBZ, at around +0.9 V. Figure 1 presents the voltammetric responses for increasing
concentrations of PQ and CBZ (A) and respective calibration curves (B).
Figure 1. SWV recordings for increasing concentrations of PQ and CBZ in 0.1 mol L −1 BR buffer solution (pH 4.0) under
optimized conditions.
The sequential measurements of PQ and CBZ provided great results for both compounds individually
when both are present in a mixture. Linear working ranges from 0.05 to 1.0 µmol L-1, and 1.0 to 50.0
µmol L-1 were obtained for PQ, and from 0.5 to 50.0 µmol L-1 for CBZ, with limits of detection of 0.17
and 0.03 µmol L-1 for PQ and CBZ, respectively. A recovery test was performed in honey, milk, juice,
and water samples after simple dilution, and recovery values ranging from 94.5 to 113.7% were
obtained, attesting the suitability of the method and 3D printed sensors.
1 Boudh S, Singh JS, Springer Singapore, 245–269, 2018.
2 Wang W, Wang X, Cheng N, Luo Y, Lin Y, Xu W, Du D, Trends Anal. Chem. 132, 2020.
3 Cardoso RM, Kalinke C, Rocha RG, dos Santos PL, Rocha DP, Oliveira PR, Janegitz BC, Bonacin JA, Richter EM,
Munoz RAA, Anal. Chim. Acta. 1118, 2020, 73–91.
[FAPESP, CAPES, CNPq]

21
USE OF SMD MICROCHIPS AS GOLD MICROELECTRODE ARRAYS IN
PHARMACEUTICAL AND ENVIRONMENTAL APPLICATIONS
Gilvana P. Siqueira (PG)a*, Lucas V. de Faria (R)a, Raquel G. Rocha (PG)a,
Tiago A. Matias (R)a, Eduardo M. Richter (R)a, Rodrigo A. A. Muñoz (R)a,
Iranaldo S. da Silva (R)b, Luiza M. F. Dantas (R)b
a Universidade Federal de Uberlândia, Instituto de Química, Uberlândia, Minas Gerais, Brazil, 38400-902
b Universidade Federal do Maranhão, Departamento de Tecnologia Química, São Luís, Maranhão, Brazil, 65080-805
*e-mail: gilpsiqueira@gmail.com.br
Microelectrode arrays (µEAs) are devices composed of microelectrodes, which operate individually
and contribute to the total measured current.1 The combination of its ultra-small dimensions and
electrochemical properties allows the monitoring of chemical species in different areas of science 2.
In this work, we investigate the effect of electrochemical treatment on the surface of low-cost and
disposable devices (microchips) containing gold microelectrode arrays (Au-µE). This procedure
involved the electrode anodization to generate nanoporous gold structures (NPAu-μE), which
contributed to (~4.4-fold) increase in the electroactive area as well as a decrease in the resistance
to charge transfer. The electrochemical response of these sensors was properly investigated using
dipyrone (DIP), cysteine (CyS) and lead(II) as target species, and for all analytes, enhanced
analytical performances were obtained under the treated surface. A batch-injection analysis with
amperometric detection (BIA-AD) was proposed for DIP and CyS and provided detection limits lower
than 1 µmol L−1, adequate precision (RSD <4.0 %) with a wide linear range (1.0 – 200.0 and 5.0 –
150.0 µmol L−1 for DIP and CyS, respectively), high analytical frequency (148 and 185 analysis per
hour, respectively) and satisfactory application in pharmaceutical samples (results similar to
spectrophotometric analyses at 95% confidence level). Moreover, the NPAu-µE sensor proved to be
suitable for Pb2+ evaluation by square-wave anodic stripping voltammetry (SWASV), with an
excellent analytical performance achieved, including a detection limit of 5 µg L−1, linear range from
40.0 to 110.0 µg L−1, and good precision (RSD = 4.3 %), which enabled the analysis of tap water
samples, with recovery close to 85%.
200 j j
3 (B)
(A) i i
150
0
I / A
I / nA
h
h
100
-3 g g
f f
e
-6 50 e
d d
Au-E b
c c
b a
a
NPAu-E
-9 0
0.0 0.4 0.8 1.2 1.6 0 300 600 900 1200 1500
E / V vs. Ag|AgCl|KCl(sat.) Time / s
Figure 1. (A) Cyclic voltammograms of Au-µE (black line) and NPAu-µE (red line) (B) determination of DIP by BIA-AD
system.
1 Huang XJ., O’Mahony AM, Compton RG, Small, 5, 2009, 776

2 Di Credico A, et al. Biosensors, 11, 2021, 450
[UFU, CNPq, FAPEMA, FAPEMIG, CAPES]

22
MINIATURIZED ELECTROCHEMICAL DEVICE COMBINED WITH GAS
EXTRACTION FOR DETERMINATION OF SULFITE
Diele A. G. Araújo (R)a,*, Ana C. M. Oliveira (PG)a, Lauro A. Pradela-Filho (R)b, Regina M.
Takeuchi (R)a, André L. Santos (R)a
a Federal University of Uberlândia, Institute of Chemistry, Uberlândia, MG, Brazil, 38408-902.
b University of São Paulo, Institute of Chemistry, Department of Fundamental Chemistry, São Paulo, SP, Brazil, 05508-
900
*e-mail: diele_cap@hotmail.com
Electroanalytical is an inexpensive and high-performance analytical technique. However, their lack

of selectivity limits their use in complex matrices such as biological, environmental, and food
samples1. The coupling of miniaturized electrochemical cells to sample pretreatment procedures is
an interesting approach able to overcome most of these limitations. Besides low consumption of
chemicals, samples, and low waste generation the selectivity and detectability can be substantially
improved by coupling extraction and/or pre-concentration steps. Thus, we proposed a new,
inexpensive, robust, durable, and simple miniaturized electrochemical device integrated with
membraneless gas extraction and preconcentration for the voltammetric determination of sulfite. A
copper wire (1.0 mm diameter) was sealed into an insulin syringe with epoxy glue and it was used
as the working electrode. A 0.3 mm thickness platinum plate and an Ag/AgCl wire used as the
auxiliary and pseudo-reference electrode, respectively, were attached with epoxy glue outside the
insulin syringe. The miniaturized electrochemical cell was filled with 500 μL of the supporting
electrolyte and acted as the SO2(g) acceptor reservoir. The sample reservoir (the donor reservoir)
was filled with 15 mL of the acidified sample solution and both compartments were connected using
a conventional air headspace configuration. After the acidification of the sample, the SO2(g) is
transported by an argon stream from the bottom part (donor reservoir) to the superior part (acceptor
reservoir) and dissolves in the alkaline supporting electrolyte being converted to 𝑆𝑂32− (aq).
Analytical curves were constructed directly in 500 μL of sulfite standard solutions at different
concentrations. As a final step, we have integrated the miniaturized device to the gas extraction
system and the extraction time was studied. Subsequently, analytical curves for sulfite using the gas
extraction system were constructed. For each sulfite concentration, voltammetric measurements
were performed in triplicate. The effect of coupling the extraction step to the voltammetric detection
of sulfite on the analytical performance of the method can be visualized in Table 1. The integrated
extraction procedure has remarkably improved all the merit figures evaluated, except the R 2 which
was almost the same in both systems.
Table 1. Analytical parameters for sulfite using the miniaturized device with and without gas extraction.
Linear range/ LD/ LQ/ Sensibilidade/
Development Method R2
µmol L-1 µmol L-1 µmol L-1 µA µmol-1 L
Miniaturized device 600 - 10000 0.99866 30.0 100.0 0.007
Miniaturized device integrated to gas

25 - 100 0.99757 4.0 13.0 0.095
extraction
Despite its simplicity and low cost, the proposed system provided good analytical performance
(LOD 4 µmol L-1), which is ascribed to the low volume of the acceptor solution (500 μL) which allowed
both the SO2 extraction and its preconcentration. The proposed system is quite versatile since it can
be applied to quantify any volatile electroactive species. A particularly attractive feature of our
analytical system is its ability to assess the real-time gas extraction curve with high temporal
resolution. These curves are crucial to studying and optimizing new gas extraction procedures since
they allow for evaluating the kinetics of the extraction process, the effectiveness of the acceptor
solution and eventual analyte loss.
1 Martins AB, Lobato A, Tasic N, Perez-Sanz FJ, Vidinha P, Paixão TRLC, Gonçalves LM, Electrochemical.
Communication, 107,2019, 106541.
[UFU, CAPES, FAPEMIG, CNPq, FAPESP]

23
NICKEL FERRITE MODIFIED ELECTRODE FOR THE ELECTROCHEMICAL
DETERMINATION OF HYDROGEN PEROXIDE
Amanda B. Nascimento (PG) a*, Lucas V. de Faria (R)a, Tiago A. Matias (R)a, Osmando F.
Lopes (R)a, Rodrigo A. A. Muñoz (R)a
a Federal University of Uberlândia, Institute of Chemistry, Uberlândia, Minas Gerais, Brazil, 38400-902
*e-mail: amanda.nascimento@ufu.br
Ferrites are materials that have a general form MF 2O4 where M is a metal like Ni, Cu, Mg, Zn and
others; and they have chemical stability, low toxicity, in addition to being able to act as
electrocatalysts. Therefore, these materials are used for electrode modification in the development
of electrochemical sensors, focusing on the enhancement of selectivity and sensitivity towards the
determination of different analytes.1 The use of ferrites in combination with carbonaceous materials,
like carbon nanotubes or reduced graphene oxide, can lead to the development of improved
electrochemical sensors due to the synergistic effect of both nanomaterials. Carbon nanomaterials
provide higher conductivity while ferrite nanoparticles their inherent electrocatalytic effect, their
combination provides ferrite stability and enhanced electrochemical sensing properties. The
determination of hydrogen peroxide is highly demanded because is a common oxidant present in
biological systems, commercial products, in the environmental and as contaminant in food. 2
Therefore, the use of ferrites or their nanocomposite with carbon nanotubes for the determination of
hydrogen peroxide by electroanalysis is feasible considering the electrocatalytic properties of ferrites
towards the electrochemical reduction or oxidation of hydrogen peroxide. The objectives of this work
are the synthesis and characterization of nickel ferrites, and their combination with multi-walled
carbon nanotubes (MWCNT) to generate the nanocomposite NiFe2O4/MWCNT for the improved
electrochemical determination of hydrogen peroxide. The nickel ferrite (NiFe2O4) was synthesized
by the hydrothermal method and the electrode modification was made using the drop-casting method
on a glassy-carbon electrode (NiFe2O4/MWCNT/GCE). For the electrode modification, suspensions
of nickel ferrites with MWCNT in dimethylformamide varying the percentage of ferrites (10%, 20%,
50% and 70% weight) in suspension were evaluated as the chemical modifier of the GCE. The
electrochemical profile of these electrodes was investigated by cyclic voltammetry in the presence
of hydrogen peroxide in KCl 0,1mmol L-1. The preliminary results showed the electrochemical
reduction of hydrogen peroxide at -0.3 V. Moreover, the current increase for this process was more
intense in the presence of nickel ferrite in comparison with MWCNT modified electrode (control). The
variation of ferrite percentage also showed differences in the electrochemical reduction of hydrogen
peroxide; the highest percentage of ferrite present the highest current values but in high percentage
of ferrite, the peak is displaced for more negative potentials. This result can be due to the lower
conductivity of ferrite in comparison with MWCNT, and this behaviour was observed when analysing
the cyclic voltammetry of the suspension containing 70% ferrite. Considering the lower reduction
potential that provides greater selectivity, the nanocomposite containing 10% NiFe2O4@MWCNT
showed the best condition with a linear range of peak current as function of hydrogen peroxide
concentration between 50µmol L-1 and 5mmol L-1. Next, the modified electrode will be evaluated by
amperometric detection using a batch-injection analysis system for further analysis of complex
samples.
1 Kurian J, et al. Journal of Magnetism and Magnetic Materials, 538, 2021, 168233
2 Zhao X, et al. Microchemical Journal, 166, 2021, 106240
[CAPES, CNPq, FAPEMIG]

24
DEVELOPMENT OF NEW COMPOSITIONS OF SILVER INK TO MANUFACTURE
REFERENCE ELECTRODES
Jéssica R. Camargoa*, Wilson S. Fernandes-Junior (UG)a, Raquel G. Rocha (PG)b, Lucas V.
Faria (R)b, Eduardo M. Richter (R)b, Rodrigo A. A. Muñoz (R)b, Bruno C. Janegitz (R)a
a Federal University of São Carlos, Department of Nature Sciences, Mathematics and Education, Araras, São Paulo,
Brazil, 13600-970
b Federal University of Uberlândia, Institute of Chemistry, Uberlândia, Minas Gerais, Brazil, 38400-902
*e-mail: jessica.10.camargo@gmail.com.br
Silver materials are known to have excellent properties such as high electrical and thermal
conductivities and chemical stability. Therefore, silver-based inks have attracted much attention1. As
it is compatible with a range of diverse substrates, this can be used in the production of uniformly
conductive sensors with low curing temperature². Another positive aspect is its low cost and its
abundance in relation to other metals, such as platinum and gold3. In this way, the combination of
these properties promotes the manufacture of effective electrodes, including as pseudo-references
within a multi-electrode electrochemical system4. To prepare the silver
inks proposed in this work, two compositions, with previously published
conductive inks polymers (colourless nail polish (NP) and shellac (SL)),
were tested. Nail polish5 is a polymer that has proven to be excellent for
the development of conductive inks, mainly because it is already
developed to come into contact with nails without causing harm to
human health. On the other hand, shellac, tested in the article by
Henrique et al.6 is another option, since this is a natural bioadhesive
polymer, which comes from a resin secreted by the female insect kerria
lacca, which is purified for commercial use. This polymer is also widely
used in the pharmaceutical industry, in the development of capsules for
Figure 1. Schematic preparation of
drugs, not showing toxicity in direct contact with humans. For testing of
reference electrodes with silver ink
the developed silver inks, three different systems were used, shown in
Figure 1. Analytical curves were made for -estradiol, showing that
even using different sensors for this purpose, the potential between the
systems was maintained. Furthermore, the developed inks were
subjected to an OCP-type stability test, using a methodology similar to
those demonstrated by Dawkins et al.7 and Søpstad et al.8. The three-
electrode system was used as follows: Ag/AgCl 3.0 mol L− KCl was
used as reference, the electrodes made with the developed inks were
used as work, and the carbon electrode as counter-electrode. In this
way, performing the OCP (Figure 2), it can be verified that in the
systems in which the developed silver inks were used, the potential was
kept close to 0.1 V. For comparison, a measurement was performed Figure 2. OCP measurements
using a carbon reference and it is noted that a variation occurred during using different inks as a work
the measurements, confirming the importance of using silver ink for the electrode.
development of pseudo-reference electrodes.
1 Nie X, Wang H, Zou J, Applied Surface Science, 261, 2012, 554.
2 Bhat KS, Nakate UT, Yoo JY, Wang Y, Mahmoudi T, Hahn Y-B, Chem. Engineering Journal, 373, 2019, 355.
3 Lee B, Kim Y, Yang S, Jeong I, Moon J, Current Applied Physics, 9, 2009, e157.
4 Araujo W, Paixao T, Analyst, 139, 2014, 2742.
5 Andreotti IAA, Orzari LO, Camargo JR, Faria RC, Marcolino-Junior LH, Bergamini MF, Journal of Electroanalytical
Chemistry, 840, 2019, 109.
6 Henrique JM, Camargo JR, Oliveira GG, Stefano JS, Janegitz BC, Microchemical Journal, 170, 2021, 106701.
7 Dawkins RC, Wen D, Hart JN, Vepsäläinen M, Electrochimica Acta, 393, 2021, 139043.
8 Søpstad S, Johannessen EA, Seland F, Imenes K, Electrochimica Acta, 287, 2018, 29.
[CAPES, CNPq, FAPESP]

25
CARBON-BLACK INTEGRATED POLYLACTIC ACID ELECTROCHEMICAL
SENSOR FOR CHLORAMPHENICOL DETERMINATION IN MILK AND WATER
SAMPLES
Marina Di-Oliveira (UG)a*, Lucas V. de Faria (R)a, Raquel G. Rocha (PG)a, Eduardo M. Richter
(R)a, Rodrigo A. A. Muñoz (R)a
a Federal University of Uberlândia, Chemistry Institute, Uberlândia, Minas Gerais, Brazil, 38.408-100
*e-mail: marina.di@ufu.br
3D-Printing or additive manufacturing is an emerging technology with several fields of application,

due to advantages such as versatility, low-cost, low material wastage, fast prototyping and the
possibility of large-scale production.1 This technology aims to produce three-dimensional physical
objects from a digital model and, over recent years, conductive thermoplastic filaments such as
carbon black integrated into a polylactic acid matrix (CB/PLA) have enabled the development of
affordable electrochemical sensors.1,2 The use of antibiotics for the treatment of bacterial infections
and prophylactic purposes is routine in veterinary practices4. In fact, chloramphenicol (CAP) is a
broad-spectrum antibiotic widely applied in veterinary medicine due to its relatively low-cost and high
efficiency against bacterial infections. Nevertheless, the overuse might leave residues, in food
products, such as milk, and in aquatic environments, leading to an accumulation of CAP in the
organism, creating serious problems to human health. Thus, in this work, we fabricate a 3D-printed
CB/PLA electrode for the CAP voltammetric determination in milk and water samples. To enable
detection of low levels of CAP, the 3D-printed electrode surface was electrochemically treated
(application of a potential of +1.4 V followed by -1.0 V vs. Ag∣AgCl∣KCl(sat.) both for 200 s) using 0.5
mol L-1 NaOH as the supporting electrolyte1. This procedure conferred increased electroactive area
and consequently lower charge transfer resistance (verified by cyclic voltammetric (CV) responses
and electrochemical impedance spectroscopy), which provided a considerable improvement of CAP
response (12-fold) when compared to the untreated surface (Fig. 1A). A method exploiting the
reduction of the nitro group at around -0.45 V (vs. Ag∣AgCl∣KCl(sat.)) was developed by differential
pulse voltammetry. A wide linear range (10 to 331 µmol L-1, r = 0.998, Fig. 1B) with a detection limit
of 0.98 µmol L-1 and a good precision (RSD = 5.1 %, n = 10), as well as selectivity against other
classes of antibiotics, were achieved. Water and milk samples were spiked with known amounts of
CAP and after a simple dilution process in supporting electrolyte (0.1 mol L-1 HCl), they were
immediately analyzed by the standard addition method. Recovery values between 88 and 93%
attested to accuracy of the method, as well as absence of matrix effect.
Figure 1. (A) Cyclic voltammetric responses for 1.0 mmol L−1 CAP in 0.12 mol L−1 BR buffer (pH = 2.0). The inset in Fig.
(A) shows EIS measurements of 2 mmol L−1 [Ru(NH3)6]2+/3+ in 0.1 mol L−1 KCl solution, before (blue line) and after
electrochemical treatment (black line) of the 3D-printed CB/PLA electrode at +0.17 V vs. Ag∣AgCl∣KCl(sat.). (B) Baseline-
corrected DPV responses obtained for increasing CAP concentrations (10 to 331 μmol L−1) using 0.1 mol L−1 HCl as
supporting electrolyte.
1 Richter EM, Rocha DP, Cardoso RM, Keefe EM, Foster CW, Munoz RA, Banks CE, Analytical Chemistry, 91, 2019,
12844.
2 Ambrosi A, Pumera M, Chemical Society Reviews,45, 2016, 2740.
3 Faria LV, Lisboa TP, Campos NS, Alves GF, Matos MAC, Matos RC, Munoz RAA, Analytica Chimica Acta, 2021, 338569.
[FAPEMIG, CAPES, CNPq, InovaLeite e Instituto Sua Ciência]

26
3D-PRINTED ELECTROCHEMICAL SENSOR USING CARBON
BLACK/POLYLACTIC ACID FILAMENT FOR THEOPHYLLINE DETECTION
Wilson S. Fernandes-Junior (UG)a*, Luiz O. Orzari (PG)a,b, Bruno C. Janegitz (R)a
aFederal University of São Carlos (UFSCar), Laboratory of Sensors, Nanomedicine, and Nanostructured Materials
(LSNano), Araras, São Paulo, Brazil, CEP 13604-900
b Federal University of São Carlos, Graduate Program in Materials Science, Department of Physics, Chemistry, and
Mathematics, Sorocaba, São Paulo, Brazil, CEP 18052-780

*e-mail: wilsonfernandes@estudante.ufscar.br
Theophylline (TP) is a bronchodilator drug, one of the most prescribed medications for asthma and
chronic obstructive pulmonary disease treatment. It is widely available in drugstores and
compounding pharmacies and has an affordable price compared to other bronchodilators1. In nature,
TP can be found in small amounts in teas and cocoa seeds, and the bronchodilator effect was
discovered in 1922 when it was introduced for asthma clinical treatment2. It is extremely necessary
to monitor TP to avoid serious toxicological effects by using simple, fast, accurate, and economic
techniques, due to its narrow safety range3. Fused Deposition Modeling is a 3D printing technique
that consists of layer-by-layer deposition of a
polymeric filament on a printing bed passing through a
heated nozzle4. This technique produces many
devices in chemistry, whether in microfluidic analysis,
energy storage, and electrochemical sensors5. In this
context, this work aims the TP detection, in biological
fluids, using a 3D printing electrochemical sensor. The
electrode was produced using a carbon
black/polylactic acid commercial filament for the
electrodes, a non-conductive thermoplastic
polyurethane filament for the base in which the
electrodes were inserted, and a bottle cap as an
electrochemical cell, as seen in Figure 1. The
proposed sensor was characterized by scanning
electron microscopy, Fourier-transform infrared
spectroscopy, and electrochemical impedance
spectroscopy, and it was applied in electrochemical Figure 1. Electrochemical sensor. (A) 3D-printed
measurements such as cyclic and square wave carbon black/polylactic acid electrodes. (B)
voltammetries. In this regard, an oxidation peak was Electrodes inserted on the 3D base. (C) 3D base
observed for TP at 0.8 V and the proposed sensor, printed with non-conductive filament. (D)
Obtention of the analytical signal
under optimized parameters, was capable to detect it
in synthetic biological fluids samples, presenting a linear range from 5.00 to 150 μmol L−1 with a limit
of detection of 0.16 μmol L−1.
1 Barnes PJ, Am J Respir Crit Care Med, 188, 2013, 901.
2 Dasgupta A, Krasowski MD, Chapter 16 - Caffeine and theophylline, in: Dasgupta A, Krasowski MD (Eds.), Therapeutic
Drug Monitoring Data (Fourth Edition), Academic Press 2020, 351.
3 Duan YC, Wang AQ, Ding YP, Li L, Duan DD, Lin JX, et al., J Pharm and Biomedical Analysis, 192, 2021, 7.
4 León A, Domínguez-Calvo A, Molina S, 182, 2019, 108044.
5 Cardoso RM, Kalinke C, Rocha RG, Santos PL, Rocha DP, Oliveira PR, Janegitz BC, Bonacin JA, Richter EM, Muñoz
RAA. Analytica Chimica Acta, 1118, 2020, 73.
[CNPq, FAPESP, CAPES]

27
VOLTAMMETRY OF IMMOBILIZED MICROPARTICLES APPLIED TO
IDENTIFICATION OF ANTARCTIC AND SUB-ANTARCTIC MACROALGAE
Ana Paula O. L. Inacio (UG)a*, Pio C. Neto (R)b, Claudio M. P. Pereira (R)c, Daiane Dias (R)a
a Federal University of Rio Grande, School of Chemical and Food, Rio Grande, Rio Grande do Sul, Brazil, 96203-900
b University of São Paulo, Institute of Chemistry, São Paulo, São Paulo, Brazil, 05508-220
c Federal University of Pelotas, Center of Chemical, Pharmaceutical and Food Sciences, Pelotas, Rio Grande do Sul,
Brazil, 96010-900
*e-mail: anapaulaoliveira1814@gmail.com
It is well known that in analytical chemistry, the direct analysis of solid-state samples is often avoided,
because of the complexity of the matrix. However, from a practical analytical viewpoint, the solid-
state study is less susceptible to loss, contamination and sample exposure.1 In this context, Scholz,
and co-workers developed the first solid-state study in electroanalytical, the voltammetry of
immobilized microparticles (VIMP)2. At once, macroalgae have a variety chemical composition,
consumed worldwide for medicinal, pharmaceuticals and other purposes. The study of its
composition is commonly performed by chromatography3, but VIMP can be studied to identify
macroalgae based on their susceptible compounds to oxidation and reduction processes. For this,
the electrochemical experiments were performed in 0.25 mol L-1 acetic acid/sodium acetate buffer at
pH 4.6. A graphite electrode impregnated with paraffin (PIGE) as a working electrode, Ag/AgCl (KCl
3.0 mol L-1) as a reference and a platinum wire as an auxiliary. Nine species of Antarctic
(Myriogramme Manginii, Iridaea Cordata, Palmaria Decipiens, Pyropia Endiviifolia, Adenocystis
Utricularis) and sub-Antarctic macroalgae (Gigartina Skottsbergii, Mazzaella Laminarioides,
Lessonia Searlesiana, Lessonia Flavicans), were lyophilized, macerated and stored without light.
The sample immobilization was performed by abrasion on the PIGE surface that was inserted into
the electrolyte. The measurements were performed by SWV (from 1.5 to -1 V, Estep = 4 mV amplitude
= 25 mV, f = 5 Hz). The differences in the obtained voltammograms made possible a classification
criterion for macroalgae. At first, based on their anodic voltammograms, the macroalgae Gigartina
Skottsbergii, Lessonia Searlesiana, and Iridaea Cordata were identified by their specific
characterization peaks (SCP) (-0.2 and 0.16 V; 0.0 and 0.7 V; 0.6 V, respectively). The remaining
macroalgae, which do not show a SCP, were identified by comparing their voltammograms. Both
macroalgae Lessonia Flavicans and Adenocystis Utricularis presented an oxidation peak at 0.5 V,
but only the second one presented a peak at 1.1 V. To the macroalgae Myriogramme Manginii and
Palmaria Decipiens, the common peak was at 1.0 V, but only the Myriogramme Manginii presented
a peak at 0.4 V. Lastly, Mazzaella Laminarioides and Pyropia Endiviifolia presented a peak at -0,3
V, but only the first one shows an oxidation peak at 0.4 V. Based on the cathodic voltammograms,
seven of the nine species presented SCP: Mazzaella Laminarioides (-0.6 V), Lessonia Searlesiana
(0.1 V), Lessonia Flavicans (0.9 V), Myriogramme Manginii (0.3 V), Pyropia Endiviifolia (-0.7 V) and
Adenocystis Utricularis (0.5 V). For others, Gigartina Skottsbergii and Palmaria Decipiens presented
a reduction peak at -0.5 V, but only the first one shows a peak at 1.0 V. The Iridaea Cordata
macroalgae was compared to Lessonia Flavicans without including their SCP. Both of them
presented a cathodic peak at -0.3 V, but only the second one shows a peak at 0.2 V. The differences
in the response of compounds in the voltammetric study are associated with the electrochemical
reactions of the three-phase system, which are hampered according to the complexity of the matrix,
especially when the study is carried out in solid-state1. However, the electrochemical profiles
obtained were efficient to identify and characterize the macroalgae in this work, through the
classification screening method developed.
1 Maciel JV, Fava EL, Silva TA, Dias D, Fatibello-Filho O, Journal of Solid-State Electrochemistry, 21, 2017, 2859.
2 Scholz F, Nitschke L, Henrion G, Naturwissenschaften, 76, 1989, 71.
3 Sasidharan S, Chen Y, Saravanan D, Sundram KM, Latha LY, African Journal of Traditional, Complementary and
Alternative Medicines, 8, 2011, 1.
[FURG, EQA, CNPq]

28
3D PRINTED SENSORS FOR DETECTION OF NEUROTRANSMISSOR
EPINEPHRINE
Vinicius A. O. P. da Silva (PG)ab*, Bruno C. Janegitz (R)b
a Federal University of São Carlos, Center for Exact Sciences and Technology, São Carlos, SP, Brazil, 13565-905
b Federal University of São Carlos, Department of Nature Sciences, Mathematics and Education, Araras, SP, Brazil,
13600-970
*e-mail: vinicius.silva@estudante.ufscar.br
3D Printing technology has been explored in different areas. Its potential in rapid and practical
prototyping and construction stands out when compared to other techniques. With the advance, it is
possible to print with micrometric resolution at a relatively low cost. 1 Studies applying 3D printing to
build electrochemical sensors with conductive thermoplastic filaments have been increasing in the
literature.2 In this work, electrochemical 3D printed sensors, using filaments based on carbon
nanomaterials3, are proposed for the detection of biomarkers generated by the stress stimulation,
acting in the early detection of diseases. The construction of the sensor involves its 3D printing
followed by chemical treatment4, dipping it in dimethylformamide and washing it in ethyl alcohol, and
a physical treatment, performing the surface sanding. The morphological characterization of the
sensor was performed before and after the treatment by scanning electron microscopy (SEM), where
it is possible to visualize the removal of Polylactic Acid (PLA) from the surface and exposure of
graphene nanofibers. This was confirmed by x-ray diffraction (XRD) analysis, where we can see the
disappearance of the amorphous peak corresponding to PLA and the appearance of crystalline
peaks, corresponding to graphene. The square wave voltammetry (SWV) technique was used, in
which the sensor reached the highest peak current for epinephrine detection. Under optimized
parameters, the analytical curve was built, comprising a linear concentration range from 4.0 to 80
µmol L−1, with r2 of 0,991, and a limit of detection of 0.22 µmol L−1. The proposed sensor was applied
for the analysis of epinephrine in human blood serum samples, obtaining recoveries between 91.3
and 110%. Therefore, the proposed voltammetric method using G-PLA as an electrochemical sensor
proved to be effective for the detection of epinephrine.
Figure 1. Square wave voltammograms obtained by G-PLA in presence of different concentrations of epinephrine, from
4.0 to 80 µmol L−1; in 0.1 mol L−1 phosphate buffer pH7; step = 5mV, frequency = 30 Hz, and amplitude = 60mV. Inset:
The analytical curve for epinephrine.
1 Chan HN, Shu Y, Xiong B, Chen Y, Chen Y, Tian Q, Michael SA, Shen B, Wu H, ACS Sensors, 1, 2016, 227
2 Foo CY, Lim HN, Mahdi MA, Wahid MH, Huang NM, Scientific Reports, 8, 2018, 1
3 Manzanares Palenzuela CL, Novotný F, Krupička P, Sofer Z, Pumera M, Analytical Chemistry, 90, 2018, 5753
4 Santos PL, Katic V, Loureiro HC, Santos MF, Santos DP, Formiga AL, Bonacin JA, Sensors and Actuators B: Chemical,
281, 2019, 837
[CNPq, FAPESP CAPES]

29
USE OF SPPE-P6LC@CHI/PANI/Fe3O4@Au ELECTRODE-TYPE FOR
ETHINYLESTRADIOL DETECTION IN CLINICAL AND BIOLOGICAL SAMPLES
Igor G. S. Oliveira (PG)a*, Adriana C. da Silva (UG)a, Fernando C. Gallina (UG)a, Willyam R. P.
Barros (R)a
a Universidade Federal da Grande Dourados, Rodovia Dourados-Itahum, Km 12, Cidade Universitária - Dourados, MS,
Brazil, 79800-000
*e-mail: igorgabrielso@hotmail.com
Ethinylestradiol (EE2) is a, bioactive estrogen, synthetic female sex hormone derived from 17β-
estradiol. This compound is used in several areas, such as, medicine1,2 being one of the most
common hormone to be found in aquatic environments. Thus, it is necessary to use sensitive and
viable techniques for the EE2 detection. The present work proposed the development of a modified
graphite-based screen-printed polyethylene electrode (SPPE). The SPPE was modified with a
mixture of chitosan solution and Printex 6L carbon (P6LC) plus polyaniline (PANI) and, a layer of a
Fe3O4@Au. In this sense, the electrode was called of SPPE-P6LC@Chi/PANI/Fe3O4@Au. The study
of the variation of graphite ink layers and variation of the proportion of graphite ink were carried out
by CV in 1.0 mmol L-1 Fe(CN)63-/4 at KCl 0.1 mol L-1 as support electrolyte, demonstrating that the
adherence of 3 layers for the proportion of graphite:varnish 70%/30% (w/v) reached the best
electrochemical responses to reversibility criteria (Fig. 1). The study of pH (5 to 10) was carried out
in acetate in 0.1 mol L-1 eletrolyte, and was observed a greater current response at 0,34 V, reaching
0,96 µA. According to the linearity (pH versus E/V), of the response obtained, it was that the EE2
oxidation mechanism corresponds to the same number of prótons and electrons. Regrading the
detection of EE2, by SWW, the study range was from 0.5 µmol L-1 to 100.0 µmol L-1, obtaining a
value of LD= 0.3 µmol L-1. The application of the SPPE-P6LC@Chi/PANI/Fe3O4@Au sensor was
performed in urine sample, with an analytical recovery of ~105%.
Figure 1. Cyclic voltammetric curves at scan rate 50 mVs−1 in 1.0 mmol L-1 Fe(CN)63-/4 at KCl 0.1 mol L-1 as support
electrolyte.
1 Duijkers IJ, Klipping C, Zimmerman Y, Appels N, Jost M, Maillard C, Bennink HJC, The European Journal of
Contraception & Reproductive Health Care, 20, 2015, 476.
2 Wiesinger H, Berse M, Klein S, Gschwend S, Höchel J, Zollmann FS, Schütt B, British Journal of Clinical Pharmacology,
80, 2015, 1399.
[CAPES, UFGD]

30
PHOTOELECTROCHEMICAL SENSOR BASED ON GCE/BIVO4/MIP-AA FOR
DOPAMINE DETECTION
Felipe A. Gorla (PG)a,b*, Caroline S. dos Santos (PG)b, Roberto de Matos (R)b, César R. T.
Tarley(R)b,c
a Federal Institute of Parana (IFPR), Campus Assis Chateaubriand, Assis Chateaubriand, Parana, Brazil, 85935-000
b State University of Londrina (UEL), Departament of Chemistry, Londrina, Parana, Brazil, 86057-970
Departament of Chemistry, Institute of Chemistry, Campinas, Sao Paulo, Brazil, 13083-970

*e-mail: felipe.gorla@ifpr.edu
Dopamine (DA) is a neurotransmitter with an essential role in the body, due to its action on the
hormonal and central nervous system.1 Abnormal levels of DA can cause some neurological,
endocrinal, and cardiovascular disorders, and even Parkinson's disease,1 making it necessary to
develop sensitive and selective methods capable of detecting DA in body fluids. In this sense,
photoelectrochemical sensors have great potential due to their low instrumental cost and high
sensitivity. These sensors are based on semiconductor materials, which when excited form an e-/h+
pair that is used to increase the analytical signal.2 In addition, the use of a molecularly imprinted
polymer (MIP) can significantly increase the sensitivity and selectivity of the sensor. 3 Thus, the
objective of this work is to develop a photoelectrochemical sensor based on the combination of
bismuth vanadate (BiVO4) with a MIP synthesized through the controlled INIFERTER strategy,
aiming at the future determination of DA in urine samples. The hydrothermal method was used to
synthesize BiVO4 from Bi(NO3)3.5H2O and NH4VO3. For MIP synthesis, MWCNT was oxidized and
modified with 3-CPTMS and DDTC. Then, the MWCNT-DDTC was dispersed in a toluene:DMF 3:1
(v/v) mixture, followed by the addition of DA, trimethylolpropane trimethacrylate, and acrylic acid
(MIP-AA), and exposure to UV radiation under magnetic stirring. Two other MIPs were synthesized
through the same procedure, changing the AA to methacrylic acid (MIP-MAA) and itaconic acid (MIP-
ItA). For each MIP, a non-imprinted polymer (NIP) was produced, removing DA from the reaction.
XRD analysis showed a predominance of the monoclinic phase for BiVO4, confirmed by SEM
images, and the band gap calculated using the Kubelka-Munk equation was 2.54, which are
desirable characteristics for excitation of BiVO4 by visible light. MIPs and NIPs were characterized
by FT-IR, TGA, SEM, TEM, and textural analysis (BET and BJH). For photoelectrochemical
measurements, a glassy carbon electrode (GCE) was first modified with a 6 µL droplet of the
synthesized BiVO4 suspended in DMF. The amount of BiVO4 in suspension was optimized at 3.5 mg
mL-1. In this step, the applied potential and pH were studied, and the optimum values were found to
be 0.6 V and pH 6. Following that, the amount of polymeric material added to the 3.5 mg mL-1 BiVO4
suspension was optimized and set at 0.1 mg mL-1. GCE/BiVO4/MIP-AA and GCE/BiVO4/NIP-AA
demonstrated high photocurrent improvements (1.45±0.04 and 1.02±0.05 µA, respectively) for DA
at 10 µmol L-1 compared to the other monomers (0.77±0.06, 0.74±0.02, 0.82±0.05 and 0.72±0.04
µA, for MIP and NIP of MAA and ItA, respectively), values 2.42 and 1.70 times higher than
GCE/BiVO4 (0.60±0.02 µA). Electroactive areas of 0.045 and 0.022 cm2 were obtained for
GCE/BiVO4/MIP-AA and GCE/BiVO4/NIP-AA, while for GCE/BiVO4 it was 0.020 cm2. The study of
electrolyte type and concentration, in addition to a new pH study, were carried out using
GCE/BiVO4/MIP-AA and the best condition was phosphate buffer 0.1 mol L-1 pH 7. The change in
pH value can be attributed to AA and DA interactions. The results obtained so far allow us to
conclude that both BiVO4 and MIPs and NIPs were successfully synthesized and characterized, and
the GCE/BiVO4/MIP-AA sensor is promising for the selective determination of DA at low
concentrations in urine samples.
1 Zhang D, Wu L, Chow DS-L, Tam, VH, Rios DR, Journal of Pharmaceutical and Biomedical Analysis, 117, 2016, 227.
2 Malathi A, Madhavan J, Ashokkumar M, Arunachalam P, Applied Catalysis A, General, 555, 2018, 47.
3 Prete MC, Santos DM, Effting L, Tarley CRT, Journal of Chemical & Engineering Data, 64, 2019, 1978.
[IFPR, UEL, INCTBio, LADEMA, LAEEM, ESPEC, LMEM, LARX, CAPES, CNPq]

31
3D PRINTED Au-BASED SENSOR FOR DETECTION OF CREATININE
Amanda Neumann (UG)a*, Luiz R. G. e Silva (PG)a, Jéssica S. Stefano (R)a, Luiz O. Orzari
(PG)a, Déborah C. Azzi (R)b, Bruno C. Janegitz (R)a
a Laboratory of Sensors, Nanomedicine, and Nanostructured Materials (LSNano), Federal University of São
Carlos, 13600-970 Araras, São Paulo, Brazil
b ADB Pesquisa e Desenvolvimento, 13600-140, Araras, São Paulo, Brasil
*e-mail: amandaneumann@estudante.ufscar.br
Creatinine (CNN) is a metabolite considered a possible biomarker for COVID-19 disease, since high
blood CNN levels indicate malfunctioning of the renal system, that is commonly associated with
COVID-191. CNN levels greater than 150 µmol L−1 in urine or blood indicate renal dysfunction, and
severe damage when values are above 500 µmol L−1, while values below 40 µmol L−1 are indicative
of decreased muscle mass 2. Therefore, CNN monitoring is extremely important to supervise the
functions of the human body, in addition to being a possible biomarker of COVID-19. Thus, this work
presents a 3D printed electrode modified with gold particles for detection of CNN in urine and human
serum. 3D printed graphene electrodes were employed for CNN detection in a low volume solution
(70 µL). To modify the surface of the 3D printed electrode, the chronoamperometry was used,
applying a constant potential of −0.6 V for 400 seconds in the presence of 5.0 mmol L−1 gold(III).
The voltametric analyzes were performed for the detection of CNN. Fig. 1 shows the cyclic
voltammogram obtained with the 3D printed electrode, modified with gold particles in the presence
and absence of 2.0 mmol L−1 of CNN. It is observed that in the presence of CNN, the analytical signal
decreases due to the strong interaction of the nitrogen groups with the gold particles (Fig. 1b) present
on the electrode surface, thus causing a partial surface blockage and reducing the analytical signal.
The quantification of CNN was performed using square wave voltammetry (SWV) technique. The
operational parameters of the SWV were optimized by 23 full factorial design and central composite
design. The optimal values chosen were 5.0 mV potential step, 20 mV modulation amplitude and 10
Hz frequency. The analytical curve was constructed in the range of 0.05 to 3.2 mmol L−1 (Fig. 2).
From the generated curve, the R2 obtained was 0.998, which provided the equation ΔI (µA) = 8.46
+ 9.318 × CCNN (mmol L−1). Limits of quantification (3 × SDintercept/slope) and detection (3.3 × LOD)
were estimated as 0.048 and 0.015 mmol L−1, respectively. An analytical recovery test was
performed using SWV. The synthetic urine sample and human serum was fortified with three different
concentrations of CNN (0.10; 0.80; and 2.10 mmol L−1) and recovery values presented results with
the range of 95.0 to 107%. Thus, attesting that 3D printing is an excellent tool for creating dispositive
capable of detecting CNN in biological samples.
Figure 1. (a) Cyclic voltammograms of blank solution (black) Figure 2. (a) Square wave voltammograms for CNN
and (red) 2.0 mmol L−1 CNN using Au/G-PLA electrode. concentrations (0.05 to 3.2 mmol L−1) in 0.5 mol L−1 NaCl.
Supporting electrolyte: 0.5 mol L−1 NaCl (b) Schematic SWV parameters: 5.0 mV (potential step l); 20 mV
illustration of the interaction of creatinine with particles of Au. (modulation amplitude); 10 Hz (frequency). (b) Analytical
curve.
1 Werion A, et al., Kidney Int. 98, 2020, 1296.

2 Chen JC, et al., Sensors Actuators, B Chem. 115, 2006, 473.

32
USE OF PRINTED DEVICES FOR DETECTION Pb ELECTROCHEMISTRY
Lucimara R. P. Ruis (UG)a*, Liriana M. Roveda (PG)a, Marcela Z. Corazza (R)b,
Bruno G. Lucca (R)c, Fábio Gozzi (R)a, Adriana E. Carvalho (R)a
a Universidade Federal da Grande Dourados, FACET, Dourados, MS, Brazil, 79825-070
b UniversidadeEstadual de Londrina, Departamento de Química, Londrina, PR, Brazil, 86038-440
c Universidade Federal de Mato Grosso do Sul, Instituto de Química, Campo Grande, MS, Brazil, 79074-460
*e-mail: Lucimara.ruis062@academico.ufgd.edu.br
The presence of contaminants in the environment, such as heavy metals, has been a constant
concern, as their impacts are harmful not only to the environment, but also to the potential risk that
it can cause to human health. Lead is one of the main metals that scientists are looking for a better
way to investigate its contamination in water, food and products consumed on a daily basis. Among
them, lead metal may be present in industrial wastewater.1 The identification of these metals can be
performed using electrochemical detection techniques, which have several advantages, such as:
portability, selectivity, high sensitivity, in addition to being of low instrumentation cost compared to
chromatographic techniques. In recent years, several studies have been reported proposing different
analytical techniques for monitoring these pollutants. Among them, the voltammetric methods of
analysis have stood out. In this sense, monitoring in different environmental matrices is a challenge
and a necessity of modern society and 3D printed electrochemical sensors have gained great
prominence in the scientific community. The present study proposes the detection of Pb2+ ions using
electrodes in a system where the three electrodes are graphite printed in a 3D printer,2 the working
electrode, the pseudo-reference electrode and the counter electrode. Analyzes were performed
using voltammetric techniques, such as cyclic voltammetry and square wave voltammetry. For the
measurements, a buffer solution BR (Britton Robinson) 0.1 mol L-1 was used to carry out an
optimization of the carbon electrode and for the other measurements, a standard solution of Pb 1000
mg L-1 in solution was used. of 0.1 mol L-1 HCl. To verify if the technique used has good selectivity,
measurements were carried out at different concentrations, obtaining a calibration curve and the
equation of the straight line with a coefficient of determination close to 1.0, as can be seen in Figure
1. From the standard deviation and the angular and linear coefficient, the detection and quantification
limit were calculated to determine the lowest concentration of the analyte that can be measured at
an acceptable level of confidence by the method performed. Addition and recovery measurements
were performed with lead ion in different water samples, namely: tap water and mineral water, thus
confirming the electrode efficiency with a good recovery percentage.
Figure 1. Square wave voltammogram (SWV) for a solution of 2.0 mg L-1 lead in 0.1 mol L-1 HCl. Conditions: conditioning
potential of +0.4 V, conditioning time of 20 s, Ei = -1.25 V and Ef = 0.0 V, potential step of 5 mV and amplitude of 70 mV.
1 Costa LM, Ribeiro ES, Segatelli MG, Nascimento DR, Oliveira FM, Tarley CRT, Spectrochimica Acta, 66, 2011, 329.
2 Petroni JM, Neves MM, Moraes NC, Silva RAB, Ferreira VS, Lucca BG, Analytica Chimica Acta, 1167, 2021, 338566.
[UFGD, UFMS, CNPq]

33
INVESTIGATIONS OF THE RELATIONSHIP BETWEEN STRUCTURE AND
ELECTROCHEMICAL PROPERTIES OF FULLERENOL NANOPARTICLES
João Paulo Vita Damasceno (R)a*, Neuryelen dos Santos Bandeira (PG)a, Lauro Tatsuo
Kubota (R)a
a Institute of Chemistry, Department of Analytical Chemistry, University of Campinas, Campinas, São Paulo, Brazil,
13084-971
*e-mail: joaopvd@unicamp.br
Fullerenols are fullerene derivatives enriched on hydroxyl groups that are usually obtained as
nanoparticles dispersed in water. These materials are used in energy-related and biological
applications, and they have been reported as radical scavengers, antioxidants, and stable radicals.
However, most syntheses demand highly toxic solvents to dissolve precursor fullerene, potentially
dangerous reagents such as hydrogen peroxide, and prolonged reactions. Herein, a novel and faster
procedure for fullerenol preparation based on the reaction between fullerene C60 and potassium
hydroxide in a polar solvent is reported,1 and the essential electrochemical characterization of the
obtained materials. Initially, fullerenol nanoparticles with diameters up to 90 nm were prepared.
These nanoparticles are composed of low-modified fullerenols, as probed by thermogravimetric
analysis and x-ray photoelectron spectroscopy, and they remain dispersed indefinitely in the organic
medium due to electrostatic and solvation colloidal interactions, both generated by functional
groups.1 These stable inks were deposited over glassy carbon electrodes (GCE) by drop casting and
dried inside a fume hood. A control electrode was prepared by drop casting pristine fullerene C60
dissolved in toluene over GCE. Both materials were analysed by cyclic voltammetry using different
potential ranges and electrolytic media. An electrochemical reduction was conducted between 0 and
-1.5 V in an aqueous KCl solution. Pristine fullerene exhibits a single and intense peak in the first
cycle at about -1.37 V, which disappears in the next cycles. This peak is attributed to the reduction
of epoxide groups present in pristine fullerene samples stored under ambient atmosphere.
Fullerenols present intense and broad peaks between -0.5 and -1.5 V, which are attributed to the
electrochemical reduction of functional groups, mainly epoxy and peroxy groups, as reported by
Chng and Pumera for the electrochemical reduction of graphene oxide.2 All fullerenols present higher
electrochemical capacitance in relation to pristine fullerene, probably due to the higher surface area
of these nanostructured electrodes. The slight change in the voltametric profile of pristine fullerene
recorded in 5 mmol L-1 [Fe(CN)6]4-/3- (in 0.5 mol L-1 KCl) after electrochemical reduction suggests
preservation of the electroactive sites. On the other hand, fullerenols exhibit entirely different
behaviour. Before electrochemical reduction, these materials almost did not respond to the redox
probe, most probably due to lower electrical conductivity and negatively charged surface. After
reduction, GCE modified with fullerenols has presented the redox couple of [Fe(CN) 6]4-/3- between
about 0.167 and 0.342 V with current intensity next to 7 μA. In comparison, the bare GCE electrode
has signals between about 0.22 and 0.30 V with current intensities close to 80 μA in the same
conditions. Even after electrochemical reduction, such absence of electrocatalytic activity is probably
related to incomplete reduction, which contributes to keeping the electrical conductivity relatively low
and preserves some functional groups on the material's surface.3 The prepared fullerenol inks are
promising for electrochemical applications since they present electrocatalytic activity towards other
redox-active species.
1 Damasceno JPV, Kubota LT, submitted.
2 Chng ELK, Pumera M, Chemistry - An Asian Journal, 6, 2011, 2899.
3 Ambrosi A et el., Chemistry - An European Journal, 17, 2011, 10763.
[FAPESP, INCTBio, CAPES, CNPq]

34
LASER SCRIBING INDUCES ALUMINA AGGREGATES AND GRAPHENE-LIKE
NANOSTRUCTURES ON ABRASIVE CELLULOSE SUBSTRATE
Shellyda G. G. Souza (UG)a,b*, Habdias A. Silva-Neto (PG)b, Danielly S. Rocha (PG)b, Thiago
R. L. C. Paixão (R) c, Wendell K. T. Coltro (R)b,d
a Universidade Federal de Goiás, Faculdade de Farmácia, Goiânia, Goiás, Brazil, 74605-170
b Universidade Federal de Goiás, Instituto de Química, Goiânia, Goiás, Brazil, 74690-900
c Universidade de São Paulo, Instituto de Química, São Paulo, São Paulo, Brazil, 05508-000
d Instituto Nacional de Ciência e Tecnologia de Bioanalítica, Campinas, São Paulo, Brazil
*e-mail: shellydasouza@discente.ufg.br
Laser induced cellulose-based materials have been successfully explored for various sensing
applications1. There are role examples of paper-based substrates commercially available with
enormous potential to be converted to graphene-like structures. Here, we describe for the first time
a laser scribing protocol aiming to create graphene-like electrodes on abrasive paper substrates.
Electrochemical paper-based analytical devices (ePADs) were manufactured by using a combination
of CorelDraw software and a CO2 laser cutter tool. As proof of concept, the proposed device was
coupled to a voltammetric technique for measuring tadalafil (TAD) in authentic tablet samples. For
manufacturing the ePAD, a sand-paper substrate was laminated against a polyester sheet and
pyrolyzed by laser beam (Figure 1). Next, the desired geometry of the electrochemical cell was
delimited (~0.2 cm2) through a glass varnish binder, incorporated the regions electrical contact trail
and reference electrode (RE) with Ag ink and exposed to electrochemical pre-treatment. For realizing
the morphological, structural, and electrochemical characterizations, high-resolution scanning
electron microscopy (SEM), energy dispersive spectroscopy, Raman spectroscopy, contact angle
(AC) and cyclic voltammetry (CV) were carefully performed. The obtained morphological analysis
indicated that the pyrolyzed paper exhibited non-oriented porous structures like popcorn anchored
with alumina oxide nanoparticles (Ø = 60 ± 5 nm). Through the Raman spectra, it was possible see
the respective band D and G at 1333 and 1593 cm-1, wherein the value of ID/IG ~ 0.79 that can be
associated to graphene-like material. Measurements in terms of CA (69 ± 3º) and electrochemical
responses in the presence of [Fe(CN)6]4–/3– also indicated a hydrophobic characteristic and diffusion-
controlled mass transfer upon the conductively surface, respectively. The electrochemical sensing
of TAD based on differential pulse voltammetry (DPV) revealed values of linear range from 25 to 250
μmol L-1 (R2 = 0.99), limit of detection ~ 9.6 μmol L-1 and acceptable reproducibility (RSD ≤ 10%).
The DPV responses involving the standard addition method in the presence of pharmaceutical
samples exhibited recovery results of TAD ranged from 93 to 108%. We successfully demonstrated
the fabrication of graphene-like sites and nanoparticles composed of alumina upon abrasive paper
and believe that the pyrolyzed electrodes can be explored for electroanalytical application.
Figure 1. (A) The pyrolyzed electrode; (B) SEM image and (C) EDS of the nanoparticle; (D) DPV responses in the
absence and presence of TAD.
1 Araujo WR et al. Angewandte Chemie, 129, 2017, 15309.
[INCTBio, CNPq, LabMic, USP, UFG, GME]

35
COMBINING CONDUCTIVE GRAPHITE-BASED INK AND PAPER SUBSTRATE
TO MANUFACTURE DISPOSABLE ELECTROCHEMICAL DEVICES
Laísa C. Oliveira (UG)a,b*, Danielly S. Rocha (PG)b, Habdias A. Silva-Neto (PG)b,
Wendell K. T. Coltro (R)b,c
a Universidade Federal de Goiás, Faculdade de Farmácia, Goiânia, Goiás, Brazil, 74605-170
c Instituto Nacional de Ciência e Tecnologia de Bioanalítica, Campinas, SP, Brazil
*e-mail: laisaoliveira@discente.ufg.br
The transfer process of conductive ink onto substrate materials can be realized through several
manufacturing techniques including the stencil-printing strategy, which is one of the simplest
methods. However, the freshly literature has been reported few examples of conductive inks to
electrochemical sensing finality. The purpose herein study aims to demonstrate the feasibility of the
development of carbon-based ink and electrodes on paper substrate for pharmaceutical application.
Electrochemical devices were constructed using the stencil-printing approach with three different ink
(graphite:polymeric binder) proportions (40:60, 50:50 and 60:40%; w/w), which were incorporated
on waterproof paper substrate. Afterwards, cyclic voltammetry (CV) measurements were carried out
in the presence of redox probe [Fe(CN)6]4-/3-. The obtained values of anodic peak current intensity
(ipa) were 30.3 ± 0.5; 39.5 ± 2 and 46.8 ± 2 μA for the proportion 40:60, 50:50 and 60:40%,
respectively. The peak-to-peak separation (ΔEp) and ratio of anodic-to-cathodic peak current signal
(ipa/ipc) ranged from 160 to 226 mV and 0.9 to 1.0, respectively. The achieved result showed that the
best electrochemical parameters were obtained in the ink proportion of 50:50% (w/w). Structural,
morphological, and electrochemical characterization upon the optimized surface was performed
through Raman spectroscopy, scanning electron microscopy (SEM), contact angle (CA) and CV,
respectively. The above-mentioned analysis indicated that the graphite ink was successfully
anchored on paper substrate with predominance of carbon hybridization sp2, well-distributed
particles flakes, CA = 79° ± 3 and heterogeneous electron transfer rate (Ks) = 0.63 x 10-3 (± 0.01)
cm s-1. Considering the electrochemical sensing, paracetamol (PAR) was selected as model analyte
and exhibited well-defined peak oxidation at 0.8 V vs Ag and diffusion-controlled electron transfer.
The square wave voltammetry (SWV) responses for the detection of PAR provided limit of detection
and linear range of 0.2 µmol L-1 and 1 – 60 µmol L-1 (R2 ≥ 0.99), respectively. The selectivity
experiment was also evaluated in the presence and absence of some potential interferences
involving the oxidation of PAR and the calculated interference ranged from 0.7 to 8%. Finally, three
different pharmaceutical products were analyzed and the achieved amount of PAR was 761, 733
and 500 mg. Comparative analysis by performance liquid chromatography (HPLC) demonstrated
agreement with the label values and statistically similar (test t; confidence level 95%). Thus, we
introduced how is simple and affordable the combination of conductive ink and paper-based
substrate for creating disposable electrodes dedicate to selective analysis PAR in pharmaceutical
samples. Future studies will be also dedicated for performing pharmacokinetic analysis.
Figure 1: (A) Sensor. (B) WE com ampliação 1000x. (C) Representation of the PAR oxidation reaction and CV of the study
of the effect of the variation of the sweep speed on the peak current intensity using PAR 1 mM in acetate buffer and plot
of the anodic peak current intensity as a function of the square root of the sweep speed. (D) SWV for different PAR standard
concentrations PAR analytical curve.
1 Silva-Neto HA, Cardoso TMG, McMahon CJ, Sgobbi LF, Henry CS, Coltro WKT. Analyst, 146, 2021, 3463.
[CNPq, CAPES, INCTbio, Labmic, UFG, GME]

36
USE OF GOLD LEAF FOR MODIFYING DISPOSABLE ELECTROCHEMICAL
SENSORS FOR L-TYROSINE DETECTION
Sabrina Cletoa*, Amanda Neumann (UG)a, Jéssica R. Camargoa, Bruno C. Janegitz (R)a
a Department of Nature Sciences, Mathematics and Education, Federal University of São Carlos, Araras, São Paulo,
Brazil, 13600-970.
*e-mail: sabrina.cleto@estudante.ufscar.br
The development of new sensory architectures using everyday materials can be a viable alternative
to carry out analyzes of interest in diverse areas1. The use of sensors for quality control of food
supplements is interesting, since only supplements containing enzymes or probiotics must be
registered by Anvisa2. The present work aimed at the preparation of a disposable sensor based on
a conductive ink, obtained from the mixture of nail polish (NP) and powdered graphite (GP), on a
substrate of acetate sheets (AS). In this three-electrode system, the working electrode was modified
with 24K gold leaf (Figure 1). Electrode modification using Au is widely studied for technology
development with a wide variety of applications in the fields of physics, chemistry, catalysis,
technology, biomedicine and materials science3. Due to its intrinsic properties such as electronic,
optical, magnetic and thermal, as well as catalytic properties, this material is considered vital for
technological advances4. However, the high cost involved with gold can be surpassed using a gold
in thin leaf. In this way, the developed sensor acquired the interesting properties of gold and still has
an affordable price to be produced on a large scale. The constructed electrode was named Au-GP-
NP/AS and electrochemical characterization measurements were performed. Given the possibilities
of analytes, L-Tyrosine was chosen as it is an amino acid used as a nutritional supplement5. In the
body, it has a primordial function in the brain, since it acts in the synthesis of dopamine, which is
responsible for mood, sleep and attention6. Therefore, with an electrode made from accessible
materials and an architecture of easy production, relatively low cost, simplified and miniaturized, it
was possible to detect L-Tyrosine in initial tests. In addition, the surface modification presented, in
the future may be used for the development of new sensors and biosensors for the detection of
analytes of interest.
Figure 1. Photograph of electrodes Au-NP-GP/AS.
1 Steinberg MD; Kassal P, Steinberg IM. Electroanalysis, 28, 2016, 1149.

2 ANVISA, 2020. Available in: <http://antigo.anvisa.gov.br/documents/33916/2810640/Suplementos+Alimentares/
a6fd2839-6d80-496a-becb-8b2122eff409>
3 Xiao T et al., 206, 2020, 120210.
4 Cobley, Claire M. et al. Chemical Society Reviews, v. 40, n. 1, p. 44-56, 2011.
5 Lütke-Eversloh T, Santos CNS, Stephanopoulos G, Applied microbiology and biotechnology, 77, 2007, 751.
6 Meeusen R et al. Applied Physiology, Nutrition, and Metabolism, 32, 2007, 857.

37
USE OF PRESSURE AND IONIC LIQUID FOR INCREASED PHOTOCATALYTIC
CO2 REDUCTION APPLYING TIO2 NANOTUBES CATALYST DECORATED
WITH GOLD NANOPARTICLES
João Angelo de Lima Perini (R), Kallyni Irikura (R), Lilian D. M. Torquato (R), Maria Valnice
Boldrin Zanoni (R)*
São Paulo State University (UNESP), Institute of Chemistry, Araraquara, São Paulo, Brazil, CEP 14800-060
*e-mail: maria.valnice@unesp.br
CO2 transformation to higher value-added compounds is a topic of great importance nowadays due
to the global warming generated by the increase of CO2 concentration in the atmosphere. Thus, the
present work aims to prepare, characterize, and evaluate the performance of TiO2 nanotubes (TNT)
catalyst modified with Au nanoparticles (NP) during the CO2 photocatalytic reduction. The deposition
of Au NPs onto TNT catalyst was performed by reduction of a solution of chloroauric acid with citric
acid from lime juice adapted from the method of Turkevich et al. 1 The reduction of CO2 was
conducted in a stainless-steel reactor, filled with 150 mL of Na2SO3 solution containing dissolved
CO2 by bubbling this gas in presence of the ionic liquid (IL) BMIM-BF4 up to 4% v/v. After that, the
reactor was sealed and pressurized with CO2 up to 5 atm. The surface of the catalyst was irradiated
using a solar simulator equipped with an AM 1.5G filter and a 300 W Xe arc lamp. The methanol
generation was monitored by GC-FID using a solid-phase microextraction fiber and methane was
monitored by GC-TCD using a gastight syringe. The results obtained showed that the generation of
methanol and methane using TNT catalyst was negligible. On the other hand, in the presence of
TNT Au NP the methanol generation was higher, achieving 57.6 μM after 180 min of reaction.
Therefore, the incorporation of the Au NPs on the surface of TNT catalyst can improve its
photoexcitation since there was a greater absorption in the visible light region. In addition, the work
function of Au is higher than TiO2, thus, when Au NPs and TiO2 are in contact, Schottky barriers is
formed, providing an effective electron trap for the photogenerated electrons, minimizing the
electron-hole recombination. With the increase of in electron life-time, the generation of CO2
reduction products was enhanced. Regarding the pressure, at 1 atm the generation of methanol was
57.6 µM. When the pressure was increased to 3 and 5 atm, the generation of methanol increased to
94.8 and 151.2 µM, respectively, due to the increase dissolution of CO2 in water. The effect of BMIM-
BF4 IL was evaluated by the increase of the percentage of IL from 0 to 4% v/v at 5 atm pressure. In
these conditions, the generation of methanol and methane using 4% v/v of IL was 320.8 μM and
159.2 μM after 180 min of reaction, respectively, which represents an increase of 112.2% (methanol)
and 142.7% (methane) compared to experiments performed in the absence of BMIM-BF4. The
quantum yields for CO2 photo-conversion to methanol and methane calculated during the best
catalytic condition (5 atm and 4% v/v IL) were 0.60% and 0.52%, respectively. An unprecedented
method using NaH13CO2 as a source of 13CO2 was also developed focusing on a deeper analysis of
the CO2 reduction route by photocatalysis. The isotope-labeled studies proved that 13CO2 is the only
source for photoproduction of 13CH3OH using the IL. In conclusion, these findings indicated that the
combination of the Au co-catalyst size in nanometric size, high pressure, and BMIN-BF4 IL can
provide an efficient modulation of CO2 conversion.
1 Turkevich MJ, Stevenson PC, Hillier J, Discuss. Faraday Soc. 11, 1951, 55.
[FAPESP, CNPq]

38
DISPOSABLE SENSOR PREPARED WITH CONDUCTIVE INK BASED CARBON
BLACK AND GLASS VARNISH FOR ELECTROCHEMICAL DETECTION OF 17Α-
ETHINYLESTRADIOL
Gabrielle Lacerda Gouveia (R)a, Jefferson H.S. Carvalho (PG)a,b*, Bruno C. Janegitz (R)a
a FederalUniversity of São Carlos, Laboratory of Sensors, Nanomedicine, and Nanostructured Materials (LSNano),
Araras, São Paulo, Brazil, CEP 13604-900
b Federal University of São Carlos, Department of Physics, Chemistry and Mathematics, Sorocaba, São Paulo, Brazil,
18052-780
*e-mail: jeffersoncarvalho@estudante.ufscar.br
17α-ethinylestradiol is a synthetic hormone, commonly used in female contraceptives, with

pharmacological and chemical characteristics similar to those of natural estrogens [1]. Among the
problems related to its handling, there is an increase in the possibility of developing vascular
diseases in the long term, and, in an environmental nature, it is directly linked to the contamination
of effluents, causing problems to the local ecosystem [2]. On the other hand, the development of
disposable sensors has shown to be increasingly advantageous in analytical applications. This is
due to its advantages compared to other systems such as versatility in compositions, the practicality
of confection, mass production, and relatively low cost [3]. The proposed electrochemical sensor
was made with conductive ink based on carbon black and glass varnish applied to detect 17α-
ethinylestradiol. Carbon black was chosen due to its almost spherical particle structure, which
aggregates to form a structure similar to graphite, corroborating the conductive character [4]. A
compositional study was carried out for a better proportion between the ink components, and their
electrochemical behavior (cyclic voltammetry) and morphology (scanning electron microscopy and
Fourier infrared spectroscopy). Figure 1 shows a cyclic voltammogram in the presence of 1.0×10-3
mol L-1 Ferrocenoethanol in 0.1 mol L-1 KCl and in the absence of the electrochemical probe. The
sensor was submitted to the choice of the best pulse technique (square wave voltammetry), studies
of its parameters (amplitude, frequency, and step), in addition to the pH of the medium.
1 Schleicher, AT. Dissertation (Master in Environmental Technology and Water Resources) - University of Brasília, 2013.
2 Hannah R, et al. Environmental Toxicology and Chemistry, 28, 2009, 2725.
3 Heba M. Mohamed, TrAC Trends in Analytical Chemistry, 82, 2016, 1.
4 Silva TA et al. Journal of Nanomaterials, 2017.

39
SIMULTANEOUS DETECTION OF CARBAMATE PESTICIDES USING AN EASY-
TO-PREPARE ELECTRODE MODIFIED WITH MORINGA OLEIFERA BIOCHAR
Jonatas O. S. Silva (PG)a*, José F. Santos (UG)a, Honnara S. Granja (PG)a,
Wandson S. Almeida (PG)a, Thiago F. L. Loeser (UG)a, Lisiane S. Freitas (R)a,
Márcio F. Bergamini (R)b, Luiz H. Marcolino-Junior (R)b, Eliana M. Sussuchi (R)a
a Federal University of Sergipe, PostGraduate Program in Chemistry, São Cristóvão, SE, Brazil, 49100-000
b Federal University of Paraná, PostGraduate Program in Chemistry, Curitiba, PR, Brazil, 80060-140
*e-mail: osjonatas@hotmail.com
Moringa oleifera is a tree of great availability and its constituent parts have been constantly applied
in the adsorption and removal of inorganic and organic contaminants, such as pesticides, making it
an excellent option for electrode modification1. The sensitive, selective and simultaneous
determination of pesticides in the application medium and environments that can be reached by
them, such as rivers and effluents, using an easy-to-prepare tool based on biochar from moringa
pods husks biomass, configures as an excellent option for minimizing costs and analysis time, in
addition to ensuring determinations of residue values closer to the real, due to the simplicity in the
treatment of samples and data collection2. In the present work we developed an electrochemical
sensor modified with biochar derived from the biomass of moringa pods husks for the simultaneous
determination of the carbendazim and carbaryl levels. Initially, the influence of the presence of
biochar from the moringa pods husks (400 °C) in 10% of the electrode composition (MOB/E) in
relation to an unmodified electrode (CPE) on the analytical signals of the carbendazim (CBZ) and
carbaryl (CBR) was evaluated. The data obtained showed that the modification was responsible for
increasing by about 30% and 70% the analytical response obtained for CBZ and CBR, respectively.
In order to increase the efficiency of the modified electrode, the pesticide analysis parameters were
optimized and under optimized conditions there was an increase of approximately 30 times for the
current intensity of the CBZ and 40 times for the CBR, evidencing the importance of optimization for
the sensitivity of the method. An analytical curve (Figure 1) was obtained in a concentration range
of 0.29 – 6.00 μmol L-1 for CBZ and 29.9 – 502 μmol L-1 for CBR. For the method, values for limits
of detection (0.08/1.41 μmol L-1) and quantification (0.25/4.28 μmol L-1) to CBZ/CBR were obtained,
highlighting the applicability of the sensor for simultaneous determination of CBZ and CBR levels.
Figure 1: (A) Representative differential pulse voltammograms (corrected baseline) obtained in the presence of different
concentrations of CBZ and CBR. Inserted: Magnification of voltammograms obtained for CBZ. (B) Variation of anodic
peak current as a function of CBZ concentration. (C) Variation of anodic peak current as a function of CBR concentration.
1 Gomes HO, Freire PTC, Nascimento RF, Teixeira RNP, Journal of Water Process Engineering, 46, 2022, 102576.
2 Kalinke C, Oliveira PR, Bonacin JA, Janegitz BC, Mangrich AS, Marcolino-Junior LH, Bergamini MF. Green Chemistry,
23, 2021, 5272.
[SEnM, LABSENSE, LAC, PPGQ, CLQM, SergipeTEc, CAPES, CNPq, FAPITEC/SE]

40
ELECTROCHEMICAL SENSOR BASED ON GRAPHENE QUANTUM DOTS FOR
DETERMINATION OF ESTRONE IN DIFFERENT SAMPLES
José F. Santos (UG)a*, Jonatas O. S. Silva (PG)a, José F. Macedo (PG)a,
José C. S. Júnior (PG)a, Wandson S. Almeida (PG)a, Eliana M. Sussuchi (R)a
a
Federal University of Sergipe, Department of Chemistry, São Cristóvão, SE, Brazil
*e-mail: jfelipequim@gmail.com
Hormones such as estrone (E1) and estradiol (E2) interact with the body's metabolism by activating
or blocking hormone receptors, hindering production and causing hormone degradation. High levels
of E1 and E2 are associated with an increased possibility of breast cancer and prostate cancer1.
Thus, it is necessary to monitor these species. Nanomaterials such as graphene quantum dots
(GQD) may present some characteristics such as high surface area and good electrical conductivity,
which makes them excellent electrode modifiers2. In this work, an electrode modified with GQD
(GQD/E) was developed for the voltammetric determination of E1. The analysis parameters were
optimized and the results obtained for the GQD/E show an increase in the sensitivity of the proposed
sensor, highlighting the importance of optimizing the parameters of the analytical method developed
for the determination of E1 in complex samples. An analytical curve was constructed with a linear
range of 0.05 – 10.00 µmol L-1 (R2=0.997), with limits of detection (LOD) and quantification (LOQ) of
28.00 nmol L-1 and 96.00 nmol L-1, respectively. The effect caused by the presence of concomitant
species in the analysis medium on the analytical response of the method was investigated to
evaluate the selectivity of the sensor to substances that may be in the matrix. Figure 1a shows that
in E1:Concomitant (1:10) for leucine (LE) and urea (UR) and in the ratio (1:100) for glycine, citric
acid (CA), Na+, K+, Fe+2, Ca+2, NH4+, Mg+2, Cu+2, NO3-2, HCO3-, CO3-2, Cl- and SO4-2, which do not
respond significantly (>10%) in the analytical signal of E1. The GQD/E was used to determine E1 in
fortified samples of synthetic urine, wastewater, sea water and tap water. Voltammetric
measurements were performed (Figure 1b) and through linear regressions (Figure 1) recovery
values were obtained at three different concentration levels for all samples with recovery values
between 95.9 - 108.1 %, which highlights the applicability of the proposed sensor to selectively
determine estrone in different samples.
Figure 1. a) Variation of the analytical signal of E1 in the presence of different concentrations of organic and inorganic
species. b) Representative cyclic voltammograms obtained for the additions of wastewater sample fortified with E1 in the
concentration range of 0.19 to 0.60 μmol L-1. c) Linear correlation between Iap and E1 concentration.
1 Wu CC, Shields JN, Akemann C, Meyer DN, Connell M, Baker BB, Pitts DK, Baker TR, Science of the Total Environment,
757, 2021,143736.
2 Sant’Anna MVS, Silva JOS, Gevaerd A, Lima LS, Monteiro MDS, Carregosa ISC, Wisniewski A, Marcolino-Junior LH,
Bergamini MF, Sussuchi EM, Food Chemistry, 373, 2022, 131521.
[CAPES, CNPq, FAPITEC/SE, UFS, SEnM, CLQM, PETROBRAS, SergipeTec]

41
ELECTROCHEMICAL DETECTION OF SARS-COV-2 USING A LOW-COST
BIOSENSOR COMPOSED OF PENCIL GRAPHITE ELECTRODES
Lucas Felipe de Lima (PG)a*, André Lopes Ferreira (R)a, William Reis de Araujo (R)a
a State University of Campinas - UNICAMP, Portable Chemical Sensors Lab, Department of Analytical Chemistry,
Institute of Chemistry, P.O. Box 6154, 13083-970, Campinas, SP, Brazil.
*e-mail: lucasflima14@gmail.com
Since its onset, the COVID-19 pandemic has led to more than 6.32 million deaths and cost to world
economy billions of dollars1. Different methods using reverse transcription-polymerase chain
reaction (RT-PCR) and serology tests have been used to diagnose SARS-CoV-2 in saliva and
nasopharyngeal/oropharyngeal (NP/OP) swab samples. RT-PCR has served as the gold standard
test during the pandemic because of its high accuracy and specificity. However, this technique is
time-intensive, costly, and requires highly skilled workers for operation, thus making it not a suitable
solution for mass testing, especially in developing countries where resources are limited.
Most rapid tests commercially available for diagnosing COVID-19, which are antigen-based, exhibit
low sensitivity, which limits their ability to accurately identify asymptomatic individuals, thus hindering
proper control of viral spread. Therefore, there is an urgent need to develop low-cost, sensitive, and
rapid tests for the early diagnosis of SARS-CoV-2 infections. In an attempt to overcome such
limitations, several portable devices have been developed offering rapid and decentralized clinical
diagnosis2,3. However, these electrochemical, colorimetric, serological, and molecular-based
technologies are limited by their relatively high cost (>$10 per test), need for materials and equipment
that are not easily accessible, the requirement of specialized labor, and the fact that they are time-
consuming and not sufficiently rapid. Here, we present a low-cost electrochemical biosensor for rapid
COVID-19 diagnosis that uses highly accessible and inexpensive materials. The do-it-yourself (DIY)
test consists of a transducer made of 0.5mm graphite leads modified with gold nanoparticles
(AuNPs) and human angiotensin-converting enzyme 2 (ACE2) receptor inserted in a plastic vial for
sample collection and analysis (electrochemical cell). This testing device enables on-site SARS-
CoV-2 detection within 6.5 min, faster than currently FDA-approved tests. Each biosensor unit can
be manufactured for a total cost of $1.50, including the vial ($0.30), graphite leads ($0.20), and all
the modifiers required to ensure high sensitivity ($1.00), including glutaraldehyde (GA), AuNPs,
cysteamine (cys), ACE2, and bovine serum albumin (BSA). The graphite working electrode was
modified with AuNPs stabilized with cys to allow anchoring of ACE24. BSA was used to block the
remaining active sites on the electrode surface to avoid non-specific interactions between the clinical
sample and the biosensor. The detection principle is based on the current suppression of a redox
probe (5 mmol L-1 [Fe(CN)6]-3/-4) induced by the SARS-CoV-2 spike protein recognition and detected
by Square Wave Voltammetry (SWV) technique, resulting in a limit of detection for the viral spike
protein of 229 fg mL-1. The proposed method was applied to 110 clinical samples and provided
100.0% sensitivity, 100.0% specificity, and 100.0% accuracy for 10 saliva samples analyzed and
88.7% sensitivity, 86.0% specificity, 87.4% accuracy for 100 OP/NP samples. In summary, we
describe a low-cost and rapid COVID-19 test that is easy to assemble and may enable high-
frequency testing. Due to its reduced cost and DIY design, our method provides an opportunity to
increase access to diagnostic testing for underserved communities.
1 Lima LF, Ferreira AL, Torres MDT, Araujo WR, Fuente-Nunez Cla, Proc. Natl. Acad. Sci. U. S. A., 118, 2021, 1.
2 Drain PK, Hyle EP, Noubary F, Freedberg KA, Wilson D, Bishai WR, Rodriguez W, Bassett IV, Lancet Infect. Dis., 14,
2014, 239.
3 Song Y, Zhang Y, Bernard PE, Reuben JM, Ueno NT, Arlinghaus RB, Zu Y, Qin L, Nat. Commun., 3, 2012, 1.
4 Yan R, Zhang Y, GuoY, Xia L, Zhou Q, Science, 80, 2020, 2762.
[CAPES, FAPESP, CNPq, University of Pennsylvania (UPENN), Cesar de la Fuente-Nunez]

42
TITANIUM DIOXIDE-COBALT MODIFIED ON CARBON BASED-ELECTRODES
FOR SELECTIVE VOLTAMMETRIC DETECTION OF AMOXICILLIN
Leyllanne K. A. Souza (PG)a*, Danielly S. Rocha (PG)b, Habdias A. Silva-Neto (PG)b, Fabiane
C. de Abreu (R)c, Wendell K. T. Coltro (R)b, Anielle C A. Silva (R)c, Paulo T. Garcia (R)a
a UNIFESSPA, institute of Science, Marabá, Pará, Brazil, CEP 68507-590
b UFG,Institute of Chemistry, Goiânia, Goiás, Brazil, CEP 74690-900
c UFAL, Departament of Chemistry, Maceió, Alagoas, CEP 57072-900
*e-mail: leylanearaujo@gmail.com
Amoxicillin (AMX) is an antibiotic broadly used around the world. In the last 7 years, ca. 800 million
antibiotic products were commercialized in Brazil, and 32% represented the consumption of AMX.
Approximately 60% of the active principle of AMX is excreted in the urine, indicating that it is
important to propose analytical methods for the quality control of AMX as well as in its environmental
monitoring.1,2,3 However, manufacturing selective electrochemical sensors to molecule AMX is
challenging. Here, we propose the combination of “do it yourself” stencil printed carbon electrodes
(SPCE) and nanocomposites based on titanium dioxide-cobalt for the selective analysis of AMX in
pharmaceutical samples. The geometry of SPCE consist to conventional three electrodes: working
(WE), reference (RE) and auxiliary (AE). Drop casting approach was successfully utilized for modify
TiO2-Co on electrodes surface. Detailed morphological, structural and electrochemical
characterization upon the electrodes surface were carefully realized. Considering the chemical and
square wave voltammetry (SWV) conditions, the detection of AMX was performed in the presence
of 0.1 mol L-1 PBS at pH 7.00 by using the parameters: frequency, step, amplitude, deposition time
and deposition potential of 5 Hz, 0.004 V, 0.04 V, 10 s and 0.0 V vs Ag, respectively. The
characterization analysis showed that the anchored TiO2-Co nanoparticles was totally distributed on
WE surface with size values of 30 ± 7 nm (Figure 1). The repeatability studies (intra-day and inter-
day; n=3) of the modified electrodes realized in the presence of 1.0 mmol L-1 of AMX exhibited values
of relative standard deviation (RSD) were 8.1 and 5.8 %, respectively. In terms of sensing
performance of AMX, the calculated values of limit of detection (LOD = 5.8 mol L-1) and
quantification (LOQ = 19.5 mol L-1) were calculated in linear range from 20 to 150 mol L-1 (R2 =
0.99, Figures H-I). The SWV experiments of selectivity indicated low interference (≤15%) to the main
compounds traditionally found in pharmaceutical formulations. Through the data obtained from the
reference analysis by using a UV-visible spectrophotometer and by the voltammetric sensor, it is
possible to infer that they did not reveal statistical difference in both analyzes at 95% confidence
level, thus indicating that the proposed modified sensor can be performed for detecting of AMX in
pharmaceutical samples. We believe that the proposed combination of carbon-based electrodes
incorporated with TiO2-Co emerging as novel and attractive sensor for selective analysis of AMX in
pharmaceutical products.
Figure 1. (A) Layout of SPCE. Obtained SEM images of WE (B), (C), (D) before and (E), (F), (G) after the modification
process. (H) SWV experiments in the absence and presence different concentrations of AMX. a) blank; b) 20; c) 25; d) 50;
e) 75; f) 100; g)125 and h) 150 mol L-1 AMX. (B) Analytical curve.
1 Wong A et al. Talanta 206, 2020, 120252.

2 Hrioua A et al. Bioelectrochemistry 137, 2021,107687.
3 Del Fiol FS et al. Frontiers in Pharmacology, 13, 2022, 1.
[CAPES, CNPq, LaSAP, LEMAN, GME laboratories]

43
ELECTROCHEMICAL SENSOR BASED-CARBON BLACK MODIFIED WITH
MIXED OXIDE SiO2/TiO2/Sb2O5 FOR DIRECT DETERMINATION OF
THIAMETHOXAM IN RAW HONEY AND WATER SAMPLES
Raísa Rodrigues Gioia (PG)a, Julia Oliveira Fernandes (PG)a, Cassiano Augusto Rolim
Bernardino (PG)b, Bernardo Ferreira Braz (PG)a, Claudio Sabbatini Capella Lopes (PG)a,
Ricardo Erthal Santelli (R)a,c, Fernando Henrique Cincotto (R)a,c*
a Universidade Federal do Rio de Janeiro, Departamento de Química Analítica, Rio de Janeiro, RJ, Brazil, 21941-909.
b Universidade Federal do Rio de Janeiro, Programa de Engenharia Civil, Rio de Janeiro, RJ, Brazil, 21941-909
c Instituto Nacional de Ciência e Tecnologia em Bioanalítica, Campinas, SP, Brazil, 13083-970
*e-mail: fernandocincotto@iq.ufrj.br
The use of neonicotinoid pesticides in agriculture has been highlighted over the years and currently
represents more than a quarter of all insecticides used globally to increase agricultural production1.
Thiamethoxam (TMX) is a 2nd generation neonicotinoid pesticide with a half-life in soil from 7 to 109
days, whose name IUPAC is 3-(2-chloro-1,3-thiazol-5-ylmethyl)-5-methyl-1,3,5-oxadiazinan-4-
ylidene(nitro)amine2. In Brazil, TMX is an agrochemical classified as moderately toxic to the
environment, and routinely used in farmlands to produce fruits, vegetables, among other edible
plants. Maximum residue limits (MRLs) are adopted to control the use of pesticides on crops. This
parameter varies according to the regulatory agency in each country. In Brazil 3, TMX in farmland
production, can vary from 0.01 to 1.0 mg kg-1. In the United States4, the values of MRLs for TMX
range from 0.02 to 20 mg kg-1 and European Union range from 0.01 to 5 mg kg-1. Currently, the
European Union Commission5 has restricted the use of thiamethoxam to seed treatment, soil
treatment, and regarding certain specific crops foliar application can only be performed before
flowering, due to strong toxicity to bees. There are extensive reports in the literature attesting the
fact that the use of TMX as farmland pesticides, can lead to its accumulation by plants, and
eventually promote the contamination of the pesticide in the pollen and nectar of the plant, resulting
in a significant reduction of the bee population. In addition, this pesticide is highly toxic to all aquatic
arthropods, except for water fleas. Electrochemical sensors can be applied to quantify traces of
contaminants from agricultural activities. However recently, nanomaterials derived from carbon black
have been used to produce sensors to quantify pesticides in environmental and food samples. The
study aimed to develop an electrochemical sensor based on glassy carbon, mixed oxide
(SiO2/TiO2/Sb2O5), and carbon black. The material was synthesized, characterized, and used to
determine thiamethoxam in raw honey and water. The morphologic structure and electrochemical
performance of the sensors were characterized by scanning electron microscopy and cyclic
voltammetry. Differential pulse voltammetry with a concentration of 0.1 mol L -1 of Britton-Robinson
buffer at pH 7.0 allowed the generation of a method to determine thiamethoxam in a linear range of
0.25 to 100.5 μmol L-1 and with a limit of detection of 0.012 μmol L-1. The system efficiently quantified
traces of thiamethoxam in raw honey and tap water samples. The modified sensor did not present
interferences for K+, Na+, Ca2+, Mg2+, glyphosate, imidacloprid and carbendazim. In addition, the
device showed good recovery values for thiamethoxam when applied directly on honey and water
samples without any treatment, which suggests the development of an electrochemical sensor to
monitor in real time hazardous substances in environmental and food matrices.
1 Sanaullah M et al., Soil and Tillage Research, 196, 2020, 104464.
2 Maienfisch P et al. Pest Management Science, 57, 2001, 906.
3 Agência Nacional de Vigilância Sanitária, T48 - Tiametoxam., 2021.
4 Code of Federal Regulations, 2014, PART 180 - Tolerances and exemptions for pesticide chemical residues in food.
5 European Food Safety Authority, 2018, Peer review of the pesticide risk assessment for bees for the active substance
thiamethoxam considering the uses as seed treatments and granules.
[CNPq, FAPERJ]

44
NEW CONDUCTIVE INK BASED ON CARBON NANOTUBES FOR THE
CONSTRUCTION OF A DISPOSABLE ELECTROCHEMICAL SENSOR
Jefferson H.S. Carvalho (PG)a,b*, Bruno C. Janegitz (R)a
a Laboratory
of Sensors, Nanomedicine, and Nanostructured Materials (LSNano), Federal University of São Carlos
(UFSCar), Araras, São Paulo, Brazil, CEP 13604-900
b Department of Physics, Chemistry and Mathematics, Federal University of São Carlos, Sorocaba, São Paulo, Brazil,
18052-780
*e-mail: jeffersoncarvalho@estudante.ufscar.br
As a complementary alternative to established analytical methods, disposable electrochemical

sensors have gained prominence over the years for their diversity of manufacture and application,
relatively low production cost, low consumption of reagents, and the possibility of miniaturization of
conventional electrochemical systems [1, 2]. In line
with the new sensory platforms with the possibility of
disposal, conductive inks based on conductive
nanoparticles have gained notoriety for the wide range
of new materials for manufacturing electrochemical
devices [3]. In this study, we present the results
obtained for the construction of a disposable
electrochemical sensor, based on a conductive ink of
carbon nanotubes and alkyd resin, deposited on a PET
(Polyethylene terephthalate) substrate of beverage
bottles are presented. An in-depth study was carried
out for the ink composition, such as the amount of ideal
conductive material and the use of solvent.
Electrochemical characterizations (cyclic voltammetry)
and morphological characterizations (scanning
electron microscopy and Fourier transform infrared
spectroscopy) were used for characterizations. Figure
1 shows the method of conductive ink preparation and
electrode proposition. 3-Nitro-L-tyrosine was used as
the analyte of interest. Elevated levels of this analyte Figure 1. production steps for the construction
in biological fluids are related to the probability of of the electrochemical sensor and its finalized
developing neurodegenerative diseases and, real image.
therefore, need to be quantified for possible early
diagnosis of diseases [4]. This study revealed a linear range from 0.70 to 100 mol L− and a
detection limit (LOD = 3×SDblank/slope) equal to 0.53 mol L− of 3-nitro-L-tyrosine, in 0.1 mol L− of
phosphate-buffered saline (PBS) (n=10). The sensor was applied to a urine sample by the addition
and recovery method, obtaining recoveries between 95.2 and 106%.
1 Guth U, Vonau W, Zosel J, Measurement Science and Technology, 20, 2009, 042002.
2 Maduraiveeran G, Sasidharan M, Ganesan V, Biosensors and Bioelectronics, 103, 2018, 113.
3 Cummins G, Desmulliez MP, Circuit world, 2012.
4 Roy E, Patra S, Madhuri R, Sharma P K, Talanta, 132, 2015, 406.

45
DEVELOPMENT OF SCREEN-PRINTED ELECTRODE BASED ON CARBON
NANOMATERIALS FOR SIMULTANEOUS ANALYSIS OF PHENOLS IN WATER
Gabriele Smanhotto Malvessi (PG)a*, Thaynara Dannehl Hoppe (PG)a, Daniela Brondani (R)a
a Universidade Federal de Santa Catarina, Departamento de Ciências Exatas e Educação, Campus Blumenau, Santa
Catarina, Brasil, 89036-256.
*e-mail: gabimalvessi@gmail.com
Screen-printed electrochemical sensors have received great attention in recent decades because of
their attractive features, such as easy to use, portable, high selectivity and sensitivity, fast results,
reduced sample volume, relatively low cost, and they can also be disposable.1 These printed devices
have been built and/or modified with different nanomaterials to improve their performance in sensing
applications.2 In this context, it was proposed to develop screen-printed sensors produced in the
laboratory using conductive inks based on carbon nanomaterials for application in the detection of
environmental pollutants. For this, the silk-screen technique was used in the construction of
electrochemical sensors, with 2 types of design: SPE (Screen-Printed Electrode) and SPETA
(Screen-Printed Electrode Type Alligator) (Figure 1). For the construction of the electrodes, 40
compositions of homemade inks were tested, varying the constituents of the formulation, the
percentage of conductive material and polymeric material, and the addition of solvent. Initially, the
conductive inks were evaluated using the ferrocyanide-ferricyanide redox couple (Fe(CN)63-/4-), and
the most promising inks were selected for characterization and analytical performance evaluation
using phenolic compounds of
environmental interest. The
characterization analysis of the
screen-printed electrodes
produced with the selected inks
were carried out by voltammetric
techniques, electrochemical
impedance spectroscopy, Fourier-
transform infrared spectroscopy
and scanning electron microscopy.
From the evaluation of the
voltammetric response to toxic
phenolic compounds, the best
results were obtained with the T26
ink, composed of powder
nanographite, graphene Figure 1. Schematic illustration of the production of screen-printed
nanoplatelets and glass varnish electrodes with different designs, using homemade conductive inks based
(40:10:50 wt%). Studies were on carbon nanomaterials for environmental applications.
carried out to optimize the
experimental conditions (supporting electrolyte and square wave voltammetry parameters) for the
detection of bisphenol A (BPA), catechol (CC) and 4-nitrophenol (4-NP). Under optimal conditions,
calibration curves were constructed (individually and simultaneously), from which linear response
ranges for BPA, CC and 4-NP were 2,5–200 µmol L-1, 10–200 µmol L-1 e 10–200 µmol L-1,
respectively, with limits of detection (LOD) of 1,7 µmol L-1, 6,9 µmol L-1 and 2,8 µmol L-1. Sensors
produced with the selected conductive ink (T26) were evaluated for their analytical performance,
including stability, reproducibility and potential interference. Finally, the proposed sensors were
applied in the analysis of water samples fortified with phenolic pollutants, presenting an acceptable
relative error (not greater than 10.0%).
1 Ambaye AD, Kefeni KK, Mishra SB, Nxumalo EN, Ntsendwana B, Talanta, 225, 2021.
2 Musa AM, Kiely J, Luxton, R, Honeychurch KC, TrAC Trends in Analytical Chemistry, 139, 2021.
[CNPq, FAPESC, UFSC-Blumenau, LAEES, CReATe]

46
DEVELOPMENT OF ELECTROCHEMICAL SENSOR USING SILVER BIOGENIC
NANOPARTICLES APPLIED TO Fe DETERMINATION
Rafaela Vendramin Comunello (UG)a*, Mônika G. Heinemann (PG)a, Daiane Dias (R)a
aFederal University of Rio Grande, School of Chemical and Food, Rio Grande, Rio Grande do Sul, Brazil, 96203-900
*e-mail: rafacomunello@gmail.com
Silver nanoparticles synthesized with C. pyrenoidosa (AgNPs/CP) have excellent chemical, electrical
and biological properties, as well as present high adsorption, reactivity and conductivity.1 Therefore,
the use of this material in the development of electrochemical sensors is of great interest and it can
be applied to the determination of Fe, which is an essential nutrient for life, since its deficiency can
cause iron deficiency anemia.2 Thus, this study proposes the green synthesis of AgNPs/CP for the
development of a sensor applied to the determination of Fe in drug using cyclic voltammetry (CV).
Different parameters were evaluated in the biosynthesis and the results were evaluated by
spectrophotometry, VC and electrochemical impedance spectroscopy (EIS). The best synthesis
conditions were pH 6.1, 100 °C, 80 minutes and using 5 mL of the aqueous extract (Rtc = 27.3 Ω).
The colloidal stability of AgNPs/CP formed according to storage time was evaluated and the material
remained stable for 50 days. The ultrastructural analysis of AgNPs/CP after defining the best
synthesis conditions was performed using TEM in order to verify their shape and size. The results
showed spherical, dispersed and homogeneous AgNPs/CP, in addition to presenting a lighter
material coating the AgNPs/CP, which corresponds to the organic material of C. pyrenoidosa used
in the biosynthesis of AgNPs/CP.3 The AgNPs/CP had an average diameter of 5.04 ± 0.78 nm, which
was estimated from the count of 350 particles present in arbitrarily chosen areas of the TEM images.
After obtaining the AgNPs/CP, they were used to prepare the sensor. First, it was dispersed with 1.5
mg of carbon black (CB) and 1.4 ml of ultrapure water and sonicated for 1 h. Afterwards, 0.1 mL of
AgNPs/CP was added to this dispersion and sonicated for 3 min. Then, 6 µL of this dispersion was
pipetted onto the surface of the glassy carbon electrode (GCE) and dried for 2 h. After preparing
sensor (AgNPs/CP-CB/GCE), it was applied to the determination of Fe in a medicament sample
using CV and spectrophotometry as a comparative method. The Fe masses quantified in the sample
by the proposed (VC) and comparative (UV-vis) method was 39.82 and 40.02 mg, respectively. The
mass of Fe specified on the tablet packaging is 40 mg and the values obtained with both methods
were in agreement (99.5%) and very close to that described by the manufacturer. Furthermore, the
LD and LQ calculated for the method proposed by VC were 5.60 and 16.98 mg L-1, respectively.
Thus, this set of results prove the precision and accuracy of the proposed method, demonstrating
the feasibility of this analytical strategy for the determination of Fe in drug samples. Therefore, from
the results obtained in this work, it was possible to conclude that C. pyrenoidosa was able to reduce
Ag+ ions to Ag0 and stabilize the NPs formed through its biomolecules. The method developed was
accurate and precise, sensitive, low cost, non-toxic and with minimal sample manipulation. Thus,
the developed sensor and the proposed method can be used for the detection of Fe in drugs, thus
helping in the quality control of the same.
1 Alghoraibi I, Soukkarieh CH, ZEIN R, Alahmad A, Walter JG, Daghestani, M. Inorganic and Nano-Metal Chemistry, 50,
2020, 895
2 Batista-nascimento L, Pimentel C, Andrade MR, Rodrigues PC. Oxidative Medicine and Cellular Longevity, 20, 2012
3 Fatimah IS. Journal of Advanced Research, 7, 2016, 961
[FURG, EQA, PPGQTA, CAPES]

47
A SIMPLE, LOW-COST AND OPEN-SOURCE PORTABLE POTENTIOSTAT FOR
WIRELESS ELECTROCHEMICAL ANALYSIS
Dagwin Wachholz Junior (PG)a*, Lourenço H. B. Vidotto (PG)a, Lauro T. Kubota (R)a
a Institute of Chemistry, University of Campinas - UNICAMP, 13083-970, Campinas, Brazil
*e-mail: dagwinjr@gmail.com
The measurements in electrochemical sensors are traditionally made by costly laboratory

equipment. Recently, many efforts have been dedicated to developing innovative portable devices
coupled with personal computers or smartphones to promote a gain in portability, versatility, and
agility in analytical procedures 1. In that way, we have developed a new open-source wireless and
battery-powered (3.7 V) potentiostat. The system relies on a CC2541-BLE MCU by Texas
Instruments for data sampling, processing, and transmission. The detector interfaces with a
smartphone using a Bluetooth protocol to receive the experimental parameters and visualize the
results in real-time. This approach simplifies the design and decreases both size and cost of the
device. An open-source Android application named “BluChem” was also developed for controlling
the device wirelessly, in addition to an API and a website that can be used in remote locations for
data processing and analysis. The developed device can perform chronoamperometry, cyclic
voltammetry, differential pulse and square wave voltammetry. Different measurements were carried
out using [Fe(CN)6]4−/3− in 0.1 M KCl solution and a three-electrode system to evaluate the device
performance. The results were compared with analogous measurements performed using a
conventional benchtop instrument (Autolab PGSTAT30 - Metrohm). In Fig. 1a, a comparison
between the data acquired by CV with the portable potentiostat and the Autolab is shown, evidencing
the same performance of both instruments. The analysis of different scanning rates for 5mM of
[Fe(CN)6]4−/3− evidence that the current outputs from both instruments had high repeatability (RSD
<10%) for all explored scanning rates. The slopes obtained by the two instruments were almost
superimposable, which values of 3.61x10−4 ± 2.72x10−5 [A/ V s−1] for the proposed device and
3.62x10−4 ± 5.94x10−6 [A/ V s−1] for Autolab resulted in being statistically equal (p > 0.05) (Fig 1.b).
 
 
I (A) = x− + x−[C](mM)
(c)
(A) (B)  
2
R = 0.9991 (C)

   
I / A

  
(b)
I / A
I / A
Ip / A

I / A

 Portable 1 

10 mV/s  I (A) = x− + x−[C](mM)
R2 = 0.9999
− 
Portable 2        
−
Portable 3 −
150 mV/s C / mM (a)
− 
−
−     
Autolab 1 ipc Autolab E vs Ag/AgCl / mV Autolab
Autolab 2 − ipa Autolab 
ipc Portable
Portable
Autolab 3
− 
ipa Portable
−
−             − −   
E vs Ag/AgCl / V v1/2 / V s−1 E vs Ag/AgCl / V
Figure 1. (A) Voltammetric profiles for comparison of the performance of a portable device and Autolab (50 mV s−1). (B)
A plot of peak current vs scanning rate (Ip/A vs v/V s −1). Inset: CV obtained (portable) by varying the potential scanning
rate (10 to 150 mV s−1) ([Fe(CN)6]4−/3− (5.0 mM), 0.1 M KCl). (C) SWV voltammograms using optimized parameters
obtained for [Fe(CN)6]4−/3− ((a) 2.0 mM, (b) 3.0 mM and (c) 5.0 mM) using the portable device (line) and Autolab (dashed).
All measurements were conducted in triplicate (n = 3).
For the validation of DPV and SWV techniques by using the portable potentiostat, three different
concentrations of [Fe(CN)6]4−/3− were investigated. The results from both instruments presented the same
performance with high repeatability for all concentrations (Fig 1c). In addition, the slope values were statistically
equal (p>0.05), evidencing the applicability of the proposed device for quantification purposes using pulse
techniques. The portable device was also validated in other circumstances. The measured currents were
statistically equivalent to the benchtop equipment in all analyzed conditions. The operational ranges of current
and voltage (±1.5 V, ±1 mA) are smaller than most benchtop potentiostats. However, its operating range is
sufficient for most electrochemical analyses. Furthermore, considering that the device is simple, small in size
(~ matchbox), cheap (< 70 USD), and completely wireless, it offers new opportunities to develop affordable
diagnostics, sensors, and wearable devices.
1 Alves TM et al. Braz. J. Anal. Chem., 8, 2021, 22.
[FAPESP, CNPq, CAPES]

48
VOLTAMMETRIC DETERMINATION OF 17Α-ETHINYLESTRADIOL WITH A
CARBON DOTS MODIFIED CARBON PASTE ELECTRODE
Brenda R. L. Freire (PG)a, José C. S. Junior (PG)a*, Michael D. S. Monteiro (PG)a, Eliana M.
Sussuchi (R)a
aResearch Group of Nanomaterials and Electrochemical Sensors - SEnM, Graduate School of
Chemistry, Federal University of Sergipe, Postal Code: 49.100-000, São Cristovão, SE, Brazil
*e-mail: ozeca10@hotmail.com
Classified as endocrine disruptive compound, 17α-ethinylestradiol (EE2) is a synthetic hormone

derived from estradiol and commonly used in contraceptive pills1,2. However, conventional water and
sewage treatment plants are not designed to the removal or treatment of this hormone. Therefore,
both the detection of EE2 at low concentrations and the development of new analytical methods are
necessary. This study focused on the development of a modified electrode with carbon dots (B-CD)
for the voltammetric detection of EE2 in solution. The carbon dots were synthesized through the
hydrothermal route in a PTFE autoclave at 180 °C for 8 h, using citric acid (CA) as carbon source
and urea (UR) as nitrogen source. Spectroscopic characterizations like Fourier-transform infrared
(FTIR) (Figure 1), Raman and emission spectroscopy showed evidence of material formation. After
optimizations of the analysis parameters an analytical curve presented in the Figure 2, provided
information regarding the linearity of the electrochemical signal in the concentration range of 0.01–
0.80 µmol L-1 (R2 = 0.997) with a limit of detection equal to 0.59 nmol L-1 The results showed that the
developed electrode can be used for the determination of EE2 in aqueous solutions, given the great
analytical performance presented.
b) 120 5
0,7 a) 0,7 4
0,6 3
UR 0,6 100
0,5 2
h I / A 1
0,4
0,5 0
Intensity (%)
N-H 0,3 80
-1
I (A)
0,4 0,2
-2
0,1
60 -3
O-H CA
0,3 0,0 -4
a 0,0 0,1 0,2 0,3 0,4 0,5 0,6 0,7 0,8 0,9
CEE2
-5
(mol L-1)
0,2 40 -6
B-CD
-7
0,1
-8
C=O 20
C-N -9
O-H/N-H 0,0
-10
3500 3000 2500 2000 1500 1000 0,35 0,40 0,45 0,50 0,55 0,60 0,65
-1 E (V) vs Ag/AgCl/KCl 3.0 mol.L-1
Wavenumber (cm )
Figure 1. FTIR spectra of citric acid, urea and carbon dots, Figure 2. a) Calibration curve (baseline corrected) to
in KBr tablets. EE2. Inserted graph (b): Linear correlation from EE2
anodic peak in concentration range from 0.01 µmol L-
1 to 0.8 µmol L-1.
1 Nunes CN et al. J. Anal. Methods Chem, 1, 2016, 94.

2 Lima PR, Bergamasco R, Evidência, 17, 2017, 119.
[CNPq, CAPES, FAPITEC/SE, LCNT, NUPEG/PETROBRAS/UFS, CLQM, PPGQ-UFS,

LabRMN, SergipeTec]

49
HIGHLY SENSITIVE DETECTION OF CARBENDAZIM USING A STIMULATED
VISIBLE LIGHT GCE/BiVO4 SENSOR BY A ZWITTERIONIC BUFFER
Lázaro F. Silva (PG)a, Felipe A. Gorla (PG)a,b, Caroline S. dos Santos (PG)a, Roberto de
Matos (R)a, César R. T. Tarley (R)a,c*
a State University of Londrina (UEL), Department of Chemistry, Londrina, Parana, Brazil, 86057-970
b Federal Institute of Parana, Campus Assis Chateaubriand, Assis Chateaubriand, Parana, Brazil, 85935-000

*e-mail: tarley@uel.br
Carbendazim (CBZ) is one of the main fungicides of the benzimidazole class, used in agricultural
control processes as it is an effective agricultural defensive in the control of a variety of agricultural
diseases caused by fungi in grains, fruits, viticulture, and vegetables. However, CBZ is a teratogenic
compound of low-moderate toxicity, and its residues can cause liver and chromosomal diseases in
humans and animals and aquatic organisms1, which makes it necessary to develop sensitive and
selective strategies for the detection and quantification of CBZ in food samples. In the meantime,
electrochemical techniques have great potential for this task, due to their high sensitivity and low
instrumental cost. Nonetheless, CBZ can show high oxidation potentials and low electrochemical
activity when using commercial electrodes, making its determination at low concentrations an
analytical challenge.2 In this sense, the use of photoelectrochemical sensors can be interesting, as
they take advantage of the promotion of electrons from the valence band to the conduction band
when the material is excited, which generates an e-/h+ pair,3 allowing an increase in the
electroanalytical signal. Thus, the objective of this study is to develop a highly sensitive
photoelectrochemical sensor based on the modification of a commercial glassy carbon electrode
with BiVO4 for the determination of CBZ. The synthesis of BiVO4 was carried out through a
hydrothermal strategy using Bi(NO3)3.5H2O and NH4VO3 as precursors. SEM images and XRD
spectra allowed the classification of BiVO4 as 100% monoclinic, and the band gap energy of 2.54
was calculated through the Kubelka-Munk equation. It is noteworthy that these characteristics are
desirable for the construction of photoelectrochemical sensors excited by light in the visible region.
For photoelectrochemical measurements, the GCE was previously modified with 12 µL of a 2.0 mg
mL-1 BiVO4 suspension containing DHP. The influence of photoelectrochemical responses was
evaluated at different potentials (0.8 to 1.2 V) using CBZ at 30 µmol L-1 and acetate buffer 0.1 mol
L-1 at pH 4. It was observed the best photocurrent (0.216 µA) at 1.2 V. Afterwards, the influence of
pH (2.0 to 8.0) using BR buffer 0.1 mol L-1 as a supporting electrolyte was evaluated, whose optimum
value was found at pH 8.0 yielding a photocurrent of 13.68 µA. The influence of different types of
supporting electrolytes (KCl, phosphate buffer, McLlvaine buffer, and the zwitterionic buffers HEPES
and PIPES) at 0.1 mol L-1 under pH 8 were also investigated. Using CBZ at 30 µmol L-1 a substantial
increase on the photocurrent (50.33 µA) was achieved using the HEPES, justified by the antenna
effect characteristic of zwitterionic buffers.4 In order to evaluate the influence of HEPES
concentration (0.05, 0.1, and 0.3 mol L-1), the CBZ concentration was reduced to 0.1 µmol L-1.
Photocurrents of 26.00, 45.67, and 59.33 µA of CBZ were obtained for the respective concentrations
of HEPES, thus indicating the high sensitivity determination of CBZ in the presence of HEPES 0.3
mol L-1. Photocurrents at lower concentrations of CBZ (1 ng L-1) were also recorded and mensurable
when compared to blank measurements. The results obtained by the GCE/BiVO4 sensor associated
with the antenna effect promoted when using HEPES is a promising strategy for the determination
of CBZ at very low concentrations.
1 Wang Y-S., Huang Y-J., Cheng W-C., Yen J-H, Journal of Hazardous Materials, 172, 2009, 84.
2 Tian C, Zhang S, Wang H, Chen C, Han Z, Chen M, Zhu Y, Cui R. Zhang G, Journal of Electroanalytical Chemistry,
847,2019,113243.
3 Malathi A, Madhavan J, Ashokkumar M, Arunachalam P, Applied Catalysis A, General, 555, 2018, 47.
4 Xiao X, Zhu W-W, Lei Y-B, Liu Q-Y, Li Q, Li W-W, RSC Advances, 6, 2016, 35449.
[UEL, IFPR, INCTBio, LADEMA, ESPEC, CAPES, CNPq]

50
SYSTEM FOR BUTYLPARABEN ANALYSIS
Maria Letícia M. Soares (UG)a,b, Lucas M. Silva (PG)a,b, Carolina V. Uliana (R)a,b, Paulo C. F. L.
Gomes (R)a,b, Hideko Yamanaka (R)a,b*
a UNESP, Instituto de Química, Departamento de Química Analítica, Inorgânica e Físico-química, Araraquara, São
Paulo, Brazil, 14800-900.
b UNESP, National Institute for Alternative Technologies of Detection, Toxicological Evaluation & Removal of
Micropollutants and Radioactives (INCT-DATREM), Araraquara, São Paulo, Brazil, 14800-900.

*e-mail: hideko.yamanaka@unesp.br
Parabens are a chemical class used as preservatives in food, pharmaceutical and cosmetic products, due to
their broad antimicrobial activity, high stability to temperature and pH changes, as well as their low cost 1.
However, studies point out that parabens have estrogenic activities, and when bound to estrogen receptors in
the human body they cause toxic effects. There are few studies on the electrochemical quantification of
paraben in the literature. However, the junction between the electroanalytical method and the microfluidic
analysis system presents several advantages, among them, the significant reduction of residues and the
possibility of automating the detection and sampling system. In this way, a microfluidic cell for a printed
electrode was built, with the purpose of detecting and quantifying butylparaben (BPA), Figure 1a. The
performance of a microfluidic system was evaluated using a commercial printed carbon electrode Metrohm
DropSense. After optimizing the operating conditions of the analysis system, the electrochemical behavior of
BPA was evaluated at different pH values in Britton-Robinson (BR) 0.04 M buffer solution, thus the best pH
value for BPA determination was pH 8.0 and a linear relationship between Ep x pH can be observed, where
the oxidation of BPA involves 1H+/1e-. Due to the absorptive characteristic of BPA in conventional electrodes,
therefore, the reuse of the used electrode was evaluated, indicating that the microfluidic system minimizes
absorptive effects on the surface of the working electrode, Figure 1b. In (-) cyclic voltammograms are observed
in electrolyte support, buffer solution BR 0.04 M pH 8; in (-) the oxidative signal of BPA can be observed, with
Ep = 0.62 V and in (-) cyclic voltammograms in electrolyte support buffer solution BR 0.04 M pH 8 after washing
the surface with H2O milliQ, followed by 95% ethanol and finally with milliQ H 2O and dried in N2, showing that
there is no BPA on the work surface. Analytical curve was obtained by using amperometric technique from
0.50 to 50.0 µM, Figure 1c.
10
b 3,5 c 4,5
4,0
8 3,0 3,5
3,0
I / A
2,5 2,5
6 2,0
I / A
1,5
2,0
I / A
1,0
4 blank 0,5
BPA 1,5 0,0
blank after cleaning 0 10 20 30 40 50

2 [BPA] / M
1,0
0 0,5
a
-2 0,0
0.0 0.2 0.4 0.6 0.8 1.0 0 25 50 75 100 125 150 175 200 225
E/V Tempo / s
Figure 1. a) Microfluidic device coupling to graphite printed electrode. b) Cyclic voltammograms of the 50.0 µM
butylparaben solution in 0.04 mol L-1 BR buffer and the blank, both at pH 8 with a scanning speed of 50.0 mV.s-1. c) Multiple
amperograms of BPA solutions from 0.50 to 50.0 µM. Insert: analytical curve.
The system presented a sensitivity of 0.077 µA. µM-1, limit of detection 0.284 µM, limit of quantification 0.861
µM, repeatability 2.1% and reproducibility 2.5%. It was observed that the microfluidic system is effective for
the detection and quantification of BPA, minimizing the absorptive effect of the analyte on the surface of the
working electrode and allowing a high repeatability and reproducibility system.
1 GOMES FER et al. Journal of Electroanalytical Chemistry, 769, 2016, 124.

51
LASER-SCRIBING FABRICATION OF A DISPOSABLE ELECTROCHEMICAL
DEVICE FOR FORENSIC DETECTION OF CRIME FACILITATING DRUGS
Mayra V. Paschoarelli (UG)a*, Mathias S. Kavai (UG)a, Lucas F. Lima (PG)a,
William R. de Araujo (R)a
a State University of Campinas - UNICAMP, Portable Chemical Sensors Lab, Department of Analytical Chemistry, Institute
of Chemistry, P.O. Box 6154, 13083-970, Campinas, SP, Brazil.
*e-mail: mayra.paschoarelli@gmail.com
Benzodiazepines (Bz) are prescription-restricted psychotropic drugs, commonly used as sedatives

and hypnotics for treating anxiety, epilepsy, and muscular spasms1. However, their role in Drug-
Facilitated Crimes (DFCs) has gradually increased due to their effects on the central nervous system,
such as confusion, sedating, and amnesic effects, making the victim highly susceptible to criminal
activities, such as sexual assaults.2-3 Given the problems concerning the illicit use of drug-facilitated
sexual assaults and poisoning, which is increasing lately, new portable and disposable devices are
required for rapid forensic analysis of these substances at crime scenes and from toxicological
perspectives. Therefore, the laser-scribing graphene (LSG) electrodes have emerged in recent years
as an alternative electrochemical system for developing accessible, portable, disposable, and high-
performance sensors, since they combine simplicity, low cost, easy manufacturing, and high
sensitivity and do not require sophisticated laboratory instrumentation for manufacturing and
analyses.4 Here, we reported a novel LSG electrochemical sensor fabricated by controlled and
localized CO2 laser carbonization of polyetherimide (PEI) substrate for diazepam (Dz) and
midazolam (Mz) detection in alcoholic beverages, such as whisky and sugarcane spirits (cachaça).
Morphological and structural characterizations were performed by scanning electron microscopy and
Raman spectroscopy, demonstrating that the electrochemical device presents a highly porous and
disordered carbon material, which contributes to high active area and electron-transfer properties.
In addition, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS)
demonstrate the superior electrochemical performance of the PEI-LSG device after a simple
cathodic treatment step (apply -2.0 V for 120 s in 0.1 mol L-1 H2SO4). Both Dz and Mz presented
irreversible and pH-dependent reduction processes involving a 2:1 ratio between electrons and
protons. The electrochemical detection of Dz and Mz was carried out using the square wave
voltammetry (SWV) technique under optimized experimental conditions using 0.1 mol L -1 Britton–
Robinson buffer (pH=2.0 for Dz and pH=4.0 for Mz). Analytical curves were built for both drugs
ranging from 2.5 to 100 μmol L-1, with linear behavior in the range of 2.5 to 50.0 μmol L-1. The limit
of detection (LOD) was calculated according to the equation LOD = 3×SDintercept/Slope, providing
0.0017 μmol L-1 for Dz and 0.61 μmol L-1 for Mz. Meanwhile, the limit of quantification (LOQ) was
calculated as LOQ=3.3 x LOD, resulting in 0.0058 μmol L-1 for Dz and 2.01 μmol L-1 for Mz. The
accuracy and robustness of the developed electrochemical sensor were evaluated through the
addition and recovery protocol, by spiking two different concentrations (10 and 50 µmol L-1) of Dz
and Mz in commercial whisky and cachaça samples to simulate a common sample/evidence of a
drug-facilitated crime case. The recoveries ranged from 97.1% to 117.3% for Dz and 96.4% to
114.3% for Mz, highlighting the adequate accuracy of the proposed method. Based on these
analytical features, easy manufacturing method, and low cost, we can point out the proposed method
as an excellent alternative approach for rapid and in-field analysis of drug-facilitated sexual assaults
and other relevant criminal applications.
1 Gentili S et al., Journal of Analytical Methods in Chemistry, 1, 2016, 1234581.
2 Griffin CE et al., The Ochsner Journal, 13, 2013, 214.
3 Rocha DS et al., Talanta, 232, 2021, 122408.
4 Zhu B et al., Electrochimica Acta, 378, 2021, 138132.
SAE/UNICAMP, CNPq, FAPESP, CAPES]

52
SPECIATION OF CHROMIUM AND THALLIUM ALONG THE AMAZON AND
PARÁ ESTUARIES AND MIXING PLUME (AMAZON-GEOTRACES, 2018)
Igor Tessele (PG)a*, Leandro Carvalho (R)a
a Federal University of Santa Maria (UFSM), Department of Chemistry, Santa Maria, RS, Brazil, 97105-900
*e-mail: igortessele@hotmail.com
The transport of elements from rivers to the ocean is governed by different physicochemical
processes. Thus, generally a small fraction of the total element reaches the ocean due to the
aggregation and flocculation processes during estuarine mixing.1 The Amazon River has the largest
drainage basin in the world, making it a major source of trace elements to the Atlantic Ocean.2
However, few studies exist quantifying trace elements and their species in this region. The aim of
the study was to analyze the input and removal processes involved in the the transport of Cr and Tl
species in the Amazon estuarine mixing zone in North Brazilian continental shelf. The studied
samples were collected during the scientific expedition M147 on board of the Meteor research vessel
(Amazon – GEOTRACES process study GApr11) in the Amazon and Pará River outflow regions.
Here we present the results for particulate (>0.2 µm), labile and total dissolved (<0.2 µm) fractions
of Cr and Tl in surface waters by using adsorptive cathodic and anodic stripping voltammetry (AdCSV
and AdASV). The measurements were based on the sequential determination with
diethylenetriamine pentaacetic acid (DTPA) as a complexing reagent (6.25 mM) for Cr at pH 6.2 and
Tl after addition of acetate buffer at pH 4.6 as the working electrolytes. Total and total dissolved Cr
and Tl were determined after UV digestion of the samples at pH 2.0 for 2h using an UV digestor (Hg
lamp 250W). In general, we observed that all Cr and Tl species decreased with increasing salinity
within the estuarine mixing zone, as shown in Figure 1.
Figure 1. Distribution of Cr ant Tl within the estuarine mixing zone in Amazon regions.
For both Cr and Tl species (total, particulate and dissolved), a non-conservative distribution was
observed within the whole salinity gradient, probably due to biological uptake and flocculation
processes. In addition, dissolved and particulate Cr ant Tl also decreased with increasing salinity.
We also observed unique source signatures in dissolved and labile particulate Cr ant Tl species from
the Amazon River outflow regions. In addition, elevated levels of Tl in Mangrove belt were observed,
probably due to a possible influx of groundwater and/or desorption process from particulate material.
Lastly, this work in contributing to the understanding of the complex interaction of processes that
controls trace metal–DOM input into the Atlantic Ocean.
1 Boyle EA., Edmond JM, Sholkovitz ER, Geochimica et Cosmochimica Acta, 41, 1977, 1313.
2 Carvalho LM, Hollister AP, Trindade CR, Gledhil M., Koschinsky, A, Marine Chemistry, 236, 2021, 104019.
[CNPq, DFG]

53
USE OF GRAPEHENE-BASED FREE-STANDING FILM IN SIMULTANEOUS
DETERMINATION OF CONTAMINANTS OF EMERGING CONCERN
Layne T. S. Zanon (PG)a, Edson H. P. Arruda (PG)a, Vitor H. N. Martins (PG)a, Victor H. R.
Souza (UG)a, Magno A. G. Trindade (R)a*
a University Federal of Grande Dourados, FACET, Dourados, MS, Brazil, 79804-970
*e-mail: magnotrindade@ufgd.edu.br
The increasing consumption of pharmaceutical and personal care products (PPCPs) has
considerably increased the concentration of contaminants in aquatic environments. These products
are part of a group of chemicals that are considered contaminants of emerging concern (CEC).
Electrochemical techniques are among the alternatives that can be used in the determination of
CECs.1,2 These techniques present high sensitivity, low cost, quick execution, and do not require
complex pre-treatments of the sample before analysis. Thus, we present an electrochemical cell
using a free-standing film based on reduced graphene oxide-polyaniline mixture (rGO 2% / PAni
0.25%) as a working electrode (WE).3 A newly designed electrochemical cell, built with low-cost
materials and with easy handling, was assembled to fit the WE at the bottom of the cell. This cell
configuration enables some improvements such as the mass transport of electroactive species to
the electrode surface as well as being easy to avoid the electrode fouling by removing (using a
homemade stirring rod) the electrogenerated products from its surface.1 As a proof of concept, we
tested the featured device in the determination of acetaminophen (ACP), salicylic acid (SA), and
norfloxacin (NOR) in tap and Lake water samples. After optimizing the best experimental and
instrumental conditions, the performance of the alternative electrochemical cell was tested by
detecting target analytes (detection peaks at +0.75V for ACP, +1.2V for SA, and +1.35V for NOR),
showing suitable recovery values. Finally, we validated this method using high-performance liquid
chromatography (HPLC), which showed acceptable values of recovery. As well, the intra-day and
inter-day study using ten measurements (for 60 µmol/L of ACP and SA and 90 µmol/L NOR) were
found to be highly reproductive.
1 Rosa TM et al. Talanta, 196, 2019, 39.
2 Oliveira LH, Trindade MAG. Analytical Chemistry, 88, 2016, 6554.
3 Martins VHN et al. ACS Applied Nano Materials, 4, 2021, 5553.
[PPGQ-UFGD, CNPq, CAPES, INCT-DATREM, INCT-Nanocarbono, FUNDECT]

54
ELECTROCHEMICAL DETECTION OF THE SYNTHETIC TRYPTAMINE 5-MEO-
MIPT USING CARBON ELECTRODES: A SIMPLE SCREENING TEST FOR
FORENSIC ANALYSIS
Larissa M. A. Melo (PG)a*, Luciano C. Arantes (R)b, Dilton M. Pimentel (R)a,
Wallans T. P. dos Santos (R)a
a Universidade Federal dos Vales do Jequitinhonha e Mucuri, Departamento de Química, Diamantina, Minas Gerais,
Brazil, 39100-000
b Instituto de Criminalística, Polícia Civil do Distrito Federal, Seção de Perícias e Análises Laboratoriais, Brasília, Distrito
Federal, Brazil, 70610-200

*e-mail: larissamamelo@gmail.com.br
The 5-methoxy-N-methyl-N-isopropyltryptamine (5-MeO-MiPT) is a synthetic tryptamine, also known

as “moxy”, used as a stimulant drug for recreation. The 5-MeO-MiPT is an illicit drug that belongs to
the group of new psychoactive substances (NPS). The NPS have widely been consumed and
reported in last years in forensic analysis, which can cause a great problem of public health in many
countries. Furthermore, there are not efficient methods for a preliminary identification of these NPS
in seized samples as for example for the 5-MeO-MiPT, generating legal problems to draw up of
arrest warrant when the dealers are caught in the act. Electroanalytical methods can offer an
attractive technique for screening tests in forensic samples due to their simplicity of application in
on-site analysis with low-cost and portable sensors. In this context, this work presents, for the first
time, the electrochemical behavior of 5-MeO-MiPT using carbon electrodes in order to develop a fast
and simple eletroctroanalytical screening method of this drug in forensic analysis. Although, the 5-
MeO-MiP showed only one irreversible oxidation process at a glassy carbon electrode, many
electrochemical processes were exhibited at a carbon graphite screen-printed electrode (SPE-Gr).
The oxidation reaction of 5-MeO-MiPT on both carbon electrode surfaces involves an adsorption-
controlled process with the loss one-electron and one-proton. The voltammetric screening method
of 5-MeO-MiPT for application in forensic samples was performed in 0.1 mol L-1 phosphate buffer
solution pH 7.0 at the SPE-Gr using adsorptive stripping differential pulse voltammetry (AdSDPV).
The proposed method showed a wide linear range (0.05 and 30.0 µmol L-1) with low LOD (0.015
µmol L-1), which is sufficiently low to access seized forensic samples containing 5-MeO-MiPT. A
good stability of the electrochemical responses for 5-MeO-MiPT detection at SPE-Gr was obtained,
using the same or different (N = 3) electrodes (RSD < 4.0 %). Interference studies for other drugs
demonstrated that the proposed sensor is selective for an electrochemical screening of 5-MeO-
MiPT. In addition, real seized samples were identificated with 5-MeO-MiPT by proposed method and
confirmed by HPLC-MS. The analytical performance of SPE-Gr with AdSDPV for 5-MeO-MiPT
detection demonstrates great potential for application of a simple, fast and selective screening
method in forensic analysis.
[UFVJM, CNPq, CAPES, FAPEMIG]

55
RISK ASSESSMENT OF TOXIC METALS AS CONTAMINANTS OF DIETARY
SUPPLEMENTS BY USING STRIPPING VOLTAMMETRY
Gabriel Reis (PG)a, Luiza Garcez (UG)b, Luis Ferraz (R)b, Leandro Carvalho (R)b
a Federal University of Santa Maria (UFSM), Graduate Program in Pharmaceutical Sciences, Santa Maria, RS, Brazil,
97105-900
b Federal University of Santa Maria (UFSM), Department of Chemistry, Santa Maria, RS, Brazil, 97105-900
*e-mail: g.mreis@yahoo.com.br
Medicinal plants can absorb toxic substances depending on the conditions of cultivation through
water, soil, and pollution. Once used in the form of powder or extracts to the production of food
supplements, they can endanger the health of consumers of these products. Furthermore, the
exposure time and concentration of these harmful substances can be aggravating factors due to the
heterogeneity of the use of supplements. The ingestion of excess micronutrients such as copper
(Cu), cobalt (Co), and zinc (Zn) can cause damage to health. In addition, cadmium (Cd) and lead
(Pb) present kidney and liver toxicity, and exposure to small amounts and short durations are
necessary to pose health risks. In this work, Zn, Cd, Pb, Cu, Ni and Co were determined in different
samples of dietary supplements (n=67) marketed in Brazil. The dietary supplements were purchased
from online stores and local commerce. A known amount of ~0.3 g sample was used for the total
oxidative digestion (HNO3/H2O2) in a microwave oven (Anton Paar Multiwave Pro®). After digestion
and dilution to 50 mL water, a fixed volume of each sample was added to the voltammetric cell
containing the supporting electrolyte. All the voltammetric measurements were carried out in a 746
VA Trace Analyzer coupled to a 694 VA Stand using a hanging drop mercury electrode (HMDE) as
a working electrode, a 3 mol L-1 Ag/AgCl reference electrode and graphite pin auxiliary electrode.
The metals were simultaneously determined by the standard addition method (n=3) as shown in
Figure 1.
Figure 1. Simultaneous and sequential voltammetric determination of metals in a dietary supplement sample by ASV abd
AdCSV after HNO3/H2O2 digestion in a microwave oven.
The method allowed the determination of the metals in all the 67 studied samples. Zn concentrations
varied from 0.22 to 7,166.83 g g-1 with an estimated daily mean intake of 2.79 mg g bw-1 day-1. Cd
concentrations varied from 0.036 to 3.75 g g-1 with an estimated daily mean intake of 0.070 mg g
bw-1 day-1. Pb concentrations varied from 0.017 to 16.22 g g-1 with an estimated daily mean intake
of 0.055 mg g bw-1 day-1. Cu concentrations varied from 0.25 to 622.36 g g-1 with an estimated daily
mean intake of 0.428 mg g bw-1 day-1. Ni concentrations varied from 0.005 to 12.44 g g-1 with an
estimated daily mean intake of 0.094 mg g bw-1 day-1. Co concentrations varied from 0.003 to 2.26
g g-1 with an estimated daily mean intake of 0.007 mg g bw-1 day-1. Lastly, the permitted daily
exposure was calculated considering the recommended dosage from the manufacturer and
compared with maximum limits according to ICH Q3D(R2)1.
1 ICH guideline Q3D (R2) on elemental impurities. Current step 5 version, dated 02 May 2022.
[CNPq]

56
EXTRACTION INDUCED BY MICROEMULSION BREAKING FOR
VOLTAMMETRIC DETERMINATION OF Cd, Pb AND Cu IN BIODIESEL
Cristian H. Krause (PG)a*, Alexandre B. Schneider (R)a, Márcia M. da Silva (R)a, Leandro
Kolling (PG)a, Marina B. Mazzei (UG)a, Fernando N. Leal (UG)a
a Universidade Federal do Rio Grande do Sul, Instituto de Química, Porto Alegre, RS, Brazil, 91501-970
*e-mail: crizzkrause@gmail.com
There is an increasing demand for the evaluation of determining specifications and the quality control
of biodiesel. Among these emerging requirements, it is necessary to create alternative procedures
for fast and simple determinations of trace metal such as Cd, Pb and Cu since they are known to
interfere with the motor performance, contribute to the formation of gums and sediments in vehicle
and fuel tanks, form insoluble salts, induce corrosion and accelerate the deterioration of this biofuel.
Moreover, metal contamination affects the biodiesel stability against oxidation and the combustion
of metal-containing fuels can be a source of pollution, releasing the contaminants Cd and Pb to the
atmosphere.1,2 Different approaches using different analytical techniques, mainly atomic absorption
spectrometry (AAS), have been proposed to determine Cd, Pb and Cu and other trace metals in
biodiesel using different sample preparation methods.3,4 On the other hand, square wave anodic
stripping voltammetry (SWASV) is one of the most sensitive analytical techniques in the
determinations of metals,5,6 being a simple and low-cost alternative for this purpose. Extraction
induced by microemulsion breaking (EIMB) is a recent and interesting alternative for extracting trace
metals from fuel samples to an aqueous media.7 Here, a new approach involving SWASV was
developed to simultaneously determine cadmium and lead and, in the same cell, sequentially copper,
in biodiesel after EIMB. The composition of the water-in-oil microemulsion (ME) involved 10.50 mL
biodiesel, 4.20 mL n-propanol and 0.30 mL of the 6.0 mol L-1 HNO3 solution. The EIMB was carried
out by adding 1.10 mL ultrapure water resulting in two well separated phases: an upper organic
phase, and a lower aqueous phase containing the analytes. The latter was further transferred to a
cell containing 1.0 mol L-1 acetic acid/acetate buffer at pH 4.0 and 10 mg L-1 Hg(II). The apparatus
comprised a portable potentiostat and a cell with an in situ plated mercury film glassy carbon
electrode (HgGCE) as working electrode (WE). The experimental parameters involved deposition
potential of -1.20 V fixed for 200 s, for Cd and Pb and -0.80 V fixed for 100 s, for Cu. Potential
amplitude of 50 mV, potential step of 6 mV and a frequency of 30 Hz were applied. The LOD for Cd,
Pb and Cu were 0.33 µg L-1, 0.48 µg L-1 and 0.66 µg L-1, respectively. The accuracy of the method
was evaluated by recovery assays of spiked samples and by analyzing a standard reference
material. The use of EIMB together with SWASV proved to be a low-cost alternative to carry out
simple, sensitive, selective and accurate measurements of Cd, Pb and Cu in biodiesel samples.
1 Martiniano LC et al., Fuel, 103, 2013, 1164.
2 Yaakob Z, Narayanan BN, Padikkaparambil S, Renewable and Sustainable Energy Reviews, 35, 2014, 136.
3 Silva JSA., et al., J Braz Chem Soc, 21, 2010, 620.
4 Antunes GA et al., Energy and Fuels, 31, 2017, 2944.
5 Mirceski V et al., Electroanalysis, 25, 2013, 2411.
6 Borrill AJ, Reily NE, Macpherson JV, Analyst, 144, 2019, 6834.
7 Vicentino PO, Cassella RJ, Talanta, 162, 2017, 249.
[CAPES, CNPq, CECOM-UFRGS]

57
SCREEN PRINTED NON-ENZYMATIC ELECTROCHEMICAL SENSOR BASED ON
CARBON AND HYDROXIDE/OXIDE COPPER FOR GLUCOSE DETECTION
Neuryelen dos Santos Bandeira (PG)a, João Paulo Vita Damasceno (R)a*, Lauro Tatsuo
Kubota (R)a
a Institute of Chemistry, Department of Analytical Chemistry, University of Campinas, Campinas, São Paulo, Brazil,
13084-971
*e-mail: joaopvd@unicamp.br
Carbon-based materials have increased in numerous applications, including electrochemical

sensors and biosensors, due to the low cost, high sensitivity and high intrinsic electrical conductivity
of graphitic materials1. Some recent studies report graphitic materials, especially graphene oxide
and reduced graphene oxide combined with copper oxides/hydroxides, as electrochemical glucose
sensors, classified as non-enzymatic sensors2,3. These sensors have several advantages over
enzymatic sensors like cost-effectiveness, good thermal and chemical stability, and long-term
stability. Another type of sensor that stands out for its low cost, abundance of material, sustainability,
and disposables is the paper-based printed sensor. Furthermore, due to its versatility, paper allows
the construction of innovative devices by integrating different analytical procedures4. The screen
printing of these paper devices stands out for their simple and equipment-free manufacturing. In this
work, the electrodes were manufactured in chromatographic paper using a mask to design the
electrodes. Therefore, an Ag/AgCl reference electrode and a platinum counter electrode were used.
A commercial carbon slurry was diluted in acetone with the addition of different concentrations of a
CuCl2 solution. In addition, a little oxidized graphene oxide was added to increase the conductivity
of the prepared ink. In the first stage of preparation of these sensors, the ready ink was deposited
on the mask, silkscreened to deposit the ink uniformly, and taken to the oven for curing at 100 ºC. In
the second part, the space between the working electrode and its connector was enameled to leave
this space hydrophobic, making it protect it from coming into contact with the solution. Finally, silver
ink was deposited on the connector and retaken to the oven to cure the silver paint at the same
temperature. The results showed a redox pair at -0.10V (cathodic peak) and 0.12V (anodic peak),
indicating the presence of copper oxide/hydroxide. Furthermore, an increase in the anodic peak
current was observed with the addition of glucose in a phosphate buffer solution. The operation
range of the sensor is at the mmolar level, similar to those observed with enzymatic biosensors. It
can be attributed to the chemical reaction between glucose and copper. Thus, it is highlighted that
non-enzymatic electrodes made of paper and carbon material containing copper oxide/hydroxides
are very promising to be used as disposable electrochemical sensors for glucose.
1 Damasceno JPV, Kubota, LT. Materials Today Chemistry, 21, 2021, 100526.
2 Phetsang S et al. Sci Rep, 11, 2021, 9302.
3 Barragan JTC, Kogikoski, J, Silva, ETSG, Kubota, LT. Analytical Chemistry, 90, 2018, 3357.
4 Deroco PB et al. Chapter Four - Paper-based electrochemical sensing devices, Comprehensive Analytical Chemistry,
Elsevier, 89, 2020, 91.
[INCTBio, CNPq, IQ-UNICAMP]

58
APPLICATION OF DEEP EUTECTIC SOLVENTS IN ANALYTICAL CHEMISTRY
Orlando Fatibello-Filho*
Federal University of São Carlos, Department of Chemistry, São Carlos-SP, Brazil, P. O. Box 676, 13560-970
New trends in analytical chemistry encourage the development of methods aligned with Green
Chemistry. A new class of green solvents called deep eutectic solvent (DES) was prepared almost
two decades ago by Andrew Abbott and co-workers1. A DES is frequently defined as a binary or
ternary mixture of compounds that can associate mainly via hydrogen bonds and, when mixed at a
certain molar ratio, result in a eutectic mixture, a liquid that has a freezing point lower than those of
the individual components. In this keynote, different approaches for the preparation of DES and
natural deep eutectic solvents (NADES) and their applications in analytical methods will be
presented. Special attention will be devoted to environmentally friendly alternative for liquid-phase
microextraction (LPME) of colorants in food products and their spectrophotometric determination 2.
In electroanalysis, the modification of carbon paste and film electrodes with different DES and
NADES has been showing promising alternatives3,4. Finally, the future of this promising area will be
addressed.
1 Abbott AP, Capper G, Davies DL, Rasheed RK, Tambyrajah V, Chem. Commun. 1, 2003, 70
2 Piton GR, Augusto KKL, Santos DJA, Fatibello-Filho O, J. Braz. Chem. Soc. 32, 2021, 564
3 Piton GR, Augusto KKL, Wong A, Moraes FC, Fatibello-Filho O. Electroanalysis 33, 2021, 2351
4 Augusto KKL, Piton GR, Gomes-Jr PC, Patelli G, Moraes FC, Fatibello-Filho O. Analytical Methods, 14, 2022, 2003
[FAPESP, CNPq]

59
3D-ELECTRODE FOR SULFANILAMIDE MONITORING IN BREAST MILK,
SYNTHETIC URINE, AND PHARMACEUTICAL FORMULATION SAMPLES
Thalles P. Lisboa (R)a, Guilherme F. Alves (PG)a, Lucas V. de Faria (R)b, Cassiano C. de
Souza (PG)a, Wallace B. V. de Oliveira (PG)a, Maria Auxiliadora C. Matos (R)a,
Renato C. Matos (R)a*
a Federal University of Juiz de Fora, Chemistry Department, Juiz de Fora, MG, Brazil, 36036-900
b Federal University of Uberlândia, Chemistry Institute, Uberlândia, MG, Brazil, 38400-902
*e-mail: renato.matos@ufjf.br
The development of low-cost sensors has received much attention from the scientific community in
recent years. Associated with this issue, additive manufacturing technology, also known as 3D-
printing has proven to be a powerful tool that allows wide design freedom, revolutionizing several
areas of the sciences1. In addition, the current demand for low-cost devices coincides with the need
to monitor antibiotics in complex matrices have been motivating the development of 3D-printed
electrodes with carbon-based materials has become an attractive alternative. In general, materials
such as carbon nanotubes, carbon black (CB), and graphene are mixed with thermoplastic materials
such as polylactic acid (PLA) and acrylonitrile butadiene styrene (ABS) to form conductive filaments.2
Therefore, this work describes a simple and cost-effective method for manufacturing a 3D-printed
electrode. This electrode presented a similar design to commercial electrodes, where a
stereolithography printer was used to build the electrode body using an acrylic resin. The
electroactive surface was filled by a 3D-pen using a carbon black integrated polylactic acid (CB/PLA)
conductive filament. After a simple and fast (400 s) surface treatment, the 3D-printed CB/PLA
electrode was combined with Differential Pulse Voltammetry (DPV) technique for sulfanilamide
(SAA) determination. The influence of pH on the SAA electro-oxidation process was evaluated using
a BR buffer solution (10 mmol L-1) with values ranging from 2.0 to 10.0. An irreversible oxidation
signal can be observed only in a direct scan with a peak potential at around +1.15 V, in an acidic
medium. Furthermore, there was no significant potential shift to more cathodic regions, pH 2.0 was
chosen as the optimum condition, considering that provided higher analytical sensitivity (higher
current signal). The mass transport regime was studied using an SAA solution with a concentration
of 200 µmol L-1. A linear correlation was observed between the square root of the scan rate and the
current, indicating that the mass transport of SAA species is a diffusion-controlled process, similar
to the behavior observed for other carbon-based electrode surfaces. A method based on differential
pulse voltammetry was optimized, which presented a wide linear range from 1 to 39.2 µmol L -1
(Figure 1) and good precision (RSD = 1.8%, n = 10. The developed electroanalytical method was
applied to breast milk, synthetic urine, and otologic solution samples, showing excellent analytical
performance with a detection limit of 12 nmol L-1. In addition, the sensor provides fantastic selectivity
towards other antibiotic classes, and when applied in spiked samples, recovery values between 93
and 108% were obtained, which demonstrated good accuracy as well as the absence of matrix
effect. It is highlighted that no laborious sample preparation steps were required (simple dilution in
supporting electrolyte). Thus, the proposed 3D-printed device proves to be a promising analytical
tool for routine analysis.
Figure 1. DPV voltammogram of the analytical curve for SAA ranging from 1
to 39.2 μmol L-1 (a-o). Inserted graph showing the linear adjustment with line
equation Ip (μA) = (0.056 ± 0.001)[SAA] (μmol L-1) - (0.104 ± 0.022) and
correlation coefficient R2 = 0.994. Conditions: reference electrode - Ag/AgCl,
KCl(saturated), supporting electrolyte BR buffer (10 mmol L-1, pH = 2.0),
amplitude 90 mV, step potential 5 mV.
1 Lisboa TP, Alves GF, de Faria LV, de Souza CC, Matos MAC, Matos RC.
Talanta 247, 2022, 123610.
2 Hamzah HH, Shafiee SA, Abdalla A, Patel BA. Electrochemistry
Communications 96, 2018, 27.
[UFJF, FAPEMIG, CAPES, CNPq]

60
LOW-COST PAPER-BASED ELECTROCHEMICAL SENSOR FOR THE
DETECTION OF CIPROFLOXACIN IN HONEY AND MILK SAMPLES
Cassiano C. de Souza (PG), Guilherme F. Alves (PG), Thalles P. Lisboa (R), Maria
Auxiliadora C. Matos (R) and Renato C. Matos (R)*
Federal University of Juiz de Fora, Chemistry Department, Juiz de Fora, MG, Brazil, 36036-900
*e-mail: renato.matos@ufjf.br
Honey and milk contain a complex matrix of components, which presents a considerable analytical
challenge. They are animal products rich in nutrients, broadly consumed worldwide, and part of the
processed food production chain. The management and welfare of food-producing animals are
essential to ensure quality control. In this sense, healthy animals are directly linked to an appropriate
product. Due to many bees and cattle being manipulated daily, it becomes challenging to separate
and treat only those which are not in good physical form. So, there are prophylactic practices adopted
by farmers to prevent bacterial outbreaks, such as adding antibiotics to livestock feed or applying it
to beehives beforehand. Therefore, the present work aimed to develop and optimize a disposable,
easy, affordable, and accurate electrochemical sensor based on a laboratory-made graphite
conductive ink supported on filter paper. Furthermore, an electroanalytical method by differential
pulse voltammetry (DPV) was optimized for the sensitive quantification of ciprofloxacin (CIP) in milk
and honey samples. The sensor was fabricated employing a low-cost conductive ink, composed of
nail polish and powdered graphite, supported by filter paper. Scanning Electron Microscopy (SEM)
was applied to investigate the paper-based electrode (PBE) surface morphology before and after
the polishing. It was feasible to observe that the surface before the polishing displays an irregular
coating with graphite sheets in multiple sizes and after the polishing, the surface became more
homogenous, with slight grooves provoked by the mechanical exfoliation. The cyclic voltammogram
[Fe(CN)6]3-/[Fe(CN)6]2- (1 mmol L-1) showcases a current increment of 33% of the polished electrode
in comparison to the non-polished electrode. The electroactive area of the PBE was estimated using
Randles-Sevcik equation, with scan rates varying from 50 to 300 mV s-1. The geometric area of the
working electrode was 12.62 mm2. The area estimated for the non-polished electrode was 2.29 mm2,
whereas the electroactive area estimated for the polished electrode was 4.90 mm2. It indicates that
the mechanical treatment enhances the electroactive area by 113 % and it agrees with the current
increase. The pH effect on the electrochemical oxidation process of CIP (333 µmol L−1) was
investigated with a BR buffer solution ranging from 2.0 to 10.0, according to the slope of this
regression (44.7 mV pH-1), it can be inferred that one electron and one proton are involved in the
CIP oxidation process when PBE is employed, which agrees with other studies reported in the
literature.1 In this sense, it was decided to use the pH 10 buffer solution, since it provides a faster
electronic transfer process (potential anticipation) with the highest analytical response among the
pH values studied. The mass transport regime was assessed with a CIP concentration of 333 µmol
L-1, which indicates a diffusional process of the electroactive species onto the PBE surface. A method
based on differential pulse voltammetry was optimized, which presented a wide linear range from
9.90 to 220 µmol L-1, adequate sensitivity (4.96 µmol L-1), and selectivity among other classes of
antibiotics (CIP analytical response variation was less than 4%).2 The method was applied for
monitoring CIP in spiked samples of honey, whole, and skimmed milk, with adequate precision (RSD
< 3.1 %) and accuracy, recovery values ranged from 80 to 95%. The PBE showed sensitivity in the
same order of magnitude as other electrochemical sensors reported in the literature. Furthermore,
the simple fabrication process allied to minimal sample treatment, only a simple dilution, makes the
paper sensor a promising tool for food control analysis.
1 Alves GF, Lisboa TP, de Faria LV, de Farias DM, Matos MAC, Matos RC. Electroanalysis 33, 2021, 543.
2 Souza CC, Alves GF, Lisboa TP, Matos MAC, Matos RCM. Journal of Food and Composition and Analysis 104700,
2022.

61
ELECTROCHEMICAL SENSING AT YOUR FINGERTIPS: WEARABLE GLOVE-
BASED SENSORFOR FORENSIC AND ENVIRONMENTAL APPLICATIONS
Paula Cristine Rocha Corsatoa*, Lucas Felipe de Lima (PG)a, William Reis de Araujo (R)a
a State University of Campinas – UNICAMP, Institute of Chemistry, Department of Analytical Chemistry, Campinas, SP,
Brazil, 13083-970
*e-mail: pcorsato15@gmail.com
The integration of chemical sensors within wearable platforms possibilities the continuous
monitoring of health and the well-being of the users aiming at the prevention, monitoring, and
management of diseases, which enables laboratory-based chemical analyses directly on the
body1,2. In addition, wearable devices can perform portable analyses to assess the user's
surroundings to investigate its exposure to toxic or hazardous materials. The glove-embedded
electrochemical sensors allow analyses directly in liquid and solid samples by dipping the finger-
based sensors intothe aqueous sample or swiping the fingertip onto the contaminated surface for
sampling the solid microparticles3,4. In this work, we developed a wearable electrochemical sensor
manufactured by a screen-printing method on nitrile gloves using carbon and Ag/AgCl inks.
Following, the working electrode was modified with multiwalled carbon nanotubes (MWCNTs) and
poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) for sensitive detection of
nitro explosive picric acid (PA) and the emerging pollutant 4-nitrophenol (4-NP) in tap and river
water samples. Morphological characterizations by scanning electron microscopy (SEM) revealed
that the MWCNTs appear entangled in bundles with a uniform diameter and micrometric length
dispersed in the PEDOT:PSS matrix, which contributes to the increased active area and high
electron transfer properties. The redox characterization of PA and 4-NP was performed by cyclic
voltammetry and demonstrates that both species present irreversible and pH-dependent redox
processes involving a 1:1 electron:protonratio. The electrochemical quantification of PA and 4-NP
was performed by the square wave voltammetry (SWV) technique under optimized experimental
conditions using 0.1 mol L-1 Britton- Robinson buffer (pH=2.0 for PA and pH=7.0 for 4-NP). An
analytical curve was built for PA using thepeak current values of the reduction process close to -
0.6 V and exhibited a linear behavior in the concentration ranging from 5 µmol L-1 to 100 µmol L-1,
with a limit of detection (LOD) of 1.74 µmol L-1 and limit of quantification (LOQ) of 5.73 µmol L-1.
The analytical curve for 4-NP was built using thepeak current values from the oxidation process
close to 0.7 V and exhibited a linear response in theconcentration ranging from 1 µmol L-1 to 100
µmol L-1, resulting in a LOD and LOQ of 0.33 µmol L-1 and 1.11 µmol L-1, respectively. The
robustness and accuracy of the developed wearable electrochemical sensor were evaluated
through addition and recovery protocol, by spiking different concentrations (5 µmol L-1 to 50 µmol
L-1) of PA in tap water and 4-NP in tap and river water samples.The recoveries ranged from 96.5%
to 112.7% for PA and 98.9% to 114.2% for 4-NP, highlighting theadequate accuracy of our method.
Based on these analytical parameters, we can point out the proposed electrochemical lab-on-a-
glove sensor as an excellent alternative for rapid, simple, and in-field analyses of nitro explosives
and emerging pollutants samples.
1 Bandodkar AJ et al., Analyst, 138, 2013, 5288.
2 Jong MD et al., Chem. Sci., 7, 2016, 2364.
3 Ferreira PC, et al., Trends Anal. Chem., 2019, 119, 1–16.
4 Mishra RK, et al., ACS Sensors, 2017, 2, 553–561.
[CNPq, CAPES, FAPESP]

62
ASSEMBLY OF AN IMPROVED HYBRID CASCADE FOR ETHYLENE GLYCOL
OXIDATION: ENHANCED CATALYTIC PERFORMANCE FOR A BIOFUEL CELL
Jefferson H. Franco (R)a, João Victor Bonaldo (R)a, Adalgisa R. de Andrade (R)a*,
Shelley D. Minteer (R)b
a University of São Paulo, Department of Chemistry, Ribeirão Preto, São Paulo, Brazil, 14040-901
b University of Utah, Department of Chemistry, Salt Lake City, Utah, United States, 84112
*e-mail: ardandra@usp.br
In the last years, enzymatic fuel cells (EFCs) have been investigated for application in small devices, such as
biosensors 1 and wearable electronic 2. Nevertheless, introducing a large group of enzymes on an electrode surface
leads to poor stability, decreasing the catalytic activity and power density 3. Notably, one way to increase electrode
stability is to prepare simple films containing the smallest number of components possible. Therefore, an alternative
route has been to prepare hybrid films 4. Recently, hybrid electrodes containing at least two catalysts capable of
harvesting the maximum number of electrons per fuel molecule have been proposed 5: these promising electrodes
combine just one organic catalyst and an enzyme on the electrode surface. Therefore, we report an Enzymatic Fuel
Cell (EFC) combining an enzyme that can cleave carbon-carbon bonds (oxalate oxidase (OxOx)) with an organic
catalyst (Pyrene-TEMPO (TEMPO = 2,2,6,6-tetramethyl piperidinyl-N-oxyl)) immobilized on the surface of modified
carboxylated multi-walled carbon nanotubes (MWCNT-COOH). This combination gave a hybrid bi-catalyst electrode
for complete ethylene glycol (EG) oxidation. The hybrid electrode provided nine-fold enhanced catalytic activity (0.17
mA cm-2) in the presence of EG as compared to the electrode in the absence of EG (0.018 mA cm-2), indicating that
the enzyme combined with the organic catalyst improved energy generation through deep EG electrooxidation.
Electrochemical impedance spectroscopy reveals that the addition of the enzyme in the electrode containing
MWCNT-COOH-Pyrene-TEMPO increased the charge transfer resistance (𝑅𝑐𝑡 ) and the capacitance of the double
layer (Table 1). Long-term electrolysis for 15 hours showed that the hybrid electrode presented outstanding current
density and stability (Figure 1). The EG oxidation products were identified and quantified by high-performance liquid
chromatography (HPLC-UV/RID). The results confirmed complete EG oxidation in the presence of CO 2 in the
solution, allowing 10 electrons to be collected from the fuel (Scheme 1). Overall, this study illustrates the
development of a simple and improved hybrid bi-catalyst electrode for promising applications in small electronic
devices.
Table 1. Evaluation and comparison of 𝑅𝑐𝑡 (Ω) and capacitance (nF) values for the different electrode architectures measured at 0.850 V
vs Ag/AgCl,Cl-.
Electrode MWCNT-COOH MWCNT-COOH/Pyrene-TEMPO Hybrid system
Absence of EG Rct (Ω) 60.9 178 90.1
Capacitance (nF) 220 370 400
N 0.928 0.882 0.89
𝑅𝑐𝑡 (Ω) 193 92 124
Presence of EG
Capacitance (nF) 230 780 523
N 0.934 0.875 0.831
B (𝑠 1/2) 1.23 1.58 1.61
Figure 1. Current versus time measurement for long-term electrolysis at Scheme 1. Proposed mechanism for complete EG
the MWCNT-COOH/Pyrene-TEMPO (solid black line) or MWCNT- oxidation catalyzed by the MWCNT-COOH/Pyrene-
COOH/Pyrene-TEMPO/OxOx (solid red line) electrode in the presence TEMPO/OxOx hybrid bi-catalyst electrode.
of 100 mmol L-1 EG. The supporting electrolyte (150 mmol L-1 citric
acid-phosphate buffers (pH = 5.2) is represented by the dashed black
line. Applied potential: 0.850 V vs Ag/AgCl.
1 Franco JH, Minteer SD, De Andrade AR, Biosensors, 11, 2021, 41

2 Huang X, et al. Biosensors and Bioelectronics, 124-125, 2019, 40
3 Aquino Neto S, Minteer SD, De Andrade AR, Journal of Electroanalytical Chemistry, 812, 2017, 153
4 Macazo, FC, Minteer SD, Current Opinion in Electrochemistry 5, 2017, 114
5 Franco JH, et al. Biosensors and Bioelectronics, 121, 2018, 281
[FAPESP, CAPES Army Research Office MURI]

63
Au SENSOR BASED ON A NEW PORPHYRAZINE TO NITRITE
DETERMINATION
Leonardo M. A. Ribeiro*a, Hiago N. Silvaa, Marcos M. Toyamab, Henrique E. Tomaa, Mauro
Bertottia
a University of São Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São Paulo, SP, Brazil, 05508-
000
b Mauá Institute of Technology, Chemistry Department/IMT, São Paulo, SP, Brazil, 09580-900
*e-mail: leonardo.ma.ribeiro@usp.br
The nitrite ion (NO2-) is a common chemical in our routine, recurrently used on processed meat as a curing
salt1. In 2015, the International Agency for Research on Cancer (IARC) classified NO 2- as “probably
carcinogenic to humans”. Moreover, NO2- can be toxic to the aquatic environment, fish, and crustaceans 2;
hence, strategies to monitor such compound easily, quickly, and inexpensively are welcome. The aim of this
work is to fabricate a new Au microfiber (d = 25 µm) electrochemical sensor whose surface was modified with
a Cobalt (II)-tetra(3,4-pyridyl)-porphyrazine (TRP) film. A similar porphyrazine containing Cu(II) as a metallic
center has already demonstrated catalytic activity towards nitrite oxidation 3, suggesting that this new
compound can also mediate electron-transfer reactions. A 10-4 mol L-1 TRP solution (prepared in methanol)
was dip-coated onto the electrode surface of a homemade Au microsensor 4. The microelectrode was
fabricated in a micropipette tip by inserting the metallic fiber, which was then fixed with epoxy resin. A Ni-Cr
wire was used to make the electronic contact5,6. Figure 1 shows CVs in a PBS solution containing NO 2- at
optimized pH (from 4 to 12) and a shift in the onset potential towards less positive values, and a remarkable
anodic current increase can be noticed using the modified microelectrode compared to the bare one. The TRP
leaching was also evaluated, and the modified electrode was subjected to 350 CVs in the 0.0 to 0.95 V potential
range (PBS, pH = 7.0). A slight current decrease (at E = 0.85 V) was observed after the experiment (around
3%), demonstrating the film was stable under the experimental conditions. Preliminary experiments with cyclic
voltammetry indicate a correlation between limiting current and NO2- concentration at low concentrations (µmol
L-1 range), and further experiments are in progress to reach sub-µmol L-1 levels range.
40
35
30
25
i / nA
20
15
10
-5
0.0 0.2 0.4 0.6 0.8 1.0
E / V vs Ag/AgCl(KCl sat)
Figure 1. Cyclic voltammograms recorded in PBS Figure 2. Molecular structure of TRP.
(pH = 7) before (blue curve, modified Au microelectrode)
and after the addition of nitrite (1 mmol L-1) using a bare Au
(black curve) and a modified microelectrode (red curve).
Scan rate: 100 mV s-1.
1 Hartman PE, Chem. Mutagen, 7, 1982,211.

2 Kroupova H, Machova J, Svobodova Z, Vet. Med. (Praha), 50, 2005, 461.
3 Matsumoto MY et al. J. Braz. Chem. Soc. 20, 2009, 728.
4 Huang XJ, O’Mahony AM, Compton RG, Small 5, 2009, 776.
5 Li H, Wu, N.; Nanotechnology 19, 2008, 6.
6 Ordeig O. et al. Electroanalysis 19, 2007, 1973.
[FAPESP, CNPq]

64
ELECTROCHEMICAL SENSOR BASED ELECTROPOLYMERIZED
MOLECULARLY IMPRINTED POLYMER POLY(ANILINE) ON
NANOSTRUCTURED SURFACE FOR LINOLEIC ACID DETECTION
Max Fabricio Falone (PG)a*, Edervaldo Buffon (PG)b, Nelson Ramos Stradiotto (R)a,b
a São Paulo State University (UNESP), Institute of Chemistry, Araraquara, SP, Brazil. 14800-060.
b São Paulo State University (UNESP), Bioenergy Research Institute, Araraquara, SP, Brazil. 14800-060.
*e-mail: max.falone@unesp.br
The agro-industrial wastes from the processing of guava are equivalent to 10 - 15% of the total mass
of the fruit, consisting of peel, macerated pulp, and seeds. However, they are irregularly discarded.
These wastes are rich in fatty acids such as Linoleic Acid (LA). LA has rich anti-cancer, anti-obesity,
and anti-diabetic properties. Thus, the literature reports several analytical methods for the
determination of LA based on chromatographic and spectroscopic techniques. On the other hand,
electrochemical sensors stand out for bringing together excellent advantages for analytical
determinations, such as: portability, sensitivity, selectivity, low detection and quantification limits.
However, this work reports for the first time the determination of LA in guava agro-waste on an rGO-
modified surface decorated with FeNPs coated with molecularly imprinted poly(aniline). Initially, a
GCE was modified with rGO using a suspension of graphene oxide 0.50 mg mL-1 in Na2SO4 0.10
mol L-1, where a potential of –1.4 V was applied for 500 s. Then, a 5.0×10-3 mol L-1 solution of FeCl3
in KCl 0.10 mol L-1 was used to carry out the electrodeposition of FeNPs on the GCE/rGO. The
formation of FeNPs on the electrode surface occurred by applying a constant potential of –1.4 V until
reaching an electrodeposition charge of 30 mC. The molecularly imprinted film was prepared from a
0.1 mol L-1 aniline solution containing 3.0×10-6 mol L-1 LA in 0.5 mol L-1 H2SO4. The
electropolymerization process was carried out using CV, where 15 voltammetric cycles were applied
in a potential range of –0.2 to 1.0 V (50 mV s-1). The experimental factors that affect the analytical
performance of the sensor were optimized: monomer concentration, number of cycles for
electropolymerization, extraction time and rebinding time. Subsequently, the electrode was
characterized by scanning electron microscopy, X-ray energy spectroscopy, electrochemical
impedance spectroscopy and cyclic voltammetry. The electrode exhibited a wide concentration
range from 1.0×10-12 mol L-1 to 1.0×10-10 mol L-1, with a LOD 3.0×10-13 mol L-1 and excellent
sensitivity (3.4×107 L A mol L-1). Furthermore, tests were made with interferents that composes the
guava waste sample and the results show a high capacity of the sensor for the selective recognition
of LA. Sensor repeatability tests were performed, through intra-day and inter-day measurements
showing current values of 1.1 (± 0.5) × 10−4 µA (RSD = 4.9 %; n = 15) and 3.8 (± 0.4) × 10−4 µA (RSD
= 1.1%; n = 15), respectively. The device stability was monitored for 15 days through alternating
measurements, in this way, the sensor preserved 92% of the current magnitude obtained initially.
The determination of LA in the sample was performed using the standard addition method. The
sample was enriched with known concentrations of LA in the range 1.0×10−12 mol L-1 to 5.0×10 −12
mol L-1. The concentration of LA found in a sample of guava agro-waste was (1.3 ± 0.1) mol L-1 (n =
3). These results were validated through recovery trials, where the recovery rates obtained ranged
from 93.5% to 111% (RSDs ≤ 5%). The results indicate that it can be successfully applied in the
detection of LA in guava agro-waste exhibiting high sensitivity and selectivity, excellent analytical
performance, good repeatability, reproducibility and stability.
[FAPESP]

65
PEPTIDE-BASED ELECTROCHEMICAL BIOSENSING FOR USE IN COVID-19
ANTIBODY TESTS
Ana C. H. Castro-Kochi (R)*a, Ítalo R. S. Bezerra (PG)a,b, Aline M. Pascon (R)a,b, Gabriela H.
Silva (PG)b, Eric A. Philot (R)g, Vivian L. de Oliveira (R)f, Rodrigo S. N. Mancini (PG)a, Gabriel
R. Schleder (R)h, Carlos E. Castro (R)a, Murilo Santhiago (R)a,b, Ives Charlie-Silva (R)c, Diego
S. T. Martinez (R)b, Ana L. Scott (R)g, Renato S. Lima (R)a,b,d,e, Wendel A. Alves (R)a
aFederal University of ABC, Center for Natural and Human Sciences, Santo André, SP, Brazil, 09210-580
b Brazilian
Center for Research in Energy and Materials, Brazilian Nanotechnology National Laboratory, Campinas, SP,
Brazil, 13083-970
c University of São Paulo, Institute of Biomedical Sciences, São Paulo, SP, Brazil, 05508-000
d University of Campinas, Institute of Chemistry, Campinas, SP, Brazil, 13083-970
e University of São Paulo, São Carlos Institute of Chemistry, São Carlos, SP, Brazil, 09210-580
f University of São Paulo, Heart Institute, São Paulo, SP, Brazil, 05508-000
g Federal University of ABC, Center for Mathematics, Computing and Cognition, Santo André, SP, Brazil, 09210-580
h Harvard University, School of Engineering and Applied Sciences, Boston, Massachusetts, USA, 02134
*e-mail: castro.achonorato@gmail.com
The methods used to detect SARS-CoV-2 contributed to slowing the spread. While valuable, these
have proven to be limited in massive applications. Therefore, we need to develop portable, scalable,
rapid, and field-deployable methods for COVID-19 detection.1 Thus a peptide (PEP) (Asn-Asn-Ala-
Thr-Asn-COOH), which mimics the SARS-CoV-2 spike protein, was synthesized and validated by
bioinformatics and ELISA and used to create electrochemical biosensors based on carbon electrode,
gold nanoparticles (AuNPs) and electrochemical impedance spectroscopy (EIS) for screening
COVID-19 in patient sera (ethics comitte nº CAAE:43139921.2.0000.5594). AuNPs with different
diameters (18 nm and 28 nm) were synthesized. AuNPs (18 nm) increased the surface area but
decreased the Rct values by 58% due to the smaller curvature, which limits the binding sites. It was
observed that regardless of the size of AuNPs, the presence of PEP allows distinguishing the positive
control (PC; convalescent patients) from the negative control (NC; pre-pandemic patients) (Figure
1A). The other tests were performed with AuNPs (28 nm). Molecular docking studies have shown
that hydrogen bonds and hydrophobic interactions drive the binding of PEP with anti-Spike
antibodies (IgGS).2 The biosensor showed good sensitivity (LOD 1.28 µgmL-1) (Figure 1B) and
stability (95.1%, 20 days). In addition, it was specific in assays using antibodies and complex real
samples (Figure 1C and 1D). The Rct values obtained for NC and PC could be easily classified,
considering the cut-off point (black line; 8.0 kΩ)(Figure 1D). The heterogeneity in PC samples was
due to age, sex and genetics, which influence the humoral response. With the application of a simple
equation adjusted by machine learning, this biosensor provided COVID-19 screening of 39 biological
samples in healthy and infected groups. 100.0% accuracy.2 This platform promises to provide direct,
fast, and accurate COVID-19 biosensors with significant economic and social impacts at the point of
care.
Figure 1. Biosensor for COVID-19 (A) Bar graphs with Rct values on the effect of size AuNPs on immobilization and
detection. (B) Calibration curve using IgGS. (C) Bar graphs of Rct values obtained by Randles plot of specific IgG (IgGS)
and non-specific IgG (IgGR1: Anti-Receptor Binding Domain (RBD), IgGR2: Anti-RBD-mFc, IgGE: Anti-Guinea Pig CD3,
IgGT: Anti-Guinea Pig T-Bet). (D) Bar graphs with Rct values obtained from real samples. n=3. Electrolyte: Solution of 5
mM K4Fe(CN)6/K3Fe(CN)6 in 0.1 M KCl.
1 Nicoliche, CYN. et al. ACS Appl. Mater. Interfaces, 14, 2022, 2522–2533.
2 Castro, ACH. et al. ACS Nano, 2022 – submitted
[INCTBio, CAPES, CNPq, FAPESP, UFABC]

66
ELECTROCHEMICAL SENSOR BASED ON MOLECULARLY IMPRINTED
POLYAMINOACID CONTAINING RGO AND MNNPS FOR THE
DETERMINATION OF PALMITIC ACID
Bruna Gabrielle Olsen Pinto (UG)a,b*, Max Fabricio Falone (PG)a,b, Edervaldo Buffon (PG)a,b,
Nelson Ramos Stradiotto (R)a,b
a São Paulo State University (UNESP), Institute of Chemistry, Araraquara, SP, Brazil. 14800-060.
b São Paulo State University (UNESP), Bioenergy Research Institute, Araraquara, SP, Brazil. 14800-060.
*e-mail: bruna.olsen@unesp.br
The guava cultivation in Brazil has increased considerably in the last few years, showing a great
economic importance for the country. However, the agroindustrial waste of this fruit, composed of
peel, pulp and seed, is discarded irregularly, contributing to waste of compounds of interest and
environmental contamination1. This way makes it necessary to find an efficient alternative to reuse
this waste. A class of compounds that this waste constitute is the fatty acids, mainly in the form of
linoleic, oleic and palmitic acid (PA), that have great importance in the pharmaceutical and food
industry, specially. Thus, the development of analytics methods for the determination of PA brings
great relevance to the fruit refinery and it can promote an environmentally favorable destination2.
The PA determination methods found in the literature are based on chromatographic and
spectroscopic techniques. Although they achieve good analytical performance, they also have high
instrumental cost and a long time for the analysis. Meanwhile, electrochemical sensors have
advantageous analytical characteristics such as selectivity, low limit of detection and quantification
and lower cost. This is the first electrochemical sensor for the determination of PA containing
reduced graphene oxide (rGO) and manganese nanoparticles (MnNPs) coated with L-Serine (L-
Ser), an aminoacid that adds great aspects to the sensor due to its compatibility with other molecules,
being also a biodegradable polymer since it is soluble in water, contributing for an environmentally
friendly device3. Initially a suspension of 0.4 mg mL-1 of rGO in 0.1 mol L-1 of Na2SO4 solution was
electrodeposited on the GCE surface through chronoamperometry, applying a potential of -1,4V for
500s. Then, the MnNPs were electrodeposited from a 5.0×10-3 mol L-1 MnSO4 in a 0.1 mol L-1 Na2SO4
solution onto the GCE/rGO through chronoamperometry in which a potential of -0.7 V was applied
until the charge reached 0.9 mC. Next, in the electropolymerization, a solution containing 1.0×10-5
mol L-1 L-Ser and 1.0×10-6 mol L-1 PA in PBS (pH 7.0) was used. The GCE/rGO/MnNPs electrode
was then immersed in this solution for 10 consecutive voltammetric cycles, applied in the potential
range from -0.3 V to 1.3 V (50 mVs-1). The template extraction process involved the application of 6
consecutive voltammetric cycles in a 0.1 mol L-1 NaOH solution in a potential range from 0 V to 1.5
V (50 mV s-1). For the rebinding process, the electrode was placed for 10 minutes in different
concentrations of PA in PBS (pH 7.0). All experimental factors that influence the analytical
performance of the device were optimized, such as L-Ser concentration, electropolymerization and
extraction cycles, electropolymerization and rebinding pH and rebinding time. The electrode was
characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy,
electrochemical impedance spectroscopy and cyclic voltammetry. The analytical curves were
obtained in a linear range from 2.0×10-12 mol L-1 to 1.0×10-11 mol L-1, represented by the linear
regression equation ∆I (µA) = 7.8× 105 C PA + 1.1× 10-5 (R2 = 0.993), showing a LOD of 6.5×10-13 mol
L-1. Furthermore, tests were made with interferents that composes the guava waste sample. The
proposed device showed high selectivity, repeatability, reproducibility, and excellent stability, being
successfully applied in the selective determination of PA obtained from agroindustrial guava waste.
1 Sousa JR et al., Química Nova, 37, 2014
2 Hayes DG; Smith G A, Biobased surfactants, 2019.
3 Alhedabi T et al., Materials Chemistry and Physics, 185, 2016.
[FAPESP]

67
AN ELECTROCHEMICAL CALIBRATION-FREE METHOD
FOR ASCORBIC ACID SENSING
Valdomiro S. Conceição (PG)a*, Douglas P. M. Saraiva (PG)a, Mauro Bertotti (R)a
a University of São Paulo, University of São Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São
Paulo, SP, Brazil, 05508-900
*e-mail: valldomiro@usp.br
Ascorbic acid (AA) is added to several food products, such as juices and dairy drinks, due to the
antioxidant property that prevents the proliferation of microorganisms [1]. In addition to food
preservation, ascorbic acid - or vitamin C - is a drug marketed as a vitamin supplement because it
scavenges free radicals in the body, acting on the immune system [1]. Analytical methodologies for
quantifying this molecule so present in everyday life are essential, but quantitative analyzes require
calibration curves, which implies excessive use of reagents, waste production, extended periods for
the analyst to build an analytical plot, and errors due to interfering matrix effects. An attractive
strategy would be direct methods, such as coulometry, but in cases of complex samples, the difficulty
is accentuated since it is necessary to guarantee that only the analyte undergoes the reaction of
interest and that it is entirely consumed in the analytical process. Given the characteristics of the
current quantitative methods, the main goal of this work is to develop an electrochemical method to
analyze ascorbic acid without using a calibration plot. Conceição et al. have demonstrated that it is
possible to get information on the concentration of a well-known electrochemical probe (potassium
ferricyanide) through a proof of concept study using chronoamperometry at a microelectrode [2]. We
aim now to extend the work by determining the concentration of a chemical species with irreversible
electrochemical behavior. AA has such a feature, and the use of a disk microelectrode, in addition
to allowing miniaturization, has a series of characteristics in which it is worth highlighting the
presence of a steady-state current (iL) = 4nFDCr, where F is the Faraday constant, D is the diffusion
coefficient, n is the number of electrons involved in the redox reaction, C is the concentration of the
electroactive species, and r is the radius of the microelectrode. The radius value is precisely found
by optical microscopy, and the n value can be obtained through knowledge of the electrochemical
reaction. Information on the diffusion coefficient is difficult to obtain since D depends on the sample
medium. However, Figure 1 shows that such value can be extracted from the chronoamperogram,
according to a methodology proposed by Denuault and coauthors [3]. Taking into account the limiting
current (iL), the concentration can be determined from the equation given above. Samples of known
concentrations and vitamin supplements of AA were used
in this study, and concentration values of (3.6±0.2) mM for
the 4.0 mM AA solution, and (2.19±0.03) mM for the 2.0
mM AA solution, and (24.7±0.9) mM for the 22.7 mM
drugstore supplement were found, i.e., the deviations
were around 10%, 9.5% and 8.8%, respectively. In view
of the satisfactory results, further experiments are in
progress using the same methodology for samples with
low concentrations, such as milk samples.
Figure 1. Chronoamperogram recorded with a

carbon fiber disc microelectrode in 4 mM ascorbic
acid solution. E = 1.2 V. Inset: Linearization of the
transient-state.
1 Motshakeri M, et al., Electrochimica Acta,265, 2018.

2 Conceição VS, et al., Reunião Anual da Sociedade Brasileira de Química, 45, 2022.
3 Denuault G, Mirkin MV, Bard AJ, 308, 1991, 1-2.
[FAPESP, CNPq]

68
EVALUATION OF NANOPOROUS GOLD BASED SENSORS FOR
ELECTROCHEMICAL DETECTION OF NITRITE
Daniel V. Braga (UG)a, Bruna B. Nascimento (UG)a, Douglas P. M. Saraiva (PG)a Mauro
Bertotti (R)a*
a University of São Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São Paulo, SP,
Brazil, 05508-900
*e-mail: daniel12veloso@usp.br
Nitrite is widely used as a preservative in beverages, processed foods, and meat, as it has
antimicrobial effects against hazardous bacteria and can be dangerous for human health and the
environment. The maximum allowed value for nitrite is 0.9 ppm for drinking water and 200 ppm for
processed meat. In large amounts in the human body, nitrite can oxidize the iron from the hemoglobin
nucleus, generating methemoglobin and impairing oxygen transport. The interaction of nitrite with
amines can also lead to carcinogenic compounds such as N-nitrosamines. Usually, nitrite is detected
by capillary electrophoresis, chemiluminescence, spectrophotometry, chromatography, and
electrochemical techniques. The high sensitivity, low cost, and fast response have increased the
interest in electrochemical methods. Many nanomaterials have been used as electrochemical
sensors to improve the signal generated by nitrite oxidation. In this work, we evaluated the production
of nanoporous gold-based sensors for nitrite electrochemical detection. Nanoporous structures with
high electroactive surface area were prepared via one step anodization-electrochemical reduction
method using a gold microelectrode. The chosen NPG formation technique was based on the
evolution of oxygen during anodic corrosion and subsequent reduction of the oxide formed in 0.5
mol L-1 sulfuric acid. Briefly, a sweep is performed from a double layer potential up to a potential for
nanopores formation, generally less positive
than 2.0 V at a slow scan rate of 20 mV s-1.
Under these conditions, oxygen evolution and
electrode surface corrosion occur, generating
nanopore structures. The applied potential is
maintained for a given time, initially 900 s,
under vigorous agitation, in order to prevent
bubbles from obstructing the electrode
surface. After the anodizing step, a reduction
is performed by linear sweep, again at 20 mV
s-1, from the fixed applied potential to 0.3 V.
During this process, the electrode´s metallic
surface is restored. Studies regarding the
electrochemical characterization of the
surfaces obtained by changing the anodization
potential and time show a strong influence of
such parameters on the electroactive surface
area and the response towards nitrite Figure 1. Voltammetric curves recorded with bare (in black)
oxidation. At optimized conditions, a slight and nanoporous gold microelectrodes produced at 2.0 V (in
red) and 2.4 V (in blue) in a 2.0 mmol L-1 NaNO2 solution in
oxidation potential shift towards less positive PBS. Scan rate: 0.1 V s-1. Microelectrode radius: 25 µm.
values and an increased current response
compared with the bare gold microelectrode
were noticed.
1 Liu Z, Manikandan VS; Chen, A. Curr. Opin. Electrochem., 16, 127-133, 2019.
2 Kumar A; Gonçalves JM, Sukeri A, Araki K; Bertotti M, Sens. Actuators B Chem., 263, 237-347, 2018.
[CNPq, ENQA, LSEME, SBQ, FAPESP]

69
GRAPHITE-BASED ADDITIVELY MANUFACTURED ELECTRODE AS
PLATFORM FOR THE BIOSENSING OF SARS-COV-2
Jéssica S. Stefano (R)a*, Luiz Ricardo G. Silva (PG)b, Raquel G. Rocha (PG)b, Laís Canniatti
Brazacac, Eduardo M. Richter (R)b, Rodrigo A. A. Muñoz (R)b, Bruno C. Janegitz (R)a
a Federal University of São Carlos, Department of Nature Sciences, Mathematics and Education, Araras, SP, Brazil,
13600-970.
b Federal University of Uberlândia, Institute of Chemistry, Uberlândia, Minas Gerais, 38400-902, Brazil.
c University of São Paulo, Institute of Physics, São Carlos, SP, 13560-970, Brazil.
*e-mail: jessica.s.stefano@gmail.com
The 3D printing technology has gained ground due to its wide range of applicability.1 Recently, 3D
printing has played a significant role in the fight against the global pandemic event COVID-19 through
decentralized and on-demand manufacturing of different personal protective equipment and medical
devices.2 On the other hand, 3D printing technology is still considered recent and with great potential
to be explored for the most diverse purposes, including the manufacturing of analytical devices.3 The
development of new conductive filaments contributes significantly to the production of improved
electrochemical devices. In this sense, the fabrication of 3D printed devices for the development of
electrochemical (bio)sensors becomes an almost unexplored field of great interest. Therefore, this
work has presented the use of a new electrochemical 3D printed sensor, based on a lab-made
graphite conductive polylactic acid (PLA) filament (Gpt-PLA) for the detection of SARS-CoV-2 spike
protein. A ready-to-use lab-made conductive filament with a high graphite loading (40% wt.) on a
PLA matrix, was used for 3D printing the counter, reference, and working electrodes, and a simple
base constructed using non-conductive PLA was responsible for the electrodes coupling. The
biosensor was prepared using the electrode simply polished mechanically in sandpaper, and
assembled by covalently bonding EDC:NHS (10.0 and 20.0 mmol L-1, respectively), directly on the
electrode surface (1 h). Next, 1 µg mL-1 SARS-CoV-2 antibody was incubated for 1 h, and finally, the
blocking with BSA solution (1% w/v) was performed for 30 min. Fig. 1A schematizes the biosensor
preparation process. For the analysis, a redox probe (1.0 mmol L-1 ferrocene methanol - FcMeOH)
was used before and after the incubation (30 min) of the SARS-CoV-2 spike S1 protein, in increasing
concentrations (from 5.0 to 75.0 nmol L-1), and the decrease in the redox probe response by cyclic
voltammetry, using the biosensor, was monitored (Fig. 1B and C).
Figure 1. (A) Representative scheme of the biosensor fabrication, (B) CVs obtained for increasing concentrations of spike
S1 protein (5.0 to 75.0 nmol L-1), in presence of 1.0 mmol L-1 FcMeOH in 0.1 mol L-1 KCl. Scan rate: 100 mV/s. (C)
respective calibration curve.
LOD value of 1.36 nmol L−1 (0.10 μg mL−1) and sensitivity of 0.17 μA nmol−1 L (0.01 μA μg−1 mL)
were obtained. The biosensor was tested in the analysis of S1 protein in synthetic saliva after
fortification with four known concentrations (5.0; 10.0; 20.0 and 50.0 nmol L−1 S1). Recovery values
ranging from 86.3 ± 2.7% to 96.1 ± 10.2% attested to the biosensor's suitability for saliva tests.
Therefore, the use of lab-made conductive filaments is promising and can become an alternative
route for the obtention of electrochemical biosensors. In addition, the development of 3D printed
electrochemical biosensors capable of detecting the SARS-CoV-2 virus can be used as a tool of
great assistance in the rapid testing of patients and providing monitoring of infected people.
1 Cardoso RM, Kalinke C, Rocha RG, et al., Anal. Chim. Acta. 1118, 2020, 73.
2 Choong YYC, Tan HW, Patel DC, et al., Nat. Rev. Mater. 5, 2020, 637.
3 Munõz J, Pumera M, Chem. Eng. J. 425, 2021, 131433.
[FAPESP, CAPES, FAPEMIG, CNPq]

70
A NO SENSOR BASED ON A PORPHYRAZINE Pt MODIFIED ELECTRODE
Bruna Bossarda*, Leonardo M. A. Ribeiro (PG)a, Hiago N. Silva (PG)a, Marcos M. Toyama
(R)b, Henrique E. Toma (R)a, Mauro Bertotti (R)a
a Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP, Brazil, 05508-
000
b Mauá Institute of Technology, Chemistry Department/IMT, São Paulo, SP, Brazil, 09580-900
*e-mail: bruna.bossard.nascimento@usp.br
Since the ’90s, NO has been seen as a molecule of great biological importance and responsible for
numerous biological processes such as vasodilation, immune response, neurotransmission, and
some pathologies. However, the direct in vivo NO detection is problematic owing to several factors
such as NO concentration range, selectivity, and calibration1. The construction, development, and
characterization of a NO microsensor based on a platinum microelectrode modified with TRP, a
cobalt (II)-tetra(3,4-pyridyl)-porphyrazine (TRP), is presented in this work. The platinum microwire (d
= 25 µm) was inserted into a micropipette tip filled with epoxy resin for fiber isolation. The external
electrical contact was made with a Ni-Cr wire and carbon black2. The platinum fiber was dip-coated
on a 10-4 mol L-1 TRP solution and then let dry before use. Cyclic voltammetry (CV) was performed
in 5.4 mmol L-1 [Fe(CN)6]3- with 0.5 mol L-1 KCl as supporting electrolyte using the bare and the
modified microelectrode. The typical sigmoidal shape was obtained for both microelectrodes,
demonstrating that the immobilized TRP has no influence on the electron transfer process involving
[Fe(CN)6]3-. Similar behaviour was noticed by repeating the experiments with both microelectrodes
in 0.5 mol L-1 H2SO4. Figure 1 A shows CVs recorded with the modified microelectrode in a 0.6 mmol
L-1 NO solution in PBS, and a remarkable current increase can be noticed compared to the bare
microelectrode. The sensor's analytical response to changes in NO concentration was examined
through amperometry at 0.9 V during successive additions of the target analyte in PBS, as shown in
Figure 1 B. A current response clearly distinguishable from that of the background is noticed for 4.5
nmol L-1 NO, demonstrating the sensor is very sensitive toward NO detection. The TRP leaching
was evaluated by running 100 CVs (range from 0 to 0.95 V, scan rate 500 mV s-1) in a 0.6 mmol L-1
NO solution prepared in Phosphate Buffer Solution (PBS), with no significant changes being
observed. These preliminary studies suggest that a microsensor with such features can be applied
to investigate biological processes related to the release of NO in tissues or cells.
Green and red curves correspond to CVs recorded with both microelectrodes in supporting electrolyte. Scan rate: 100 Mv
S-1. (B) Amperometry performed at 0.9 V with the modified electrode during successive additions of NO solution (0.3, 1.0,
4.5, 22, 39, 57, 110, 462 nmol L-1 and, 1.3, 4.9, 9.4, 15, 33, 51 µmol L-1) (inset: start of additions with current response).
Figure 1. (A) CVs recorded in PBS + 0.6 mmol L-1 NO with a bare (blue line) and a modified microelectrode (black line).
1 Hall, C. N.; Garthwaite, J.; Nitric Oxide - Biol. Chem. 2009, 21, 92.
2 Paixão, T. R. L. C.; Bertotti, M.; Quim. Nov. 1306, 32.
[CNPq, FAPESP, PRG-USP]

71
EVALUATION OF NANOPOROUS GOLD SENSORS FOR NITRIC OXIDE
DETECTION
Douglas P. M. Saraiva (PG)a* Daniel V. Braga (UG)a, Mauro Bertotti (R)a
a Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP, Brazil, 05508-
900
*e-mail: dpm.saraiva@gmail.com
Nitric oxide is a nonionic radical that can easily permeate cell membranes. Inside the body, it acts
as a mediator for countless processes, such as wound healing, vasodilation, inflammation, immune
response, and long-term memory potentiation. It is produced at low concentrations ranging from pM
to nM in the endothelial and neuronal tissue, usually initiating enzymatic cascades through cyclic
GMP. In contrast, NO can be found at higher concentrations in inflammatory processes, ranging
from nM to µM, creating nitrosative and oxidative environments that assist the body´s defenses in
killing pathogens. With so many effects and even antagonizing ones happening in different
conditions, the presence of nitric oxide inside the body heavily depends on its concentration and
location. Numerous methods and techniques can be used for nitric oxide detection, and
chemiluminescence and EPR measurements are commonly used for real-time measurements.
However, none of these methods can easily be used for inside-the-body measurements. Moreover,
nitric oxide has a very low half-life as it is very reactive with oxygen, so ex-vivo measurements are
usually done by one of its byproducts. On the other hand, electrochemical sensors are easy to
miniaturize for implantation and are typically used for in-vivo quantification, providing an efficient way
to explore the nitric oxide function in biological
processes. In this work, we bring the
construction and evaluation of nanoporous
sensors for nitric oxide detection for later use
in in-vivo measurements. Nanoporous
structures with high electroactive surface
area were prepared by a dynamic hydrogen
bubble template method in acidic medium.
Briefly, in a 5 mM HAuCl4 + 0.5 M H2SO4
solution, the microelectrode is polarized at a
very negative potential, usually less negative
than -1.0 V, to achieve vigorous hydrogen
evolution for a given time. Under these
conditions, the chloroaurate anion is
simultaneously reduced at the electrode
surface with the hydrogen bubble evolution,
creating the nanoporous structure. The nitric
Figure 1. Voltammetric curves recorded in PBS with a bare
oxide electrochemical behavior was Au electrode and a nanoporous Au microelectrode before (in
examined by varying the metal precursor black and grey, respectively) and after the addition of NO (0.6
concentration, deposition potential, and mM) (in red and blue, respectively). Scan rate: 0.1 V s -1.
deposition time in the sensor fabrication. The Microelectrode radius: 25 µm.
experiments were made using a 25 µm of
radius gold microelectrode. At the optimized conditions, a nine-fold increase in oxidation current
response and a peak potential shift towards a less positive potential of about 80 mV were found
compared with the bare electrode. The gold electroactive area was determined by measuring the
charge under the gold oxide monolayer in sulfuric acid. The electroactive area increase under these
conditions, of about 150 times, is much more significant than the increase in the oxidation current,
and we are still looking for plausible explanations.
1 Liu, Z., Manikandan, V. S.; Chen, A. Curr. Opin. Electrochem., 16, 127-133, 2019.
2 Brown M.D.; Schoenfisch M. H. ACS Sens. 4, 7, 1766–1773, 2019.
[CNPq, ENQA, LSEME, SBQ, FAPESP]

72
LABEL-FREE IMPEDIMETRIC BIOSENSOR
FOR DETECTION OF SARS-CoV-2 USING DNA SEQUENCES
Tayane A. Freitas (R)a, Ilanna C. Lopes (R)b, Mayara I. S. Lima (R)c, Auro A. Tanaka (R)a,d*
a Universidade Federal do Maranhão, Departamento de Química, São Luís, MA, Brazil, 65080-805
b Universidade Federal do Maranhão, Departamento de Tecnologia Química, São Luís, MA, Brazil, 65080-805
c Universidade Federal do Maranhão, Departamento de Biologia, São Luís, MA, Brazil, 65080-805
d Instituto Nacional de Ciência e Tecnologia em Bioanalítica, C.P. 6154, Campinas, SP, Brazil, CEP 13083-970
*e-mail: tanaka.auro@ufma.br
An acute febrile illness with severe respiratory distress syndrome emerged in December 2019. This
illness progressively evolved, being an infection caused by a new coronavirus called severe acute
respiratory syndrome coronavirus 2 (SARS-CoV-2).1 This virus causes COVID-19, leading to a
pandemic that still considerably affects the socioeconomic and public health sectors worldwide. The
development of various technological solutions to combat the COVID-19 outbreak is rapidly taking
shape across the globe. In the last years, the use of biosensors as viable tools for medical diagnosis
and other applications has grown significantly due to their high sensitivity, specificity, precision, low
cost, and potential for large-scale production.2 Thus, a label-free DNA biosensor to detect SARS-
CoV-2 was developed using screen-printed gold electrodes (SPGE) modified with SARS-CoV-2
oligonucleotide sequences. A thiolated single-stranded capture probe (CP1-thiol) was immobilized
by chemisorption on the surface of SPGE, which was then hybridized with a sequence of the
complementary DNA strands of SARS-CoV-2 (CT1, biological sample simulation). According to the
experimental results obtained, it was observed that the recognition of the CT1 target was based on
the variation of the charge transfer resistance (ΔRct) of the redox probe ([Fe(CN)6]3−/4−), showing a
decrease in Rct after hybridization. This behavior was attributed to the new conformation of the
secondary structure of the double strand. Label-free detection was performed using electrochemical
impedance spectroscopy (EIS). To obtain the best performance, experiments were conducted to
characterize the DNA biosensor surface and optimize the analysis conditions such as capture probe
immobilization, probe immobilization time, and hybridization time. The developed biosensor showed
good linearity in the range of 5.0 × 10−10 to 7.5 × 10−8 mol L−1 (r = 0.993) and a detection limit of
0.42 nmol L−1 (Figure 1), analysis time of approximately 35 minutes, and low sample consumption
(requires a maximum of 15 μL). Thus, the developed DNA biosensor for SARS-CoV-2 was viable for
further application in diagnosing COVID-19.
500
400
400
300
300
Rct ()
-Z" ()
200
200
-10 100
100 5.0 x 10
r = 0.993
0
-8
7.5 x 10 mol L of CT1
-1 0
Rct () = 1544.3 - 161.8 log [CT1]
0 200 400 600 800 1000 1200 1E-9 1E-8 1E-7

-1
Z' () [CT1] (mol L )
Figure 1. Nyquist plots of the biosensor after hybridization with different concentrations of CT1: 5.0 × 10-10 to 7.5 × 10-
8 mol L-1. Standard deviations (n = 3).
1 Narita F, Wang Z, Kurita H, Li Z, Shi Y, Jia Y, Soutis C, Advanced Materials, 33, 2021, 1.
2 Bahl S, Bagha AK, Rab S, Javaid M, Haleem A, Singh RP, Journal of Industrial Integration and Management, 6, 2021,
175.
[CNPq]

73
EVALUATION OF NANOPOROUS GOLD-COBALT PORPHYRAZINE SENSOR
FOR ASCORBIC ACID DETERMINATION
Pedro H. A. Damasceno (PG)a Douglas P. M. Saraiva (PG)a Gilberto J. Silva Junior (PG)a
Leonardo M. A. Ribeiro (PG)a, Hiago N. Silva (PG)a, Marcos M. Toyama (R)b,
Henrique E. Toma (R)a, Mauro Bertotti (R)a
a Department of Fundamental Chemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP, Brazil, 05508-900
b Mauá Institute of Technology, Department of Chemistry /IMT, São Paulo, SP, Brazil, 09580-900
*e-mail: pedrodamasceno@usp.br
Vitamin C (ascorbic acid (AA) or ascorbate, its ionized form) is a water-soluble vitamin with many
functions. Among them, antioxidizing is one of the most important and known. Easily sharing
electrons, it is oxidized to dehydroascorbic acid. In the immunologic system, ascorbic acid is a strong
agent against free radicals produced endogenously and exogenously, and humans can get it by
alimentation1. It prevents scurvy, assists the organism´s defense system, and is fundamental to
forming collagen fibers that compound human tissues such as dermis, cartilage, and bones. Ascorbic
acid can scavenge singlet oxygen or act as a chelating agent and is primarily used as an antioxidant
in foods and drinks. Studies using wine showed that the benefit of ascorbic acid as an antioxidant is
associated with its capacity to sequester molecular oxygen. Ascorbic acid reacts rapidly with
radicals, like hydroxyl, showing strong trends in its use as an efficient antioxidant 2. In this work, the
fabrication of a cobalt (II) – porphyrazine/nanoporous gold sensor for ascorbic acid electrochemical
detection was evaluated. The microelectrode is homemade using a 25 μm diameter gold fiber. The
bare gold microelectrode sensor was first modified with nanoporous structures prepared by the
dynamic hydrogen bubble template (DHBT)3 method and then modified with a cobalt-porphyrazine.
Briefly, a - 3.0 V potential is applied for 150 s in a 5 mmol L-1 HAuCl4 + 0.5 mol L-1 H2SO4 solution.
During this step, the chloroaurate anion is reduced at the electrode surface simultaneously with the
hydrogen bubble evolution, creating the nanoporous structure (NPG). Then, the microelectrode
surface is modified by dip coating in a 100 µmol L-
1
cobalt (II)-tetra(3,4-pyridyl)-porphyrazine (TRP)
solution prepared in methanol. Compared to the
bare microelectrode, the anodic oxidation of
ascorbate exhibited a peak potential shift of about
850 mV to less positive potential using the
TRP/NPG modified electrode and a 100-fold
current increase, as shown in Figure 1. Such a
remarkable performance can be attributed to the
synergic effect caused by the presence of
numerous active sites in the NPG structure and the
catalytic activity of the immobilized porphyrazine.
Morphologic and electrochemical studies will be
performed to characterize the distribution of the
catalyst particles into the NPG film and to
understand the mechanism involving the electron Figure 1. Cyclic voltammograms (second cycle) recorded in
transfer step. The sensor will be then employed as phosphate buffer solution (pH ≈ 7) using a bare microelectrode
a miniaturized tool to get information on the role of (green curve) and the NPG-TRP modified microelectrode
ascorbate against cellular disorders caused as a (black curve), and in phosphate buffer solution (pH ≈ 7) + 4
mmol L-1 AA with the NPG modified microelectrode (blue
result of oxidative stress induced by UV irradiation curve) and the NPG-TRP modified microelectrode (red curve).
in epithelial cells (intra and extracellular). The inset shows a magnified CV recorded with the bare
microelectrode in phosphate buffer solution (pH ≈ 7) + 4 mmol
1Pisoschi, A. M.; Pop, A.; Serban, A. I.; Fafaneata, C.; L-1 AA. Scan rate: 100 mV s-1.
Electrochim. Acta 2014, 121, 443.
2Manela-Azulay, M. et al.; An. Bras. Dermatol. 2003, 78, 265.
3Regiart, M. et al.; ChemElectroChem 2020, 7, 1558.
[CNPq, LSEME, FAPESP]

74
FABRICATION OF AN ELECTROCHEMICAL SENSOR FOR THE DETECTION OF
PHOSPHATE IONS IN WATER SAMPLES
Luiz F. Z. Felipe*a,b, Gilberto J. Silva Junior (PG)a, Mauro Bertotti (R)a
a University of São Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São Paulo, SP, Brazil, 05508-
900
b Faculdades Oswaldo Cruz, São Paulo, SP, Brazil, 01151-000
*e-mail: nandozavatti@gmail.com
Phosphorus is an essential nutrient for the maintenance of aquatic ecosystems and its most common
inorganic form in water is the phosphate ion (PO43-). Such anion is mostly formed by rock weathering,
being liberated in the ground and transported to water resources. However, excess phosphate ions
in water can cause eutrophication, with concurrent change in important parameters for water quality,
such as pH, turbidity and dissolved oxygen concentration [1]. In this sense, the development of a
device that allows monitoring the concentration of these ions in water becomes highly desirable.
Electrochemical sensors stand out due to advantages such as portability, low cost, ease of
miniaturization and environmental compatibility. Hence, a nanoporus gold (NPG) electrode was
developed through a well-known electrodeposition method (Dynamic Hydrogen Bubble Template
(DHBT)) [2] applying -2 V for 150 s in a 1 mM HAuCl4 + 0.5 M H2SO4 solution. The electrochemically
active area of the NPG electrode was 3-fold higher compared to that of the bare Au electrode. As
phosphate is a non-electroactive ion, its detection was based on the reaction between phosphate
and molybdate ions, according to the following equations:
(1) 7 PO43- + 12 Mo7O246- + 72 H+ → 7 PMo12O403- + 36 H2O
(2) PMo12O403- + n H+ + n e- → [HnPMo12O40]3-
Fig. 1 shows the electrochemical behavior of PO43- ions in the presence and absence of excess
molybdate.
Figure 1A shows no Faradaic
process in a solution containing
only phosphate. On the contrary,
two reduction peaks at E = 0.1 V
and -0.1 V are noticed in the
presence of molybdate,
corresponding to the electron-
transfer processes involving the
phosphomolybdate complex
(equation 2). The current
increases with increasing the
Figure 1. Cyclic voltammograms recorded using a gold bare electrode in a phosphate concentration,
0.1 M KCl solution (pH 1) containing 30 mM MoO42- and different demonstrating that phosphate
concentrations of PO43- (A) and with different electrodes in a 2 mM PO43- + 30
2-
mM MoO4 solution (B). Scan rate: 50mV/s.
can be electrochemically
detected if molybdate is in
excess. Attempts to enhance the sensitivity were based on the use of the NPG electrode, and Figure
1B shows the cathodic current is higher compared to that of the bare Au electrode, even though such
an increase is not remarkable. However, studies to optimize parameters of the electrode surface
modification step are still being carried out, with the objective of verifying the best conditions for
manufacturing the NPG electrode. Electrode surface morphology will be evaluated through Scanning
Electron Microscopy images and optimization studies of parameters such as deposition potential
and time will be carried out to guarantee the formation of a highly porous structure, in order to
improve the sensitivity and/or selectivity of the analytical method.
1 Warwick, C. et al. Biosensors and Bioelectronics. 41, 2013, 1.
2 Sukeri, A.; Bertotti, M. Journal of Electroanalytical Chemistry. 805. 2017, 18.
[FAPESP]

75
3D-PRINTED SUPPORT COMBINED WITH COMPOSITE MATERIAL FOR
ELECTROCHEMICAL DETERMINATION OF DOPAMINE
Wallace B. V. de Oliveira (PG), Thalles P. Lisboa (R), Cassiano C. de Souza (PG), Guilherme
F. Alves (PG), Maria Auxiliadora C. Matos (R), Renato C. Matos (R)*
Federal University of Juiz de Fora, Chemistry Department, Juiz de Fora, MG, Brazil, 36036-900
*e-mail: wallacemnj@gmail.com
Dopamine (DOP) (4-(2-aminoethyl)-benzene-1,2-diol) is one of the most important natural

catecholamines biologically produced, originating from the decarboxylation of 3,4-
dihydroxyphenylamine. It is a fundamental neurotransmitter in the functioning of biological systems
such as renal, cardiovascular, and hormonal, because it acts as a chemical messenger, allowing
communication between different nerve cells¹. The dysregulation of this transmitter can lead to the
development of several diseases in the nervous system, such as Parkinson's and schizophrenia².
Therefore, this work sought the development of a disposable electrochemical sensor, simple and
low cost, using composite material (conductive graphite paint and nail polish) immobilized in 3D
printed support (3Ds-NPGE). Composite material compositions and electrochemical treatments
using KOH were optimized, and the best composition of the material was defined as the equal ratio
of graphite mass to polymer mass. The 3Ds-NPGE was combined with the differential pulse
voltammetry technique for DOP determination in water and pharmaceutical formulations samples.
Scanning electron microscopy (SEM) technique was applied to investigate the surface morphology
of 3Ds-NPGE before and after electrochemical treatment. The cyclic voltammogram of [Fe(CN)6]3-
/[Fe(CN)6]2- (5 mmol L-1) showcases a significant increase in current density of 143%, as well as the
electroactive area of the sensor of 8.64 mm² to 21.26 mm². The influence of pH on the DOP electro-
oxidation process was evaluated using a BR buffer solution (10 mmol L-1) with values ranging from
2.0 to 10.0, the pH 4.0 was chosen as the optimum condition, considering that provided higher
analytical sensitivity (higher current signal). The mass transport regime was studied using a DOP
solution with a concentration of 500 µmol L-1. A linear correlation was observed between the square
root of the scan rate and the current, indicating that the mass transport of DOP species is a diffusion-
controlled process. A method based on differential pulse voltammetry was optimized, which
presented a wide linear range from 0.02 to 10 µmol L-1 (Figure 1). The developed electroanalytical
method was applied to water and pharmaceutical formulations samples, showing excellent analytical
performance with adequate sensitivity (LOD = 5.7 nmol L-1), accuracy adequate (RSD < 8.7%), and
recovery values between 92 to 107 %. The 3Ds/NPGE showed similar sensitivity to other
electrochemical sensors reported in the literature. In addition, a fast and simple electrochemical
treatment method makes the sensor an optimal and promising tool for drug control.
Figure 1. DPV voltammograms of the analytical curve for DOP by

the electrochemically treated electrode ranging from 0.02 to 10
µmol L-1. The inserted graphs showing the zoom of the first linear
range, and the three linear adjustments. Conditions: Reference
electrode – Ag/AgCl, KCl (saturated), supporting electrolyte BR
buffer (10 mmol L-1, pH = 4.0), amplitude 110 mV and step
potential 4 mV.
1 Beitollahi H, Safaei M, Tajik S. Analytical and Bioanalytical Chemistry Research 6, 2019, 81.
2 Alothman ZA, Bukhari N, Wabaidur SM, Haider S. Sensors and Actuators, B: Chemical 146, 2010, 314.

76
DEVELOPMENT OF A DUAL ELECTROCHEMICAL SENSOR FOR MONITORING
THE RELEASE OF CALCIUM IONS FROM DENTAL COMPOSITES
Paula C. Falcoswki*a, Bruna M. Fronza b, Roberto R. Braga b, Mauro Bertotti (R)a
a University of São Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São Paulo, SP, Brazil
b University of São Paulo, Department of Biomaterials and Oral Biology, School of Dentistry, São Paulo, SP, Brazil
*e-mail: paula.calli@usp.br
Recent studies have shown that ions donor materials may contribute for dentin and enamel
remineralization by donating calcium and phosphate ions1. The Ca2+ ions release is an important
parameter in the material characterization, and the monitoring of Ca2+ concentration is usually
performed by spectroscopic methods such as ICP-OES. Measurements are carried out at different
times after the sample is immersed in deuterated water2. However, the use of electrochemical
methods for this kind of analysis is not much explored once Ca2+ ions are not
electroactive. Accordingly, this work aims at developing a dual-sensor capable of positioning and
detecting Ca2+ released from dimethacrylate-based light curing resin containing silanated barium
glass and dicalcium phosphate dihydrate particles at different ratios: 0%, 50% and 100%.
Measurements can be made in real-time using the Scanning Electrochemical Microscopy (SECM)
technique, and the sensor is capable of collecting information that can be associated with Ca2+
release in the interface of dentine and the composite. The Ca2+ detection was performed using a
potentiometric ion-selective sensor (ISE), which was fabricated with an ion-selective membrane fixed
into a borosilicate micropipette tip, an internal solution of 100 mmol L-1 calcium chloride solution, and
a Ag|AgCl wire that operates as an internal reference electrode. The calibration against a Ag|AgCl
reference electrode (Figure 1) indicates a linear response between 3 x 10-5 and 0.3 mol L-1. The
sensor positioning above the substrate was carried out by an amperometric microelectrode. The
device consisted of a 50 µm diameter platinum fiber sealed in a borosilicate micropipette, and the
electrical contact was made with silver glue. Both electrodes, potentiometric and amperometric, were
constructed in a Theta micropipette, giving rise to a dual device (Figure 2). The electrode can be
moved on the z-axis above the sample, and the precise positioning (20 µm) can be performed
through an approach curve recorded at -0.1 V using ferricyanide as an electroactive probe in the
negative feedback mode of SECM.
Figure 1. E vs pCa2+ Calibration plot using the Figure 2. Scheme of the dual microsensor. Left:
ISE calcium selective sensor. amperometric sensor. Right: potentiometric sensor.
1 Ires PM et al. British Dental Journal, 229, 2020, 450.

2 Aponso S et al. Journal of Dental Research, 00, 2018, 1.
[CAPES, FAPESP]

77
SCREEN-PRINTED PAPER ELECTRODE MODIFIED WITH DEEP EUTETIC
SOLVENT FOR DETECTION AND QUANTIFICATION OF PROGESTERONE
Camila G. Neves (PG)a, Igor G.S Oliveira (PG)a, Guilherme C. Paveglio (R)a,
Willyam R.P Barros (R)a*
a Universidade Federal da Grande Dourados, Cidade Universitária, Dourados, MS, Brazil, CP 364, 79804-970
*e-mail: willyambarros@ufgd.edu.br
The electrochemical sensors have been used to control the dosage of emerging contaminants,
mainly in endocrine compounds, including Progesterone (P4) [1]. Thus, the present work proposed
the development of a graphite-based screen-printed paper electrode (SPE) doped with a mixture of
Printex 6L carbon (SPE-P6LC) and deep eutectic solvent (DES) (SPE-P6LCDES). The electrodes
were applied for detection of P4 in mineral water, synthetic urine and artificial saliva samples. The
electrochemical measurements were performed in a system composed of a single compartment cell
(20 mL), containing a reference electrode (Ag/AgCl/KClsat.), Pt wire as counter electrode and SPE
as working electrode. For studies of the variation of graphite ink layers and variation of the proportion
of graphite ink were carried out by VC in KCl 0.5 mol L-1 as support electrolyte containing 1 mmol L-
1
Fe(CN)63-/4-. From these studies, it can be verified that the adherence of 2 layers for the proportion
of graphite:varnish 50%/50% (w/v) had better electrochemical responses to reversibility criteria. The
electroactivity of P4 was evaluated by differential pulse voltammetry in PBS 0.1 mol L -1 at pH 4.0.
From this study, oxidative signals were observed at +0.54 V. The linear range of P4 was 0.0090 –
0.812 μmol L-1 (LD= 0.00582 μmol L-1) and 0.00080 – 0.050 μmol L-1 (LD= 0.00075 μmol L-1) for the
non-doped SPE and SPE-P6LCDES, respectively.
0.8
0.8
0.6
i / A
0.4
0.6
ipa= 0.1827 + 13.191[P4]
0.2 2
R = 0.999
i / A
0.00 0.01 0.02 0.03 0.04 0.05

0.4 [P4] / mol L
-1
0.050 mol L-1
0.2
0.00080
0.0
-0.3 0.0 0.3 0.6 0.9 1.2

E / V vs. Ag/AgCl/KClsat.
Figure 1. Differential pulse voltammograms on the electrochemical response for different concentrations of P4 in 0.1 mol
L−1 PBS solution (pH 4.0) analyzed in SPE-P6LCDES. Inset: analytical curve to P4.
P4 recovery studies were performed in mineral water, synthetic urine and artificial saliva samples,
obtaining recoveries between 92% - 110%. Thus, the use of DES + Printex 6L carbon as a modifier
of the SPE, allowed strong interactions intramolecular with the analyte molecules, increasing the
intensity of the current responses, being a sensor with greater sensitivity, selectivity and
reproducibility.
1 Shamsipur M, Pashabadia A, Taherpoura AA, Bahramia K, Shargic H, Electrochimica Acta, 225, 2017, 292.
[CAPES, UFGD]

78
LOW-COST AND ECO-FRIENDLY ELECTROCHEMICAL SENSORS BASED ON
PAPER SUBSTRATES FOR NITRITE DETERMINATION
Juliana L. M. Gongoni (PG)a*, Lauro A. P. Filho (R)a, Iana V. S. Arantes (PG)a, Davi M. De
Farias (PG)a, Thiago R. L. C. Paixão (R)a
a Departamento de Química Fundamental, Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brazil,
05508-000
*e-mail: juliana.gongoni@usp.br
Nitrite (NO2-) is widely used in sausages and cured meat as a stabilizing agent due to its capability
to inhibit bacterial growth1. NO2- can also be found in fertilizers and, consequently, in water and soil.
Also, it is present in saliva and is normally used as a marker for periodontal diseases. Higher nitrite
concentration can be harmful to the human body since it can react with amines in acid conditions
and generate nitrosamines2, which can cause intestine cancer. Therefore, the continuous monitoring
of NO2- is necessary. In this context, paper-based electrochemical sensors can be a useful tool to
determine NO2-, providing low cost, portability, and easy-to-use. The devices were fabricated using
paper substrates and laser-scribing technique. Two layers of kraft paper were glued together and
carbonized with a CO2 laser, generating a fully integrated three-electrode electrochemical cell. The
sensors were electrochemically treated by applying -1V vs Ag for 60s in 1 mol L-1 KCl. The treated
sensors provided a better voltammetric profile (Figure 1A) toward nitrite oxidation, demonstrating
that the electrochemical treatment is an excellent alternative to improve the analytical performance
of the sensors. A calibration curve was constructed for nitrite (Figure 1B), and a good linear
correlation was obtained from 40 to 200 µmol L-1. The limits of detection and quantification were
estimated at 4.314 and 14.371 μmol L-1, respectively. The analytical method was applied to quantify
nitrite in soil, water, and synthetic saliva samples. The recovery percentages varied from 98 to
105%, indicating satisfactory accuracy. Acceptable RSD values (<8%) were also obtained for all
samples, showing good precision. Therefore, the proposed sensor is a great analytical platform for
NO2- determination.
40
120 160
30
I / A
120 20
I / A
80
I / A
10
80
40 0
50 100 150 200
CNitrite / M
40
0
0
-40
-0.2 0.0 0.2 0.4 0.6 0.8 1.0 0.0 0.2 0.4 0.6 0.8 1.0
E / V vs. Ag E / V vs Ag
Figure 1. A) Cyclic voltammograms for non-treated (black lines) and treated paperboard electrodes (blue lines) recorded
in PBS buffer pH 7 (dashed lines) + 1.0 mmol L-1 nitrite (solid lines).  = 20 mV s-1. B) Linear sweep voltammetry for treated
paperboard electrodes recorded in PBS buffer pH 7 in different nitrite concentrations. Inset: calibration curve
1 Sanz Y, Vila R, Toldrá F, Nieto P, Flores J, International Journal of Food Microbiology, 37, 1997, 225.
2 Bozkurt H, Erkmen O, Journal of the Science of Food and Agriculture, 84, 2004, 279.
[CAPES, FAPESP, CNPq]

79
DISPOSABLE AND LOW-COST CARBON INK PAPER-BASED DEVICE FOR
PARAQUAT DETERMINATION
Rodrigo M. Ramos (PG)a*, Vanessa N. Ataide (PG)a, Bruno Ferreira (PG)a, Thiago R. L. C.
Paixão (R)a
a Instituto de Química da USP, Departamento de Química fundamental, São Paulo, SP, Brazil.
*e-mail: rmr@iq.usp.br
In recent years, research in environmental area has grown in interest, especially when it is aimed at
creating new ways of monitoring and analyzing the environment, and, consequently, at the
development of sophisticated instrumentation for those purposes. This instrumentation has, as a
great disadvantage, the high cost and the need for highly specialized labor for its operation. On the
other hand, there has also been a growing interest in the development of low-cost methods that
present results similar to those of more sophisticated instrumentation1, making this a very promising
field of research. The use of sensors as a tool to monitor the environment has been an area of
particular interest, since these devices allow the development of solutions for a wide range of
analyzes and monitoring. In this work, we used a conductive carbon ink made with low-cost
materials2 to transfer conductive tracks to a paper support for application in samples of
environmental interest, such as paraquat. The carbon conductive ink was formulated from a mixture
of nail polish and graphite in a ratio of 50/50 mass3, using acetone as solvent. This ink was
transferred to the kraft paper substrate using a sticker paper mask, and the layout was cut into the
CO2 laser machine. This layout contains an arrangement of three electrodes (figure 1 – left). The
electrochemical behavior of ferricyanide potassium and hexamine ruthenium chloride (III), two redox
probes with known behavior, was evaluated using cyclic voltammetry. After the device
characterization, studies with paraquat4 showed excellent electrochemical response with the cyclic
voltammetry in KCl medium. However, the optimal current signal was obtained in the square wave
voltammetry technique with simple optimization steps, such as potential window (-1.5 to -0.2 V),
amplitude (170 mV), frequency (2 Hz), deposition potential (-1.5 V) and deposition time (30 s). This
allowed the construction of a calibration curve for paraquat going from 10 µmol L -1 to 500 µmol L-1,
demonstrating that it is possible to use the proposed device in environments where technical and
financial resources may be scarce, such as small rural areas where water contamination by paraquat
can represent a risk for the local population.
200
-200
i/A
-400
-600
-800
-1,6 -1,2 -0,8 -0,4 0,0

E/V vs Ag
Figure 1. (left) Constructions steps of the low-cost disposable electrochemical device; (right) Cyclic voltamgram recorded
in solution of 1 mmol L-1 K3[Fe(CN)6] (solid red line) and 0.1 mol. L-1 KCl medium (traced black line).
1 Cate D. M. et al. Analytical Chemistry. American Chemical Society, 6 Jan. 2015.

2 Pradela-Filho, L. A. et al. Electrochimica Acta, V. 258, P. 786–792, 20 Dec. 2017.
3 Aparecida, I. et al Journal of Electroanalytical Chemistry, V. 840, N. Feb. P. 109–116, 2019. Elsevier.
4 Xiao L. et al New Journal of Chemistry. 2008;32(9):1628-1633.

80
PROBING THE FORMATION OF ALCOHOL AND CO2 ADDUCTS USING GOLD
ELECTRODES MODIFIED WITH 2-MERCAPTOETHANOL
Berlane Gomes Santos (PG)a*, Fernanda Pereira Carli (PG)a, Claudimir Lúcio do Lago (R)a,
Ivano Gebhardt Rolf Gutz (R)a and Lúcio Angnes (R)a
a University of São Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São Paulo, São Paulo, Brazil,
05508-000
*berlanegsantos@gmail.com
Adducts of alcohol and CO2 (hemiesteres of carbonic acid or monoalkyil carbonates) are compounds
formed spontaneously in the presence of amount of water [1]. Although the poor description of these
adducts in literature, they are widely present in the most diverse mediums, including foods [2].
However, these species go unnoticed either because of the lack of knowledge of their existence or
because of their instability. The aim of this study was to show the formation of these compounds on
the surface of a gold electrode modified with 2-mercaptoethanol, as shown in the Fig. 1 (A).
1.8
0 min
A 1.6
B
5 min
10 min
1.4
15 min
20 min
1.2
25 min
1.0 30 min
I / µA
35 min
0.8 40 min
45 min
0.6 50 min
55 min
0.4
60 min
0.2
0.0
0.0 0.1 0.2 0.3 0.4
E/V vs Ag/AgCl/KClsat.
Figure 1. (A) Representation of electrode modification and gold with 2-mercaptoethanol and hemiester formation. And (B)
DPV recorded as a function of time in the presence of K3[Fe(CN)6] and HCO3-.
After the modification of the electrode, a conventional electrochemical cell of three electrodes was
equipped containing electrolyte sodium acetate pH 8.2. After this 0.5 mM K3[Fe(CN)6] and 1.0 mM
sodium bicarbonate were added to the cell as a source of CO2. The hypothesis tested was that if
adducts were formed on the electrode surface it could cause some change in the electrochemical
signal of the ferricyanide. For this differential pulse voltammograms were registered as a function of
time Fig 1 (B). The results indicated that hemiester was formed because there was a decrease in
the electrochemical signal of the ferricyanide due to the formation of the hemiester which has a
negative charge causing an electrostatic repulsion of the ferricyanide which has no negative charge.
The proof of adduct formation in membrane condition as done here is important because it is possible
for some biological process to occur through this mechanism.
1 Vidal DT, Nogueira T, Saito RM, do Lago CL. Electrophoresis, 32, 2011, 850.
2 Rossi MR, Vidal DT, do Lago CL. Food chemistry, 133, 2012, 352.
[FAPESP]

81
EVALUATION OF A HYPOTHETICAL PROTEIN VIA CONDUCTING
POLYMER/AUNPS – BASED IMPEDIMETRIC IMMUNOSENSOR FOR THE
DIAGNOSIS OF VISCERAL LEISHMANIASIS
Adriano de Souza Silva (PG)a*, Jaqueline Volpe (PG)a, Marcio E. V. Miyata (R)b,
Dênio E. P. Souto (R)a
a Federal
University of Parana (UFPR), Department of Chemistry, Laboratório de Espectrometria, Sensores e
Biossensores, Curitiba, Paraná, Brazil, 81531-980
b Federal University of Parana (UFPR), Department of Chemistry, Grupo de Pesquisa em Macromoléculas e Interfaces,
Curitiba, Paraná, Brazil, 81531-980

*e-mail: adrianosilva@ufpr.br
Neglected tropical diseases (NTDs) are a set of infectious diseases that primarily affect tropical and
subtropical countries. Due to the lack of investment in public policies for prevention and control, ends
up contributes to greater social inequality significantly affecting populations who live in a context of
expressive poverty. Visceral Leishmaniasis (VL) is one of these diseases with a high incidence,
which, even in the 21st century, represents a serious public health problem. 1,2,3 Currently, the most
employed methods in clinical laboratories for the serological diagnosis of VL are the enzyme-linked
immunosorbent assay (ELISA), and the direct and indirect immunofluorescence essays. However,
these methods have still presented low sensitivity to detect asymptomatic cases, and low specificity,
which generates a considerable number of false-negative results and cross-react with other
parasites. In addition, several recombinant proteins have been evaluated and revealed
improvement, especially in terms of specificity in VL diagnosis. In order to develop a portable, low-
cost, and highly sensitive method for detecting VL antibodies, in this study, a novel impedimetric
immunosensor based on conducting polymer polypyrrole (PPy) doped with poly(sodium-4-styrene
sulfonate) (NaPSS) and decorated with gold nanoparticles (AuNPs) was developed. As a sensor
recognition element a new recombinant protein, namely C1 protein, was used. Protein C1 has shown
a high immunogenic character with a strong binding affinity. However, the function of this protein in
the protozoan L. infantum is still unknown, constituting a very interesting material for scientific
research purposes. Then, a comparison in terms of analytical responses of the proposed C1 protein
with a well-established recombinant protein in VL immunodiagnostics, rK39, was realized. 4
Regarding the experimental steps necessary for the development of the impedimetric
immunosensors, firstly disposable stainless-steel mesh electrodes were functionalized towards the
electrodeposition of PPy:PSS and subsequent incorporation of AuNPs, followed by
mercaptopropionic acid (MPA) binding to obtain a self-organized monolayer (SAM). Afterwards,
each impedimetric immunosensor was constructed by covalently linking the respective recognition
element, C1 protein and rK39 protein, onto the polymeric platform. Scanning and transmission
electron microscopies, IR-spectroscopy, and electrochemical techniques were used for the
characterization of each step involved in the immunosensor construction process. From the
proposed immunosensors, it was possible to establish a comparison profile between the proteins
(C1 and rK39) when compared to their reactions against specific antibodies (anti-C1 and anti-rK39).
The results showed linear response for different polyclonal antibody concentrations, good sensibility,
and very satisfactory limits of detection (in the order of femtomolar) and quantification. In addition,
the proposed impedimetric immunosensors were evaluated toward canine serum samples,
presenting good analytical performance, which allowed for discrimination between positive and
negative samples in a wide range of antibody concentrations. In summary, the proposed platform in
this work presented a satisfactory performance, both in terms of sensitivity and selectivity without
the necessary electrochemical probe, as a disposable impedimetric immunosensor proposing a new
alternative diagnostic tool for VL.
1 Mitra AK, Mawson AR. Tropical Medicine and Infectious Disease, 3, 2017, 36.
2 Ibarra-Meneses AV, Moreno J, Carrilo E, Trends in Parasitology, 36, 2020, 1.
3 Bélard S, Strata E, Zhao A, Ritmeijer K, et a. Travel Medicine and Infectious Disease, 39, 2021, 101924.
4 Souto DEP, Volpe J, Gonçalves CC, Ramos CHI, Kubota LT, Talanta, 205, 2019, 120122.

82
MODIFIED PENCIL ELECTRODE WITH GOLD FOR ELECTROANALYTICAL
APPLICATIONS
Larissa Leffa Fernandes (UG)*, Manoelly O. Rocha (PG), Vladimir Lavayen (R), Jacqueline
Arguello da Silva(R)
Universidade Federal do Rio Grande do Sul, Chemistry Institute, Porto Alegre, Rio Grande do Sul, Brazil
*e-mail: larissa.leffa@ufrgs.br
Electrodeposition is a simple and cost-effective method widely used to obtain different surface
coatings. Plated jewelry is an example of coated objects with precious metals such as gold and
silver. The process is carried out by bathing the piece of jewelry in a solution containing the desired
metal and applying a current. Electrodeposition methods are also used to modify electrodes to
increase the electrochemically surface area, enhance selectivity, and promote electrocatalytic
activity1,2. Graphite pencils have become attractive for electroanalytical applications due to their low
cost, commercial availability, easy renewable surface, and eco-friendly resource. This work
demonstrates that pencil graphite recovered by gold nanostructures through the electrodeposition
technique improves its electroanalytical performance. Electrochemical measurements were
performed in a conventional three-electrode cell with pencil graphite as the working electrode, a
platinum wire as the counter electrode, and a saturated calomel electrode as the reference electrode,
using a potentiostat/galvanostat Autolab PGSTAT128 (Ecochemie, Holland) interfaced to a
computer and controlled by NOVA 2.1 software. The electrolyte solution consisted of a solution of
0.05 mol L-1 sodium sulfate and 15 mmol L-1 of chloroauric acid solution (HAuCl4). The
electrodeposition was performed by applying a constant potential of -0.6 volts for 200 seconds.
Potassium hexacyanoferrate was used as a redox probe to evaluate the effect of the pencil
modification in 0.1 mol L-1 of potassium chloride. Figure 1 shows the cyclic voltammograms
registered at a scan from 25 mV s-1 to 300 mV s-1 before and after electrodeposition. As can be
noticed that the modification significantly enhanced the current intensity of the ferro/ferricyanide
redox pair due to increased active surface. The Randles-Sevcik equation allows the calculation of
the area before and after electrodeposition, equal to 0.29 cm² and 0.42 cm², respectively. In addition,
there was a decrease in the peak potential difference, which denotes a significant improvement in
electron transfer.
(a) (b)
Figure 1. Cyclic voltammograms at different scan rates before (a) and after (b) gold electrodeposition.
1 Pu J, Shen Z, Zhong C, Zhou Q, Liu J, Zhu J, Zhang H, Adv. Mater. 32, 2020, 1903808.
2 Lai L, Li H, Sun Y, Ding G, Wang H, Yang Z, Appl. Sci. 9, 2019, 3824.
3
[INCTBio, PPGQ/UFRGS, PROPESQ/UFRGS, CAPES]

83
VOLTAMMETRIC DETERMINATION OF EDOXABAN, AN ANTICOAGULANT
PHARMACEUTICAL DRUG
Manoelly Oliveira Rocha (PG)a*, Martin Steppe (R)b, Jacqueline Arguello (R)a
a Universidade Federal do Rio Grande do Sul, Chemistry Institute, Porto Alegre, Rio Grande do Sul, Brazil
b Universidade Federal do Rio Grande do Sul, Faculty of Pharmacy, Porto Alegre, Rio Grande do Sul, Brazil
*e-mail: rochamanoelly@gmail.com
Edoxaban (EDX) is a selective factor Xa inhibitor of the coagulation cascade, approved by the FDA
in 2015 and by ANVISA in 2018 as an anticoagulant therapy. EDX also proves to be effective in
treating pulmonary embolism (PE) in hospitalized COVID-19 patients1. This study conducts the
electrooxidation of EDX using a carbon paste electrode (CPE) and cyclic voltammetry.
Electrochemical measurements were performed using an Autolab potentiostat controlled by NOVA
2.0.1 software. The electrochemical cell used was a conventional cell containing three electrodes:
CPE as the working electrode, a platinum wire as the counter electrode, and a saturated calomel
electrode as the reference electrode. Figure 1A shows the chemical structure of edoxaban and the
cyclic voltammograms registered at different pH values (5-10) in the Britton-Robinson buffer solution.
The voltammograms show an irreversible peak that shifted to less positive potential as the pH
increased. The linear relationship of the peak potential with the pH, Ep = 1,70 -0,061pH, R² = 0,99,
revealed a slope close to the theoretical value of 59 mV/pH, indicating that the oxidation process of
EDX occurred with the involvement of an equal number of electrons and protons2. Tafel slope value
also revealed that one electron is exchanged in the electrochemical reaction3. The effect of the scan
rate was also evaluated, where the analysis of peak current versus the square root of the scan rate
revealed a diffusion-controlled process. More studies are being conducted to acquire more
information about the electrooxidation process of EDX. On the other hand, the linear relationship
between the peak current and EDX concentration, shown in Figure 1B, indicates that the
electroanalytical method using CPE represents a simple, low-effective, and suitable tool for
analyzing this drug.
(A) (B)
Figure 1. A) EDX cyclic voltammograms at different pH values (A) and increasing concentrations of EDX (B).
1 Langella V et al., Expert Rev. Clin. Pharmacol. 14:10, 2021,1289.

2 Brett, AMO. Electrochemistry. Principles, Methods, and Applications Oxford University Press Inc, 1994.
3 Rizh M, Sultan MA, Taha EA, Attia AK, Abdallah, YM. Analytical Methods,9, 2017, 2523.
[INCTBio, PROPESQ-UFRGS, PPGQ-UFRGS, CAPES]

84
ELECTROCHEMICAL APTASENSOR WITH 3D DNA ORIGAMI STRUCTURE
FOR THE EARLY DIAGNOSIS OF HEPATITIS B
Rafael G. Pontes (PG)a*, Lauro T. Kubota (R)a
a Universidade Estadual de Campinas, Instituto de Química, Campinas, São Paulo, Brazil, 13083-970
*e-mail: r165087@dac.unicamp.br
Hepatitis B (HB) is still a disease that demands global attention as it is one of the leading causes of
morbidity and mortality worldwide. Diagnosis is primarily performed by detecting the HBsAg surface
antigen in serum samples using the ELISA1. However, ELISA is complex and time-consuming,
making its wider application difficult. Thus, developing new, highly sensitive, easy-to-perform, cost-
effective assays is urgently needed. DNA has attracted attention in the construction of
electrochemical biosensors. The Watson-Crick base pairing principle uses DNA as a building block
in nanoscale programmable materials technology. In addition, the ability to bind to specific targets
with sensitivity and specificity combined with intrinsic electrochemical activity makes DNA capable
of acting as a recognition element2. The tetrahedral nanostructures (TDN) were built according to
the previously reported method3. Briefly, equimolar amounts (1 µL of each strip) were mixed with 5
µL of TCEP (30 mM) and 41 µL of TM buffer, followed by heating at 95 °C for 10 min and then cooling
to 4 °C for 10 min. This way, TDN with the aptamer at one corner and three thiol groups at the other
three corners were constructed (Figure 1a). Then, 5 µL of this solution was placed on the Au
electrode surface and kept for 17 h at 4 °C in a humidity saturation condition. As a result,
tetrahedrons are immobilized on the electrode through the Au-S bond. Finally, different
concentrations of HBsAg were deposited on the modified electrode and kept at 4°C for 2 h. Detection
was performed by electrochemical impedance spectroscopy.
a) b) c)
Figure 1. a) Schematic representation of the electrode modification; b) Electrochemical impedance spectra of each
electrode modification step (inset – cyclic voltammograms of the modification steps); c) Analytical curve.
The bare gold electrode, as expected, showed the lowest resistance to charge transfer (Rct) (150
Ohms). However, the Rct values increased with the incorporation of TDN on the electrode surface
(746.5 Ohms) due to the repulsion between the negative charges of the DNA structure and
[Fe(CN)6]3-/4-. After the interaction of HBsAg with the aptamer, a further increase in Rct was observed
(894.7 Ohms), suggesting that the formation of the aptamer/antigen complex difficult the interfacial
electron transfer. The logarithm of the HBsAg concentration showed a linear relationship with the
∆Rct with R2 = 0.9968 and a linear response range from 10 fg mL-1 to 1000 fg mL-1. This operational
range in fg mL-1 demonstrates the good sensitivity of the developed aptasensor and reinforces the
possibility of using the DNA origami in the aptasensor development.
1 Ho, Joan Kha-Tu; Jeevan-Raj, Beena; Netter, Hans-Jürgen. Viruses, 12, 2, 2020, 126.
2 Huang, Ying et al. Sensors, 17, 10, 2017, 2375.
3 Wei, Min; Zhang, Wenyang. Sensors and Actuators B: Chemical, 276, 2018, 1.

85
GLYCEROL OXIDATION THROUGH BIOELECTROCHEMICAL SYSTEMS (BES)
Alex Soares Castro (PG)a,c*, Jefferson H. Franco (R)a,c, Valeria Reginatto (R)b, Adalgisa
Rodrigues de Andrade (R)a,c
a Universidade de São Paulo, Departamento de Química, Laboratório de Eletrocatálise e Eletroquímica Ambiental
(LEEA), Ribeirão Preto, São Paulo.
b Universidade de São Paulo, Departamento de Química, Laboratório de Biotecnologia Ambiental e Energias Renováveis
(LABIORE), Ribeirão Preto, São Paulo.

c UNESP, Instituto Nacional de Tecnologias Alternativas para Detecção, Avaliação Toxicológica e Remoção de
Micropoluentes e Radioativos, Araraquara, SP, Brazil, 14800-900

*e-mail: alex.soares.castro@usp.br
In recent years, bioelectrochemical processes have become an emerging technology in the

production of electrical energy and chemicals in a sustainable way. These processes include
microbial fuel cells (MFCs) and microbial electrolysis cells (MECs). With similar architecture, these
reactors contain an anode in which the biofilm is formed by exoelectrogenic bacteria that oxidize
organic matter and generate CO2, electrons, and protons and; a cathode in which O2 is reduced to
H2O (in MFCs) and H+ is reduced to H2 (in MECs). The difference between them is that: an MFC
converts chemical energy into electrical energy, being an excellent alternative, it can eliminate part
of the effluents and residues released into the environment, and, in a MEC, an extra voltage or
current is applied for the production the desired products – and the potential to be applied is linked
to the potential of the investigated substrates. Thus, the purpose of this work is to evaluate the
efficiency of the same reactor in two modus operandi to oxidize glycerol in the anodic chamber, for
later application in the generation of biohydrogen (bio-H2). After biofilm growth and microbiota
acclimatization to the glycerol substrate. We tested the efficiency of modus operandi reactors (i)
MFC and (ii) MEC. The power (Fig. 1A) test showed that, when operated as an MEC, the reactor
reached a power 42% higher (P = 94 mW.m2) than the initial value, while after operating as an MFC
the power dropped by 45% (P = 36 mW.m2). In quantitative terms, the oxidation of glycerol with the
reactor operating as MEC became 5x more efficient when presenting the oxidation of 40% of glycerol
(1.5 g/L) against 8% in the MFC mode in the same time interval (6h) (Fig. 1B). Chromatographic
analysis (HPLC) confirmed the possibility of the reactors forming value-added by-products, such as
1,3-propanediol, glycolic acid, glyceric acid, and butyl acid (Fig. 1C). Therefore, the MEC reactor
developed showed good efficiency when oxidizing glycerol, so the next step will be analysis of the
yield of bio-H2 production, and evaluation of the MEC reactor developed in this work as a
sustainable/renewable way to produce bio-H2.
(A) (B) (C)
Figure 1. (A) – Reactor power density curve (- -) after feeding with glycerol and stabilization of the system; (red) after
operating in MEC mode, and (blue) after operating in MFC mode. Polarization curve measured under pseudo-potentiostatic
condition, v = 1 mV/s, A = 231 cm2; (B) Concentration in percentage (%) of glycerol and its oxidation products in the MEC
reactor over 6h of electrolysis; (C) Consumption given in percentage (%) after oxidation of glycerol in the reactor operating
as MFC and as MEC.
[FAPESP, CAPES, CNPq INCT]

86
USE OF ZEOLITE AND NANOTUBES AS AN ENZYMELESS SENSOR FOR
VOLTAMMETRIC ANALYSIS OF HYDROQUINONE IN PHARMACEUTICALS
Roberta Castro Martins (PG)a, Thawan Gomes de Oliveira (PG)b, Zuy Maria Magriotis (R)c,
Adelir Aparecida Saczk (R)d, Lúcio Angnes (R)b, Fabiana da Silva Felix (R)d*
a Federal University of Lavras (UFLA), Postgraduate Program in Biomaterial Engineering/School of agricultural sciences
of Lavras (ESAL), Lavras, Minas Gerais, Brazil, 37200-900
b University of São Paulo (USP), Institute of Chemistry, São Paulo, São Paulo, Brazil, 05508-900
c Federal University of Lavras (UFLA), School of Chemical Engineering, Lavras, Minas Gerais, Brazil, 37200-900
d Federal University of Lavras (UFLA), Departament of Chemistry, Lavras, Minas Gerais, Brazil, 37200-900
*e-mail: fabiana.felix@ufla.br
Biosensor can be defined as a device capable of producing analytical information using a biological
recognition element held on the surface of the transducer element1. The bioreceptors can be
enzymes, antibodies, microorganisms and nucleic acids, being the first one the most commonly
used1. However, due to the low stability of enzymatic biosensors, for instance, new approaches have
been studied2. Nanoenzymes are nanomaterials that are able to mimic the behaviour of the natural
enzyme2. The ones that are able to mimic peroxidases can act in two ways: the Fenton reaction or
the electron transfer reaction3. These nanoenzymes, when compared to the natural enzymes, are
more stable and have a better durability. However, it is necessary to continue promoting the same
reaction conditions as for natural enzymes because of their way of mimic4. To try to circumvent this
demand, a new class of sensors, the enzymeless sensors, have been studied since the 2010s in
different applications such as detection of hydrogen peroxide and simultaneous determination of
parathion and paraoxon in environmental samples. These sensors are able to identify and quantify
a given analyte as natural enzymes, but without the need to synthesize a material that has a structure
similar to that of the natural enzyme4. In this work we use carbon paste mixed with zeolites (CP-
ZEO), in which they are crystalline aluminosilicates with three-dimensional microporous structures
and have high surface areas, in construction of voltammetric sensors. CP-ZEO electrodes were
modified with carbon nanotubes, mainly, for improving the electron transfer properties during flow
injection analysis (FIA) in determination of hydroquinone in pharmaceutical products. Hydroquinone
is used in skin whitening treatments, and since August 2021 it has been on ANVISA's banned
substance list5. There are no published studies, so far, using CP-ZEO modified with nanotubes for
analyses of pharmaceuticals. Amperometric experiments in associated with FIA are being performed
and its main parameters of sampling loop, flow rate and applied potential have already been
optimized in 100 μL, 5.38 mL min-1 and 0.15 V, respectively. Studies with cyclic voltammetry were
carried out in 1.0 x 10-3 mol L-1 potassium ferrocyanide in 0.1 mol L-1 KCl, indicating an increase in
the effective area of approximatelly 95 % and 22 % of the electrode modified with zeolite and carbon
nanotubes when compared with the electrodes of carbon paste and carbon paste with zeolite,
respectively. Other voltammetric studies carried out in 5.0 x10-4 mol L-1 hydroquinone solution in
0.1 mol L-1 phosphate buffer at pH 6.5, indicated an increase in the reduction current of 98 % when
the CP-ZEO was compared with an enzymatic biosensor. Future studies will be held for the
quantification of hydroquinone in pharmaceutical creams.
1 Thévenot D.R. et al. Biosensors & Bioelectronics 16, 2001, 121.
2 Zhang et al., Nano-Micro Lett. (2022) 14:95
3 Jin S. et al., Sensors & Actuators: B. Chemical 283, 2019, 18.
4 Ahmadi et al., RSC Adv., 2020, 10, 23457.
4 ANVISA. Resolução da diretoria colegiada- RDC nº 529, de 04 de agosto de 2021.
5
[FAPEMIG, CNPq, CAPES, FAPESP, EMBRAPA-CAFÉ]

87
LASER-INDUCED GROWTH OF GOLD NANOPARTICLES IN 3D-PRINTED
ELECTRODES SURFACES: A NEW APPROACH IN SENSING AND BIOSENSING
PLATFORMS
William B. Veloso (PG)a*, Iana V. S. Arantes (PG)a, Gabriel N. Meloni (R)a, Wilson A. Ameku
(R)a, Lucio Angnes (R)a, Rodrigo A. A. Muñoz (R)b, Thiago R. L. C. Paixão (R)a
a Universidade de São Paulo - USP, Instituto de Química – IQ/USP, São Paulo, SP, Brazil, 05508-000
b Universidade Federal de Uberlândia - UFU, Instituto de Química, Uberlândia, MG, Brazil, 38400-902
*e-mail: william.veloso@usp.br
3D printing has become a popular strategy for fabricating electrochemical sensors and analytical
devices. This growing popularity results from the many advantages 3D printing presents when
compared to traditional manufacturing methods, particularly low production cost, minimal material
waste, and design freedom.1,2 However, 3D printed electrodes often present high electrical
resistance and poor electron transfer kinetics and electrocatalytic properties.3 These can be
overcome by incorporating metallic nanostructures at the electrode surface, improving its charge
transfer kinetics and catalytic properties. Here we report a simple and quick (4 step) method for
fabricating Au nanoparticles (AuNPs) decorated 3D printed electrodes using an “off-the-shelf”
conductive polylactic acid (PLA) filament and a CO2 laser to induce the formation of AuNP at the
electrode surface. The fabrication process involves: 1- printing the desired electrode geometry from
the commercial carbon black /PLA filament; 2- Pre-treating the electrode surface by pyrolyzing the
electrodic surface with the CO2 laser; 3- Drop-casting HAuCl4 solution (chemical precursor for the
AuNP) at the treated region; 4- Photothermal reduction of Au(III) at the electrode surface, forming
AuNP, using the CO2 laser.4 Formation of AuNP on the electrode surface was investigated by
voltammetry in acidic media, scanning electron microscopy and energy dispersive X-ray
spectroscopy. The electrochemical behavior of the decorated electrodes was investigated by cyclic
voltammetry and electrochemical impedance spectroscopy in 1:1 [Fe(CN)6]3−/ [Fe(CN)6]4− solution
(Figure 1). Modified electrodes will be employed as an enzymatic biosensor for H2O2 detection using
horseradish peroxidase. This “proof-of-concept” application will highlight the potential of our method
for quickly fabricating biosensors, which is extremely useful for developing point-of-care/action
analytical devices.
Figure 1. (A) Cyclic voltammograms and (B) electrochemical impedance spectroscopy of CB/PLA (black trace), laser
treated (pyrolyzed)-CB/PLA (blue trace), and AuNPs decorated CB/PLA (red trace) electrodes in 1:1 5.0 mmol L -1
[Fe(CN)6]-3/-4 containing 1.0 mol L-1 KCl. Voltammetric scan rate of 20 mV s-1. Impedance spectroscopy performed with a
10 mV sinusoidal perturbation around the open circuit potential with a frequency range from 0.05 Hz to 5 kHz.
1 Ambrosi A, Pumera M. Chem. Soc. Rev. 45, 2016, 2740.

2 Cardoso RM, Kalinke C, et al. Anal. Chim. Acta, 1118, 2020, 73.
3 Kumar, KP Akshay, et al. Electrochem. Commun. 120, 2020, 106827.
4 El Hamzaoui H, et al., Mater. Res. Bull. 46, 2011, 1530.
[USP, IQ/USP, CNPq, CAPES, FAPESP]

88
SIMULATION OF THE EFFECTS OF ACID RAIN AND LANDFILL LEACHATE ON
THE MOBILITY OF ELEMENTS FROM FLUORESCENT LAMPS
Laís N. Vianaa* (PG), Tatiana D. Saint’Pierrea (R)
a Pontifícia Universidade Católica do Rio de Janeiro, Department of Chemistry, Rio de Janeiro, RJ, Brazil, 22453-900
*e-mail: laisnviana@yahoo.com.br
Fluorescent lamps (FL) are known for the presence of toxic metals in their composition. Despite the
gradual replacement by light emitting diode (LED), FL are still consumed and are quite present in
urban wastes, especially in developing and underdeveloped countries, where recycling rates are still
low, and the lamps are often discarded without proper care.1,2 This results in an increased risk of
environmental contamination. In this work we evaluate the mobility of potentially toxic elements from
the fluorescent lamp phosphor material (PM) by leaching it with synthetic solutions of acid rain (SAR)
and landfill leachate (SLL). These solutions were prepared following the literature, with the pH
adjusted to 4.73 and 7.99 to simulate respectively the pH of the acid rain and landfill leachates
reported for Rio de Janeiro.3,4 The PM of 70 used FL units were collected and homogenised. A
method of combined replicates was applied to allow further homogeneity and increase working
volume in the leaching experiment. For each replicate, 1.0 mL of the synthetic solution was added
to 0.6 g of the PM, as defined by an experimental design. The procedure was performed in duplicate
to allow half the tubes to be placed into a mechanical shaker for 3 h (active study) while the other
tubes were standing for the same period (passive study). Samples were centrifuged, the supernatant
was collected, diluted, and analysed by inductively coupled plasma mass spectrometry (ICP-MS).
For comparison purposes, 100 mg of the PM were also submitted to a microwave-assisted quasi-
total extraction (MAE) with 1.2 mL of HNO3, 3.5 mL of HCl and 0.5 mL of H2O.5 The solutions were
centrifuged to remove the suspended solids and were analysed by ICP-MS. The concentrations of
Al, Ba, Ce, Cd, Cr, Eu, Hg, La, Mn, Ni, P, Pb, Sr, and Y were above the limits of quantification, which
varied from 0.01 µg kg-1 (La) to 2.8 µg kg-1 (Al), in all samples. MAE concentrations varied from 8.27
± 0.44 mg kg-1 (Cr) to 161 ± 7 g kg-1 (P). To evaluate the accuracy of the method, MAE solutions
were fortified in two levels with analytical standards, and the recoveries were within 84 and 110%.
The concentrations leached by the SLL varied from 0.36 ± 0.03 µg kg-1 (Pb) to 14.4 ± 0.8 mg kg-1
(Hg) in passive study, and from 0.50 ± 0.04 µg kg-1 (Pb) to 34.0 ± 1.0 mg kg-1 (Sr) in active study. As
for SAR, leached concentrations in passive study varied from 0.29 ± 0.03 µg kg -1 (Cd) to 22.9 ± 0.5
mg kg-1 (Hg), and from 2.6 ± 0.3 µg kg-1 (Cd) to 18.8 ± 0.9 mg kg-1 (Al) in active study. Leaching with
SAR was more efficient for most investigated elements (Al, Ce, Cd (only in active study), Cr, Eu, Hg,
La, P, Pb, and Y). The agitation usually increased the metal leaching, except for Hg and,
occasionally, Cd (SLL) and Ce (SAR), which presented higher concentrations in passive leaching
solutions. The SLL was especially the most effective leaching solution for the Sr extraction, which
reached up to 42x the leached concentration achieved with SAR in passive study. Significant
differences were observed in the leaching of Y compared to the other REE with both SLL and SAR.
It can be explained by the presence of the metal in the red phosphor, which is easier to solubilize.
The single leaching experiment employed in this study resulted in lower extraction percentages when
compared to MAE concentrations. Nevertheless, some concerning metals, such as Hg, Cd, and Pb,
were mobilized by the synthetic solutions. Considering the real disposal scenario, where variations
in pH and composition of leach solutions and successive leaching events can occur, the extraction
of metals can increase significantly, potentially causing local contamination. Additionally, REE and
Sr, which are some of the most leached metals, are still unregulated in the environment. Considering
the composition of FL, and the mobility observed for some toxic and potentially toxic elements,
stricter measures must be taken regarding their disposal and recycling.
1 Dos Santos TS, Batista MC, Pozza AS, Rossi LS, Eng. Sanit. Ambient., 20 (4), 2015, 595.
2 Kadam AR, Nair GB, Dhoble SJ, J. Environ. Chem. Eng., 7, 2019, 103279.
3 De Mello WZ, Environ. Pollut., 114, 2001, 235.
4 Guyonnet D et al., Geotext. Geomembr., 27, 2009, 321.
5 Hobohm K et al., Talanta 147, 2016, 615.
[CNPq, FAPERJ, CAPES, PUC-Rio]

89
DISPOSABLE ELECTROCHEMICAL SENSORS PRODUCED BY INKJET
PRINTING AND SILVER INK
Lauro A. Pradela-Filho (R)a*, Juliana L. M. Gongoni (PG)a, Iana V. S. Arantes (PG)a, Davi M.
De Farias (PG)a, Thiago R. L. C. Paixão (R)a
a Universidade de São Paulo, Departamento de Química Fundamental, Instituto de Química, São Paulo, SP, Brazil,
05508-000
*e-mail: lauropradela@usp.br
Inkjet printing is a promising approach to producing conductive tracks due to its automated nature
based on an ink deposition system with high precision. Compared to other printing technologies, this
technique does not depend on the masks to print the desired pattern. Consequently, this approach
allows the deposition of thin films with versatile designs, including size and formats1. This work aims
to understand the inkjet printing process to produce electrochemical sensors for analytical
applications. Silver ink was used to print the electrodes with a Dimatix Printer. The geometry of the
sensors was designed with the CorelDraw software. The ink deposition process was optimized by
printing only the working electrode. The best parameters were selected considering the
electrochemical response of the electrodes and the resistivity of the conductive tracks.
Hexaammineruthenium ion was used as the electrochemical probe for the voltammetric
characterization. Different materials were evaluated as substrates, including paper, Con-tact®
adhesive tape, and polyimide film. Con-tact® adhesive tape was the best substrate to produce the
electrodes. The printing parameters included the substrate temperature (40ºC), drop spacing (17
µm), number of layers (1 layer), and length of the working electrodes (13 mm). Once optimized, a
three-electrode electrochemical cell was engineered (Figure 1A), and its signal for Ru(NH)63+/Ru(NH)62+
was compared with the off-chip reference electrode (Ag/AgCl/KCl 3 mol L-1), representing a shift in
the reduction potential of approximately -190 mV. In addition, the sensors showed batch-to-batch
reproducibility (RSD = 3 % for Ipa). The analytical applicability of the electrochemical sensors was
then evaluated by constructing calibration curves for hydrogen peroxide (Figure 1B) and picric acid
(Figure 1C). Good linear correlations were obtained for these species, demonstrating the proposed
system are attractive platforms for analytical applications. Moreover, inkjet printing showed to be a
great tool to fabricate electrochemical sensors with consistent responses, opening possibilities for
large-scale production.
A 0 0
B C
-20 -50
-40
I / A
I / A
-100
-60 80 140
Ip / A
-150
Ip / A
-80
40 70
-100 -200
0 0
-120 0 700 1400
-250 0 500
CHP / mol L-1
CPA / mol L-1
-140
-300
-1.5 -1.2 -0.9 -0.6 -0.3 0.0 0.3 -1.5 -1.2 -0.9 -0.6 -0.3 0.0 0.3
E / V vs. Ag E / V vs. Ag
Figure 1. A) image of the inkjet-printed silver electrodes. Linear sweep voltammograms of inkjet-printed silver electrodes
recorded in 0,1 mol L-1 acetate buffer with A) 25 µmmol L-1 to 1600 mmol L-1 hydrogen peroxide (HP) and B) 25 to 600
µmol L-1 picric acid (PA).  = 100 mV s-1. Insert calibration curves.
1 Singh M, Haverinen HM, Dhagat P, Jabbour GE. Advanced Materials, 22, 2010, 673.

90
SQUARE WAVE VOLTAMMETRIC DETERMINATION OF MERCURY AND
SELENIUM IN WATER SAMPLES
Carlos Eduardo Costa Lopes (UG)*, Joao Pedro de Carvalho Silva (UG), Lizandra
Nascimento Castro (UG), Charbel Nagib Mouchrek (PG), Luiza Maria Ferreira Dantas (R),
Iranaldo Santos da Silva(R)
Federal University of Maranhão, Chemistry Technology Department - CCET, São Luís. Maranhão, Brazil, CEP 65080-
805
*e-mail: lopes.carlos@discente.ufma.br;
Mercury is one of the most toxic metals to humans and animals1, and it is used in several industrial
applications. In addition to this, it has the capacity to bioaccumulate and biomagnify in the organism
of humans and animals, easily becoming from a trace level concentration to a trophic level. Despite
this, selenium is an essential nutrient for most living beings, however, it is toxic in high concentration
levels2. In this context, the development of a simultaneous determination method for these elements
is extremely important to evaluate the risk associated with contaminated environments, food, water,
and medicaments ingestion3. So, the proposed method presents an alternative to quantify mercury
and selenium simultaneously in water samples on gold electrode, through subtractive anodic
stripping voltammetry, by using square wave voltammetry technique (SWSASV). Evaluating the
effects of anions (CH3COO-, PO43-, SO42-, NO3- and Cl-) and acids (CH3CHOOH, H3PO4, H2SO4,
HNO3 and HCl), the analytical signal provided the selected electrolyte and the chloride importance
for simultaneous analyses due to the potential peak separation, as illustrated in Fig 1. The
optimization of the electrolyte and chloride concentrations, potential and deposition time, and the
calibration curves were also approached. The influence of analytes selectivity was made by fixing
one analyte concentration while increasing the other and, due to this, it was established a simple
way to achieve the simultaneous voltammetric determination of mercury and selenium. The
proposed method showed satisfactory figures of merit, such as detection limits of 0.23 and 0.09 ppb
and limits of quantification of 0.77 and 0.28 ppb for mercury and selenium, respectively. This work
was applied for several water samples, like mineral, potable and river water using the standard
addition method.
Figure 1. SWSASVs containing 50 ppb of mercury and 50 ppb of selenium in H 2SO4 50 mmol/L without (-) KCl and with
(-) 5,0 mmol/L. Conditions: Deposition potential: -0.4 V; Deposition time: 60 s; Amplitude= 25 mV; Frequency=10 Hz; Step
= 5 mV.
1 Simião DC, de Andrade FP, Lima WG, de Jesus ML, Dorim PHG, Paiva MC, Water Supply, 22, 2022, 5535.
2 Silva DLF, da Costa MAP, Silva LOB, dos Santos WNL, Food Chemistry, 24, 2019, 273.
3 Gajdosechova Z, Mester Z, Feldmann J, Krupp EM, TrAC - Trends in Analytical Chemistry, 104, 2018, 95.
[UFMA, CNPq, CAPES, FAPEMA]

91
A NANOPOROUS GOLD MICROELECTRODE FOR MERCURY DETECTION BY
SQUARE-WAVE ANODIC STRIPPING VOLTAMMETRY
Renato Soares de Oliveira Lins (PG)a*, Mauro Bertotti (R)a
*e-mail: renatolins@usp.br
Illegal gold mining is still an issue affecting the Amazon. Mercury is a metal normally used in the
amalgamation process for gold extraction. This activity can release mercury into soils and water,
contaminating the environment. Consequently, developing new strategies to quantify mercury in
rivers and sediments in remote locations is important. Electrochemical techniques are promising
alternatives for this purpose since such measurements can be performed with portable
instrumentation and disposable devices. In addition, the electrochemical system can be miniaturized
without compromising sensitivity. These aspects are advantageous compared to traditional
spectroscopic methods. Accordingly, this work aims to develop a green method to determine Hg(II)
by using square-wave anodic stripping voltammetry (SWASV) and a nanoporous gold
microelectrode (µE-NPG). The gold microelectrode (µE-Au) was prepared with a gold fiber (diameter
= 25 µM) immobilized into a micropipette tip with epoxy glue. To obtain the µE-NPG sensor, the µE-
Au was modified through an anodization method by applying 2.0 V for 15 min in 0.5 mol L -1 sulfuric
acid solution. The µE-Au and the µE-NPG were characterized by cyclic voltammetry (Figure 1 A)
and field-emission scanning electron microscopy (Figure 1B) to obtain the active surface area and
morphological information. The microelectrodes were used for Hg(II) determination, and the results
are shown in Figure 1C. The nanoporous gold microelectrode showed a higher analytical signal (~5
fold) for this analyte compared to the bare Au microelectrode. The main advantages of the gold
nanostructures include the higher surface area and better mass transport (radial diffusion),
increasing the mercury deposition onto the electrode surface. In addition, it is essential to highlight
that the proposed system has a miniaturized size, opening possibilities for the fabrication of portable
analytical platforms for point-of-need applications.
Figure 1. (A) Cyclic Voltammograms recorded at bare Au and NPG microelectrodes in 0.5 mol L -1 H2SO4, Electrochemical
Surface Area (ECSA) for Au-µE (6,5 x 10-6 cm2) and NPG-µE (9.4 x 10-5 cm2). Scan rate: 0,1 V s-1; (B) FE-SEM images of
the bare Au and NPG surface; (C) SWASVs in the presence and absence of 2 µmol L -1 Hg(II) recorded at bare Au and
NPG microelectrodes. Conditions: Edeposition 0.0 V; tdeposition 60 s; Econditioning: 0.7 V; tconditioning 30 s; Step potential: 5 mV;
Amplitude: 20 mV and Frequency 20 Hz.
1 Bernalte E, Arévalo S, Pérez-Taborda J, Wenk J, Estrela P, Avila A, Di Lorenzo M, Sensors and Actuators B: Chemical,
307, 2020,127620.
2 Jaramillo DXO, Sukeri A, Saravia LPH, Espinoza-Montero PJ, Bertotti M, Electroanalysis, 29, 2017,2316.
[CNPq, FAPESP, CAPES

92
LASER-SCRIBED ELECTROCHEMICAL SENSORS COMBINED WITH BATCH
INJECTION ANALYSIS FOR SULFANILAMIDE DETERMINATION
Davi M. de Farias (PG)a*, Juliana L. M. Gongoni (PG)a, Iana V. S. Arantes (PG)a, William
B.Veloso (PG)a, Lauro A. Pradela-Filho (R)a, Thiago R.L.C. Paixão (R)a
a University of São Paulo, Institute of Chemistry, Department of Fundamental Chemistry,
São Paulo-SP, Brazil, 05508-900.
*daviimarques@usp.br
The demand for analytical devices became evident with the pandemic scenario caused by the
SARscoV-2 virus. The characteristics of the devices must cover central points including versatility,
portability, low cost, and sensitivity. Electrochemical sensors are promising tools that enable
miniaturization without compromising the system sensitivity. Sulfanilamide (SULF) is an antibiotic
with antimicrobial properties used in human and veterinary medicines.¹ Despite its benefits, SULF is
considered an emerging pollutant. Consequently, it is important to develop sensors able to monitor
the SULF levels in environmental samples. Hence, this work aims to fabricate disposable and
versatile electrochemical devices to determine this compound. The electrochemical sensors were
fabricated using the laser scribing technique and a phenolic substrate. This technique consists of
using a CO2 laser source to induce the formation of conductive carbon tracks onto the desired
substrate.² After fabricating, the sensors were modified by drop-casting functionalized multi-walled
carbon nanotubes onto the electrode surface. Figure 1 shows the modified electrodes provided a
better voltammetric response for sulfanilamide determination. Once characterized, the sensors were
combined with batch injection analysis (BIA) using amperometric detection. Calibration curves were
constructed by increasing and decreasing the sulfanilamide concentrations with the proposed
system. The transient signals and calibration curves are shown in Figures 1B and C, respectively.
This system requires only 2 mL of supporting electrolyte and 15 µL of samples, reducing the waste
produced throughout the analysis. In addition, this set promotes an enhancement of analytical
frequency and system automation. As a result, an analytical method was developed with the
proposed device and used to determine sulfanilamide in pharmaceutical, environmental and
biological samples. The analytical method was validated by using the labeled concentrations and
recovery studies, demonstrating accuracy and precision. Therefore, the proposed system is a
promising tool for the rapid monitoring of sulfanilamide.
400 25
A 350
1.0
60
B 25 150  150  C
0.8
J (/cm-2)
E/ V. vs Ag
50
300
0.6 20
0.4
40
20 100 
J (/cm-2)
30
250 0.2 100 
20
15 15
I / A
I / A
0.0
200 Unmodified Modified
50 
150 10 50  10
100 25 
5 25 
50
10  10  5
0
0
0
0.3 0.4 0.5 0.6 0.7 0.8 0.9 1.0 1.1 1.2 0 300 600 900 1200 1500 1800 0 20 40 60 80 100 120 140 160
E/V. vs Ag t/s CSULF / mol L-1
Figure 1. A) Cyclic voltammograms of unmodified (black lines) and modified sensors (red lines) recorded in KCl 1.0 mol
L-1 (dashed lines) in the presence of sulfanilamide 1.0 mmol L -1 (solid lines).  = 20 mV s-1. B) Transient signals are
obtained by combining the proposed sensors with the BIA system. The signals represent the injection of 15 μL of different
sulfanilamide concentrations in buffer BR 0.04 mol L-1 pH 10. Eapplied: +0.9 V vs. Ag. C) Respective calibration curves.
1 De Faria LV et al. Journal of Electroanalytical Chemistry, 892, 2021, 115298.

2 Mendes LF et al., TR. Carbon, 159, 2020, 110–118.

93
SANDWICH-LIKE ELECTROCHEMICAL APTASENSING OF HEAT SHOCK
PROTEIN 70 KDA (HSP70): APPLICATION IN DIAGNOSIS/PROGNOSIS OF
CORONAVIRUS DISEASE 2019 (COVID-19)
Masoud Negahdary (R)a, Lucio Angnes (R)a
a University of São Paulo, Institute of Chemistry, Department of Fundamental Chemistry,
São Paulo, São Paulo, Brazil, 05508-900.
The coronavirus disease 2019 (COVID-19) is challenging, infected all countries, and created a
terrible pandemic in recent years. Therefore, efforts to design new tools for the molecular diagnosis
of this disease are critical so that the knowledge and experience gained building aptasensors can
be applied in controlling and managing similar diseases. In this study, using aptamer-conjugated
gold nanoparticles (aptamer-AuNPs) and a modified glassy carbon electrode (GCE) with reduced
graphene oxide (rGO) and Acropora-like gold (ALG) nanostructure, a sandwich-like system has been
provided for sensitive detection of heat shock protein 70 kDa (HSP70), which applied as a functional
biomarker in diagnosis/prognosis of COVID-19. The surface of the GCE was modified with rGO and
ALG nanostructures, respectively. Then a thiol-functionalized aptamer sequence as the first part of
the biorecognition element was immobilized on the surface of the GCE. After adding different
concentrations of analyte, the second part of the biorecognition element (aptamer-AuNPs) was
immobilized on the electrode surface to improve the diagnostic performance. The designed
aptasensor detected HSP70 in a wide linear range, from 5 pg mL-1 to 125 ng mL-1, with limit of
detection (LOD) ~ 3 pg mL-1. The designed aptasensor was stable for 3 weeks, and applicable in
detecting 40 real plasma samples of COVID-19 patients while the diagnostic sensitivity and
specificity were 90 % and 85%, respectively, compared with the reverse transcription-polymerase
chain reaction (RT-PCR) method. Using different nanomaterials (AuNPs, ALG nanostructure, and
rGO) increased the sensitivity, the linear diagnostic range, and the stability of the desired aptasensor.
Using aptamer strands in two positions (immobilized on the surface of the GCE and conjugated with
AuNPs) provided the most optimal conditions for analyte detection. Since the applied aptamer
sequence was thiol-functionalized in the 5’ end, it could bind tightly to the gold surface (Au-S covalent
bond). The presence of rGO sheets on GCE provided a specific and stable surface for molecules of
ALG nanostructure. The immobilizing of aptamer strands as the biorecognition element equipped
with the mentioned gold nanomaterials could provide special diagnostic performance. In particular,
the capability of the designed diagnostic platform in the assays of reproducibility, regeneration,
stability, selectivity, and application of real plasma samples of COVID-19 patients was evaluated,
and the results encountered were excellent. The role of the HSP70 biomarker in diagnosing other
important diseases such as various cancers, heart diseases, and diabetes should also be
investigated. In addition, in the subsequent efforts, other nanomaterials, other signal transducers,
and other biorecognition elements can be investigated in the presence of aptamers.

94
ENHANCED PERFORMANCE OF SCREEN-PRINTED ELECTRODES TOWARDS
FORENSIC SAMPLES BY FAST AND REAGENTLESS CO2 LASER TREATMENT
Bruno Ferreira (PG)a*, Iana V.S. Arantes (PG)a, Rodrigo M. Ramos (PG)a, Thiago R.L.C.
Paixão (R)a
a University of São Paulo, Department of Fundamental Chemistry - / Chemistry Institute, São Paulo, São Paulo, Brazil,
*e-mail: bruno_ferreira@usp.br
Screen-printed electrodes (SPEs) became an important tool for detecting and determining several
species through electroanalytical chemistry. Their main advantages include simple design,
portability, adequate selectivity, and relatively low cost compared to other commercially available
electrodes1. Nonetheless, the literature shows several works reporting structure modifications of the
SPE surface to improve the limits of detection and quantification of these sensors or to enable a
simple detection that was not possible in the bare electrode2. However, these modifications are
usually time and reagent-consuming. Hence, we propose a fast and reagentless CO2 laser-scribing
treatment of a commercial carbon SPE (DropSens) aiming at the electroanalytical detection of
forensic targets. Thus, for the laser treatment, the working electrode was submitted to a laser power
of ~2.7 Watts, with a 15 mm s-1 scan rate and an 11 mm distance from the laser beam. The treated
electrodes were then characterized by scanning electron microscopy (SEM), Raman spectroscopy,
and electrochemical techniques using ferri/ferrocyanide ([Fe(CN)6]-3/-4) as a redox probe. Fig. 1 a)
shows the cyclic voltammograms (CVs) of the [Fe(CN)6]-3/-4, where better performance is observed
for the treated SPE compared to the bare SPE with a significant current increase. Electrochemical
impedance spectroscopy (EIS) was also performed, exhibiting a decrease in the charge transfer
resistance (RCT) of 240 Ω to 4.00 Ω after the laser treatment. Furthermore, the treated SPEs
presented enhanced performance over the detection of lysergic acid diethylamide (LSD) and cocaine
hydrochloride (COC), as shown in the CVs in Fig. 1 b) and c), respectively, where an increase in the
peak current response and a decrease in the peak potential can be observed, attesting the
applicability of the treated devices over forensic samples. A calibration curve was also constructed
using the linear sweep voltammetry (LSV) technique from 1 to 100 mol L-1 COC, showing promising
analytical performance with good sensitivity (0.070 A mol-1 L) and good linearity (R=0.9909),
exhibiting great potential for analytical applications.
Figure 1. Cyclic voltammetry responses of a) 5 mmol L-1 [Fe(CN)6]-3/-4 in 1.0 mol L-1 KCl at a scan rate of 20 mV s-1; b) 0.5
mmol L-1 LSD in 0.1 mol L-1 KClO4 at a scan rate of 100 mV s-1; and c) 0.5 mmol L-1 COC in 0.04 mol L-1 BR buffer (pH 9)
at a scan rate of 100 mV s-1 in the bare SPE (black color) and the laser-treated SPE (red color).
1 Liu G, Lin YY, Wu H, Lin Y., Environmental Science and Technology, 41, 2007.
2 Tasić N, Cavalcante L, Deffune E, Góes, MS, Paixão, TRLC, Gonçalves, LM., Electrochimica Acta, 397, 2021.

95
ATOMIC SPECTROMETRY
96
METHOD DEVELOPMENT FOR Ca, Fe AND Zn DETERMINATION IN SEWAGE
SLUDGE BY LASER-INDUCED BREAKDOWN SPECTROMETRY
Fernanda F. Thiengoa*, Aline de C. Eliasa, Maciel S. Luzb, Danielle P. Intimac, Ivanise
Gaubeurd, Juliana Naozukae, Cassiana S. Nomuraa
a University of Sao Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São Paulo, SP, Brazil.
b Center for Technology in Metallurgy and Materials, Institute for Technological Research, São Paulo, SP, Brazil.
c Basic Sanitation Company of the State of São Paulo, São Paulo, SP, Brazil.
d Center for Natural and Human Sciences, Federal University of ABC, Santo André, São Paulo, SP, Brazil.
e Federal University of São Paulo, Department of Chemistry, Diadema, SP, Brazil.
*e-mnail: thiengo@usp.br
Sewage sludge is defined as a semi-solid organic waste, containing a range of inorganic

components, including nutrients (e. g. Ca, Fe and Zn)1. The final disposal of residual sludge from
sewage treatment stations is a serious environmental issue worldwide and the concern about its
management is real2. The use in agriculture shows a set of advantages such as the improvement of
physical, chemical and biological properties of soil resulting in the increase of productivity 3. But for
that, quality control is required. Analysis of this kind of sample is not an easy task, especially due to
the need of prior conversion into solution by acid dissolution and/or digestion procedures before
analysis. In those cases, direct solid samples analysis with minimum or none sample treatment is
an interesting alternative. Laser-induced breakdown spectrometry (LIBS) deserves attention, and is
one of the most promising and powerful analytical methods for the direct analysis of solids samples
of different compositions. On the other hand, quantitative measurement in complex matrices has still
been considered the "Achilles heel" of this method due to the strong matrix interference caused by
a complex nature of laser-sample interaction5. In this context, this research work proposes the
method development for direct quantitative measurements of Ca, Fe and Zn in sludge sample by
LIBS. Sewage sludge sample was air-dried to constant mass, ground in an automatic agate mortar
and pestle mill, and sieved through a mesh size of 50 µm. Pressed pellets were prepared using an
automatic hydraulic press (Xpress 3635) by transferring 0.35 g of the ground sample and 0.15 g of
cellulose binder to a 13 mm die set and applying 8 ton cm-2. LIBS measurements were carried out
with a commercial J 200 Tandem LIBS system (Applied Spectra) and its instrumental parameters
have been optimized: delay time (0.25 µs), spot size (50 µm), laser pulses (270) and laser energy
(20 mJ). Ca, Fe and Zn mass fractions were determined by LIBS using two calibration approaches,
external calibration (EC) and internal standard calibration (ISC), both using certified reference
materials (CRM). EC method resulted in high relative errors (> 50%), probably due to matrix effect,
which was minimized using carbon emission wavelength at 193.160 nm since accurate results were
found. Limits of detection and quantification of ISC proposed method were 0.007 Wt.% and 0.022
Wt.%, 0.21 Wt.% and 0.69 Wt.% and 0.02 Wt.% and 0.07 Wt.%, for Ca, Fe and Zn respectively.
Using ISC, no significant differences were found at 95% confidence level (t-test) between found
(20.9, 3.7, 0.17 Wt.% for Ca, Fe and Zn) and reference values (19.7, 3.0, 0.14 Wt% for Ca, Fe and
Zn), showing this approach is feasible for quantitative analysis of sewage sludge.
1 Tamanini, C. R.; Motta, A. C. V.; Andreoli, C. V.; Doetzer, B. H. Brazilian Archives of Biology and Technology, v. 51,
2008, 643.
2 Costa, V. R. Ciência Hoje, v. 18, 1995, 8.
3 Pedroza, J. P.; Haandel, A. C.; Beltrão, N. E. M.; Dionísio, J. A.; Duarte, M. E. M. Revista Brasileira de Engenharia
Agrícola e Ambiental, v. 10, 2006, 586.
4 Machado, R. C.; Andrade, D. F.; Babos, D. V.; Castro, J. P.; Costa, C. C.; Sperança, M. A.; Garcia, J. A.; Gamela, R.
R.; Pereira-Filho, E. J. Anal. At. Spectrom., 35, 2020, 54.
5 Martinez, M; Baudelet, M. Analytical and Bioanalytical Chemistry, 412, 2020, 27.
[CAPES, CNPq, FAPESP, INCTAA, SABESP]
––––––––––––––––––––– Atomic spectrocopy –––––––––––––––––––––

97
CHROMIUM LEVELS IN TOBACCO, FILTER AND ASH OF ILLICIT BRANDS
CIGARETTES MARKETED IN BRAZIL
Thalles P. Lisboa (R)a*, Aparecida M. S. Mimura (R)b, Júlio C. J. Silva (R)a, Renato C. Matos
(R)a, Rafael A. Sousa (R)a*
a Federal University of Juiz de Fora, Chemistry Departament, Juiz de Fora, MG, Brazil, 36036-900
b Federal University of Vale do São Francisco, Chemistry Graduation Collegiate, São Raimundo Nonato, PI, Brazil,
64770-000
*e-mail: thallespl_jf@hotmail.com and rafael.arromba@ufjf.br
Smoking is a public health problem and an important source of human exposure to toxic metals. This
habit has been related to more than 50 diseases and has caused a large number of premature
deaths worldwide1. The tobacco plant, Nicotiana tabacum L., is known for absorbing metallic
elements from its culture environment and accumulating them in its leaves2. So, by investigating how
the toxic metals present in tobacco are partitioned into the different parts of the cigarette and how
the metals release is, during the burning of the cigarette, one can estimate the average consumption
of these contaminants1. Regarding the mineral chromium, it presents two states of oxidation, the
trivalent and hexavalent forms (CrIII and CrVI, respectively). CrIII is considered essential for some
metabolic processes in the body, while CrVI is defined as toxic and a potential carcinogen. Based in
Graphite Furnace Atomic Absorption Spectrometry (GF AAS), this work describes an efficient and
comparable analytical method with other spectrometric techniques for the determination of chromium
in different constituent parts of cigarette samples (tobacco, filters, and ashes). The method
developed was evaluated using 12 samples and the results showed recovery values between 83
and 107% for the addition and recovery experiments. The accuracy was also evaluated using a
reference sample of tomato leaves (NIST SRM 1573a), which proved the efficiency of the method.
The method limits of detection were 20.4, 75.8 and 80.7 ng g-1 for tobacco, filter, and cigarette ash
samples, respectively. The average chromium values found among the samples analyzed were in
the range of 0.96 to 3.85 and from 0.32 to 0.80 μg/cigarette for tobacco and ashes, respectively.
Meanwhile, for most pre-burn and post-burn filter samples, the values of chromium concentration
remained below limits of detection. The developed method presented adequate results about
precision and accuracy, demonstrating its applicability in the determination of chromium in cigarette
samples. Regarding the samples investigated, high rates of chromium transfer from tobacco to ash
were observed, while the filters showed low absorption after the simulated burning process. This
suggests that significant amounts of this metal, may reach the airways of smokers and, in addition,
the ash may act as an environmental contaminant. Additionally, since illicit brands are inserted in
the Brazilian market, it becomes important to develop control tools that allow rapid, sensitive and
selective characterization of these contaminants in order to protect the health and safety of
consumers.
1 Lisboa TP, Mimura AMS, Silva JCJ, Sousa RA, Journal of Analytical Toxicology, 44, 2020, 514
2 Lisboa TP, Faria LV, Matos MAC, Matos RC, Sousa RA, Microchemical Journal, 150, 2019, 1.
[UFJF, FAPEMIG, CAPES, CNPq, RQ-MG]

98
EVALUATION OF IN VITRO BIOACCESSIBILITY OF MINERALS IN SPIRULINA
SAMPLES
Ingrid K. I. Salgado (PG)*a, Milena P. Ferreira (PG)a, Marcela Z. Corazza (R)a, Cesar R. T.
Tarley (R)ab
a State University of Londrina, Department of Chemistry, Londrina, PR, Brazil, 86057-970
b National Institute of Science and Technology in Bioanalysis (INCTBio), State University of Campinas (UNICAMP),
*e-mail: ingridiagla@hotmail.com
In recent years, the consumption of food supplements combined with a healthy and balanced diet
has been encouraged. For this, the use of natural supplements, rich in minerals and bioactive
components, which help in the maintenance and proper functioning of the body, stands out. In this
context, Spirulina, marketed in the form of capsules and granules, has been much consumed for
being considered a food supplement that numerous nutritional and therapeutic properties for human
health, in addition to relevant concentrations of minerals. However, knowing the total content of
minerals as well as other nutrients is not enough to provide the true nutritional value of a food.
Therefore, several research groups have dedicated efforts to the study of in vitro bioaccessibility of
essential nutrients, such as minerals. In these studies, the bioaccessible fraction (BF) of the nutrient
released from the matrix during gastrointestinal digestion is estimated, a fraction that is available to
be absorbed by the body.1 Thus, this study evaluated the bioaccessibility of macro (Ca, Mg) and
microminerals (Zn, Fe, and Mn) in Spirulina powder samples, using the unified bioaccessibility
method (UBM) developed by BARGE.2 For this, three different commercial brands of Spirulina were
purchased from health food stores, and for each brand three packets were purchased and
homogenized. The total mineral concentration was determined by flame atomic absorption
spectrometry (FAAS) after microwave-assisted digestion, as well as the bioaccessible and residual
gastric and gastrointestinal fraction after in vitro tests. Centesimal composition (proteins, ash, lipids
and moisture) was also performed. The moisture in the samples ranged from 2 to 5%, and the
percentage for proteins ranged from 52 to 67%, which classifies spirulina as a food with high protein
content, according to ANVISA.3 Ash content varied from 7 to 10%, and the values obtained for lipids
ranged from 1 to 7%. The highest total concentrations of minerals were obtained for Mg (2407.1 -
3150.7 mg kg-1), Ca (637.2 - 976.3 mg kg-1) and Fe (631.9 - 833.5 mg kg-1). The total mineral content
obtained for Mn ranged from 26.2 to 61.4 mg kg -1 and for Zn from 8.0 to 19.8 mg kg-1. The highest
gastrointestinal BF (%) were observed for Zn and Mg (>80), which can be explained by the high
protein content, which favors bioaccessibility for these elements.4 The BF observed for Mn ranged
from 64 to 77%, and for Ca were less than 58%. The lowest percentage of bioaccessibility was
observed for Fe with values lower than 45%, can be attributed to the presence of calcium in spirulina,
which can decrease the availability of iron in the intestine.5 Tokalıoğlu et al.6 investigated the mineral
bioaccessibility of 10 commercial nutritional supplements, which contained a variety of vitamins and
minerals and, the range found for the gastrointestinal phase was 3-35% for Zn, 3-14% for Fe, 34-
91% for Mg, 53-62% for Mn. Thus, comparing the results obtained for spirulina with commercial
supplements, spirulina, as a natural food supplement, can provides better bioaccessibility of some
minerals such as Zn, Mn, Fe and Mg. In this sense, the ingestion of spirulina powder is a viable
alternative to a good nutritional diet.
1 De Lima ACS, Soares DJ, Da Silva LMR. Food Chemistry, 161, 2014, 142.
2 Wragg J et al., Science of The Total Environment, 409, 2011, 4016.
3 ANVISA. Resolução RDC nº 54 de novembro de 2012.
4 Lönnerdal B, The Journal of Nutrition, 130, 2000, 1378.
5 Stelmach E, Szymczycha-Madeja A, Pohl P. Food Chemistry, 197, 2016, 388.
6 Tokalıoğlu S, Clough R, Foulkes M, Worsfold P. Food Chemistry, 150, 2014, 327.
[CAPES, CNPq, INCTBio, UEL]

99
USE OF MULTI-ENERGY CALIBRATION IN THE QUANTIFICATION OF
METALS FROM NICKEL METAL HYDRIDE BATTERIES LIQUOR
Kaíque A. M. L. Cruz (PG)a, George L. Donati (R)b, Maria C. Hespanhol (R)a*
a Grupo de Análises e Educação para a Sustentabilidade (GAES), Departamento de Química, Centro de Ciências Exatas
e Tecnológicas, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil, 36570-900
b Department of Chemistry, Wake Forest University, Winston-Salem, North Caroline, USA
*e-mail: mariacarmo@ufv.br
With the advances in technologies, researchers can count on the use of increasingly modern and
precise instruments to carry out their analyses. However, choosing an inefficient calibration method
can lead to inaccurate results, even if adequate instruments are used. Multi-energy calibration
(MEC)1 has been presented as an alternative to calibrations by external standard (EC), internal
standard (IS) and standard addition (SA), since MEC can correct severe matrix effects, in a strategy
similar to SA calibration, which has been shown to be the best choice until now when it comes to
samples that can cause more severe matrix effects. Meanwhile, only two solutions are used in MEC,
reducing considerably the cost and sample preparation time in comparison to SA. In this work, MEC
was applied to quantify Ce, La, Ni, Co and Mn, present in nickel metal hydride (NiMH) batteries
liquor, obtained from dissolution with H2SO4 3,00 mol L-1, using microwave-induced plasma optical
emission spectrometry (MIP-OES). The results obtained from MEC were compared with those
obtained when EC e SA were used. The selection of leaching liquor as matrix in this study is due to
the growing interest in the use of secondary resources to obtain critical elements. Since NiMH
batteries are produced in large quantities due to high consumption and contain high contents of
cobalt and rare earth elements, they are an interesting secondary resource. The use of MIP-OES
deserves to be highlighted as this technique has proved to be be attractive and an alternative to well-
established instrumental techniques, such as inductively coupled plasma optical emission
spectrometry (ICP-OES) and flame atomic absorption spectrometry (FAAS). In MIP-OES, the gas
used to sustain the plasma is nitrogen, making this technique safer and economically better, as it
eliminates the use of flammable gases such as acetylene and nitrous oxide used in FAAS for an
example. In addition, the nitrogen gas that sustains the plasma can be readily removed from the
atmosphere using a nitrogen generator, which significantly reduces the costs of analysis when
compared to the use of argon employed in the ICP-OES for an example. In this work, the accuracy
was evaluated by addition/recovery studies performed for SA and MEC, which, as a result, showed
good agreement with each other. Recoveries between 90-110% were obtained for the analysis of
Mn, Co, Ni, La and Ce. For the MEC, coefficients of variation of 2.5, 1.8, 2.0, 2.3 e 5.8 % were
obtained for Ni, Mn, Co, La and Ce, respectively. The determination of Mn and La using EC did not
show agreement with those obtained from SA and MEC, even if high dilution factors were used (600
and 1800 times for Mn and La, respectively). This good correlation between MEC and SA and poor
correlation in the EC is due to the similarity in the preparation of the analytical curves (AC) used in
MEC and SA, when the liquor sample itself, used in the construction of the AC, tends to suppress
the matrix effects, which is not observed in the EC. Limits of detection (LOD) equals to 0.01, 0.02,
0.001, 0.4 and 0,06 mg kg-1 were obtained for Ni, La, Mn, Ce and Co, respectively, using the MEC.
This work showed that it is possible to successfully perform the precise and exact quantification of
metals present in NiMH battery liquor in H2SO4 3 mol L-1 using MEC with the aid of a more
economical and environmentally safe technique, MIP-OES.
1 Virgilio, A., Gonçalves, D. A., McSweeney, T., Gomes Neto, J. A., Nóbrega, J. A., Donati, G. L, Anal. Chim. Acta, 982,
2017, 31.
[INCTAA/CNPq, CAPES/FCT, CAPES, FAPEMIG, CNPq]

100
EVALUATION OF MINERAL COMPOSITION IN JUICES BY MIP-OES USING
MULTIVARIATE ANALYSIS
Mariane G. Nabiça (PG)a, Patrícia O. Nunes (PG)a, Alexsandro S. Martins (PG)a, Allan da S.
Cruz (PG)a, Fábio I. M. Carvalho (R)b, Heronides A. Dantas Filho (R)a, Kelly G. F. Dantas(R)a*
a Federal University of Pará, Faculty of Chemistry, Institute of Exact and Natural Sciences, Belém, Pará, Brazil, 66075-
110.
b Federal Rural University of the Amazon, Parauapebas, Pará, Brazil.
*e-mail: kdgfernandes@ufpa.br
The manual preparation of fruit juices has become an inconvenience for a majority, due to the fast
pace of life in society, and therefore, the Brazilian consumer has shown growing interest in
consuming "ready-to-eat" products as industrialized juices. Due to the importance of monitoring and
evaluating the mineral composition of industrialized juices, the objective of this study was to
determine aluminum, copper, calcium, iron, manganese, potassium, and sodium in industrialized
juice by MIP-OES using multivariate analysis. Pearson correlation, principal component analysis
(PCA) and hierarchical cluster analysis (HCA) were used in order to conduct a more thorough
evaluation of the original data. Twenty-five juice samples from 11 different brands (brands A; B; C;
D; E; F; G; H; I; L and M) acquired in local supermarkets (Belém, Pará, Brazil) were digested in a
cavity microwave oven using a combination of HNO3 (14 mol L-1), H2O2 30% (v/v) and ultrapure
water. The efficiency of the sample preparation procedure and the accuracy of the measurements
by MIP-OES were evaluated by the method of addition and recovery of analyte. For all elements,
the recoveries obtained were within the acceptable range of 80% to 120%. In relation to the
elements, Ca, K, and Na presented the highest contents in the juices in box and can when compared
to the other elements studied. The concentrations of Al, Ca, Cu, Fe, K, Mn, Na and Zn in the juices
were 0.30 mg L-1 to 5.20 mg L-1, 9.0 mg L-1 to 370.0 mg L-1, 0.09 mg L-1 to 1.00 mg L-1, 0.20 mg L-1
to 2.5 mg L-1, 1.0 to 385 mg L-1, 0.05 mg L-1 to 22.38 mg L-1, 6.0 mg L-1 to 5299.0 mg L-1, and 0.20
mg L-1 to 0.23 mg L-1, respectively. These values found for Ca, Cu, Fe, K, Na, Mn, and Zn not
exceeded the minimum recommended values for the recommended dietary reference intake (DRI)
of 300 mL/day of juice. Aluminum was found in three juices canned of different flavors and from the
same manufacturer. The levels found for Na were higher than the values reported on the juice labels.
This shows the importance of greater control by inspection agencies, since in only 300 mL of some
samples of juice to provid above 100% recommended for Na in a full day. Through the Pearson
correlation, PCA and HCA was possible to observe that the separation between the industrialized
juices samples was not directly correlated with the type of packaging, but with the mineral
composition of the juice. The PCA and HCA revealed that the main contributions to the formation of
the two distinct groups (A and B) were the concentrations of Ca, Cu, Fe, K, Mn, Na and Zn. The data
obtained in this study increases the understanding of the mineral composition of industrialized fruit
juices and can be of great interest in terms nutritional value and toxicological studies.
1 Otten JJ, Hellwig JP, Meyers LD, Dietary reference intakes, The essential guide to nutrient requirements, The National
Academies Press: Washington, USA, 2006.
[CAPES]

101
HR CS GF AAS AND DIRECT SOLID SAMPLING FOR THE DETERMINATION OF
HEPATIC COPPER IN THE DIAGNOSIS OF WILSON'S DISEASE
André N. Silva Filho (PG)a, Bernardo F. Braz (R)b, Jennifer Lowe (R)c, Maria C. Chindamo
(R)d, Fernando H. Cincotto (R)b,e, Ricardo E. Santelli (R)b,e*
a Universidade Federal do Rio de Janeiro, Programa de Pós-Graduação em Química, Rio de Janeiro, RJ, Brazil, 21941-
909.
b Universidade Federal do Rio de Janeiro, Departamento de Química Analítica, Rio de Janeiro, RJ, Brazil, 21941-909.
c Universidade Federal do Rio de Janeiro, Instituto de Biofísica Carlos Chagas Filho, Rio de Janeiro, RJ, Brazil, 21941-
170
d Universidade Federal do Rio de Janeiro, Departamento de Clínica Médica, Rio de Janeiro, RJ, Brazil, 21941-590
e Instituto Nacional de Ciência e Tecnologia em Bioanalítica, Campinas, SP, Brazil, 13083-970
*e-mail: santelli@iq.ufrj.br
Wilson’s disease (WD) was first described in 1912 by neurologist Samuel Alexander Kinnier Wilson
as a progressive degeneration and invariably fatal disease, characterized mainly by neurological
deficits and liver cirrhosis 1. WD is an autosomal recessive disorder caused by a P-type ATPase
mutation that prevents copper from binding to ceruloplasmin (protein that is the copper transporter
in the blood serum). Thus, copper deposition can occurs in hepatocytes, in the brain, in the periphery
of the cornea and in the kidneys. The diagnosis of WD is challenging, especially in the absence of
neurological changes and/or Kayser-Fleisher rings. Low serum concentrations of ceruloplasmin are
observed, as there is no incorporation of copper in apoceruloplasmin for its formation, but some
patients have normal concentrations. The gold standard for diagnosis of WD is the liver copper
concentrations. A hepatic copper content higher than 250 mg/kg (dry basis) is considered as the
best available diagnosis test 2. Otherwise, if the hepatic copper concentrations is below 50 mg/kg it
is indicative that the patient does not have the disease. However, in a recent paper, Duncan and co-
workers in a pilot study for quality assurance for copper (and iron) in liver biopsies, done over two-
year period with twenty-six trace element laboratories in seven countries, conclude that accuracy
and precision was not good for the majority of laboratories. Also, accuracy was found to be
statistically poorer if sample weights less than 3 mg were analyzed. In the present study, an analytical
method was developed for the direct determination of copper by solid liver sampling using High
Resolution Continuous Source Graphite Furnace Atomic Absorption Spectrometry (SS HR CS GF
AAS). A temperature program was optimized including a drying step of the hepatic tissue on to the
platform to allow the collection of the dry mass of the sample to be analysed. The method was
validated using DOLT-2 certified reference material and lyophilized livers from laboratory animals,
which were also analyzed by ICP-MS for comparison. Due to the large range of copper concentration
that can be found in the liver, three different absorption lines were used (217.894 nm, 249.2146 nm
and 244.164 nm) in order to obtain a greater dynamic range of concentration. The detection and
quantification limits obtained, in mg/kg, for these lines were respectively: 0.082 and 0.272; 10.8 and
35.8; and 49.3 and 164 for copper. Several samples from bovine liver and Wistar rat liver were
analysed by the developed method and by ICP-MS as a reference method. The results found are in
good agreement between these techniques. A liver biopsy sample from a patient suspected of
having WD was analyzed and the results indicated that the patient is a carrier of the disease, due to
having a concentration greater than 250 mg/kg of Cu in dry liver mass, value higher than the cut-off
concentration established in the literature.
1 Wilson SAK, Brain, 34, 1912, 295.
2 Ferenci P, et al., Clinical Gastroenterology and Hepatology, 3, 2005, 811.
3 Duncan A, Harrington CF, Catchpole A, Taylor A, Journal of Trace Elements in Medicine and Biology, 52, 2019, 288.
[CNPq, FAPERJ]

102
METHOD DEVELOPMENT FOR Nd DETERMINATION IN THE PRODUCTION
CHAIN OF PERMANENT MAGNETS BY LIBS AND WD XRF
Aline de C. Elias (PG)a*, Gilmar A. de Almeidab, Maciel S. Luz (R)b, Juliana Naozuka (R)c,
Cassiana S. Nomura (R)a
a University of Sao Paulo, Department of Fundamental Chemistry, Institute of Chemistry, Sao Paulo, SP, Brazil.
b Center for Technology in Metallurgy and Materials, Institute for Technological Research, Sao Paulo, SP, Brazil
C Federal University of Sao Paulo, Department of Chemistry, Diadema, SP, Brazil.
* e-mail: allinne-carvalho@hotmail.com
Rare earth elements (REEs) have received attention in recent years due to their high demand in the
production of technological and electronic devices, including REE magnets, which have been widely
used in high value-added products such as hybrid and electric vehicles. Super-magnet is made from
the electrochemical reduction of didymium oxides with subsequent incorporation of iron and boron,
followed by grinding the alloy in a jet mill, compacting the metal powder and aligning the magnetic
field. The quality of this material depends on the purity of the REEs and the concentration of
didymium and for this reason, quality control is imperative1,2. Spectroscopy techniques have been
used for this purpose over the years, however, most of them requires the conversion of the samples
into solution, which is time-consuming and improve the risk of contamination. Alternatives that allow
direct solid analysis are X-ray fluorescence (XRF) and laser-induced breakdown spectroscopy
(LIBS)2,3. Besides all advantages associated to direct solid samples analysis methods, some
challenges have to be overcome. Heterogeneity and the strong matrix effect damage the precision
and accuracy of analytical results, especially in LIBS. In this case, the improvement of sample
homogeneity associated with the use of appropriate calibration strategy is imperative4. In this way,
this work proposes a simple method to allow the determination of Nd in didymium oxide samples
and in (Nd, Pr)-Fe-B alloys by LIBS and WD XRF using fusion process to improve the sample
homogeneity. External calibration (EC) was used by preparing standards spiked with increasing
masses of Nd2O3 over Al2O3 used as diluent. Fused glass beads was prepared by mixing 7 g of flux
(49.75% LiBO2, 49.75% Li2B4O7 and 0.5% LiBr) to 1 g of sample and heating the mixture in a
automated fusion furnace (Katanax, SpexSamplePrep) using the following heating program (T/°C,
ramp/°C min-1, hold/min): step 1 (1050, 30, 6) and step 2 (1100, 50, 2). Analysis were performed
with a J 200 Tandem LIBS system (Applied Spectra, USA), with Nd:YAG laser operating at 266 nm.
The most intense line (Nd (II) 401.23 nm) was monitored, and the instrumental parameters were
optimized (delay time = 0.50 µs; pulse energy = 20 mJ; number of shots = 401; spot size = 65 μm).
The analytical parameters were estimated and linear responses were obtained between 10 to
80Wt% of Nd2O3 and LD 0.5%. For analysis by WD XRF (Supermimi 200, Rigaku, Japan), the Nd-
Lα angle (72.101º) was chosen for the calibration. The measurements were carried out under
vacuum, X-Ray Tubes (Pd target), analyzer crystal: LiF (220), 30 mm diameter of radiation in 20 s
and spin of 30 rpm. Results obtained by reference method5 (ICP OES after acid decomposition) for
alloy and for didymium oxide were in accordance for both proposed methods (Table 1), showing the
applicability of those methods for Nd determination in alloy and didymium oxide samples.
Table 1. Results for Nd mass fraction in alloy and didymium oxide obtained LIBS and XRF.
Reference value (Wt%) LIBS (Wt%) WD XRF (Wt%)
Nd Alloy 24.3  1.3 26.6  3.0 25.9  1.0
didymium oxide 64.3  0.9 60.4  3.9 58.4  3.0
1 Rosental, S. Cetem. Cap 30, 2005, 62
2 Santos, C. A. L; Panossian, Z. Revista IPT: Tecnologia e Inovação. v. 1, 2017, 4.
3 Pinto, F. G; Junior, R. E; Saint’Pierre, T. D. Analytical Letters, v. 45, 2012, 1537.
4 Carvalho, A. A. C; Alves, V. C; Silvestre, D. M; Leme, F. O; Oliveira P. V; Nomura, C. S, Geostandards and
Geoanalytical Research, v.41, 2017, 585.
6 Papai, R; Fonseca, K. T; Almeida, G. A; Silva, A. L. N; Nagasima, T. P; Jabes, E. G; Santos, C. A. L; Landgraf,
F. J. G; Luz, M. S. Brazilian Journal of Analytical Chemistry, v.9, 2022, 124.
7
[CNPq, CAPES, FAPESP, IPT]

103
PHYSICOCHEMICAL CARACTERIZATION AND DETERMINATION OF MACRO
AND MICRONUTRIENTS IN FRUIT FUNCTIONAL FLOURS
Karoline Rayzel Rodrigues de Campos (PG), Alex Virgilio (R)*
University of São Paulo, Center for Nuclear Energy in Agriculture, Analytical Chemistry Laboratory, Piracicaba, SP,
Brazil, 13416-000
*e-mail: alexvirgilio@cena.usp.br
The functional flour market has been growing in the recent years, mainly motivated by consumer’s
search for healthier food alternatives. These products are usually obtained from fruits, vegetables
and grains through drying, milling and/or other technological processes. Many of these flours can be
manufactured from products with low consumer appeal (eg. unripe fruits or visually damaged
vegetables) and by-products from food processing industries. Its use is important from the
environmental, economic and social point of view by promoting the reduction of residues generation,
minimization of food waste and expansion of the food supply.1 Despite the high demand for these
flours, the knowledge about their physicochemical characteristics and mineral composition is quite
scarce. Most functional flours are sold in bulk at local markets and online commerce, so labeling and
information regarding the manufacturer, origin and nutritional content are often missing. Thus, the
development of simple, fast and reliable analytical methods for quality control, food safety and
traceability of these products is extremely important. In this work, eight different fruit flour samples
(açaí, blackberry, green banana, coconut, orange, apple, passion fruit and grape) were analyzed
aiming their physicochemical characterization, sample digestion by microwave-assisted digestion
using diluted acids and the determination of total Ca, Fe, K, Mg, Mn, P, S and Zn by ICP OES. A
wheat flour CRM (NIST SRM 1567a) was analyzed to check the accuracy of the procedure.
Parameters as granulometry, pH, titratable acidity and moisture were determined using official
methods for food analysis recommended by the Adolfo Lutz Institute (ALI) and the Brazilian ministry
of agriculture, livestock and food supply (MAPA), adapted from AOAC methods.2,3 Digestions were
performed in a single reaction chamber (SRC) microwave system Milestone Ultrawave, using 250
mg of sample, 6.0 mL of HNO3 (3 mol/L) and 2.0 mL of H2O2 (30%). The heating program consisted
of: i-) 5 min ramp to 160ºC, ii-) 2 min plateau at 160ºC, iii-) 5 min ramp to 230ºC and iv-) 15 min
plateau at 230ºC. After cooling down, the final volumes were made up to 14 mL with ultrapure water.
Digested samples (n=3) were analyzed using a Thermo ICAP 7400 Duo ICP OES for the
determination Ca, Fe, K, Mg, Mn, P, S and Zn. Results for granulometries were generally around
64%, with minimum and maximum of 3% (açaí) and 86%(apple), respectively. The average values
for pH and acidity were 4.3 and 18.2%, respectively, with minimum and maximum pHs (3.19 and
6.15 for blackberry and coconut, respectively) and acidities (3.22% and 43.4% for açaí and
blackberry, respectively). Moisture contents were between 6.45% (açaí) and 23.4% (apple). The
determination of macro and micronutrients in NIST SRM 1567a shown recoveries in the 92-107%
range, with precision as relative standard deviations better than 8.7%. Results for elemental
determinations in green banana, grape, coconut, passion fruit, orange, blackberry, açaí and apple
flours shown concentrations ranges (in mg/kg) of 312-5199 (Ca), 1428-26868(K), 161-2774(Mg),
247-4912(P), 300-1705(S), 7.9-134(Fe), 5.1-49.8(Mn) and 3.2-42(Zn). Grape flour samples
presented the highest contents for S, Fe and Mn and coconut flours were related to the highest levels
of Mg, P and Mn. Orange and passion fruit flours were the samples where Ca and K were more
abundant, respectively.
1 Comunian TA, Silva MP, Souza CJF, Trend. Food Sci. Tech., 108, 2021, 269–280.
2 Instituto Adolfo Lutz. Métodos físico-químicos para análise de alimentos, 2008.
3 AOAC - Association of Official Analytical Chemists. Official methods of Analysis. 18a ed., AOAC, 2006.
[FAPESP]

104
SLOPE RATIO CALIBRATION FOR ANALYSIS OF REFRACTORY MATERIALS
BY LASER-INDUCED BREAKDOWN SPECTROMETRY
Matheus F. Nunes (PG)*a, Aline C. Elias (PG)a, Lidiana C. Nunes (R)b, Juliana Naozuka (R)c,
Fábio R.P. Rocha (R)b, Cassiana S. Nomura (R)a
b University of São Paulo, Nuclear Energy in Agriculture Center, Piracicaba, SP, Brazil
c Federal University of São Paulo, Department of Chemistry, Diadema, SP, Brazil.
*e-mail: matheus.nunes@usp.br
Geological materials such as rocks, minerals and sediments are used as raw materials in
metallurgical, agricultural and construction sectors. As their use depends on the composition,
chemical analysis is imperative1. Analysis of those kind of sample, however, is not an easy task.
Most of the analytical methods requires sample dissolution before analysis but even using different
acid mixtures and modern microwave systems that combine high pressures and temperatures,
quantitative decomposition cannot be achieved2. In this case, the use of laser-induced breakdown
spectroscopy (LIBS) which allows direct solid sample analysis with none or minimum sample
handling, shows to be a great alternative3. On the other hand, quantitative analysis is a challenge in
LIBS due to matrix effects and the difficulties in finding compatible solid standards for calibration. In
this context, this work proposed the method development for direct determination of Fe and Al in
refractory sample by LIBS using slope ratio calibration (SRC). This approach uses a single standard
for calibration and the emission intensity from increasing number of accumulated laser pulses 4.
Certified reference materials SRM679 and SRM2703 from NIST were used as standard and sample,
respectively. Analysis was carried out in the pressed powder pellets of samples by pressing a mixture
containing sample and binder for 5 min at 8 ton.cm-2 (X-Press 3635 hydraulic press, Spex Sample
Prep, Metuchen, EUA). The use of cellulose and paraffin in different proportion were investigated
and the best condition were found in the pellet prepared with 60% sample and 40% paraffin.
Measurements were carried out using a J200 LIBS system (Applied Spectra, Fremont, CA, USA).
Instrumental parameters were optimized: laser energy (5 to 20 mJ), laser spot size (35 to 140 μm)
and delay time (0.25 to 1 μs) and the best condition were laser energy of 20 mJ per pulse, a delay
time of 0,35 μs and laser spot size of 50 μm. The slope ratio calibration was carried out with 5,10,
20, 30 and 40 laser pulses per pellet site. Analytical calibration curves showed R 2 > 0,95, high
precision (RSD < 5%) and accuracy (relative error of +7% and -3% for Al and Fe, respectively)
showing the applicability of the proposed method.
1 Harmon, R. S.; Russo, R. E.; Hark, R. R.; Spectrochimica Acta Part B: Atomic Spectroscopy; v. 87, 2013, p. 11.
2 flores, E. M. M.; Microwave-assisted sample preparation for trace element determination. Elsevier, 1 st Edition, 2014.
3 Carvalho, A. A. C.; Cozer, L. A.; Luz, M. S.; Nunes, L. C., Rocha, F. R. P.; Nomura, C. S., J. Anal. At. Spectrom. v. 34,
2019, p. 1701.
4 Nunes, L. C., Rocha, F. R. P.; Krug, F. J.; J. Anal. At. Spectrom, v. 34, 2019, p. 2314.
[CNPq, FAPESP, CAPES, INCTAA]

105
OVERCOMING INTEREFERENCES ON RARE EARTH ELEMENTS
DETERMINATION BY ICP-MS USING APEX SYSTEM AND DGT
Raquel C. Gradwohl (UG)a*, Luiz Felipe P. P. Moreira (PG)a, Melina B. T. Zanatta (R)a, Amauri
A. Menegário (R)a, Johanna Irrgeher (R)b, Shaun Lancaster (R)b, Ulrike Moser (R)b
a São Paulo State University (UNESP), Environmental Studies Center, Rio Claro, SP, Brazil, 13506-900
b Montanuniversitaet Leoben, Leoben, Austria, 8700
*e-mail: raquel.gradwohl@unesp.br
The use of different materials with specific properties to increase the functionality of a given product
has increased the demand for many elements that are important components in the development of
new technologies. They are defined as Technology Critical Elements (TCE) and include metals of
the platinum group, rare earths elements (REE) and others such as Nb, Ta, Ge and Te1. The
occurrence of REE in natural water samples is at concentration levels on the order of ng L -1, which
would be below the detection limits obtained from various technique. The inductively coupled mass
spectrometry (ICP-MS) is the most promising technique for the determination of REE due to its low
detection limits, selectivity, wide linear range and multi-element capability. However, the analysis of
REEs by ICP-MS is not always straightforward for quadrupole resolution instruments (QICPMS),
where many spectral interferences from oxide ions caused by the presence of Ba and lower mass
REE analytes in the sample. The use of a desolvating nebulizer system (APEX) can remarkably
reduce oxide formation, increasing the sample transport efficiency via obtaining more effective
aerosols2. Furthermore, the DGT technique also appears as an alternative for interferences
reduction due to its ability to selectively sample the chemical species of interest in their labile forms
(Ba ions in sample solution is partially sampled when Chelex-100 resin is used). The present study
systematically evaluated the use the APEX system and the DGT technique to overcome interference
in the determination of REE in river water samples. The water samples from the Paraíba do Sul River
were collected. DGT devices were immersed in triplicate in the river for 24 hours. Chelex® was
employed as binding layer and the polyacrylamide hydrogels were used as diffusive layers. In the
laboratory, the devices were disassembled and each disk of Chelex-100 was inserted into centrifuge
tubes for analytes elution with 2 mL of 1 mol L-1 of HNO3. DGT diffusion coefficient and elution factor
values used for each REE were based in a previous work3.To evaluate the total content, the samples
were digested in a microwave oven following the adapted EPA 3015A protocol before determination
by ICP-MS. For conventional determination using the QICP-MS technique, the MO+/M+ ratio was
around 0.02, wile, for APEX system, the MO+/M+ ratio was lower than 0.003. Comparison of data
obtained without and with APEX show that concentration of Ce, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm
were significatively (5% level) overestimated by direct measurement, by factors of 1.05, 7.6, 9.1, 9.5,
1.7, 1.9, 8.7, 2.3 and 3.2, respectively. The use of DGT noticeably reduced the interferences from
BaO+ on 146Nd+, 147Sm+, 153Eu+ by factors from (without DGT) 7.6, 9.1 and 9.5 to factor of 1.1, 1.2
and 1.6 (with DGT). Additionally, sampling by using DGT provided preconcetration factor from 10-
14 (considering a 24 hour deployment time). Therefore, APEX desolvation system and the DGT
technique proved to be efficient in reducing spectral interference in the determination of REE by ICP-
MS. Combination of both approaches can significatively remove interference and improve the limit
of detection in determination REE by QICP-MS.
1 Romero-Freire A, Santos-Echeandia J, Neira P, Cobelo-Garcia A, Front. Mar. Sci. 6:532(2019)
2 Dressler VL, Pozebon D, Matusch A, Becker JB, Int. J. Mass Spectrom. 266 (2007)
3 Huang J, Hills J, Teasdale PR, Panther JG, Wang F, Welsh DT, Talanta. 230 (2021)

106
DETERMINATION OF ESSENTIAL, NON-ESSENTIAL AND POTENTIALLY
TOXIC ELEMENTS IN BUFFALO MEAT BY ICP-OES
Maria N. D. Tourinho (UG)a, Mariane G. Nabiça (PG)a, Kelly G. F. Dantas (R)a*
a Federal University of Pará, Faculty of Chemistry, Institute of Exact and Natural Sciences, Belém, Pará, Brazil, 66075-
110.
*e-mail: kdgfernandes@ufpa.br
In Brazil, the growing demand and the challenges in obtaining good quality animal protein in a short
term and at low costs have motivated the producing class to invest in the creation of buffaloes aiming
at the slaughter of the animals. Buffalo meat fulfills the demands of the current consumer market,
which seeks healthy food, and at the same time, tasty, with good appearance and very juicy,
therefore, more suitable for human health. Considering the importance of animal protein
consumption for the human organism, the objective of this study was to determine the levels of As,
Ba, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Pb, Se, and Zn in different cuts of buffalo meat by inductively
coupled plasma optical emission spectrometry (ICP-OES). In this study, 8 samples of buffalo meat
(female and male) of different cuts acquired on a farm on the island of Marajó (Northern region of
Brazil) were used. The samples were lyophilized and then digested in a cavity microwave oven using
a combination of nitric acid (14 mol L-1), 30% H2O2 (m/m) and ultrapure water. The efficiency of the
sample preparation procedure and the accuracy of the measurements by ICP-OES were performed
using the analyte addition and recovery method. The recoveries obtained ranged from 80.0% to
102%. Higher levels of Ca (122.06 mg kg-1 to 563.20 mg kg-1), Fe (45.66 mg kg-1 to 112.26 mg kg-1),
K (9303.18 mg kg-1 to 14744.18 mg kg-1), Mg (629.68 mg kg-1 to 1175.17 mg kg-1), Na (1489.27 mg
kg-1 to 2775.73 mg kg-1, and Zn (77.60 mg kg-1 to 251.73 mg kg-1) were found in the samples when
compared to the other elements studied. As, Ba, Cd, Cr, Cu, Mn, Pb, and Se were below the
detection limit. Based on the recommended dietary reference intake (DRI) values for individuals
aged 9 to 70 years, the amounts for Ca, Fe, K, Mg, Na, and Zn ingested in 100 g of meat per day
were 12.21 to 56, 32 mg/day, 4.57 to 11.23 mg/day, 930.32 to 1474.42 mg/day, 62.97 to 117.52
mg/day, 148.93 to 277.57 mg/day and 7.76 to 25.17 mg/day, respectively. In this study, 5 types of
meat cuts exceeded the recommended value per day (8 to 11 mg/day) for Zn. This study showed
that the buffalo meats studied can be considered a good source of Ca, Fe, K, Mg, Na, and Zn. On
the other hand, the samples are not sources of Ba, Cr, Cu, Mn, and Se and potentially toxic elements
(As, Cd, and Pb).
1 Madella-Oliveira AF, Quirino CR, Adona PR, Pacheco A, Revista Brasileira de Reprodução Animal, 29, 2005, 53.
2 Otten JJ, Hellwig JP, Meyers LD, Ingestão dietética de referência. O guia essencial para as necessidades
nutricionais. The National Academies Press: Washington, EUA, 2006.
[FAPESPA, CAPES]

107
IN SITU SAMPLING AND DETERMINATION OF Cr, Cu, Mn, Ni, Pb AND Zn
USING DGT AND PORTABLE XRF SPECTROMETER
Leonardo H. Zoia (UG)a,*, Amauri A. Menegário (R)a, Hendryk Gemeiner (R)a,b, Eduardo de
Almeida (R)c, Luiz F. P. P. Moreira (PG)a, Hudson W. P. de Carvalho (R)c
b São Paulo State University (UNESP), Institute of Geosciences and Exact Sciences, Basin Studies Laboratory (LEBAC),
Rio Claro, SP, Brazil, 13506-900
c São Paulo University (USP), Center for Nuclear Energy in Agriculture, Piracicaba, SP, Brazil, 13416-000
*e-mail: leonardohenrique829@gmail.com
Diffusive gradients in thin films (DGT) have shown a promising potential for labile metal
determination in freshwater due to its pre-concentration, time-integrated, matrix interference removal
and speciation analytical features. Basically, a DGT device consists of a polypropylene piston, on
which is placed a layer of a ligand conventionally consisting of a Chelex-100 resin (immobilized in
an agarose-polyacrylamide hydrogel), a polyacrylamide-agarose hydrogels diffusion layer and a
membrane filter. These layers are fixed to the piston by a polypropylene sleeve. After deployments
of the DGT devices in an aquatic system, the masses of the analytes accumulated in the ligand is
eluted and related to the analyte concentration in the aquatic system. Typically, analyte masses are
obtained in laboratory by elution with HNO3 (taking about 12-24 h) and then analyzed by expensive
techniques, such as, inductively coupled plasma mass spectrometry or inductively coupled plasma
optical emission spectrometry1. Few alternatives have been proposed to eliminate the laborious and
time-consuming procedure of analyte elution2. Also, until now, no in situ procedures for analyte
determinations in the DGT eluate have been proposed resulting in a gap for using DGT in remote
areas where no clean laboratories are available. In this work, a new method based on the association
of DGT and a portable X-ray fluorescence spectrometer (PXRF) is proposed for in situ sampling and
determination of Cr, Cu, Mn, Ni, Pb and Zn. The method consists of basically three steps: 1) an initial
calibration (in Lab) of measurements by PXRF; 2) in situ deployment of DGT devices; 3) in situ
measurements of the analytes by using PXRF after drying the binding phase of the DGT devices.
The first step was performed by deploying the DGT device for 24 hours in a standard solution, drying
the binding phase with a clean paper (quantitative filtration paper) and silica gel, and establish a
relationship between the mass accumulated in the binding gel (mac calib) and the counts (CPS)
obtained by measurements using PXRF: mac calib /CPS. After in situ deployment (second step), in the
third step, the relationship mac calib /CPS was used to find the mass accumulated in field (mac) from
PXRF measurements (CPS). Thus, the relationship Cfield = (mac Δg)/(DtS) can be used to calculate
the analyte concentrations in the aquatic system (Cfield), since the analyte diffusion coefficient (D,
cm2s-1), the diffusion layer thickness (Δg, cm), the effective area (S, cm2) and deployment time (t, s)
are known. DGT devices were assembled with Chelex-100 resin (immobilized in an agarose-
polyacrylamide hydrogel), a polyacrylamide-agarose hydrogels diffusion layer and a membrane filter
(acetate membrane). The proposed approach was validated by determining the D values for Cr, Cu,
Mn, Ni, Pb and Zn from an immersion curve (from 8 to 48 h). Immersion curves follow the first law of
Fick of diffusion showing linearity (r2 from 0.78 to 0.99) and obtained D values (in 10-6 cm2s-1) were
very close to the expected values: 3.85 (4.00), 6.52 (6.60), 3.14 (5.10), 7.52 (6.30), 10.5 (10.3) and
3.75 (3.80) for Cr, Cu, Mn, Ni, Pb and Zn, respectively. Precision of measurements were
characterized by relative standard deviations from 8 to 20% (12 h deployment time, n=3). Limits of
detection (for 48 h deployment time, n=3) were 18, 4, 26, 4, 10 and 6 ng m L-1 for Cr, Cu, Mn, Ni, Pb
and Zn, respectively.
1 Menegário, A. M., et al. Analytica Chimica Acta, 983, 2017, 54.
2 Almeida, E. et. al., Spectrochimica Acta Part B, 71–72, 2012, 70.
[CNPq CAPES, FAPESP]

108
APPLICATION OF A PHOTOCHEMICAL VAPOR GENERATION SYSTEM FOR
THE DETERMINATION OF CONTAMINANTS IN IMITATION JEWELRY BY ICP-
OES
Fernanda P. Braga (PG)a, Aderval S. Luna (R)a,b, Diego B. Batista (PG)a,b, Jefferson S. Gois
(R)a,b*
a Graduate Program in Chemical Engineering, Rio de Janeiro State University, Rio de Janeiro, RJ, Brazil, 20550013
b Department of Analytical Chemistry, Rio de Janeiro State University, Rio de Janeiro, RJ, Brazil, 20550013
*e-mail: jefferson.gois@uerj.br
The global market of jewelry moves around 350 billion dollars annually (about 100 million dollars in
Brazil). Due to the high prices of jewelry, imitation jewelry has gained attention and it is being
consumed worldwide due to its affordable prices, however, most of the time chemical composition
is unknown. Due to the problems related to toxic elements in imitation jewelry, it is necessary to
investigate the quality and safety of these products when they reach the final customer. Imitation
jewelry with an undetermined combination of metals can result in health and environmental
problems, toxic elements such as Mn, Cd, Pb, Ni, Sb, As and Hg can be found as part of their
composition1,2. Therefore, analytical techniques for monitoring the concentration of contaminants in
imitation jewelry is a current and important issue, however, it is challenging due to the complexity of
the sample matrix (mostly composed of metals in high concentration). Photochemical vapor
generation (PVG) coupled to inductively coupled plasma optical emission spectrometry (ICP-OES)
is a straightforward approach that can be applied for the determination of trace-elements in matrices
such as those of imitation jewelry3. Thus, an analytical method for the determination of Hg, Mn, and
Pb in imitation jewelry samples by PVG-ICP-OES was developed. The PVG system was built with
two UV-C germicidal lamps (254 nm), a quartz capillary with 250 cm length, 2.5 mm internal
diameter, 4 mm external diameter, and 11 mL internal volume, and a glass gas-liquid separator
(GLS). The levels of the factors acetic acid concentration and UV exposure time were optimized
using a central composite design (CCD) for the PVG system, as well as the carrier gas flow rate and
the radiofrequency (RF) power for the ICP-OES. The optimum conditions were obtained at 30% v v-
1
acetic acid, 175 s radiation exposure time, 0.05 L min-1 carrier gas flow rate, and 1300 W for RF
power. The sample preparation for the imitation jewelries was carried out by simply dissolving the
samples in 2.0 mL of HCl 12 mol L-1 in a water bath at 70 oC. The influence of the inorganic matrix
of the sample was studied, where it was found that the presence of the major constituents in imitation
jewelry may cause the suppression of the analytical signal for the analytes, probably due to the
suppression of the formation of volatile species. To circumvent this effect, chemical modifiers were
studied. The accuracy of the proposed method was evaluated by recovery tests, which ranged from
88 to 102%. The detection limits ranged from 1 to 3 mg g-1.
1 Guney M, Zagury GJ, Environmental Science & Technology 48 (2), 2014, 1238.
2 Salles FJ, Dissertação de Mestrado, Universidade de São Paulo, 2017.
3 Sturgeon RE, Journal of Analytical Atomic Spectrometry, 32, 2017, 2319.
[CNPq, FAPERJ, UERJ]

109
SIMULTANEOUS DETERMINATION OF As AND Hg IN DIETARY
SUPPLEMENTS BY CHEMICAL VAPOR GENERATION COUPLED TO
MICROWAVE INDUCED PLASMA OPTICAL EMISSION SPECTROMETRY
Gustavo R. Bitencourt (PG)a, Valderi L. Dressler (R)a, Fábio A. Duarte (R)a, Rodrigo C.
Bolzan (R)a, Érico M. M. Flores (R)a, Paola A. Mello (R)a*
a Departamento de Química, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil.
*e-mail: paola.mello@ufsm.br
The consumption of dietary supplements has been widely growing. This brings concern about
elemental contaminants that can be present, which are introduced from raw materials,
manufacturing, transport, and storage of the final products, as well as can be impaired by growing
conditions (e.g., soil and pesticides used) in case of botanicals-based dietary supplements.1 Among
these elemental contaminants, As and Hg have been controlled due to their toxicity, mainly
carcinogenicity. The United States Pharmacopeia (USP) limited the concentration of both elements
in dietary supplements to 1.5 µg g-1.2 Atomic spectrometry techniques are commonly used for
monitoring these elements,1 but the suitability of microwave induced plasma optical emission
spectrometry (MIP-OES) has not yet been demonstrated. In this work, a method for the simultaneous
determination of As and Hg in dietary supplements by chemical vapor generation coupled to
microwave induced plasma optical emission spectrometry (CVG-MIP-OES) was developed. General
CVG conditions, the influence of the MSIS (multimode sample introduction system) setup, and pre-
reduction methods for conversion of As(V) to As(III) were evaluated. After optimization, the proposed
method provided limits of quantification (LOQs) for As and Hg from 0.005 to 0.016 and 0.029 to
0.077 µg g-1, respectively, complying the maximum concentration for these elements in dietary
supplements limited by USP (1.5 µg g-1). The method was used for the simultaneous determination
of As and Hg in dietary supplements of different classes (vitamins and minerals, minerals, amino
acids, and botanicals), which were previously decomposed by microwave-assisted wet digestion.
The concentration of both elements was below 1.5 µg g-1 in all samples. Spike recoveries were from
93 to 108% for both analytes in all samples. The accuracy of the proposed method was evaluated
by using certified reference materials (CRMs) of citrus leaves (NIST 1572) and pine needles (NIST
1575a), and no statistical difference was observed between the results and reference values (t-test,
confidence level of 95%). No difference was also observed for the results obtained using the
proposed method with those using a comparative method. Based on the results, the proposed
method using CVG-MIP-OES can be considered simple, accurate, and robust, and brings an
alternative for the quality control of As and Hg in dietary supplements, being suitable for the
determination of these elements in agreement with USP requirements.
1 Smichowski P, Londonio A, Microchemical Journal, 136, 2018, 113-120.
2 USP, General Chapter <2232> Elemental Contaminants in Dietary Supplements, 2012.
[CAPES, CNPq, UFSM]

110
IMAGING OF TRACE ELEMENTS DISTRIBUTION IN RICE BY LASER
ABLATION INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY
Cristiane Pappis (PG)a, Cristian K. Pereira (UG), Dylan M. Hoffmann (UG)a, Valderi L.
Dressler (R)a*
a Federal University of Santa Maria, Department of Chemistry, Santa Maria, Rio Grande do Sul, Brazil, 97105-900
*e-mail: vdressler@gmail.com
Rice is a staple food in many countries, making up 30% of calories intake in an average diet and up
to 70% in some low-income countries1. The characteristics of the rice grains regarding nutrient and
toxic elements concentrations, are known to be dependent of the rice variety, soil where the rice is
cultivated, milling processes and storage2. Elemental determination of rice grains usually involves
sample decomposition, which is a time-consuming procedure. On the contrary, direct sample analysis
is much faster and does not involve the use of reagents to decompose the sample. Laser ablation
inductively coupled plasma mass spectrometry (LA-ICP-MS) was employed in the present work for
determination and imaging of Mg, Mn, Cu, Co, Fe, Zn, As, Sr, Sb, Ba and Pb in grains of white,
parboiled, and whole (brown) rice. The elements quantification was achieved by using external
calibration whereas filter paper discs served as support for the reference solutions and matrix
matching. Accuracy was ensured by analysing two certified reference materials (CRMs) pressed into
pellets. Investigations were carried out to evaluate the distribution of the elements on the surface and
external layers of the grains. For that purpose, the surface of rice grains fixed on a glass support was
ablated line-by-line and the ablation repeated three times. The images generated from laser ablation
revealed that the investigated elements were not homogeneously distributed in the analysed parts of
the grains. Except for Pb, the investigated elements were detected in three layers of the rice grains,
but it was observed that elements concentrations decreased from the surface to more internal layers.
Lead was detected only in the first layer of white and whole rice and in the first and second layers of
parboiled rice. Figure 1 shows how the 66Zn and 63Cu concentrations varies on the surface and as a
function of the depth in the rice grain. In general, the concentration decreases from the first ablation
to the last one in the three types or rice. This behavior was also observed for the other analytes (not
shown in Figure 1 to avoid repetition). By using LA-ICP-MS is possible to show how these elements
are distributed on the surface and external layers of parboiled, whole and white rice grains. It was
observed that the elements distribution changes spatially, at the surface and from one layer to
another. In general, the element concentrations decrease from the surface to more internal layers
and the lowest decrease was observed for parboiled rice. Agreement in the range typically observed
in LA-ICP-MS analyses was observed for the results obtained of the CRM.
Figure 1. Photograph (bottom) of a rice grain;
the dashed red line corresponds to the place
where three repeated lines (1st, 2nd and 3rd)
were ablated. The upper figures correspond to
the concentrations found for each ablation
line.
1 Fukagawa NK, Ziska, LH, Journal of Nutritional Science and Vitaminology, 65, 2019, S2.
2 Hensawang S, Lee BT, Kim KW, Chanpiwat P, Journal of the Science of Food and Agriculture, 100, 2020, 4029.
[CAPES, CNPq, UFSM]

111
CHEMICAL FRACTIONATION AND IN VITRO BIOAVAILABILITY STUDY OF
Cu, Fe, Mn AND Zn IN BARU (DIPTERYX ALATA VOG.)
Mirian C. Santosa*, Nicole M. M. C. Cotrima, Thiago L. C. Rochaa, Luiz F. Malmongeb
a Department of Analytical, Physical-Chemistry and Inorganic Chemistry (DQAFQI), Institute of Chemistry, São Paulo
State University (UNESP), Araraquara, São Paulo, Brazil. 14800-900
b Department of Physics and Chemistry (DFQ), São Paulo State University (UNESP), Ilha Solteira, São Paulo, Brazil,
15385-000
*e-mail: mirian.cristina@unesp.br
The baru (Dipteryx alata Vog.), typical fruit of the Cerrado biome, is a seed characterized as rich in
minerals. However, the determination of the total content of an element is limited information about its
use in the absorption process and, therefore, in the individual's nutrition. Through chemical
fractionation or analysis of speciation of elements, which aim to determine the concentration of the
physicochemical forms of the elements, its absorption can be predicted. The objective of this work
was to evaluate the chemical fractionation of the metals Fe, Cu, Zn and Mn in baru seeds applying
the sequential extraction method. The samples were treated with a succession of extractors in
progression of chemical action and the bioavailable portion of these metals was evaluated by in vitro
assays. Sequential extraction was performed using the following extractors: 1 mol L-1 calcium chloride
solution; 0.5 mol L-1 acetic acid solution in 5% ammonium acetate medium (with pH 5.0); 0.5 mol L-1
acetic acid solution and 0.5 mol L-1 hydrochloric acid solution. The in vitro bioavailability assays were
performed according to the IVG method (In Vitro Gastrointestinal Model), using a solution that mimics
gastric and intestinal solutions. The entire process was performed in triplicate and the concentrations
of the elements in all fractions were determined by FAAS. The total contents of Cu, Fe, Mn and Zn
present in the baru were 1.48; 4.74; 3.56 and 4.82 mg/100g respectively, and the percentages of in
vitro bioaccessibility in the gastric phase were 53.33%; 17.80%; 29.85% and 49.61% and in the
intestinal phase 17.20%; 71.03%; 25.07% and 10.92% for Cu, Fe, Mn and Zn, respectively. From the
results, it is observed that Cu, Mn and Zn are in at least 5 different chemical species in baru seeds,
while Fe is in the form of at least 4 species.
1 Rufino M S M, Propriedades funcionais de frutas tropicais brasileiras não tradicionais. 2008. Universidade Federal
Rural do Semi-Árido, 2008.
2 Cozzolino S M F, Biodisponibilidade de Nutrientes. 5. ed. Barueri: Manole, 2016.
3 Andrade É C B; Alves S P, Takase, I. Extração sequencial de cobre, ferro e zinco em ervas medicinais. Ciência e
Tecnologia de Alimentos, 25, 2005, 844.

112
EMPLOYMENT OF STANDARD DILUTION ANALYSIS AS A POWERFUL TOOL
FOR ELEMENTAL DETERMINATION IN BIODIESEL BY ICP OES WITHOUT
OXYGEN ADDITION
Vitor C. P. Marrocos (PG)a, Jefferson R. de Souza (R)b, Tatiana D. Saint’Pierre (R)a,*
a PontificalCatholic University of Rio de Janeiro, Department of Chemistry, Rio de Janeiro, RJ, Brazil, 22451-900
b North Fluminense Darcy Ribeiro State University, Departament of Chemistry, Campos dos Goytacazes, RJ, Brazil,
28013-603
*e-mail: tatispierre@puc-rio.br
Direct introduction of organic solvents into inductively coupled plasma (ICP) spectrometers causes
several problems that increase equipment maintenance costs and reduce its lifetime.1 In addition,
they may impose matrix effects, which requires further efforts to be accounted for. To minimize these
problems, it is proposed to employ standard dilution analysis (SDA) as the calibration strategy to
minimize matrix effects, also reducing reagents consumption and waste, when compared to
traditional calibration methods.2 Calibration by SDA combines the fundamentals of the standard
addition and internal standardization methods and employs only two analytical solutions. The first
one containing 50 % w/w of sample + 50 % w/w of analytes and internal standards, and the second
containing 50 % w/w of sample + 50 % w/w of blank. By keeping the same amount of matrix, a
calibration curve is constructed from the mixing of both solutions while data is collected over time.
After careful optimization of ICP OES operational parameters for introduction of biodiesel samples
diluted in xylene, Miramist CETM micronebulizer was chosen under conditions of 0.2 L min-1 for Ar
nebulizer flow rate, 0.3 mL min-1 for sample flow rate and 1500 W for RF power without oxygen
addition. One of the main drawbacks of the SDA method relies on the dataset selection that best fits
the linear model,3 which has been overcome when a second internal standard (Y) was applied thus
allowing more data obtained from the SDA region to be used for the calibration curve. Figures of
merit obtained using different approaches for calibration (external calibration (EC), standard addition
(SA), and internal standardization (IS)) were compared to those obtained by the SDA for the
determination of Na, K, Ca, Mg, P, Si, and S. All quantitative approaches had comparable limits of
quantification (LOQ) for most studied elements (ranging from 0.02 mg kg-1 (Mg) to 3.7 mg kg-1 (K)).
However, the LOQ obtained for S determination by SDA was four times better (0.6 mg kg-1) than that
obtained by IS (2.5 mg kg-1). While precision (RSD < 6 %) and accuracy (recovery 89 - 115 %) of
the results proved SDA analytical performance, its high sample throughput and matrix effects
compensation are the main advantages over traditional calibration methods.
1 Duyck C, Miekeley N, Porto da Silveira C L, Szatmari P, Spectrochimica Acta Part B: Atomic Spectroscopy, 57, 2002,
1979.
2 Jones W B, Donati G L, Calloway C P, Jones B T, Analytical Chemistry, 87, 2015, 2321.
3 Sloop J T, Bonilla H J B, Harville T, Jones B T, Donati G L, Talanta, 205, 2019, 120160.
[CNPq, CAPES, FAPERJ]

113
PHOTOCHEMICAL VAPOR GENERATION IN ORGANIC MEDIUM: AN
ASSESSMENT BASED ON THE DETECTION OF AS AND HG IN BIODIESEL AND
GLYCERIN USING AAS
Bárbara Souza Soares (PG)a, Graciany Matias (UG)a, Daniel L. G. Borges (R)a
a Federal University of Santa Catarina, Chemistry Departament, Florianópolis, Santa Catarina, Brazil, 88040900
e-mail: barbara.souzasoares@yahoo.com
Photochemical vapor generation (PVG) is a relatively recent strategy to assist on the quantification
of trace elements. Applications aiming at the analysis of organic samples are notoriously scarce, so
there is little information on the effect of the organic matrix on the effectiveness of the photochemical
process. Under this assumption, the goal of this work was to provide an in-depth investigation of
parameters that affect the vapor generation efficiency of As and Hg (as CH3Hg+ and Hg2+) in organic
samples, using biodiesel and glycerin as test samples and detection by atomic absorption
spectrometry. The photochemical reactions were triggered by photons emitted from an ultraviolet
(UV) low-pressure Hg lamp with emission at 253.7 nm. Sample solutions were introduced in the
photochemical system by flow injection and exposed to UV radiation for periods that ranged from 12
to 70 s. The generation of gas-phase Hg was established following sample dilution with n-propanol
and formic acid as the organic precursor. The PVG of As was carried out using glycerin samples
diluted with water and higher vapor generation efficiency was achieved using acetic acid or its
mixture with formic acid. Attempts to obtain a calibration function revealed significant decrease in
sensitivity associated with the photochemical process in sample solutions containing the organic
sample at concentrations as low as 10% (v/v). In order to further investigate the sensitivity loss, the
viscosity of solutions containing biodiesel was determined. Increased viscosity was determined upon
the reduction in the sample dilution factor, with subsequent loss in sensitivity associated to PVG. An
analysis of the solutions containing biodiesel by UV molecular absorption spectroscopy showed that
biodiesel samples absorbed strongly at 253.7 nm, which implies in photon competition between the
sample and the organic precursor inside the PVG reactor, with immediate effect on the sensitivity of
the photochemical process. The results obtained in this study stand to demonstrate that regardless
of the analyte, analysis of high viscosity and/or organic samples will result in efficiency loss
associated with the photochemical process. Hence, in cases where signal suppression is not
complete, standard addition calibration will likely become the technique of choice to carry out
quantitative analysis. The use of photocatalysts, which is intended as the next step in this study, is
probably important to overcome the sensitivity issues associated with the irradiation of organic
samples.
[UFSC, CAPES, CNPq, Petrobras]

114
QUANTIFICATION OF RARE EARTH ELEMENTS IN SUSPENSIONS OF
GEOLOGICAL SAMPLES USING HR-CS GF AAS
Júlia C. L. Zimmermann (PG)*, Daniel L. G. Borges (R)
Universidade Federal de Santa Catarina, Departamento de Química, Florianópolis - SC
*e-mail: julia.clz@posgrad.ufsc.br
Rare earth elements (REE) are highly useful metals applied in the manufacture of a variety of modern
technological devices, as catalysts in industrial processes and in biochemistry, as components of
non-invasive tests and as biological tracers and markers1, for instance. The high added value
classifies the REE as a strategic resource, similar to water, iron ore and petroleum 2. Quantification
of REE is frequently complex, since these elements have similar physical and chemical properties
and are prone to spectral interference in optical and mass spectrometric techniques. In addition,
REE are usually found in geological samples, which are typically difficult to process for chemical
analysis. This work intended to develop a simple strategy to quantify these elements with minimal
sample preparation using high resolution continuum source graphite furnace atomic absorption
spectrometry (HR-CS GF AAS). The samples were prepared as aqueous suspensions in 5% v/v
HNO3. Ytterbium and Samarium were investigated and method development was carried out using
a diabase rock certified reference material (CRM). The pyrolysis and atomization temperatures were
optimized for each analyte, with and without coating the graphite furnace with Zr as a permanent
modifier, based on wall atomization. The use of Zr-coated tubes resulted in shorter transient signals,
which is likely a consequence of the diminished contact between the analytes and the graphite
surface. Considering the refractory nature of the analytes, several calibration approaches were
investigated, consisting on the use of aqueous standards, calibration against different CRM prepared
as suspensions and standard addition calibration. Quantification of Yb based on calibration against
aqueous standards resulted in determined values below the certified values in up to 91%. This is
clearly due to the higher atomization efficiency achieved in the absence of a matrix, which is typical
to refractory elements. The determined values were improved following standard addition calibration
and further improvement was achieved by the adoption of a calibration curve obtained from the
insertion of increasing volumes of a CRM suspension in the graphite furnace. A similar outcome was
observed for Sm, with good agreement between determined and certified/informed values based on
calibration against SARM-19 coal suspension. The limit of detection for both elements was
established as 0.2 g g-1, considering a suspension containing the sample at 0.6% (m/v). The
detection capability may clearly be enhanced upon the adoption of more concentrated suspensions,
i.e., sample preparation is flexible and may be adjusted according to the expected analyte
concentration. Overall, Yb and Sm could be efficiently quantified in a series of geological materials,
including rocks, minerals and coal, following sample preparation as suspensions. The methods were
proven fast and reliable and are an attractive alternative to the adoption of more extensive sample
preparation approaches, such as acid digestion or alkaline fusion.
1 Martins TS, Isolani PC, Terras raras: aplicações industriais e biológicas, Quím. Nova 28, 2005.
2 Lima PCR, Terras-raras: elementos estratégicos para o Brasil, disponível em <https://www2.camara.leg.br/a-
camara/estruturaadm/altosestudos/pdf/EstudoMineraisEstratgicoseTerrasRaras.pdf> acesso em 29 JUL 22.
[CNPq, CAPES]

115
COMBINATION OF ULTRASONIC EXTRACTION, DISTILLATION, AND
NANOPARTICLE-MEDIATED (GRAPHENE QUANTUM DOTS-TIO2)
PHOTOCATALYTIC DEGRADATION FOR MERCURY SPECIATION IN
OFFSHORE PRODUCED WATERS BY COLD VAPOR ATOMIC ABSORPTION
SPECTROMETRY
Jarol R. Miranda-Andrades (R)a*, Marlin J. Pedrozo-Penãfiel (R)a, Joseane A. Mendes(R)a,
Ricardo Q. Aucelio (R)a
a Department of Chemistry, Pontifical Catholic University of Rio de Janeiro (PUC-Rio), Rio de Janeiro 22451-900, Brazil.
*e-mail: jmirandaa83@live.com
Mercury (Hg) is one of the most dangerous environmental pollutants due to its toxicity and ability to
accumulate in the environment 1. In produced water (PW), associated with offshore oil production,
Hg can be present as “inorganic mercury” (Hg2+), but it can also exist as organomercury species,
predominantly methylmercury (CH3Hg+)2. This work describes a new method for the speciation and
determination of mercury in samples of PW by CV-AAS, after separation of the mercury species,
combining the processes of: (i) Ultrasonic extraction in the presence of surfactant; (ii) Assisted
distillation under flow of argon; (iii) Photocatalytic degradation of mercury species (in the distilled
samples), using graphene quantum dot hybrid nanoparticles supported on titanium dioxide
nanoparticles (GQDs-TiO2) as photocatalytic agents. PW are characterized by having dispersed
crude oil, high levels of organic compounds and dissolved salts, which makes a direct analysis and
mercury speciation difficult. Therefore, before submitting the PW samples to the photocatalytic
reaction, the mercury species present in the samples were separated from the complex sample
matrix using the extraction and distillation protocol recently developed in our laboratory2. For
preliminary analyses, volumes of 30 mL of simulated PW samples were used, containing 5 µg L-1 of
(CH3Hg+, CH3CH2Hg+, and Hg2+) mixed with oil contents (20) mg L–1 and (30) g L-1 NaCl respectively.
After distillation (about 80% of sample), 2 mL of the samples were transferred to 5 mL volumetric
flasks, then the final volume was adjusted with aliquots of (H2O2 30%; H2SO4 0.1 mol L-1; formic
acid (1% m/v); a small volume (in μL) of the GQDs−TiO2 dispersion and ultrapure water.
Subsequently, these mixtures were transferred to quartz tubes (15 mL) connected to the CV-AAS
system. The best recoveries were found at pH 3.0; in the presence of formic acid (1% m/v), 100 μL
of the GQDs-TiO2 dispersion and after 13 min of UV irradiation, thus, it was possible to perform the
temporal separation of the released form of Hg due to the different mercurial species determined by
the CV-AAS using a dedicated system with a multiple optical path length cell. In addition, the possible
effects of the interference of oil (0.5; 1.0, 1.5 and 2.0 mg L -1) and NaCl (1.0; 2.0; 4.0; and 6.0 mg L-
1
) levels in the photocatalytic degradation step were evaluated using GQDs-TiO2 under UV. No
significant interference was found showing the reliability of the mercury speciation results using
photocatalytic degradation. The limits of detection were 12 ng L-1 for Hg2+; 17 ng L-1 for CH3CH2Hg+
and 15 ng L-1 for CH3Hg+.The precision of the method obtained by replicate analyzes of simulated
PW samples was better than 4% (RSD). The method can potentially replace tedious procedures
based on chemical derivatization of mercurial species that use toxic and expensive reagents, being
an economical alternative for the determination and ultratrace mercury speciation.
1 Y.M. Liu, F.P. Zhang, B.Y. Jiao, J.Y. Rao, G. Leng, Journal of Chromatography A, 1493, 2017.
2 J.R. Miranda-Andrades, S. Khan, M.J. Pedrozo-Penãfiel, K. de C.B. Alexandre, R.M. Maciel, R. Escalfoni, M.L.B. Tristão,
R.Q. Aucelio, Spectrochimica Acta Part B: Atomic Spectroscopy, 158, 2019.
[FAPERJ, CNPq, CAPES]

116
COMPARATIVE STUDY OF THE INORGANIC COMPOSITION OF YELLOW AND
RED CASHEW APPLE
Tiago L S Coelho (PG)a*, Darlisson S N Silva (PG)a, Ivero P. Sá (PG)b, Julymar M. Higuera
(PG)b, Cícero A Lopes Júnior (R)a, Ana R A Nogueira (R)c, Edivan C Vieira (R)a
a Federal University of Piauí, Departament of Chemistry, Teresina, Piauí, Brazil, 64049-550
b Federal University of São Carlos, Departament of Chemistry, São Carlos, São Paulo, Brazil, 13565-905
c Embrapa Pecuária Sudeste, São Carlos, São Paulo, Brazil, 13560-970
*e-mail: tiagolinusquimico@gmail.com
The cashew tree (Anacardium Occidentale L.) is widely cultivated in Brazil, which produces the
cashew apple, the cashew pseudofruit, which can be found in orange, yellow, and red colors.
Although the cashew apple has a high nutritional content, it is often discarded without any use1-2.
According to our searches, no recent studies were found that seeks to determine and compare the
mineral composition present in yellow and red cashews3-4. In this context, the determination and
comparison by ICP OES of Ca, Cu, Fe, K, Mg, Na, S, P, and Zn in yellow and red cashew apples
were carried out. Cashew apples were collected in the city of Valença do Piauí and freeze-dried.
After this step, the samples and CRM 1515 (Apple Leaves) were microwave-assisted digested in an
Ethos 1 system. For that, ca. 130 mg of sample were transferred to PFA vessels, and 6.0 mL of
diluted HNO3 (7 mol L-1) and 2.0 mL of 30% (m m-1) of H2O2 were added to the samples. The samples
were submitted to the following heating program: (1) 15 min to reach 200 °C; (2) 20 min maintained
at 200 °C. In both steps, the applied power was 1000 W. The values of the limits of quantification
LOQ 15.4, 0.2, 1.8, 1.9, 55.9, 2.1, 2.2, 4.8, and 4.7 were sufficiently low to determine the analytes
studied. All elements showed good linearity for the calibration curves obtained with r > 0.99. The
method validation was performed by calculating the accuracy of each element determined by the
CRM reference values. The recovery values
results were 82-113%, showing that the
concentrations found are in accordance with
the certified values and the precision
expressed. in relative standard deviation (%
RSD), < 6.5% (n = 3). Validated conditions
were applied in yellow and red cashews to
determine Ca, Cu, Fe, K, Mg, Na, S, P, and
Zn. For comparison purposes, Student's t-
test at 95% confidence level was used. The
results are shown in Figure 1 and show that
red cashews have a higher concentration of
the nutrients Ca, Fe, K, Cu, Zn, and S that
have average contents of 222.38 ± 0.10,
Figure 1. Concentration in mg kg-1 of Ca, Fe, K, Mg, S, Na, Cu, 9077.02 ± 564.99, 14.30 ± 0.25, 12.12 ±
P and Zn in yellow and red cashew samples.
0.34 and 1349.26 ± 31.31 mg kg-1. For the
yellow cashew apple, Mg, Na, and P were
higher than red at 800.37 ± 24.17, 250.13 ± 30.06, and 970.21 ± 29.14 mg kg -1. Among the
determining elements, Na, Ca, Mg, K, and P present high concentration values in all samples, while
Fe, Cu, and Zn were found at low concentration levels. Therefore, red cashew has a higher
concentration in most of the analyzed elements and can be an essential source of minerals. These
important nutrients perform different activities on the excellent development of the human body.
1 Brito ES, Araújo MC, Lin LZ, Harnly J, Food Chemistry, 105, 2007,1112–1118.
2 Honorato TL, Rodrigues S, Food Bioprocess Technol, 3, 2010, 105–110.
3 Preethi P, Mangalassery S, Shradha K, Pandiselvam R, LWT - Food Science and Technology, 20, 2020, 31527.
4 Reina LJC, Duran-Aranguren DD, Forero-Rojas LF, Tarapuez-Viveros LF, Heliyon, 8, 2022, 09528.
[UFPI, CNPq e Embrapa Pecuária Sudeste]

117
SEQUENTIAL AND SIMULTANEOUS DETERMINATION OF CD, NI AND FE IN
EDIBLE SEEDS BY HR-CS GF AAS USING DIRECT ANALYSIS
Leandro dos Santos (PG)a,b, Natália K. V. dos Santos (PG)a, Maria G. R. Vale (R)a,c, Morgana
B. Dessuy (R)a,c*
a Universidade Federal do Rio Grande do Sul, Instituto de Química, Porto Alegre, RS, Brazil, 91501 - 970
b InstitutoFederal de Educação, Ciência e Tecnologia Sul-rio-grandense, Pelotas, RS, Brazil, 96015-360
c Instituto Nacional de Ciência e Tecnologia do CNPq - INCT de Energia e Ambiente, Universidade Federal da Bahia,
Salvador, BA, Brazil, 40170-115

*e-mail: mbdessuy@ufrgs.br
The Food and Agriculture Organization of the United Nations (FAO) has established the promotion
of healthier food systems as a priority for 2020 and 2021. In response to this demand, some edible
seeds such as pumpkin, amaranth, chia, sunflower and quinoa have been gaining great popularity
in the human diet as they contribute to healthy eating behavior and have a high nutritional value.
Although they are source of elements of nutritional importance, they may also contain potentially
toxic elements. Thus, the accurate determination of essential and / or potentially toxic trace elements
in human health plays an important role for their evaluation. The high-resolution continuum source
graphite furnace atomic absorption spectrometry (HR-CS GF AAS) technique, employing direct
analysis of solids, stands out for allowing the sequential and / or simultaneous multi-element
determination of some elements1. This work aimed to develop an analytical method, using HR-CS
GF AAS to first quantify Cd and, in sequence, Fe and Ni, simultaneously, from a single sample
aliquot. A high-resolution spectrometer, contrAA 700 model (Analytik Jena, Jena, Germany),
equipped with graphite furnace as atomizer and a manual solid sampling accessory was used for all
measurements. The seeds were ground in a micro-mill and macerated to ensure their micro-
homogeneity. A combination of Pd/Mg and H2O2 was used as a chemical modifier. A mass up to
0.65 mg of sample was weighed onto the platform, then the chemical modifier was pipetted over the
sample, and it was inserted into the furnace. First, Cd was determined, at 228.802 nm, using a
pyrolysis and atomization temperatures of 400 °C and of 1600 °C, respectively. After, a second
atomization temperature of 2500 °C was applied and Fe (232.036 nm) and Ni (232.003 nm) were
determined. The use of H2O2 eliminated the spectral interference of smoke, allowing the use of lower
pyrolysis temperatures. Under these conditions, it was observed that Cd volatilizes at pyrolysis
temperatures above 400 ºC. Calibration curves with aqueous multi-elemental standard solutions
were used to quantify the three elements in the samples, and trueness was accessed by analyzing
the certified reference material of tomato leaves (NIST 1573a). The detection limits were 3.1 μg kg -
1
Cd, 3.4 mg kg-1 Fe and 0.062 mg kg-1 Ni. Ten seed samples were analyzed and presented a
concentration range of 39 - 254 μg kg-1 Cd, 1.1 - 8.0 mg kg-1 Ni and 38 - 147 mg kg-1 Fe. The values
found for the certified reference material were in agreement with the certificate values based on a
Student t-test on a 95% confidence level (tcritical: 2.78, tcalculated Cd: 0.42, Ni: 0.40 and Fe: 0.22). The
results shown that the method is robust and could be applied for the sequential and simultaneous
determination of Cd, Ni and Fe in other seed samples, providing more information about the mineral
composition of seeds consumed by the population.
1 Boschetti W, Orlando M, Dullius M, Dessuy MB, Vale MGR, Welz B, de Andrade J, J. Journal of Analytical Atomic
Spectrometry, 31, 2016, 1269.
[INCT E&A, CAPES, CNPq, PROPESQ]

118
A NEW BIOANALYTICAL STRATEGY USING (IPD)ICP-MS FOR THE STUDY OF
IRON INCORPORATION IN ERYTHROCYTES: A PROPOSAL FOR A BETTER
EVALUATION OF IRON ABSORPTION IN SICKLE CELL PACIENTS
Bernardo F. Braz (PG)a,b, Juliana Omena (R)c,d, Vanessa M. Voll (PG)d, Marta Citelli (R)d,
Cláudia S. C. Rodrigues (R)d, Fernando H. Cincotto (R)b,e, Ricardo E. Santelli (R)b,e*
a Universidade Federal do Rio de Janeiro, Programa de Pós-Graduação em Química, Rio de Janeiro, RJ, Brazil.
b Universidade Federal do Rio de Janeiro, Laboratório de Desenvolvimento Analítico, Rio de Janeiro, RJ, Brazil.
c Universidade Federal do Rio de Janeiro, Instituto de Nutrição Josué de Castro, Rio de Janeiro, RJ, Brazil.
d Universidade Estadual do Rio de Janeiro, Instituto de Nutrição, Rio de Janeiro, RJ, Brazil.
e Instituto Nacional de Ciência e Tecnologia em Bioanalítica, Campinas, SP, Brazil.
*e-mail: bernardobraz@pos.iq.ufrj.br
Iron (Fe) is the fourth most abundant chemical element in the earth's crust. In human life, Fe has an
essential function, acting as a cofactor of several enzymes and, mainly, in respiration. Therefore, it
is extremely important to know and better understand your metabolism. The nutritional status of Fe
in the human body is directly linked to the health1 of the individual. We have that almost 25 % of the
world population is anemic and around 800,000 deaths per year are attributed to lack of Fe. Sickle
cell disease (SCD) is a severe, multisystemic, single-gene disease that has a high occurrence in the
world. Erythrocyte rigidity and the consequent vaso-occlusion events are the main points in the
pathophysiology of this disease. These recurrent events lead to progressive organ failure, such as
the brain and lungs. The United Nations (UN) estimates that one in every 275,000 people born each
year have SCD. It is known that the highest occurrence of this disease occurs on the African
continent (and the most serious clinical conditions are also found there). Here in Brazil, SCD is the
most prevalent hereditary disease2. One of the most used forms of treatment in SCD is blood
transfusions. Each unit of transfused blood has about 250 mg of Fe and the use of this treatment,
although it prevents complications of the disease, can lead patients to Fe overload and may manifest
clinical symptoms after 10 transfusions. Several studies suggest an increase in the gastrointestinal
absorption of Fe in sickle cell patients due to the chronic hemolysis of these individuals; however,
they are frequently transfused and therefore receive large amounts of Fe 3. Clinical practice with
these patients is to restrict the intake of Fe in their diet as much as possible, which has a great impact
on the lives of these individuals. However, in several cases, iron deficiency anemia is also diagnosed
in these patients, which was not expected. ICP-MS is an analytical technique that allows the
determination of elements, through their isotopes and isotopic ratios, using mass spectrometry (MS)
of ions produced through an inductively coupled plasma (ICP)4. The Isotope pattern deconvolution
(IPD) is a mathematical tool that allows relating (and quantifying) the composition, in terms of isotopic
profiles, of a sample that has suffered interference in its isotopic signature, therefore, this tool
provides the understanding of how a tracer with isotopic composition known influence on the isotopic
signature of the sample, and using the concept of isotopic dilution it is possible to know the
concentrations of each of the constituent isotopic profiles. In the present study, a protocol was
developed for the administration of 57FeSO4 to patients treated at HEMORIO and later a method for
the use of (IPD)ICP-MS as a way of evaluating Fe absorption in several patients. With the method
developed, the groups studied (SCD patients; SCD patients with Fe overload; and healthy groups -
each of the three for children and adults, totaling 6 groups) were differentiated by the levels of Fe
absorption and the results showed are quite promising in terms of their use for the assessment of
the nutritional status of Fe.
1 Muckenthaler, M. U. et al. Cell, 168, 2017, 344-361.
2 Zago, M. A; Pinto, A. C. S. Revista Brasileira de Hematologia e Hemoterapia, v. 29, n. 3, p.207-2014, 2007.
3 BRASIL. Conselho Nacional de Saúde. Resolução nº 466, de 12 de dezembro de 2012. Diretrizes e normas
regulamentadoras de pesquisas envolvendo seres humanos. Diário Oficial [da] República Federativa do Brasil, Poder
Executivo, Brasília, DF, 12 de dezembro de 2012, p.59-62.
4 Ammann, A. Inductively coupled plasma mass spectrometry (ICP MS): a versatile tool. Journal of Mass Spectrometry, v.
42, n. 4, p. 419-427, 2007.
[CNPq, FAPERJ]

119
COMPARATIVE STUDY OF THREE ATOMIC SPECTROMETRIC TECHNIQUES
FOR HG DETERMINATION IN MARINE SEDIMENT SAMPLES
Carolina S. Provete (PG)a, Bruna M. Dalfior (PG)a, Rafael Mantovaneli (PG)a, Eldis Maria
Sartori (PG)a, Maria Tereza Weitzel Dias Carneiro (R)a, Geisamanda Pedrini Brandão (R)a*
a Federal University of Espírito Santo, Chemistry Departament, Vitória, Espírito Santo, Brazil, 29901-291
*e-mail: geisamanda@gmail.com
The determination of mercury (Hg) in marine sediment is an analytical challenge due to toxicity of
this element even at low concentrations (content generally found in nature) and complex matrix. 1,2
Therefore, it is necessary to use analytical techniques that have high sensitivity, selectivity and low
limits of quantification (LoQ).3 In this study, methodologies were evaluated using ICP-MS (inductively
coupled plasma mass spectrometry), CV-ICP OES (inductively coupled plasma optical emission
spectrometry with cold vapor generation) and TDA AAS (atomic absorption spectrometry with
thermodecomposition and amalgamation) for Hg determination in marine sediment samples. These
techniques vary from each other due to differences on the obtained analytical signal. Also, the ICP
techniques are multielemental and there is a need for the sample be in solution for its introduction
for analyses; on the other hand, the TDA AAS technique can be used with direct sampling of solids
when applied with the Milestones spectrometer DMA-80 (Direct Mercury Analyzer). In order to
analyze the efficiency of ICP-MS, CV-ICP OES and TDA AAS, it was compared the analytical figures
of merits. Accuracy was verified through CRM (certified reference material) NIST 2702 Marine
Sediment analysis, and no statistical difference between the determined and the certified value was
found with the different methods.4 The LoQ obtained were 1.9, 163 and 0.35 µg∙kg−1 for ICP-MS,
CV-ICP OES and TDA AAS, respectively. Comparing marine sediment samples analysis, Hg
concentrations had no statistical difference when determined by ICP-MS and TDA AAS. It was not
possible to determine Hg in marine sediment samples by CV-ICP OES technique due to high LoQ
obtained (163 µg∙kg−1). Although the methodology that used the CV-ICP OES technique was not
sensitive for Hg determination in the samples, it could be used to indicate the classification according
to the criteria established by CONAMA (Brazils National Environment Council) wich is 300 µg∙kg−1
for Level 1 and 1000 µg∙kg−1 for Level 2.5 ICP-MS and TDA AAS methodologies, in addition to being
suitable for verifying CONAMA criteria, were also suitable for monitoring low levels of Hg in marine
sediment samples and comparing them with international sediment quality guides from National
Oceanic and Atmospheric Administration (NOAA) in the USA6 and Canadian Council of Ministers of
the Environment (CCME) in Canada7 that established the limits of TEL (Threshold Effect Level) and
PEL (Probable Effects Level) for marine sediments at 130 and 700 µg∙kg −1, respectively.
1 Beckers F, Rinklebe J, Critical Reviews in Environmental Science and Technology, 47, 2017, 693–794.
2 Zahir F, Rizwi SJ, Haq SK, Khan RH, Environmental Toxicology and Pharmacology, 20, 2005, 351–360.
3 INMETRO, DOQ-CGCRE-008 – Orientação sobre validação de métodos analíticos, 2020.
4 Linsinger TPJ, Institute for Reference Materials and Measurements, 2010.
5 CONAMA, RESOLUÇÃO No 454, DE 01 DE NOVEMBRO DE 2012.
6 Buchman MF, NOAA OR&R Report 08-1, 34, 2008.
8 CCME, Environmental Protection, 2001, 1–3.
9
[CAPES, CNPq, FAPES, NCQP/DQUI/UFES, PPGQUI/UFES]

120
ELEMENTAL DETERMINATION IN FISH SAMPLES FROM AMAZON REGION
BY MIP OES
Lady L. P. P. de Carvalho (UG)a*, Alessandro N. Rodrigues (UG)a, Allan da S. Cruz (PG)a,
Michelle S. Lemos (R)a
a Federal University of Pará, Faculty of Chemistry, Group of Analytical Spectrometry Applied, 66075-110, Belém, Pará,
Brazil.
*e-mail: mslemos@ufpa.br
Fish consumption in the Amazon is quite high, reaching average value of 169 kg/person /year among
riverside populations1, which exceeds the value of 12 kg/person /year recommended by WHO2. Fish
is associated with healthy diet, but it is also a source of metals3. Considering that certain elements
are vital to health, while others in excess pose risk of contamination4, the investigation of inorganic
constituents present in this food is essential for food safety. This study determined the Al, Ca, Cd,
Cr, Fe, K, Mg, Mn, and Pb concentrations in 14 samples of seven fish species (Plagioscion
squamosissimus, Cynoscion acoupa, Macrodon ancylodon, Brachyplatystoma rousseauxii,
Colossoma macropomum, Brachyplastistoma vaillantii, and Arius parkeri), obtained in markets of
Belém (state of Pará, Brazil). Determinations were performed using microwave-induced plasma
optical emission spectrometry (MIP OES). Samples were analysed after digestion in cavity
microwave oven using a dilute oxidizing mixture of nitric acid and hydrogen peroxide4. The procedure
accuracy was evaluated using a certified fish protein reference material (DORM-4) and recoveries
ranged from 80% to 107%. Addition and recovery tests were performed to assess the accuracy of
measurements by MIP OES, where known aliquots of analytes were added to digested samples and
recoveries were acceptable, ranging from 80 (Al) to 115 (Cd). The limits of detection (LD) ranged
from 0.12 mg kg-1 (Cd) to 3.26 mg kg-1 (K). The results indicated the presence of elements essential
to human health such as K, Mg, Ca, and Fe in large amounts in fish samples. On the other hand,
harmful elements were also present in amounts dangerous to health, with high Pb and Cr levels.
(Table 1).
Table 1. Element concentrations in the muscles of the studied fish species.
Concentration
Al Ca Cd Cr Fe K Mg Mn Pb
(mg kg-1)
Mean 8.44 403.77 - 44.47 282.25 12073.75 1190.22 6.33 7.50
Minimum 3.33 91.47 <LD 3.33 98.27 9698.15 963.17 3.33 6.66
Maximum 18.31 1246.87 <LD 221.40 873.94 14524.90 1365.92 16.65 8.33
Standard
4.50 363.93 - 60.17 235.8 1699.3 119.834 5.074 0.84
deviation
ANVISA* 0.05 0.30
* Normative Instruction nº 88 – March 26, 2021.
High Pb (above the reference value of 0.3 mg kg-1)5 and Cr levels are worrying because these metals
represent risks to human health. Current Brazilian legislation does not establish a reference Cr value;
however, the European Food Safety Authority has proposed a tolerable daily intake (TDI) equal to
0.3 mg kg−1 of Cr (III)6.
Thus, this study provides important information on the concentration of these elements in fish and
contributes to the evaluation of food quality and safety for the consumer population.
1 FAO. Statistics division, food security statistics, food consumption, 2016.

2 Isaac VJ, Almeida MC, GIARRIZZO T, DEUS CP, VALE R, KLEIN G, BEGOSSI A, Anais da Academia Brasileira de
Ciências, 87, 2015, 4.
3Barros, BCV, Pereira SFP, Palheta DC, Silva CS, HOLOS Environment, 10, 2011, 2.
4 Lemos, MS, Cruz AS, Fernandes Dantas KG, Biological Trace Element Research, 191, 2019, 1.
5 ANVISA – Agência Nacional de Vigilância Sanitária. Normative Instruction- IN N° 88, March 26, 2021.
6 EFSA panel on dietetic products, nutrition and allergies (NDA), EFSA Journal, 13, 2015, 10.
[UFPA-GEAAP]

121
TOTAL PHOSPHORUS DETERMINATION IN LUBRICATING OIL BY
EVALUATING ATOMIC AND MOLECULAR APPROACHES VIA HR-CS GF
AAS/MAS
Leandro Kolling (PG)a, Rafaela P. B. de Lima Piecas (UG)a, Maria Goreti R. Vale (R)a,b,
Márcia Messias da Silva(R)a,b*
a Universidade Federal do Rio Grande do Sul, Instituto de Química, Porto Alegre, RS, Brazil, 91501-970,
b Universidade Federal da Bahia, Instituto Nacional de Ciência e Tecnologia do CNPq - INCT de Energia e Ambiente,
40170-115, Salvador, BA, Brazil
*e-mail: mmsilva@iq.ufrgs.br
The addition of phosphorus compounds (such as ZDDPs - zinc dialkyldithiophosphates) to

lubricating oils is justified by their anti-wear and antioxidant properties.1 However, the monitoring of
total phosphorous is necessary since, in excess, it may cause negative effects on the exhaust
system of gasoline-fueled vehicles, which could increase the emission of greenhouse gases.2
Although total phosphorus determination by high resolution-continuum source graphite furnace
atomic (or molecular) absorption spectrometry (HR-CS GF AAS/MAS) have already been proposed,
its application is complex, but feasible if favorable chemical conditions are used. 3 Therefore, a
reliable method for total phosphorus determination in lubricating oil samples via HR-CS GF
AAS/MAS was developed and presented in this work. For the phosphorus monitoring, two
approaches were evaluated: (i) via atomic absorption; and (ii) via molecular absorption, in which the
PO diatomic molecule was chosen as the target molecule. The method consisted of weighting the
samples directly into the graphite platform, followed by the adding of the chemical modifier
(magnesium acetate solution), and introducing this platform in the vaporizer (graphite furnace). For
the optimization of the proposed method, parameters such as: pyrolysis and vaporization
temperatures, the working wavelengths, and the nature and amount of chemical modifiers in different
media (aqueous and ethanolic) were investigated. Uni- and multivariate optimizations were carried
out and compared for further understanding of this analyte behavior. For calibration, standard
solutions of two sources of P (H3PO4 and NH4H2PO4) were evaluated. All the obtained curves
presented equivalent sensitivities with R² greater than 0.997 and a linear range between 97 and
1067 mg kg-1 P. This working range encompassed the threshold reported by the American Petroleum
Institute (API) of 800 mg kg-1 P (0.08% in mass).2 The samples analyzed in this work consisted of
lubricating oils purchased from local establishments. The samples were analyzed following the
proposed method and the results compared with those obtained by the reference method ASTM
D1091.4 For five real samples, P concentrations were between 490 and 690 mg kg-1 (or 0.049% and
0.069% as shown in Table 1). These values were in agreement with the results obtained using the
reference method. Besides, the results for both approaches (AAS and MAS) were statistically
equivalents at 95% confidence level (paired t-test).
Table 1. Concentration of P (shown as % in mass) in lubricating oil samples (from OL1 to OL5).
Samples Reference method* via P via PO
OL1 0,067±0,002 0.067±0.004 0.069±0.006
OL2 0,049±0,003 0.048±0.005 0.052±0.003
OL3 0,056±0,005 0.056±0.006 0.054±0.004
OL4 0,068±0,003 0.069±0.006 0.067±0.009
OL5 0,055±0,002 0.053±0.005 0.052±0.004
*ASTM D1091 - Standard Test Methods for Phosphorus in Lubricating Oils and Additives;
Photometric (Molybdivanado) Method.4
1 D. Kim, T. J. Toops, K. Nguyen, D. W. Brookshear, M. J. Lance e J. Qu, Emiss. Control Sci. Technol., 2020, 6, 139.
2 American Petroleum Institute, API 1509 - Engine Oil Licensing and Certification System, Washington DC, 2021.
3 F. G. Lepri, M. B. Dessuy, M. G. R. Vale, D. L. G. Borges, B. Welz e U. Heitmann, Spectrochim. Acta - Part B At.
Spectrosc., 2006, 61, 934–944.
4 ASTM D1091-11 - Standard Test Methods for Phosphorus in Lubricating Oils and Additives, West Conshohocken, PA,
2021.
[CAPES, CNPq]

122
CALIBRATION STRATEGIES FOR RARE-EARTH ELEMENTS DETERMINATION
IN MARINE SEDIMENT BY ICP-MS
Suellen G. Cordeiro (PG)a*, Rafael Mantovaneli (PG)a Jefferson R. Sousa (R)b, Maria Teresa
W. D. Carneiro (R)a, Geisamanda P. Brandão (R)a
a UFES, Departamento de Química, Vitória, Espírito Santo, Brazil, Zip Code
b UENF, Departamento de Química, Campos, Rio de Janeiro, Brazil, Zip Code
*e-mail: suellengcq@hotmail.com
The use of sediments matrix in the aquatic ecosystem evaluation stands out in the scientific
community for adding information capable of revealing contamination process as well as its origin.
Rare earth elements have been widely used in marine environmental assessment research because
they are excellent indicators of geological processes, since they are associated in different
proportions in the minerals that make up marine sediments.1 However, considering the complexity
of marine sediment matrix and the performance of decomposition using multi-acids, the use of
adequate calibration strategies for rare earth elements determination is necessary. Therefore, the
objective of this study was to evaluate different calibration strategies in rare earth elements
determination in marine sediment by inductively coupled plasma mass spectrometry (ICP-MS). The
strategies used were External Calibration (EC), Matrix-Matched Calibration (MMC), Standard
Addition Calibration (SA), and Multi-Isotope Calibration (MICal), all performed under the same
operating conditions.2 Among the rare earth elements, Ce, Nd, Sm, Gd, Dy, Er and Yb were chosen
because they have at least 3 isotopes, thus making possible the comparative study between the
conventional calibration strategies (EC, MMC and SA) and the non conventional (MICal). The
addition of internal standards (IP) Rh and In was also tested. Analytical figures of merit were
established, and the linearity of the analytical curves obtained through the different calibration
strategies (R2) was greater than 0.999 for all elements. The working range was from 0.05 µg∙L −1 to
10 µg∙L−1 for EC, MMC, and for MICal calibration was performed with 2 µg∙L−1. The detection and
quantification limits values obtained were adequate to determine the analytes using the different
calibration strategies. The standard deviation of the slope of the analytical curves was obtained from
the matrix analysis of least squares and later the t-student statistical test was applied to compare the
sensitivities. For most elements, it was found that there is a statistical difference among the
sensitivities of the curves obtained by EC and the curves obtained by MMC and SA, with or without
the use of IP, evidencing that there is interference of the matrix in the determination of these analytes
in marine sediment. To evaluate the accuracy of the methods, the certified reference material (CRM)
of estuarine sediment – BCR 667 was analyzed. The isotopes 140Ce, 142Nd, 152Sm, 160Gd, 162Dy, 166Er
and 174Yb were determined using EC, MMC and MICal, and only using the unconventional MICal
strategy was not found statistically significant difference between the determined and certified
concentrations. Considering that there are matrix influences in Ce, Nd, Sm, Gd, Dy, Er and Yb
determination by ICP-MS, it is recommended the use of SA calibration strategy, which despite being
well consolidated in the literature, has limitations that need to be considered. Therefore, the
unconventional multisotopic calibration strategy (MICal) is a viable alternative for the determination
of rare earth elements, which present at least 3 isotopes, in complex matrices.
1 Gallelo, G; Vázquez, CF; Chenery, S; Lang, C.; Bernett, St; Kabora, T. Hodson, Me, Stump, D. Microchemical J., 2019,
148, 691.
2 Donat GL; Amaisb RS J. Anal. At. Spectrom., 2019, 34, 2353.
[CNPq, Petrobras, NCQP/DQUI, PPGQUI/UFES]

123
EVALUATION OF FS-FAAS METHODOLOGY IN THE DETERMINATION OF
RARE EARTH ELEMENTS IN (Nd,Pr)-Fe-B ALLOYS
Gilmar A de Almeida (R)a,b*, José O.V. Bustilhos (R)a, Maciel S. Luz (R)b
a Nuclear and Energy Research Institute- IPEN, Sao Paulo, SP, Brazil, 05508-000.
b Advanced Materials, Institute for Technological Research, Sao Paulo, SP, Brazil, 05508-901.
*e-mail: gilmarm@ipt.br
The Committee on Inorganic Chemistry Nomenclature of IUPAC (International Union of Pure and
Applied Chemistry) recommends the use of the expression “rare earth elements” (REE) for the series
of lanthanides added to the scandium and yttrium, which begins in Ce and ends in Lu, having atomic
numbers from 58 to 71.1 The exploration of the magnetic, optical and redox properties of REE allows
several applications in the area of advanced materials such as automotive catalysis, petroleum
cracking, glass production, alloys nickel-metal hydride batteries (NiMH), high strength ceramics, and
permanent magnets. In this scenario, the production of permanent magnets has been growing
annually.2,3 Since permanent magnets are made up mostly of Nd-Fe-B combined to different levels
of Pr, Dy, Tb, and Co, several analytical techniques have been used to determine REE, among which
the inductively coupled plasma mass spectrometry (ICP MS) stands out. However, ICP-MS may not
achieve high analytical accuracies for high levels of Nd (up to 30% w w-1) and Pr (up to 14% w w-1)
in permanent magnets.4,5 Thus, this work proposes a fast and inexpensive way to evaluate Nd, Pr
and Dy in (Nd,Pr)-Dy-Fe-B metal alloys through Fast Sequencial Flame Atomic Absorption
Spectrometry (FS- FAAS) method. The fast sequential determination of Nd (421.2 nm), Pr (495 nm)
and Dy (421.2 nm) in different metal alloys were performed by sample dissolution in an acidic
medium in the proportion of 15mL of nitric acid 25% v v-1 and 5 mL hydrochloric acid 27% v v-1 for
up to 0.5 g of sample, followed by instrumental measurements using an Atomic Absorption
Spectrometer. Potassium chloride was evaluated as an ionic suppressor. Linear calibration curves
were obtained with coefficient correlation ranging from 0.9967 to 0.9999. The accuracy of the method
for Dy was assessed by analyzing Dy-spiked (10 to 40 mg kg-1) samples. Then, recoveries
percentages ranged from 96 to 104% showing the proposed method accuracy for Dy determination.
Additionally, Pr and Nd determination by FS-FAAS were evaluated taking into account reference
values derived from analytical methods applied by a network of laboratories. The proposed method
proved to be promising and highly accurate to determine Nd in (Nd,Pr)-Fe-B alloys containing 17.38
± 0.30 and 24.30 ± 1.3% w w-1 (Nd) applying the Student’s t-test at 95% confidence limit.
1 Heslop, R. B.; Jones, H. Química inorgânica. 2. ed. Lisboa: Fundação Calouste Gulbenkian, 1976. 944 p
2 De Sousa F. P. C.; Serra, O. A. Quim. Nova, v. 37, n. 4, p. 753-760, 2014.
3 Serra, O.A; de Lima, J. F.; de Sousa F.P. C. Revista Virtual de Química, v. 7, n. 1, p. 242-266, 2014
4 Papai, R., da Fonseca, K. T., de Almeida, G. A., Luiz, A., da Silva, N., Nagasima¹, T. P., Luz, M. S.
5 Papai, R., de Freitas, M. A. S., da Fonseca, K. T., de Almeida, G. A., da Silveira, J. R. F., da Silva, A. L. N., ... & Luz, M.
S. (2019). Analytica Chimica Acta, 1085, 21-28.
[IPEN, IPT]

124
CURRENT STATUS, NEEDS AND FUTURE PERSPECTIVES FOR
PHOTOCHEMICAL VAPOR GENERATION
Ralph E. Sturgeon (R)
National Research Council Canada, Metrology, Ottawa, Ontario, Canada K1A 0R9
e-mail: Ralph.sturgeon@nrc-cnrc.gc.ca
Trace element analysis relies on the generation, manipulation and detection of free atoms / ions of
sample constituents in the gas phase to enable their quantitation by absorption / emission /
fluorescence and mass spectrometries. Selective introduction of gaseous atomic or molecular
analyte species into such sources provides potential advantages, including enhanced detection
power (1- to 2-orders of magnitude), reduction of matrix and spectral interferences, as well as
speciation capabilities1. Vapor generation (VG) has been utilized for more than 50 years and
occupies important niche roles while expanding in elemental scope, application and methodology2.
Intriguingly, our natural world is sustained by photosynthesis; synthetic organic chemists are more
frequently turning to the specificity achieved using photo-induced catalytic processes; the
biogeochemical cycles of many elements are based on photolytic reactions. It is not surprising to
realize that analytical techniques (and many environmental cycles) may also stem from parallel
photo-driven processes. For the past two decades, our laboratory has had an interest in utilizing
photochemistry to compliment other VG practices. Photochemical vapor generation (PVG) provides
much broader elemental coverage than traditional VG techniques, with currently more than 20
elements (As, Sb, Bi, Se, Sn, Tl, Te, Fe, Co, Cu, Ni, Hg, Pb, Cd, Mo, W, Os, I, Br, Cl, Fl, Ag, Pt, Ge,
Re, Rh and Ir) showing promise. PVG is typically achieved by UV-C photolysis of an aqueous
medium of (principally) carboxylic acids to drive homogeneous redox chemistry facilitated by e –(aq),
H•, H3C• and CO2•– radicals3. Reduction of aquo-metal and oxyanion species to their elemental state
is followed by reaction with abundant H•, H3C• and CO to yield predictable volatile alkylated,
carbonylated and hydride analyte species. Most recently, homogeneous catalytic systems have
proved to be indispensable for PVG of many elements. Addition of trace concentrations of selected
transition metals to the reaction medium suggest alternative processes contribute to photo-VG
yields. The possible mechanisms of PVG will be examined, as well as the analytical performance
characteristics of the technique highlighted in this presentation.
1 Bings NH, Orlandini von Niessen JO, Schaper JN, Spectrochim. Acta, Part B, 100, 2014, 14
2 D'Ulivo A, Mester Z, Sturgeon RE, Wang Q, Welz B, Pure Appl. Chem., 83, 2011, 1283
3 Leonori D, Sturgeon RE, J. Anal. At. Spectrom., 34, 2019, 636.

125
EVALUATION OF A LOW COST FLAME PHOTOMETER FOR SODIUM
DETERMINATION
Francisco C. Rosa (R)a*, Matheus F. Pedrotti (R)a, Lucas G. O. Aguiar (UG)a
a Instituto Federal de Educação, Ciência e Tecnologia do Rio Grande do Sul, Feliz, RS, Brazil, 95770-000
*e-mail: francisco.rosa@feliz.ifrs.edu.br
According to the curriculum guidelines of chemistry courses in Brazil, students must be qualified to
conduct physical-chemical analysis by classical and instrumental methods, properly knowing the
fundamentals and limitations of the techniques1. Moreover, the chemistry courses must offer in their
curricular matrices the discipline of instrumental analytical chemistry. To offer this discipline at an
experimental level, it is necessary that students be able to attend practical classes with
electroanalytical, spectroanalytical and chromatography instruments. As the cost of purchase and
maintenance of these equipment is high, most part of Brazilian education institutes do not have
laboratories with these sophisticated equipment. For this reason, the development of low cost and
simple equipment is mandatory to allow students to perform experiments related to instrumental
analytical chemistry. In this work, the development of simple method for sodium determination in
commercial aqueous samples by using a homemade flame photometer is presented. The equipment
was built from low cost and reused materials, associated with accessible technologies such as
smartphone applications. A cross-flow nebulizer was developed to introduce aqueous sample into a
3D printed cyclonic nebulization chamber coupled to a metallic burner. In order to enable the use of
system for teaching, the analytical signals were obtained by taking pictures of the flame with the
camera of a smartphone (Mi A3, Xiaomi, China) and processing with the PhotometrixPRO® free
application. The signals of intensity of the red (R), green (G) and blue (B) components were
measured at specific position of the flame. Sodium was determined in a commercial saline solution
and soft drink samples, both with informed sodium concentration. From this, an analytical calibration
curve was built with standard sodium solutions from 0.0 to 15.0 mg L-1, prepared by serial dilution of
a Na stock solution of 1000 mg L-1. The flow of gas (propane/butane) and air, as well as the
conditions in the PhotometrixPRO® application (flame capture area, focus mode, white-balance and
resolution), were optimized to obtain the best operational conditions. A linear range was observed
from 0.0 to 5.0 mg L-1 (R2 = 0.9991). The sodium concentration obtained for the commercial saline
solution was 3.76 ± 0.10 g L-1, while for the soft drink 47.6 ± 1.9 mg L-1 and were not statistically
different from the informed values (t-test, 95% confidence level). Despite of the simplicity and low
cost of the proposed method, the obtained results showed that a relatively good agreement was
obtained. Finally, the possibility of teaching the fundamentals of the flame photometric technique
was demonstrated with the developed method and equipment. Through the development of low cost
methods, many instrumental analytical experiments of emission spectrometry can be performed in
laboratories with lower income.
1 BRASIL, 2001 MEC. Parecer CNE/CES nº 1.303/2001: Disponível em:
<http://portal.mec.gov.br/cne/arquivos/pdf/CES1303.pdf>.
[IFRS]

126
MASS SPECTROMETRY
127
MALDI-TOF IMAGING: AN ANALYTICAL APPROACH FOR EXPLORING
BACTERIAL RESISTANCE
Milena C. V. Fernandesa, André L. M. Camelo*a, Pedro H. Vendraminia, Marcelo Brocchib, Ana
V. C. Simionatoa
a
State University of Campinas, Department of Analytical Chemistry/Institute of Chemistry, Campinas, São Paulo, Brazil,
13083-970
b State University of Campinas, Department of Genetics, Evolution, Microbiology and Immunology/ Institute of Biology,
Campinas, São Paulo, Brazil, 13083-862

*e-mail: a266017@dac.unicamp.br
The inappropriate and excessive use of antibiotics for the treatment of bacterial infections has led to
the increasing presence of resistant and multidrug-resistant bacteria both in hospital settings and in
the community. Thus, understanding the metabolism of resistant bacteria is essential to combat them
more efficiently. In this context, Staphylococcus aureus bacterium, and its different lineages, have a
long history in human pathology, causing endocarditis, pneumonia and septicemia. The first reports
of Methicillin-resistant Staphylococcus aureus (MRSA) date back to 1961, but the first epidemic of
MRSA in hospitals occurred in the 1980s, and in the late 1990s, when it spread over communities.1
Currently, the impact of this bacterium on public health and the economy is evident, causing an
increase in mortality, use of health resources with high-cost drugs and medical monitoring for their
administration.2 In this scenario, mass spectrometry imaging (MSI) is considered a promising
technique for understanding the resistant characteristics of such bacteria and how they can
potentially be treated. This process consists on the identification of different ions on the surface of
the colonies and the identification of potential metabolites that characterize antibiotic resistance,
upon comparison with wild type bacteria of the same lineage. This work presents matrix-assisted
laser desorption ionization-mass spectrometry imaging (MALDI-MSI) study of colonies of Methicillin-
resistant Staphylococcus aureus, as a proof of concept of the technique for obtaining images of
bacteria colonies. Analysis by MALDI-MSI requires that a matrix be uniformly deposited on the
sample surface. This process consists in the formation of matrix crystals on the sample by
sublimation.3 Afterwards, sample is inserted on the mass spectrometer ion source, and the laser hits
the sample incorporated into the matrix crystals, which are photoionized producing matrix ions. In
this phase, the charge is transferred from the ionized matrix to the neutral molecules of the analyte,
followed by introduction of analytes and matrix ions into the mass analyzer. The matrix is
fundamental in this process and must be chosen according to the analytes chemical functions and
the laser wavelength, so that optimum energy absorption occurs.4 In this work, images of methicillin-
resistant and wild colonies of Staphylococcus aureus were obtained by a sublimation process to
apply the MALDI matrix on the samples followed by MALDI-TOF analysis. Within the significant ions
that differentiate the colonies,16 of them were putatively identified with the aid of scientific literature
and databases, such as N,N-Dihydroxy-L-valine, 2-(4-Methylphenyl)ethylamine, 3,4-Dihydroxy-L-
phenylalanine, 2-Methyl-hexanoic acid, and Glycyl-D-asparagine. Thus, this study reinforces the
potential of MALDI-MSI for identification of metabolites synthesized by different strains of
Staphylococcus aureus bacteria.
1 Chambers, H. F.; DeLeo, F. R. Nature Reviews Microbiology 7, 2009, 629.
2 Shankar, P. R.; Balasubramanium, R. Australasian Medical Journal 7, 2014, 237.
3 Hankin, J. A.; Barkley, R. M.; Murphy, R. C. Journal of the American Society for Mass Spectrometry, 2007, 18, 1646.
4 Awad, H.; Khamis, M. M.; El-Aneed, A. Applied Spectroscopy Reviews, 2015, 50, 158.
[FAPESP]
––––––––––––––––––––– Mass spectrometry –––––––––––––––––––––

128
DEVELOPMENT OF ANALYTICAL DEVICE FOR NEWBORN SCREENING USING
DRIED MATRIX SPOT AND PAPER SPRAY MASS SPECTROMETRY
Eduardo L. Rossini (R)*, Emanuel Carrilho (R)
Instituto de Química de São Carlos, Universidade de São Paulo, São Carlos, SP, Brazil, 13566-590
Instituto Nacional de Ciência e Tecnologia de Bioanalítica-INCTBio, Campinas, SP, Brazil, 13083-970
*e-mail: elrossini@usp.br
Newborn screening is considered one of the most significant advances in preventive medicine and
one of the largest public health campaigns. The early diagnostic of diseases, even for the rare ones,
can reduce or eliminate irreversible sequels that could occur if they were not detected prematurely.¹
A group of diseases analyzed in newborn screening is the aminoacidopathies, which are metabolic
disorders caused by specific deficiency in determined enzymes or transporters, such as
phenylketonuria, leucinosis (maple syrup urine disease), and tyrosinemia type II.² Dried plasma spot
(DPS) is a simple and easy-to-use sampling method that reduces the risk of contamination for the
operator, eliminates the hematocrit interference, and facilitates transportation and storage.³
However, chromatographic techniques with mass spectrometry detection are frequently employed,
but they need several steps, are time-consuming, and use large amounts of solvents. In this way,
we proposed to use paper spray (PS) mass spectrometry as a tool to integrate the advantages of
paper platform (3D-μPAD), such as low cost, easy availability, flexibility, hydrophilicity, light weight,
biocompatibility, and power-free fluid transport with the reliability of mass spectrometric.
A B Figure 1. A) Illustration of the
layers used to construct the
proposed 3D-µPAD device to
collect blood, separate the
red cells, and form dried
plasma spot to analyze via
PS-MS). B) Analysis of amino
acids via PS-MS using the
detection layer.
The proposed device to collect blood and form the DPS was constructed using lamination on the top
and bottom layers to hold all layers together, a plasma separation membrane layer (Vivid GR, Pall)
to separate the red cells and promote the formation of the DPS in the detection layers (Fig. 1A). After
the sampling, the detection layer was removed, folded, and positioned in front of the mass
spectrometry inlet using an alligator clip. A 15 µL aliquot of MeOH:H2O:Formic Acid 80:19.9:0.1 was
added to the DPS to extract the analyte and generate the spray to promote the determination via
PS-MS. Transitions of valine (118 → 72), leucine (132 → 86), isoleucine (132 → 86), phenylalanine
(166 → 120) and tyrosine (182 → 136) were examined via the loss of 46 Da (CO + H2O) using a
multiple reaction monitoring in a TSQ Quantum Access mass spectrometer (Thermo Fisher/ triple
quadrupole).
The 3D-µPAD reproducibility and homogeneity to separate the red cells to form DPS was tested
using fingertip blood (~20 μL) and analyzed by an off-line spectrophotometric ninhydrin assay. No
statistical difference (p<0,05) between the DPS formed in the device's bottom and upper detection
layer demonstrates a homogenous distribution by the plasma (CV<10%). Samples were investigated
using the 3D-µPAD described above, and it was possible to identify the presence of valine,
leucine/isoleucine, phenylalanine, and tyrosine with intensities at least twenty times higher than the
blank. The results showed that the proposed device can be used for blood microsampling to form
plasma spots. The 3D-µPAD was successfully integrated with mass spectrometry analysis using
paper spray ionization to determine amino acids with no sample preparation or derivatization.
1 Howse JL, Katz M, Pediatrics, 106, 2000, 595
2 Wasim M, Awan FR, Khan HN, Tawab A, Iqbal M, Ayesha H, Biochemical Genetics, 56, 2018, 7.
3 Verplaetse R, Henion J, Analytical Chemistry, 88, 2016, 6789.
[FAPESP, INCTBio]

129
HUMAN PRIMARY ASTROCYTES FINGER- AND FOOTPRINTING
METABOLOMICS INDICATE BIOCHEMICAL ALTERATIONS UNDER
AYAHUASCA TREATMENT
Flávia S. Zandonadia*, Álex Rosinib, Emerson A. F. Santosa, Raffaela Ignarroc, Aline A. S.
Meloa, Alexandre L. R. Oliveirac, Andréia M. Porcarib, Alessandra Sussulini (R)a,d,*
a Laboratory of Bioanalytics and Integrated Omics (LaBIOmics), Institute of Chemistry, University of Campinas
(UNICAMP), Campinas, SP, Brazil, CEP 13083-970.
b MS4Life Laboratory of Mass Spectrometry, Health Sciences Postgraduate Program, Sao Francisco University (USF),
Braganca Paulista, SP, Brazil, CEP 12916-900.

c Laboratory of Nerve Regeneration, Institute of Biology, University of Campinas (UNICAMP), Campinas, SP, Brazil, CEP
13083-970
d National Institute of Science and Technology for Bioanalytics – INCTBio, Institute of Chemistry, University of Campinas
(UNICAMP), Campinas, SP, Brazil.

*e-mails: flazando@unicamp.br and sussulini@unicamp.br
Ayahuasca (Aya) is a psychotropic Amazonian beverage formulated from the combination of the
Banisteriopsis caapi vine and the Psychotria viridis leaves in a water decoction. Aya is legally used
in Latin American countries and used in Brazil for religious, cultural, and therapeutic purposes. Its
properties constitute a bio-psycho-social-spiritual model involving effects from β-carboline-derived
alkaloids, present in the vine, and N,N-dimethyltryptamine (DMT), a tryptamine-derived alkaloid
present in the leaves, which act together in the central nervous system (CNS). Few technical-
scientific studies have been conducted to understand the effects of this brew in the metabolism.
Therefore, this work aims to investigate an in vitro primary astrocyte lineage model by untargeted
metabolomics evaluations of two cellular subfractions: secretome and intracellular content after Aya
treatment, where DMT and other β-carbolines were previously quantified. Metabolomics analysis
was performed by UHPLC-MS/MS, followed by MS-Dial data processing and statistical analysis to
identify metabolites and biochemical alterations related to Aya treatment. Aya doses were applied to
the cell cultures considering DMT concentrations of 1, 10 and 20 µM, which are in agreement with
non-toxic and toxic DMT threshold assays in primary human astrocyte cells viability. Detected
metabolite features from intra and extracellular fractions were processed. Features were obtained
from positive (+) and negative (-) electrospray ionization modes; for RPLC: 610 (+) and 1,699 (-), for
HILIC: 2,444 (+) and 5,231 (-) were annotated. For RPLC, 9 (+) and 7 (-), and for HILIC, 21 (+) and
24 (-) features were identified by their accurate mass and MS2 spectra. Reaction courses attributed
to the metabolites identified (MS2-validated metabolites) with ANOVA (p<0.05) were obtained at
MetaboAnalyst and, for cellular subfractions data, the downstream assigned to putative sub-
pathways, as purine, (hypo) taurine, tyrosine-tryptophan, and metabolism cofactors, according to 10
and 20 µM DMT Aya treatments. These pathways are described as important in astrocyte activity:
repair1, mood control2, astrocytic-neural metabolic cooperation3, neuroprotection4 and
neutrotransmitter5 synthesis in the CNS. Further biochemical-molecular assays should indicate
alterations at the molecular level related to biochemical pathways associated to major depressive
disorder.
1 Ciccarelli, R., Ballerini, P., Sabatino, G., Rathbone, M. P., D'Onofrio, M., Caciagli, F., Di Iorio, P. International Journal of
Developmental Neuroscience, 19, 2001, 395.
2 Halassa, M. M., Fellin, T., Haydon, P. G. Neuropharmacology, 57, 2009, 343.
3 Vitvitsky, V., Garg, S. K., Banerjee, R. Journal of Biological Chemistry, 286, 2011, 32002.
4 Myint, A. M., Kim, Y. K., Verkerk, R., Scharpé, S., Steinbusch, H., Leonard, B. Journal of Affective Disorders, 98, 2007,
143.
5 Rose, J., Brian, C., Pappa, A., Panayiotidis, M. I., Franco, R. Frontiers in Neuroscience, 14, 2020, 536682.
[CAPES, CNPq, FAPESP, INCTBio]

130
ASSESSING CHEMICAL COMPOSITION OF HEAVY CRUDE OIL AND ITS
DISTILLATION CUTS BY ESI (-) FT-ICR MS
João Victor Ataíde Oliveira (UG)a*, Deborah Victória Alves de Aguiar (PG)a, Gesiane da Silva
Lima (PG)a, Boniek Gontijo Vaz (R)a
a Federal University of Goiás, Chemistry Institute, Goiânia, Goiás, Brazil, 74690-900
*e-mail: oliveiraataide@discente.ufg.br
Heavy crude oils are characterized by low API gravity (°API) and present high content of
heteroatomic compounds in their composition. The heteroatom-containing species are responsible
for causing several problems in refining operations and in the environment, consequently treatments
are essential for remotion of them. In this work, a heavy crude oil sample and its distillation cuts
QAV, diesel, gas oil and vacuum residue (VR) were analyzed by ESI (-) FT-ICR MS.1
For the FT-ICR MS analyses, samples were prepared by dissolving 2 mg of the crude oil in 2 mL of
toluene, leading to a 1 mg.mL-1 solution. Afterwards, 500 µL of the solution was diluted with 500 µL
of methanol containing 2.5% of NH4OH (25 µL of NH4OH in 1000 µL of methanol), for a final
concentration of 500 µg.mL-1 in toluene:methanol (1:1, v:v) mixture. The spectral data were
recalibrated using DataAnalysis software, followed by compositional assignment using Composer
software. The Thanus software was used for graphical constructions. Negative electrospray analysis
allowed the identification of acidic species. Figure 1 shows the relative abundance of all class
identified in our samples. As shown in Figure 1, crude oil, diesel and VR are predominantly
characterized by nitrogen compounds, while the samples of QAV and diesel have mostly oxygenated
compounds. Figure 2 shows that compounds with DBE 9, 12 and 15 from the N class were identified
with greater relative abundance in crude oil, diesel and RV samples. These compounds are widely
known as carbazoles, benzocarbazoles and dibenzocarbazoles, respectively. The diesel sample
presented DBE ranging from 1 to 10, for class O2, with a carbon number range from C12 to C23.
These compounds are related to naphthenic acids, which are derived from cyclohexanoic and
cyclopentanoic acids.1 The QAV sample presented DBE ranging from 1 to 4 for class O2, and a
range of carbon numbers from C6 to C20. According to the literature these compounds are
associated with carboxylic acids – class O2 (Figure 2).2,3
Figure 2. DBE versus carbon number distribution for

majority class for the crude oil sample and its distillation
cuts.
Figure 1. Relative abundances of heteroatomic classes
identified by FT-ICR MS.
These results demonstrated that Negative-ion ESI FT-ICR MS analysis allowed us to detail the
composition of polar components of a heavy crude oil sample and its cuts.
1 Arenas-Diaz J, Lozano PD, Ramirez X, Cabanzo R, Guzman A, Mejia-Ospino E, Chemical Engineering Transactions,
57, 2017, 1603
2 Dalmaschio GP, Malacarne MM, Almeida VMDL, Fuel, 115, 2014, 190
3 Ávila BMF, Vaz BG, Pereira R, Gomes AO, Pereira RCL, Corilo YE, Simas RC, Nascimento HDL., Eberlin MN, Azevedo
DA, Energy & Fuels,26,2012, 5069
[Petrobras, CNPq, CAPES]

131
USE OF CARMINIC ACID IMMOBILIZED IN AGAROSE GEL AS A BINDING
PHASE FOR DGT: A NEW APPROACH FOR DETERMINATIONS OF RARE
EARTH ELEMENTS.
Luiz F. P. P. Moreira (PG)*ª, Edson G. de O. Junior (UG)ª, Melina B. T. Zantta (R)ª, Amauri A.
Menegário (R)ª
*e-mail: luiztm94@gmail.com.br
A very important topic within the environmental sciences are emerging contaminants, compounds
that arise from human activity within several areas such as agriculture, industry, medicine, among
others and that end up flowing into the environment either into the soil or aquatic environments.
Among this class of contaminants, technologically critical elements have gained great interest, due
to their importance in several areas, from the high technology industry to medicine. Technologically
critical elements include tellurium (Te), germanium (Ge), gallium (Ga), indium (In), niobium (Nb),
tantalum (Ta), platinum group elements and also rare earths. (La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy,
Ho, Er, Yb, Lu). This work proposed a new method for determination of rare earth elements using
the inductively coupled plasma mass spectrometry technique (ICP-MS) and the diffusive gradients
in thin films (DGT) technique (for application in aquatic environments). New binding gels were tested
for the DGT device using carminic acid as binding agent. It was concluded that the acid dispersion
directly in agarose gel presented the best performance offering a simpler, faster, and greener method
for obtaining a binding phase for the DGT technique. Deployment curve (immersion test in
laboratory) shown that 14 of the 15 determined elements (with the exception of thorium) had linearity
in their retention by the developed binding agent (retention x time), confirming the main premise of
the DGT technique (obeying Fick's diffusion law1). For the first time, the diffusion coefficients (cm² s-
1
) were obtained in agarose gels (diffusion medium) and carminic acid as binding phase: for La, Ce,
Pr, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm, Yb, Lu, U of 3.94x10-6, 3.87x10-6, 3.90x10-6, 3.79x10-6, 3.71x10-
6
, 4.13x10-6, 3.75x10-6, 3 94x10-6, 3.45x10-6, 3.97x10-6, 3.25x10-6, 4.06x10-6, 3.50x10-6, 1.32x10-6
cm² s-1, respectively. Studies on the influence of pH and ionic strength on the retention of rare earths
by using the proposed approach (diffusive gel: agarose; binding gel: carminic acid immobilized in
agarose were carryout. In the tests, performed the DGT devices in solutions with different pH, 3.5,
5.0, 6.5 and 8, and ionic strength, 0,005 mol L-1, 0,01 mol.L-1, 0,05 mol.L-1 e 0,1 mol.L-1 in NaNO3
were evaluated. The results of theses studies showed the average variation in the retention of about
20% for almost all elements in the pH tests. For the ionic strength the average variation was about
30% for all elements, however the results shown that in solution with low values of ionic strength
there is a higher variation to be considered. These results indicate the possibility of a wide range of
the proposed approach for in situ developed method. In laboratory deployment, performed using a
spiked water sample, shown that the proposed approach presents satisfactory accuracy to be used
for in situ deployments.
1 Davison W, Zhang H, Environ. Chem. 9, 2012, 1.

132
GLOBAL METABOLOMICS FOR EVALUATION OF MICROCEPHALY AND
OCULAR LESION IN NEWBORNS WITH CONGENITAL ZIKA VIRUS INFECTION
Hans R. Zamora-Obandoª*, André L. M. Cameloª, Alexandre M. Limab, Regina V. Oliveirac,
Ana V. C. Simionatoª,c
a State
University of Campinas, Chemistry Institute, Campinas, SP, Brazil, 1083-970
bOswaldo Cruz Institute, Rio de Janeiro, RJ, Brazil, 21040-900
cDepartment of Chemistry, Federal University of São Carlos, São Carlos, SP, Brazil, 3351-8111
dNational Institute of Science and Technology in Bioanalytics (INCTBio), Brazil
*e-mail: h230022@dac.unicamp.br
Zika Virus (ZIKV) is an arbovirus transmitted by Aedes aegipty mosquito. Since its discovery in
Uganda in 1947, it spreaded to Pacific Asian Islands: Micronesia in 2007, and French Polynesia in
2013-2014, where more than 50% and 75% of the population was infected, respectively.1 In 2015 a
ZIKV outbreak was set in Brazil northeastern region, leading to correlation between ZIKV infections
and increased cases of microcephaly. Microcephaly is a clinical finding characterized by reduction
of the cephalic perimeter circumference in newborns compared to age-sex matched control group
because of neurogenesis failure or disruption of the normal central nervous system development.2
Furthermore, a plethora of physiological alterations are related to intrauterine ZIKV infections such
as neurological impairment, and ophthalmological findings, characterizing the congenital Zika
syndrome (CZS).3,4 Henceforth, prenatal exposure to ZIKV is considered a threat to fetus healthy
development and a disease of public importance as declared by the World Health Organization
(WHO) in 2016. Arboviruses-related pathologies (e.g., Zika, Dengue and Chikungunya) have been
studied by different omics approaches (genomics, transcriptomics, proteomics, and metabolomics),
resulting in complementary understanding of the pathological mechanisms.5 Among those research
fields, metabolomics, the comprehensive and systematic study of metabolome, allows the
understanding of altered phenotypes. Scarce metabolomics investigations of ZIKV have been
published, and few metabolomics studies covered ZIKV-microcephaly-ocular lesion relationship in
humans.6 Here, we present an untargeted metabolomic pilot study of microcephaly and ocular lesion
in newborns of Zika-infected mothers. Blood serum samples from three groups were evaluated:
positive for both conditions, microcephaly and ocular lesion, M(+)L(+), n = 6; positive for
microcephaly and negative for ocular lesion, M(+)L(-), n = 6; and control group, n = 5. Lyophilized
samples were reconstituted in ultrapure water, protein precipitation was carried out with methanol,
supernatant was dried in vacuum concentrator, and the samples were reconstituted in
methanol/water solvent. RPLC and HILIC analysis were performed in an Agilent UHPLC system
coupled with Bruker Impact HD QTOF mass spectrometry system with ESI(+) and ESI(-) ionization
modes. MS-DIAL was used for data processing, and MetaboAnalyst 5.0 for unsupervised (PCA) and
supervised (PLS-DA) multivariate analysis. VIP score and SAM analysis were used for selecting
statistically significant molecular features. Samples tend to cluster separately in all analyses with
reasonable R² and Q² cross validation values, due to limited number of samples for each group:
RPLC-ESI(+) (0.92, 0.31); RPLC-ESI(-) (0.98, 0.73); HILIC-ESI(+) (0.74, 0.40); HILIC-ESI(-) (0.99,
0.65), respectively. A total of 74 and 76 discriminant molecular features were determined for RPLC,
ESI(+) and ESI(-) respectively, with VIP scores > 1. Also, 18 and 83 molecular features were
identified for HILIC, ESI(+) and ESI(-), respectively. SAM analysis detected 14, 1, 22 and 10
molecular features with FDR < 0.05 for RPLC and HILIC, under positive and negatives ionization
modes. Metabolites annotation and biological interpretation steps are underway. Thus, this pilot
study shows the feasibility of conducting a larger metabolomics study with promising results that will
allow a better understanding of the object of this research.
1 Masmejan S, Musso D, Vouga M, Pomar L, Dashraath P, Stojanov M, Panchaud A, Baud D, Pathogens, 9, 2020, 898.
2 Woods CG, Parker A, Archives of Disease in Childhood, 98, 2013, 707.
3 Russo FB, Jungmann P, Beltrão-Braga, PCB, Cell Microbiology, 19, 2017, 1.
4 Marquezan MC, Ventura CV, Sheffield JS, Golden WC, Omiadze R, Belfort R, May W, Survey of Ophthalmology, 63,
2018, 166.
5 Peinado RdosS, Eberle RJ, Pacca CC, Arni RK, Coronado MA, Virus Research, 307, 2022, 198610.
6 Nunes EdaC, Canuto GAB, Electrophoresis, 41, 2020, 2102.
[FAPESP, CNPq]

133
INVESTIGATION OF MDGC-MS AND HEADSPACE FOR IDENTIFICATION OF
OXYGENATED COMPOUNDS IN BIO-OIL
Ana Luiza G. Mendes (PG)a, Lavínia C. Rodrigues (UG)a, Lisiane O. Diehl (PG)b, Fábio A.
Duarte (R)a, Paola de A. Mello (R)a*
a Federal University of Santa Maria, Department of Chemistry, Santa Maria, RS, Brazil, 97105-900
b Federal University of Santa Maria, Chemical Engineering Department, Santa Maria, RS, Brazil, 97105-900
Forecasting and environmental problems related to the depletion of fossil-derived fuels indicate the
need for a transition to renewable resources.1 The utilization of biomass and waste materials, such
as agricultural and forest residues and organic waste, as renewable energy sources, is a particularly
attractive alternative and a big challenge for the future.1 Biofuels are liquids or gases produced from
biomass that can be burned for obtaining energy. The liquid oil produced through thermal pyrolysis
of biomass (thermal bio-oil) is almost black up to dark red-brown, that approximates biomass in
elemental composition and thus may exhibit varying properties, depending on the stock material.
Bio-oil is a complex mixture of water and several hundreds of mainly oxygenated, polar organic
compounds that exhibit a wide range of chemical functions: carbonyl-compounds (ketones,
aldehydes, and furanoics), sugars, carboxylic acids, and aromatics (hydrocarbons, PAHs, and
phenolics).2 For this purpose, multidimensional gas chromatography mass spectrometry (MDGC-
MS) was investigated for the identification of oxygenated compounds in bio-oil from lignocellulosic
biomass. Considering that headspace (HS) extraction has great applicability for bio-oils because
there is a large number of volatile compounds in their composition, the present work aims to
investigate the use of HS-MDGC-MS for the identification of oxygenated species in bio-oil. The
identification of oxygenated compounds (acetol, 2-cyclopent-1-one, furfural, 5-methylfurfural, 2-(5H)-
furanone, phenol, guaiacol, o-cresol, m-cresol, p-cresol, eugenol, syringol, vanillin, hydroquinone
and levoglucosan) was performed using a gas chromatograph (model MDGC/GCMS-2010,
Shimadzu, Japan) equipped with a flame ionization detector and a triple quadrupole mass
spectrometer (model GCMS-TQ8040, Shimadzu, Japan). Initially, the composition of the
chromatographic column was investigated using SH-Rtx-624 (30 m x 0.32 mm x 1.8 μm), Rxi-5Sil
MS (30 m x 0.32 mm x 0.25 μm) and Rtx-1701 (60 m x 0.25 mm x 0.25 μm). The heating program
was initially that suggested by the NREL/TP-5100-658893 standard and was optimized for a better
performance for the investigated compounds, consisting of the following steps: i) at 45 °C for 10 min;
ii) 100 °C for 2 min with a heating rate of 3 °C min-1; and iii) 260 °C for 5 min with heating at 6 °C
min-1. Based on the best performance, the MDGC-MS system was set up with the Rtx-1701 column
in the first dimension (FID) and the Rxi-5Sil MS in the second dimension (MS). In this way, the use
of HS was evaluated. The conditions applied for HS were initially 5 minutes of incubation, stirrer
temperature of 95 °C, and a speed of 750 rpm based on previous works in the literature for the
investigated analytes but in other matrices. Under these conditions, it was possible to identify some
oxygenated compounds in bio-oil and further evaluation was developed using an experimental
design (DCCR 23) investigating the incubation time (3 to 37 min), incubation temperature (50 to 130
°C), and sample volume (5 to 15 mL). Preliminary results showed HS can be promising for the
determination of oxygenated compounds in bio-oils. Also, MDGC-MS proved to be an excellent tool
for the identification and separation of compounds in bio-oil, which is an extremely complex matrix.
1 Michailof CM, Kalogiannis KG, Sfetsas T, Patiaka DT, Lappas A.A, Energy Environmental, 5, 2016, 614.
2 McKendry P, Bioresource Technology, 83, 2002, 37.
3 Christensen E, Ferrell J, Olarte MV, Padmaperuma AB, Laboratory Analytical Procedure (LAP), National Renewable
Energy Laboratory (NREL), 2016.
[CAPES, CNPq, FAPERGS, BioValue/BEECOL, UFSM]

134
USING VOLATILE PROFILE TO DIFFERENTIATE PINTO BEANS (Phaseolus
vulgaris L.) FROM DISTINCT AGRICULTURAL SYSTEMS
Luan F. C. Oliveiraa,b*, Taynara S. Matosª, Thales F. D. Pereiraª,
Alexandra C. H. F. Sawayac, Alessandra Sussulini (R)a,d
a Laboratory of Bioanalytics and Integrated Omics (LaBIOmics), Institute of Chemistry, University of Campinas
(UNICAMP), Campinas, SP, Brazil, CEP 13083-970
b Yoko Civilization Research Centre, São Paulo, SP, Brazil, CEP 05508-900
c Faculty of Pharmaceutical Sciences, University of Campinas (UNICAMP), Campinas, SP, Brazil, CEP 13083-862
d National Institute of Bioanalytical Science and Technology (INCTBio), Institute of Chemistry, University of Campinas
(UNICAMP), Campinas, SP, Brazil, CEP 13083-970

*e-mail: oliveiraluan010@gmail.com
Several exogenous factors influence plant metabolism. When in stress conditions such as caused
by changes in climate, water, pests, or mechanical damage, the lipoxygenase (LOX) pathway is
activated as a protective response, triggering intense production of volatile organic compounds
(VOC) with different functions, through the oxidation of free fatty acids1. Organic agriculture system
(OAS), as well as permaculture agricultural system (PAS), have been gaining importance, especially
for those who seek healthier food and for its appeal to ecosystems balance, as they do not use
inorganic fertilizers and pesticides, unlike conventional agriculture system (CAS)2. Pinto bean
(Phaseolus Vulgaris L.) is the most consumed among the pulses and is found in every country on
Earth3, being an important source of protein and iron, has a huge nutritional value, and compounds
such as organic acids, phenolic compounds, tocopherols, and carotenoids4. The objective of this
work was to employ the foodomics approach to investigate the profile of VOCs from Pinto beans
from conventional, organic, and agroecological farms, extracted by headspace solid-phase
microextraction (HS-SPME) associated with gas chromatography coupled to mass spectrometry
(GC-MS) and chemometric tools. Extraction was performed using
divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS, 75 µm) fiber, and separation and
detection were performed on an Agilent 7890A GC system equipped with an Agilent MSD 5975C
quadrupole mass spectrometer with electron ionization (EI) source. The MassBank of North America
(MoNA) provided the library used to assign the identities of the compounds and the multi and
univariate statistical analyses were performed using the MetaboAnalyst 4.0. The best preprocessing
was achieved by variables normalization by the median, and autoscaling (mean-centered and
divided by the standard deviation of each variable). The exploratory analysis consisted of principal
component analysis (PCA) and hierarchical cluster analysis (HCA) and partial least squares-
discriminant analysis (PLS-DA) was performed for supervised analysis. Of 931 VOCs detected, 244
were identified, from different classes such as aromatics, esters, alcohols, fatty acids, phenols,
terpenes, hydrocarbons, ketones, aldehydes, and others. The previous studies identified less than
100 compounds5. The chemometrics analysis found different profiles between PAS, OAS, and CAS,
with the VOCs related to the flavor, ecological function, and biological activity being the most relevant
for the differentiation, i.e. the terpenoids. Beans from PAS presented major VOCs class variety,
suggesting a most stimulated biosynthesis, which, in turn, points out that agroecological practices
have more impact on volatile metabolome profile. The differentiation of CAS vs. OAS was less acute,
with alcohol and aromatic metabolites being the most relevant differentiating VOCs, indicating that
changing from one system to another exerts minor changes in the terpenoids profile.
1 Karolkowski, A, Guichard, E, Briand, L, Salles, C, Foods, 10, 2021, 3140.
2 de Lima, MD, Barbosa, R, Food Anal. Meth., 12, 2019, 887.
3 Food and Agriculture Organization of the United Nations (2016), Pulses: Nutritious seeds for a sustainable future, Ed.
Food & Agriculture Org., 2016, Italy.
4 Hayat, I, Ahmad, A, Masud, T, Ahmed, A, Bashir, S, Crit. Rev. Food Sci. Nutr., 54, 2014, 580.
5 Oomah, BD, Liang, LSY, Balasubramanian, P, Plant Foods Hum. Nutr., 62, 2007, 177.
[CNPq, FAPESP, INCTBio, Yokonoen movement]

135
EVALUATION OF DIRECT OIL ANALYSIS BY LA-ICP-MS USING FILTER PAPER
AS SOLID SUPPORT FOR CALIBRATION
Eduarda Q. Machado (PG)a, b*, Danielle S. Francischini (PG)a,b, Marco A. Z. Arruda (R)a,b
a Group of Spectrometry, Sample preparation and Mechanization – GEPAM, Dpt of Analytical Chemistry, Institue of
Chemistry, State University of Campinas - UNICAMP, Campinas, SP, Brazil, 13083-970
b National Institute of Science and Technology for Bioanalytics, Institute of Chemistry, State University of Campinas –
UNICAMP, Campinas, SP, Brazil, 13083-970

*e-mail: eduardaqm@gmail.com
Petroleum is a complex matrix with large economic importance. However, according to the chemical
composition, its industrial processing is hard. Nickel and Vanadium are elements its show high
concentrations. For its determination, generally is used spectrometric techniques, which need a
sample preparation step, that can cause spectral interferences and larger limits of detection and
quantification, as also external contaminations1. In this case, the laser ablation inductively coupled
plasma mass spectrometry (LA-ICP-MS) is an alternative where solid samples can be analyzed by
a laser source, producing a dry aerosol that is transferred into the ICP-MS for an inert gas flow (Ar
or He), where is atomized, vaporized and separated to the detection according with m/z ratio. This
technique present advantages such as minimum sample preparation, less contamination risk, and
high sensitivity. However, are necessary efficient calibration strategies to reduce matrix and transport
effects2. Thus, the aim of this work is the evaluation of the use of filter paper as solid support 3 for
analysis of petroleum samples by LA-ICP-MS for determination of Ni and V. The analysis were
performed by a New Wave UP-213 ablation system with Nd:YAG (213 nm) as laser source, coupled
to an inductively coupled plasma mass spectrometer (ICP-MS, PerkinElmer ELAN DRC-e). The laser
parameters such intensity (40 to 60%), spot size (50 to 110 µm) and scan speed (50 to 110 µm s-1)
were optimized through the use of multivariate data and factorial design. For the paper homogeneity
measures, were evaluated two types of solid supports with 8 mm diameter disks: (i) a qualitative
cellulose filter paper and (ii) a 0.2 mm pore size quantitative cellulose nitrate filter paper, using
sample amount of 1, 2 or 3 µL. The LA-ICP-MS calibration method was based on the addition of a
fixed volume (2.5 µL) of standard solution at an appropriate concentration on the filter paper disks,
where three ablation lines with 1.5 mm size and 0.250 mm of the distance of each center line. In the
evaluation from homogeneity of crude oil in a quantitative filter paper, present the value being higher
for the quantitative filter paper, in addition to showing a more homogeneity in the analytical signal
whit relative standard deviation (RSD) 7.48 to 10.84% for the qualitative paper and 9.94 to 53.05%
for the quantitative paper. As optimization results, the effect of laser ablation variables and the
interactions between them in the analysis shows the result for the significance of variables through
the Pareto’s Graph, considering 95% confidence level, in which it is possible to verify that intensity
and laser spot size present positive effects to the standard solution, as also to the crude oil, which
also present positively influence of laser intensity and scan speed. As calibration results, where
verified the use of 13C as internal standard (IS) by the limits of detection (LOD) and quantification
(LOQ), as showed in the Table I. Where observed that the normalization with C for Ni and V increase
the LOD and LOQ, which impact the sensitivity of the method. This work showed the possibility of
application LA-ICP-MS for determination of Ni and V with lowest LDs (ng g -1) using filter paper as
solid support, without the necessity a sample preparation steps.
Table 1. Limits of detection and quantification (ng g-1) obtained from LA-ICP-MS to Ni and V determination with and without
C as IS.
Figures of merit
Ni V Ni/C V/C
LOD 0.058 0.012 0.078 0.032
LOQ 0.193 0.039 0.261 0.105
1 Mello PA, Pereira JSF, Mesko MF, Barin JS, Flores EMM, Anal. Chim. Acta, 746, 2012, 15.
2 Pozebon D, Scheffler GL, Dressler VL, J. Anal. At. Spectrom., 32, 2017, 890
3 Moreau P, Arruda MAZ, J. Anal. At. Spectrom., 795, 2022, 37.
[FAPESP, CNPq, Petrobras]

136
METHOD DEVELOPMENT FOR THE ANALYSIS OF ENDOGENOUS STEROIDS
IN PLASMA BY GC-MS/MS AND ITS APPLICATION IN SAMPLES OF PATIENTS
WITH PARKINSON'S DISEASE
Jéssica G. B. Fontes (PG)a*, Andressa C. S. Marques (PG)b, Bruna L. Fadel (PG)a, Gabriela
Poralla (PG)a, Ana L. Z. Rosso (R)c, Monica C. Padilha (R)b, Luciana Pizzatti (R)a
a Federal University of Rio de Janeiro, LABMOPS/LADETEC/Chemistry Institute, Rio de Janeiro, RJ, Brazil, 21941-598
b Federal University of Rio de Janeiro, LAPAA/LADETEC/Chemistry Institute, Rio de Janeiro, RJ, Brazil, 21941-598
c Federal University of Rio de Janeiro, Neurology Department/HUCFF, Rio de Janeiro, RJ, Brazil, 21941-617
*e-mail: jessica.gbf@hotmail.com
Parkinson’s disease (PD) is a neurodegenerative disorder, in which the progressive loss of

dopaminergic neurons from substantia nigra occurs.1 Nowadays, it is known there are sex
differences in PD development, an example of this is that the incidence and prevalence of the
disease in men are higher. Besides that, evidence has shown that estrogen is neuroprotective to the
dopaminergic system, which suggests possible protection for women.2 Some androgen also seems
to have a neuroprotective effect, so androgen therapy has been studied for PD treatment but still
with variable results.3 In addition, although still controversial, the levels of cholesterol, a precursor
molecule of steroid hormones, tend to be reduced in patients with PD.4 Considering these facts, the
study of PD patients’ steroid profiles is relevant for the comprehension of the disease mechanisms
that lead to new treatment strategies. Therefore, this project aims to develop and validate a method
to quantify the total fraction of 11 endogenous steroids in plasma of PD patients and healthy controls
and correlate the data obtained with the clinical characteristics of these patients. For the sample
preparation, initially, the need for deproteinization was tested and then the use of different types of
extractions and solvents were evaluated. Still in this context, three-level factorial experiments varying
in time and enzyme volume were done for the optimization of the sample hydrolysis using β-
glucuronidase from E. coli. Matrices free of the analytes were elaborated to simulate plasma and be
used for analytical curves construction. For the instrumental analysis, different injection modes,
chromatographic columns, and temperature ramps were tested in a TSQ 8000 Evo Triple
Quadrupole GC-MS/MS system operating in multiple reaction monitoring (MRM) mode in which
transitions and collision energies were optimized. Method validation was done according to the RDC
166/17 from Brazilian Health Regulatory Agency (Anvisa). Samples from PD patients (n =133) and
healthy controls (n = 39) were collected and classified by sex and following the disease’s phenotype,
severity, and age of onset. Finally, the newly developed method was applied to the samples. Data
processing was performed using TraceFinder and Xcalibur software. Statistical analyses of the data
were executed through GraphPad Prism and Metaboanalyst software. As result, the developed
method consists of the hydrolysis of 500 μL of plasma samples at 50°C for 1h using 100uL of the
enzyme followed by an LLE extraction using TBME and finally, derivatization. In splitless mode, 2
μL were injected into the system previously mentioned. Best results regarding signal intensity and
peak resolution were observed using a 5%-phenyl-methylpolysiloxane (30 m x 0.25 mm x 0.10 μm).
The obtained LD and LQ were below 0.3 and 0.5 ng/mL, respectively, in most of the analytes.
Satisfactory selectivity, linearity, precision, accuracy, and robustness were achieved, and no matrix
effect was observed in the larger part of the cases. Regarding the male cohort, all analytes could be
detected in almost 100% of the samples. This result was obtained for more than half of the analytes
considering the female cohort. Statistically significant differences were observed among different
groups of patients, mainly different ages of onset, and between patients and healthy controls in some
of the studied steroids. In conclusion, the method proved capable of quantifying the proposed
analytes in plasma samples following the pre-established criteria. Differences observed in the levels
of some steroids between groups of patients are being more widely investigated but may indicate
changes in metabolic pathways.
1 Emamzadeh, F. N.; Surguchov, A., Frontiers in Neuroscience,12, 2018, 1.
2 Smith, K. M.; Dahodwala, N. Experimental Neurology, 259, 2014, 44.
3 Bianchi, V. E, et al. Journal of the Endocrine Society, 4, 2020, 1.
4 Paul, R.; Choudhury, A; Borah, A. Neurochemistry International, 90, 2015, 125.

137
IONOMIC STUDY IN TRANSGENIC SOYBEANS CULTIVATED IN SOILS DOPED
WITH MERCURY AND EVALUATION OF SILICA NANOPARTICLES AS
NANOREMEDIATOR
Vinnícius H. C. da Silva (PG)a,b*, Marco A. Z. Arruda (R)a,b
a Spectrometry, Sample Preparation and Mechanization Group-GEPAM, Institute of Chemistry, State University of
Campinas – UNICAMP, Campinas - SP, Brazil, 13083-970
b National Institute of Science and Technology for Bioanalytics, Institute of Chemistry, State University of Campinas –
UNICAMP, Campinas - SP, Brazil, 13083-970

*e-mail: vinnishenrique@gmail.com.br
The application of nanoparticles (NPs) in agriculture has been constantly expanded, such as the use
of nanofertilizers, nanopesticides, as well as a potential remediation agent of contaminated soils1. In
this sense the present study aimed to perform an ionomic evaluation in different biological
compartments (leaf, stem, and root) of transgenic soybean plants (Roundup Ready and Intacta)
cultivated in a soil doped with mercury and with a potential remediation agent (silica nanoparticles,
SiO2NPs). Moreover, transgenic soybean crops conducted with sodium silicate (Na2SiO3) were
evaluated for a comparative purpose. In this context, the experiment was composed of six different
groups (Control; Na2SiO3; SiO2NPs; SiO2NPs + Hg; Na2SiO3 + Hg, and Hg). The soil previously
doped with 120 mg kg-1 de Hg, while the concentration of SiO2NPs and Na2SiO3 was 200 mg kg-1.
Also, NPs used in this study were characterized by dynamic light scattering, where it was verified an
average size of 25 nm. Therefore, parameters involving Hg mitigation were addressed and the
homeostasis of micronutrients (Cu, Fe, Zn, and Mn) was assessed using ICP-MS. Figure 1 shows
the graphics of scores and loadings for Fe, Cu, Zn, Mn, and Hg contents in the groups of soybeans.
Figure 1. Micronutrients and Hg for PCA scores and PCA loadings graphics for soybeans groups.
The Figure 1 indicates a tendency to separate the groups containing Na2SiO3 from the groups with
SiO2NPs, as well as, a separation between the control groups of different biological replicates from
the group with only Hg added. In addition, a translocation of Hg from the roots to the upper parts of
the plants (e.g., leaf) was observed in the groups cultivated with SiO2NPs and Na2SiO3 in the soil.
The determination of Fe, Cu, Mn and Zn concentrations allowed the evaluation of possible
disturbances in the uptake of micronutrients due to the presence and uptake of Hg by the plants.
Moreover, changes were observed in all compartments, especially in the roots, where lower Fe
concentrations were found in the Hg-grown groups, with reductions of 36, 38, and 77 % for RR; 76,
51, and 70 % for Intacta, in the groups, SiO2NPs + Hg, Na2SiO3 + Hg and Hg, respectively. This
behavior was also observed in a test performed with sulfur nanoparticles aiming at reducing Hg
toxicity in Brassica napus L.2. The order of concentration of the micronutrients was Fe > Zn > Mn >
Cu, and they were found at higher concentrations in the root, leaf, stem, respectively, being a factor
observed in both transgenic types evaluated in this study. The ionomic evaluation together with the
determination of Hg in the biological compartments of the soybean plant allowed the assessment of
the behavior of SiO2NPs and Na2SiO3 as possible remediation agents. Besides, the results can
indicate the influence of Hg on the uptake of micronutrients by transgenic soybean plants.
1 Rastogi A, Tripathi DK, Yadav S, Chauhan DK, Živčák M, Ghorbanpour M, El‑Sheery NI, Brestic M, Biotech, 9, 2019, 90
2 Yuan H, Liu Q, Guo Z, Fu J, Sun Y, Gu C, Xing B, Dhankher OP, Journal of Cleaner Production, 318, 2021, 128589.
[FAPESP, CAPES, CNPq, INCTBio]

138
ADDRESSING THE SILVER NANOPARTICLE EFFECTS IN SOYBEAN CELLS
EMPLOYING DIFFERENT IN VITRO TECHNIQUES, SINGLE-CELL, SINGLE-
PARTICLE, AND LASER ABLATION ICP-MS
Ana B. S. da Silva (R)a,b*, Danielle S. Francischini (PG)a,b, Marco A. Z. Arruda (R)a,b
a Spectrometry, Sample Preparation and Mechanization Group-GEPAM, Institute of Chemistry, University of Campinas –
Unicamp, Campinas - SP, Brazil, 13083-970
b National Institute of Science and Technology for Bioanalytics, Institute of Chemistry, University of Campinas – Unicamp,
Campinas - SP, Brazil, 13083-970

*e-mail: anabs30@unicamp.br
The plant in vitro techniques have been considered a powerful tool to propagate essential plant
species as well as to perform genetic improvement. On the other hand, this approach has also
allowed the investigation of plants at a cellular level.1 Moreover, the nanotoxicology cannot be
misplaced, and deep studies involving nanoparticles-plant cells interactions are required.2 In this
sense, this work aims to employ two different plant in vitro techniques (somatic embryogenic and
plant cell suspension culture) to investigate the silver nanoparticle (AgNPs) effects in soybean cells.
In addition, mass-based techniques such as laser ablation inductively coupled plasma mass
spectrometry (LA-ICP-MS), single-particle (spICP-MS), and single-cell ICP-MS (scICP-MS) were
employed to establish an ionomic profile of soybean callus using element spatial distribution,
nanoparticle characterization after the in vitro culture, silver fractionation, and the AgNPs cellular
bioavailability. Firstly, the somatic embryogenic was performed using a soybean stem as an explant,
and the medium used for callus induction and propagation was composed of Murashige and Skoog
basal medium, agar, sucrose, and 2,4-Dichlorophenoxyacetic acid. During the callus induction, 200
µg L-1 of AgNPs (40, 60, and 80 nm) were added to the culture medium and after 30 days the callus
samples were collected for the following analysis. Instead, the plant cell suspension culture was
prepared from the chopped callus samples and placed into a liquid medium under agitation. After
the establishment of this culture, an incubation step (4 hours) with soybean cells and AgNPs with
200 µg L-1 (40, 60, and 80 nm) was carried on, and the uptake amount of silver nanoparticles by the
cells was addressed by employing single-cell ICP-MS. Concerning the callus culture in the presence
of AgNPs, it was observed that the nanoparticle with 80 nm led to a reduction of oxidation due to the
final appearance. Also, the AgNPs internalized in the callus were characterized by spICP-MS, and
the results found indicated a low translocation from the medium to the callus for all sizes, being
absorbed approximately 0.01%, and almost 98% of silver detected was as ionic form. Regarding the
AgNPs in plant cell suspension culture, using the single-cell ICP-MS, it was possible to estimate the
amount of silver tracked by a single-soybean cell. The values of silver amount varied from 1 to 10
pg per cell when they were incubated with AgNPs (40 nm), while for 60 and 80 nm, the range was
0.05 – 10 pg cell-1, indicating a high cell heterogeneity. In addition, the mean of mass internalized
into the soybean cells was 2.7, 1.2, and 0.9 pg per cell for the suspension culture exposed to 40, 60,
and 80 nm, respectively, indicating a higher cellular bioavailability for AgNPs with 40 nm. Also, the
spatial distribution of several macro and micronutrients (Ba, Cu, Zn, Mn, Ca, P, and Fe) was obtained
using LA-ICP-MS, and the effect of ionomic profile is dependent on the nanoparticle size. For
example, comparing the distribution of Ca, Mn, Ba, P, and Mg in callus cultured in the medium with
silver nanoparticles, an opposite behavior was observed among them. Within this perspective, the
ICP-MS operated in a single particle and a single cell mode can provide important information about
nanoparticle-plant cell interactions, as well as the LA-ICP-MS is a potential complementary
technique. In addition, a size-dependence effect was noted, therefore this property needs to be
considered depending on the application in the plant biotechnology field.
1 Guan Y, Li SG, Fan XF, Su ZH, Front. Plant Sci. 7 (2016) 1.
2 Galazzi RM, Arruda MAZ, J. Trace Elem. Med. Biol. 48 (2018) 181.
[FAPESP, CAPES, INCTBio]

139
EVALUATION OF THE EFFECT OF AESTHETICAL HAIR TREATMENTS IN THE
MINERALOGRAM
Natália R. Silva (PG)a*, Cristiane R. Mauad (R)b, Tatiana D. Saint’Pierre (R)a
a Pontifícia Universidade Católica do Rio de Janeiro (PUC-Rio), Department of Chemistry, Rio de Janeiro, RJ, Brazil, Zip
Code: 22453-900
b Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ), Rio de Janeiro, RJ, Brazil, Zip Code:
20270-021
*e-mail: nataliares10@gmail.com.br
A capillary mineralogram is a complementary multielement analysis that provides information on the

concentrations of essential, non-essential, and toxic elements that may be present in the body. This
exam gives information about the patient's health and assists in diagnosing various diseases related
to intoxication or deficiency of chemical elements. However, aesthetic treatments can affect the
results of the analysis since the products used can incorporate or extract hair elements, making it
difficult to interpret the results. The aim of this research was to identify patterns in the hair mineral
composition, and consequently, in the results of the mineralogram, due to the use of aesthetic
treatments such as dyeing and straightening. This work is part of a project to encourage girls in the
science, technology, engineering, and mathematics (STEM) area, called "Girls in Science", with
financial support from the Brazilian government and the participation of 5 high schools in the
metropolitan region of Rio de Janeiro. About 180 hair samples were voluntarily provided by students
from the schools participating in the project. The hair samples were analyzed by inductively coupled
plasma mass spectrometry (ICP-MS) and the mineralogram results were compared by statistical
analysis. Concerning the samples, there was a predominance of natural hair, corresponding to 52%
of the interviewees, straightening was the most used aesthetic treatment (25%), dyeing was adhered
to by only 16% of the interviewees and the combination of both treatments was the least expressive,
accounting for only 7% of the samples. In the statistical analysis, initially, the sums of concentrations
of essential and toxic elements were compared through box plots, for natural and each kind of treated
hair. Subsequently, the sum of concentrations of nutrients and toxic elements was screened to
observe whether aesthetic treatments affect the general composition of the analyzed hairs. In this
case, significant differences were observed for some elements. The elements that showed
differences in capillary concentrations, when comparing natural and hair with different aesthetic
treatments (dyeing, straightening, or both) are Ca, Mg, Ni, and Sr. However, it was still possible to
observe a significant difference between the results of the schools that participated in the project. In
conclusion, the results indicated that aesthetic treatments, especially dyeing, significantly affect the
concentrations of some elements in the hair, hindering diagnoses based on capillary mineralogram,
corroborating the medical recommendation to let the hair grow free of aesthetic treatments for at
least 3 months before sample collection for the mineralogram.
1 Carneiro, M. et al., Quím. Nova, 25, 2002, 37.
2 Miekeley N, Dias Carneiro MTW, Porto da Silveira CL, Sci Total Environ 218, 1998, 9.
3 Miekeley, N. et al., J. Trace Elem. Medic. Biol., 15, 2001, 46.
4 Kosanovic, M.; Jokanovic, M., Environ. Monit. Assess. 174, 2011, 635.
5 Pozebon D., Dressler V.L., Curtius A.J., Quím. Nova, 22, 1999, 6.
[FAPERJ, CAPES, CNPq, PUC]

140
SIO2-NANOPARTICLES AS CONTAMINATION MITIGATING AGENT IN
TRANSGENIC SOYBEAN CULTIVATION
Daniel C. Freitasa,b and Marco A. Z. Arrudaa,b*
a Spectrometry, Sample Preparation and Mechanization Group, Institute of Chemistry, University of Campinas –
Unicamp, P.O. Box 6154, Campinas, SP 13083-970, Brazil
P.O. Box 6154, Campinas, SP 13083-970, Brazil.

*e-mail: zezzi@unicamp.br
Silicon oxides have outstanding features, such as good stability, ease of functionalization, and
biocompatibility, among others. In this way, silica nanoparticles have been widely applied to plants
cultivation, to evaluate their mitigating effect1. In this work were used silica nanoparticles (400 mg
kg-1) and Cd2+ as CdCl2 (140 mg kg-1). After performing the sample preparation (utilizing HNO3 +
H2O2) assisted by microwave technology, the samples were diluted to a residual acidity of 1% v/v.
Micronutrients and Cd2+ determinations in the samples were performed by ICP-MS and showed in
Table 1.
Table 1. Determination of concentrations (mg kg-1) of micronutrients and Cd2+ in the compartments of RR and Intacta
soybeans of different groups by ICP-MS.
Element Root Stem Leaf Root Stem Leaf
RR control Intacta control
56Fe 2052 ± 124 215 ± 20 135 ± 5 4539 ± 120 88 ± 3 161 ± 5
65Cu 11.0 ± 0.6 12.3 ± 0.6 11.0 ± 0.6 11.4 ± 0.8 11.3 ± 0.3 11.4 ± 0.4
60Ni 51 ± 3 5.0 ± 0.3 5.1 ± 0.3 85 ± 12 4.0 ± 0.5 5.0 ± 0.5
114Cd 1.9 ± 0.1 1.1 ± 0.1 0.9 ± 0.1 0.6 ± 0.1 0.5 ± 0.1 0.4 ± 0.1
RR SiO2-NPs Intacta SiO2-NPs
56Fe 4757 ± 182 53 ± 3 139 ± 3 3210 ± 149 70 ± 2 87 ± 1
65Cu 12.3 ± 1.1 6.4 ± 0.2 10.8 ± 0.8 9.5 ± 0.4 6.3 ± 0.2 7.6 ± 0.1
60Ni 81 ± 2 4.0 ± 0.3 4.4 ± 0.1 55 ± 5 4.0 ± 0.1 4.0 ± 0.4
114Cd 0.7 ± 0.1 0.4 ± 0.1 0.4 ± 0.1 0.6 ± 0.1 0.4 ± 0.1 0.4 ± 0.1
RR SiO2-NPs + Cd2+ Intacta SiO2-NPs + Cd2+
56Fe 4810 ± 161 54 ± 3 156 ± 5 3058 ± 46 59 ± 3 74 ± 2
65Cu 14.8 ± 0.4 16.3 ± 0.1 9.8 ± 0.9 14.1 ± 0.7 16.8 ± 0.8 9.4 ± 0.5
60Ni 68 ± 3 4.0 ± 0.2 5.1 ± 0.3 53 ± 2 4.5 ± 0.6 3.7 ± 0.1
114Cd 100 ± 1 4.4 ± 0.2 1.2 ± 0.1 109 ± 5 5.1 ± 0.2 0.7 ± 0.1
RR Cd2+ Intacta Cd2+
56Fe 4954 ± 129 51 ± 1 171 ± 3 3041 ± 26 57 ± 3 69 ± 1
65Cu 13.2 ± 0.9 7.7 ± 0.2 10.8 ± 0.3 13.1 ± 0.6 8.6 ± 0.5 9.5 ± 0.4
60Ni 95 ± 3 5.4 ± 0.7 4.7 ± 0.3 53 ± 1 5.2 ± 1.7 4.1 ± 0.5
114Cd 127 ± 3 4.4 ± 0.3 2.6 ± 0.1 141 ± 10 5.4 ± 0.1 1.0 ± 0.1
* All significant differences observed were based on ANOVA statistical test for n=3 and p = 0.05.
From the results shown in Table 1, differences were observed in the concentrations of micronutrients
and Cd2+. For Fe, in the roots an increase concentration > 130% was observed between different
Intacta groups, while for the leaf, a lower Fe translocation was detected (the highest value of 26%
only). Among the RR soybeans, a decrease in the Fe uptake between the roots of up to 33% was
observed, and, in the leaves, ca. 57%. Among different transgenic plants it was observed that RR
soybeans absorbed more Fe than Intacta, except for the control group soybeans. Another important
micronutrient is copper, and no differences were observed in the copper uptake between the
compartments in the same transgenic group, as well as nickel, except for the roots, where significant
differences were observed. For Cd, it was observed that the presence of SiO2-NPs provided its lower
absorption (ca. 30%) when compared to the group where no NPs were added. Therefore, differences
in both uptake and translocation of micronutrients were observed, and the stress conditions are being
evaluated by adding the NPs and Cd.
1 Yunyun L, Silica nanoparticles alleviate mercury toxicity via immobilization and inactivation of Hg(II) in soybean (Glycine
max), Environmental Science Nano, 2020.
[UNICAMP, GEPAM, CAPES, CEPETRO, FAPESP]

141
METABOLOMICS ANALYSIS AND EFFECTS OF NPS ON SOYBEAN CALLUS BY
UPLC-MS/MS
Lilian S. Katoa*, Ana B. Santosa, Vinnícius H. C. da Silva (PG)a, and Marco A. Z. Arruda ®a,b
a Spectrometry, Sample Preparation and Mechanization Group – GEPAM, Institute of Chemistry, University of Campinas
– Unicamp, P.O. Box 6154, Campinas, SP 13083-970, Brazil;
b National Institute of Science and Technology for Bioanalytics, Institute of Chemistry, University of Campinas –
Unicamp, P.O. Box 6154, Campinas, SP 13083-970, Brazil

*e-mail: lskato@unicamp.br
Brazil is the world’s largest soybean producer and second-largest exporter. Soybean is an essential source
of protein and oil, also used in the fuel sectors and as animal feed. Somatic embryogenesis is a
propagation technology recognized as the future generation of plant regeneration, providing a unique
opportunity for studying many aspects of plant growth and development under stressful conditions.
Nanoparticles (NPs) have been widely used to improve human life. They can enter plant cells, modulate
the metabolism of reactive oxygen species (ROS), and alter secondary metabolites. Untargeted
metabolomics is an approach employed to identify as many metabolites as possible, which can be used
to understand plant-NPs interaction. Thus, the present study aims to optimize the sample preparation
procedure for metabolites determination from transgenic soybean callus by UPLC-MS/MS and investigate
the expression of metabolites in the samples. Finally, to understand how NPs affect the metabolomics in
soybean callus and whether it is possible to determine a link between the NPs and classes of metabolic
compounds. The soybean plants were used to excise expanded stems sliced into 0.3 cm-long pieces. The
stem pieces were placed in callus induction media and kept at 25 °C under white light and a 16-h day/8-h
night cycle. After four weeks, callus was excised from the stem pieces, plated on proliferation media, and
kept in an incubator at 25 °C in the dark, subculturing every three weeks to a fresh medium. A first test to
evaluate the contribution of ultrasound in the extraction of metabolites was carried out. Then, 200 mg of
sample previously frozen with liquid nitrogen and ground in a porcelain crucible were weighed directly in
Eppendorf tubes, and 100 mL of extracting solution containing methanol:chloroform (80:20 v/v) was added
and vortexed for 4 min. After that, the samples were submitted to Cup-horn ultrasound for 60 s, 50%
amplitude, 10 s on-off, centrifuged at 8500 rpm for 10 min, filtered, and stored in a refrigerator until reading.
Each sample was analyzed on a UPLC-MS/MS in the positive ion mode, using a C18 column, mobile
phase consisting of a linear gradient including acetonitrile + 0.1% formic acid and ultrapure water + 0.1%
formic acid, 22 min. of total running. LC-MS/MS raw data were processed and compared using
Metaboanalyst. The samples subjected to ultrasound had a higher number of peaks (2239 ± 1.5) than the
control samples (2075 ± 16); thus, a second test was performed with six different extractor solutions,
following the same analysis method already described, including ultrasound treatment. The results in Table
1 showed that the number of peaks recovery was higher using 50% acetonitrile + 50% ultrapure water as
extracting solution; however, there was only a statistical difference (p<0.05, n=3) in the average number
of peaks recovered between extractor solution four and the others. Future tests will include the application
of extractor solution six and different amplitudes and duration times in the ultrasound. After defining these
parameters by the number of peaks, the soybean callus samples will be analyzed using the HILIC and
C18 columns to cover an ample range of polarity of the metabolites. Metabolites will be identified with the
assistance of metabolomics libraries. Finally, soybean callus will be directly contaminated with SiO 2NPs
(10 and 100 ppm), and the effects on metabolomics will be investigated.
Table 1. Summary of peaks information of all spectra after processing.
Retetion Mean peak Missing
Samples Extraction solution m/z
time (RT, s) number (n = 3)* (%)
1A, 1B, 1C, 1D, 1E 1 - 100% MeOH 1432ª 29.37
2A, 2B, 2C 2 - 70% MeOH, 30% H2O 1502ª 25.85
3A, 3B, 3C 3 - 70% MeOH, 30% CHC3 33-1290 100-1291 1524ª 24.74
4A, 4B, 4C 4 - 33% MeOH, 33% CHC3, 33% H2O 979b 51.64
5A, 5B, 5C 5 - 100% ACN 1378ª 33.14
6A, 6B, 6C 6 - 50% ACN, 50% H2O 1533ª 24.28
* Results with the same letters in the column do not differ statistically (p>0.05).

142
IDENTIFICATION AND QUANTIFICATION OF NAPHTHENIC ACIDS DIRECTLY
FROM PRODUCED WATER BY PSI-MS
Gabriel Henry Morais Dufrayer (UG)a, Igor Pereira da Silva (R)b, Marcella Ferreira
Rodrigues (PG)a, Rafael Oliveira Martinsa, Jussara Valente Roque (R)a, Gabriel Franco dos
Santos (R)a, Rosineide Costa Simas (R)a,c, Andrea Rodrigues Chaves (R)a, Boniek Gontijo
Vaz (R)a
aUFG - Universidade Federal de Goiás
b VIU - Vancouver Island University
c Mackenzie - Universidade Presbiteriana Mackenzie
In the petroleum prospecting process, the water emerges next to the crude oil, denominated in the
produced water (PW). Due to the high temperatures, pressure, and storage time in the reservoirs,
the naphthenic acids (ANs) present in the crude oil can migrate to PW 1,2. With the large volume of
PW generated concerning the amount of oil prospected and the level of toxicity for the marine flora
and fauna, it is essential to control the NAs through their identification and quantification2. The
present study aimed to characterize and semi-quantify NAs in PW using the Paper Spray Ionization
Mass Spectrometry (PSI-MS) technique3. However, due to the high concentration of saline
compounds in the PW, there is suppression of the analytical signals of the ANs, requiring a superficial
modification of the paper through an ethanolic solution of 3-aminopropyltriethoxysilane (APTS) made
according to Koga, H.; Kitaoka, T.; Isogai, A. (2011)4 for direct analysis. With the papers modified,
the ANs were extracted and analyzed by the LTQ XL mass spectrometer (Thermo Scientific,
Massachusetts, USA). For semi-quantification, a global calibration curve was constructed using the
standard solution of NAs (Sigma Aldrich, Missouri, USA). From this curve, it was possible to study
the semi-quantification of total NAs in the analyzed samples, presenting very accurate results with a
correlation coefficient, R2 = 0.9953, a low coefficient of variation (CV), CV < 13 %, and recovery
values close to 100%. Similarly, the surface modification on the paper made it possible to minimize
the effect of ionic suppression by the saline compounds, improving the ionization of NAs and
enabling their identification. Because of the results, it was possible to conclude that the PSI-MS
technique, together with the surface modification of the paper had auspicious results with a quick
and selective analysis for NAs, contouring the problem of ionic suppression through saline
compounds, avoiding laborious preparation of the sample, and reducing the use of organic solvents
which consequently allow the low cost of the analysis.
1 Clemente JS, Fedorak PM, Chemosphere, 60(5), 2005, 585-600.
2 Headley JV, Peru KM, Barrow MP, Mass spectrometry reviews, 28(1), 2009, 121-134.
3 Li A, Wang H, Ouyang Z, & Cooks RG, Chemical communications, 47(10), 2011, 2811-2813.
4 Koga, H, Kitaoka, T, & Isogai, A, Journal of Materials Chemistry, 21(25), 2011, 9356-9361.

143
ENHANCED DETECTION OF AMINO ACID NEUROTRANSMITTERS USING THE
KATRITZKY REACTION AND PAPER SPRAY IONIZATION MASS
SPECTROMETRY
Lanaia Í. L. Maciel (PG)a, Ruver R. R. Feitosa (PG)a, Raul I. Ribeiro (PG)b, Mauro C. X. Pinto
(R)b, Igor Pereira (R)a* and Boniek G. Vaz (R)a*
a Federal University of Goiás 1, Chemistry Institute, Goiânia, Goiás, Brazil, 74690-900.
b Federal University of Goiás 2, Institute of Biological Sciences, Goiânia, Goiás, Brazil, 74690-900
*e-mail: ipsigorpereira@gmail.com and boniek@ufg.br
This study describes the development of a derivatization protocol based on the Katritzky reaction
with paper spray ionization mass spectrometry (PSI-MS) for the enhanced detection of amino acid
neurotransmitters in mouse brain tissues. The protocol consists of spraying the derivatization
solution on a section of a mouse brain mounted on a glass slide, applying a small volume of solvent
to wet the sample, and pressing a triangular paper onto the sample surface to transfer the sample
constituents for paper surface, and using the paper as a substrate for PSI-MS analysis (Figure 1).
Figure 1. System overview: the derivatization solution is sprayed onto the sample; a small volume of solvent is added to
moisten the sample; a triangular paper is pressed onto the sample surface to transfer the sample constituents to the paper
surface; the paper is used for conventional PSI-MS analysis.
The Katritzky reaction facilitated the ionization of amino acids by reacting a pyrylium salt with the
amino group of the analytes, forming very stable pyridinium cations1, which greatly increased the
sensitivity of the PSI-MS analysis. Most of the intensities of the amino acids modified by the Katritzky
reaction2 were more than 10 times higher than the non-derivatized ones. The system was applied to
the analysis of brain sections obtained from mice with Parkinson's disease, and the amino acids
gamma-aminobutyric acid (GABA) and glycine (Gly), two well-known compounds in Parkinson's
disease studies, were promptly detected. The results suggest that the Katritzky reaction combined
with PSI-MS can offer a significant advance in the knowledge of protocols that improve the sensitivity
of detection of crucial biological compounds.
1 Waliczek, M. et al. Scientific Reports 6, 37720 (2016).
2 Shariatgorji, R. et al. Journal American Society Mass Spectrometry (2020).
[CAPES]

144
SPECIATION OF NAPHTHENIC ACIDS AND THEIR OXYACID-EXTRACTABLES
ANALOGS FROM PRODUCED WATER BY ORBITRAP MASS SPECTROMETRY
Marcella F. Rodrigues (PG)a, Rosineide C. Simas (R)a, Rosana C. L. Pereira (R)a, Gabriel H.
M. Dufrayer (UG)a, Nerilson M. Lima (R)a, Rogério M. de Carvalho (R)b, Iris Medeiros Júnior
(R)b, Gabriel F. dos Santos (R)a, Boniek G. Vaz (R)a,*
a Chemistry Institute, Federal University of Goias, Goiania, GO, 74690-900, Brazil
b CENPES, PETROBRAS, Rio de Janeiro, RJ, 21941-915, Brazil.
*e-mail: boniek@ufg.br
Produced water (PW) is an unresolved complex mixture that includes various chemical constituents
with distinct chemical and physical properties from metals to organic contaminants, it is water
trapped in underground formations brought to the surface together with oil and gas during the
production activities of these fluids1. This complex saline mixture is constituted mainly of naphthenic
acids (NAs) and presents toxicity to various aquatic organisms. Therefore, the terminology oxy-NA,
naphthenic acid fraction component (NAFC), water-soluble organic (WSO), and acid-extractable
organics (AEOs) has been introduced to replace NAs, since this class of compounds may contain
more than two oxygen atoms and heteroatoms such as nitrogen or sulfur in the structure 2,3. The
molecular complexity of NAs is a great challenge because it is a mixture of compounds with many
isomers that are difficult to separate and characterize, related to their toxicity to the environment.
The concern with these species has increased due to their potential disposal in the environment.
Several types of research have studied NAs in oil sand process-affected water (OSPW); however,
few studies report the characterization of these chemical constituents in produced water from
offshore oil platforms. Thus, to properly dispose of the produced waters, it is necessary to
comprehensively characterize their organic-extractables composition. In this context, this work
describes the application of non-sequential liquid-liquid extraction using the solvents n-hexane,
cyclohexane, toluene, ethyl ether, dichloromethane (DCM), and ethyl acetate at pH 2 followed to ESI
(-) Q-Orbitrap mass spectrometry analysis to verify the response of these solvents regarding the
number of formulas assigned to the NAs from produced water. The solvents DCM and ethyl acetate
showed greater extraction efficiency and, therefore, they were selected to perform the sequential
(using single solvent and a pH sequence per sample) and non-sequential (using a single solvent and
a single pH per sample) liquid-liquid extraction at pH 2, 7, and 10, whose objective was to evaluate
the effect of pH and the effectiveness of extraction methodologies in the extraction of NAs and their
Ox-NAs analogs. Our findings showed that the extractor solvents with a higher dielectric constant
(ethyl acetate and DCM) demonstrated the highest extraction efficiency. Concerning the extraction
methodology using these two solvents, more molecular formulas were observed in the sequential
extraction. The evaluation of the extraction efficiency of NAs at pH 2, 7, and 10 for the class O2NA
showed a greater number of ions at pH 10 and NAs with higher carbon numbers and a larger DBE
range. NAs with higher carbon numbers and a larger DBE range were extracted in this condition.
1 Samanipour S, Reid MJ, Rundberget JT, Frost TK, Thomas K V. Environ Sci Technol, 54, 2020, 2707.
2 Ripmeester MJ, Duford DA. Chemosphere, 233, 2019, 687.
3 Headley J V., Peru KM, Mohamed MH, Frank RA, Martin JW, Hazewinkel RRO, et al. J Environ Sci Heal Part A, 48,
2013, 1145.
[PETROBRAS, LaCEM, UFG]

145
EVALUATION OF THE EFFECTS OF THERMAL MATURITY ON THE
DISTRIBUTION OF NSO COMPOUNDS IN HYDROUS PYROLYSIS PRODUCTS
FROM TYPE-IIS KEROGEN BY ULTRAHIGH RESOLUTION MASS
SPECTROMETRY
Taynara Rodrigues Covasa, André Luiz D. Spigolona, Danielle Mitze Muller Franco (PG)a,
Rosana Cardozo Lopes Pereira (R)a, Mário Ducan Rangel (R)a, Rosineide Costa Simas (R)a,b,
Ygor dos Santos Rocha (R)c, Joelma Pimentel Lopes (R)c, Igor Viegas Alves Fernandes de
Souza (R)c, Boniek Gontijo Vaz (R)a
aUFG - Universidade Federal De Goiás
b
Mackenzie - Universidade Presibeteriana Mackenzie
c CENPES - Geochemistry Laboratory of the Petrobras Research and Development Center
Among the variables that control the oil generation process, the type and the quality of organic matter
present in the source rock associated with the history of time and temperature are the most
relevant.1 One way to understand the impact of increasing thermal maturity is to describe the
changes during oil generation, and expulsion through hydrous pyrolysis (HP) experiments on
immature source rock samples. The literature is related to what Type IIS kerogen generates oil with
higher concentrations of sulfur compounds and has different oil generation kinetics than Type
II.2 This study describes the impact of thermal maturity on the distribution and concentration of
organosulfur compounds from a hydrous pyrolysis experiment, resulting in a more robust
characterization in petroleum systems where Type-IIS kerogen is predominant. In this study, the oils
expelled and retained from hydrous pyrolysis experiments were analyzed by ESI(-) and APPI(+) FT-
ICR MS. It was observed that oxygenated compounds have similar behavior to that reported for Type
I and Type II kerogen. However, poly-oxy-sulfur compounds (OxSz class group) decreased relative
abundance, and sulfur compounds (Sz class group) increased with increment thermal maturity.
Along with the atypical behavior of sulfur compounds, it was possible to correlate the high values of
the S/C ratio and low values of the H/C ratio with the increase in thermal maturity.
1 Spigolon ALD, Lewan MD, de Barros Penteado HL, Coutinho LFC, Mendonça Filho JG, Organic Geochemistry, 83-84,
2015, 27.
2 Craddock PR, Bake KD, Pomerantz AE, Energy and Fuels, 32, 2018, 4859.

146
EVALUATION OF ALPHA-ACIDS THAT CONTRIBUTE TO THE
INTERNATIONAL BITTERNESS UNIT OF BRAZILIAN CRAFT BEERS
THROUGH MASS SPECTROMETRy
Karinne T. Gonçalves (UG)a, Paola L. M. R. Almeida (R)a, Raquel M. F. Sousa (R)a*
a Federal University of Uberlândia, Chemistry Institute, Uberlândia, Minas Gerais, Brazil, 38400-902
*e-mail: rsousa@ufu.br
Beer is an aqueous beverage fermented from sugars from the starch of malts which receives hops
to balance its flavour and, in some cases, enhance the aroma. The hop added during the boiling
process is associated with bitterness and foam and, when added later in the process (dry-hopping)
it becomes associated with the aroma as well1. The IBU (International Bitterness Unit) value is used
to express the intensity of bitterness in beers2,3. The compounds responsible for bitterness are alpha-
acids isomerized during wort boiling (brewing stage). The alpha-acids are humulone, co-humulone,
and ad-humulone (Figure 1). In this work, the chemical composition of three craft beers with different
IBUs (PM1: IBU15, PM2: IBU 50; PM3: IBU 100) and hop extract were evaluated by liquid
chromatography coupled to mass spectrometry (Agilent, Q-TOF) and correlated with IBU. In the beer
analysis, 1 mL of beer (CO2 was previously removed by sonication for 2 min) was solubilized with
1 mL of methanol, the mixture was filtered (PTFE 0.22 µm), and injected into the LC/ESI-MS/MS
(negative mode). In the hop analysis, 2.46 g of hop pellets were macerated with isooctane (3 X 50
mL), the sample was left for 4 hours in the freezer, the solid was discarded, and the organic phase
evaporated, obtaining 0.43 g of extract4. A sample of hop extract (5 mg mL-1, methanol) was filtered
(0.22 µm PTFE) and injected into the LC/ESI-MS/MS. The used chromatographic parameters were:
0.2 mL min–1 of mobile phase constant flow (30:70, methanol: formic acid (0.1 % in water)). The used
ionization parameters were: nebulizer pressure of 20 psi, drying gas at 8 L min–1, 220 ºC, 4.5 KVa.
Through these data it can be seen that the degree of IBU follows the presence of α-acids, that is, as
the IBU increases (PM1< PM2<PM3) an increase in α-acid molecules present in beers is observed.
In addition, it was verified that the hop extract does not present iso-alfa-acids, since these are formed
only under heat treatment in beer. The m/z of adhumulone and humulone are the same and so far,
we have not been able to identify the difference between them. On the other hand, cohumulone is
very different from adhumulone and humulone, because they have different fragmentation patterns.
Lupulones and colupulones (beta-acids) do not appear in beer, as they have lower polarity and,
therefore, are not extracted by the wort during brewing, but their presence is observed in the hop
extract.
Figure 1. Chromatogram (LC/ESI-MS/MS, negative mode) of craft beers and hop: IBU 15 (black line), IBU 50 (red line),
IBU 100 (green line), and Hop extract (blue line). Note: a: iso-cohumulone; b: iso-ad/humulone; c: cohumulone; d:
ad/humulone; e: colupulone; and f: lupulone.
1 Kishimoto, T et al, Journal of the American Society of Brewing Chemists, 80, 2022, 53
2 American Society of Brewing Chemists. Method Beer-23A, 1992.
3 European Brewery Convention. Analytica-EBC Method 9.6, 1987; E155.
4 Cattor, K et al. Journal of Agricultural and Food Chemistry, 61, 2013, 7916.
[FAPEMIG, CAPES, CNPq]

147
CHEMICAL CHARACTERIZATION OF ACID-EXTRACTABLE ORGANICS FROM
PRODUCED WATER BY ST-DLLME COMBINED WITH UHRMS ANALYSIS
Deborah V. A. de Aguiar (PG)a*, Gabriel F. dos Santos (R)a, Rogério M. de Carvalho (R)b, Iris
M. Júnior (R)b, Rosineide C. Simas (R)a, Boniek G. Vaz (R)a
a Chemistry Institute,Federal University of Goiás, Goiânia, GO, Brazil, 59078-970
b CENPES, PETROBRAS, Rio de Janeiro, RJ, Brazil, 21941-915
*e-mail: deborah.victoria156@gmail.com
Produced water (PW) is generated as a byproduct from offshore oil production platforms and
represents a significant environmental problem due to the high volume generated and its toxicity.1,2
Therefore, new methods are necessary to characterize the produced water and, consequently, to
provide an appropriate destination. This study aims to use solvent-terminated dispersive liquid-liquid
microextraction (ST-DLLME) followed by ultrahigh-resolution mass spectrometry (UHRMS) analysis
to extract and characterize naphthenic acids (NAs) from PW. ST-DLLME is based on a ternary
mixture of solvents classified as a dispersive, extractor, and demulsifier solvents to ensure analyte
extraction efficiency.3 Seven solvent systems at three pH values (2, 7, and 10) were used to study
the influences of the sample pH and organic-phase polarity. The relationship between pKa and pH
affects the equilibrium between ions and molecules in the aqueous phase, impacting the extraction
efficiency.4 The results showed that at pH 2, the extractability of NAs is directly affected by the
dielectric constant of the dispersive solvent. Hence, when dichloromethane was employed as a
dispersive solvent, ST-DLLME presented a better performance, and NA species within an extensive
double bond equivalent (DBE) range (1 – 8) and range of carbon numbers (C-7 – C-18) were
extracted. At pH 7 and 10, the solvent system that employed cyclohexane as a nonpolar dispersive
solvent demonstrated the highest extraction efficiency. In addition, solutions at pH 7 and 10
expressed more comprehensive molecular formulas for the corresponding compounds than those at
pH 2. ST-DLLME has been shown as a valuable analytical method for extracting NAs from PW since
this extraction technique minimizes the consumption of samples and solvents.
1 Jiménez S, Micó MM, Arnaldos M, Medina F, Contreras S, Chemosphere, 192, 2018, 186.
2 Thomas KV, Langford K, Petersen K, Smith AJ, Tollefsen KE, Environmental Science Technology, 43, 2009, 8066.
3 Mansour FR, Danielson ND, Analytical Chimica Acta, 1016, 2018, 1.
4 Huang R, McPhedran KN, Sun N, Chelme-Ayala P, El-Din MG, Chemosphere, 146, 2016, 472.
[PETROBRAS, CNPq, CAPES, FAPEG]

148
DEVELOPMENT AND VALIDATION OF A METHODOLOGY FOR THE
DETERMINATION OF THE NUMBER CONCENTRATION OF GOLD
NANOPARTICLES BY SP-ICP-MS
Vania S. Oliveira (PG)a*, Thiago O. Araujo (R)b, Sandra M. Landi (R)c
aInmetro, Postgraduate Programme in Metrology and Quality, Duque de Caxias, Rio de Janeiro, Brazil, CEP 25250-020
bInmetro, Chemical and Thermal Metrology Division, Duque de Caxias, Rio de Janeiro, Brazil, CEP 25250-020
cInmetro, Materials Metrology Division, Duque de Caxias, Rio de Janeiro, Brazil, CEP 25250-020
*vsdeoliveira@colaborador.inmetro.gov.br
The study, manipulation and control of matter at the nanoscale allows the development of materials
with unique characteristics and properties not found at the macro/micro scale, which results in
chemical changes in functionality and reactivity. Therefore, those nanomaterials arouse the interest
and attention of science and industry, especially cosmetics, pharmaceuticals and food. In this
context, nanoparticles (NPs) are used in a large number of products1, with increasing progress in
consumer products2. Among metallic NPs, gold NPs (AuNPs) are of great interest for production and
research, given their ease of handling and biocompatibility. Knowledge of the dimensional properties
of NPs, such as average size, medium distribution and size concentration, is essential to understand
their effects on health and size, as well as production for quality control. One of the techniques that
can be used for these studies is an inductively coupled plasma mass spectrometry for single coupling
(sp-ICP-MS). To ensure the reliability and comparability of these measurements, it is necessary to
employ a validated method. The present work aimed to develop and validate a method for
determining the concentration in number of gold nanoparticles by sp-ICP-MS. The purpose of the
validation is verifying the suitability of the method for the intended use and was based on the
requirements established by ISO 17025:20173 standard and in the Inmetro document DOC-Cgcre-
0084. An experimental evaluation of the performance parameters of linearity, working range,
precision, selectivity and limit of detection for nanoparticle size, concentration in number of particles
and for dissolved ion was carried out. The method proved to be selective, that is, capable of
distinguishing the signal arising from the dissolved metal from the signal of the NP, and it was also
capable of differentiating populations of particles with different dimensions.
1 SANTOS, L. et al., Química Nova, 2021.
2 BAZILIO, F. et al., Química Nova, 2021.
3 ASSOCIAÇÃO BRASILEIRA DE NORMAS TÉCNICAS (2017), ABNT NBR ISO/IEC 17025. Requisitos gerais para a
competência de laboratórios de ensaio e calibração. Rio de Janeiro: ABNT. 32 p.
4 INSTITUTO NACIONAL DE METROLOGIA, QUALIDADE E TECNOLOGIA (2020b): Norma nº DOQ-Cgcre-008:
Orientação sobre validação de métodos analíticos. Inmetro. Duque de Caxias, CGCRE, 2020.
[CNPq, INMETRO]

149
QUALITATIVE IMAGING OF ELEMENTAL SPATIAL DISTRIBUTION IN
STALAGMITE THROUGH LA-ICP-MS ANALYSIS
Danielle da Silva Francischini (PG)a,*, Renata Stábile Amais (R)a, Nicolás Misailids Strikis
(R)b , Marco Aurélio Zezzi Arruda (R)a
a Group of Spectrometry, Sample preparation and Mechanization-GEPAM, Dpt. of Analytical Chemistry, Institute of
Chemistry, Unicamp, Campinas, São Paulo, Brazil, 13083-970
b Federal Fluminense University, Dept. of Geochemistry, Niterói, Rio de Janeiro, Brazil, 24020-141
*e-mail: dani_francischini@outlook.com
Speleothems (cave mineral deposits, largely formed by the carbonate precipitation) feature as of the
main natural archives used by geoscientists to reconstruct past variations of local climate. Since
speleothems are stalagmites characterized by regions distinguished by microlayers of carbonate
deposition, analysis techniques that enable high sensitivity and spatial resolution are appropriate1.
Furthermore, the improvement of high-resolution sampling analytical methods allows us to perform
annual-resolution records. The laser ablation inductively coupled plasma mass spectrometry (LA-
ICP-MS) technique is considered versatile for multi-elemental analysis and imaging because it is
easy to handle, compatible with different types of solid samples, requires minimal sample
preparation, and provides high spatial resolution and sensitivity2. One of the challenges of imaging
analysis is to obtain accurate and precise spatial information of elements distribution in the sample,
so the optimization of the laser ablation (LA) parameters is essential2. In this context, this study
aimed to optimize the LA parameters for direct analysis of speleothem samples. Laser intensity,
frequency and spot diameter were evaluated through multivariate experimental design (23) and multi-
response data3, the influence of ablation scan speed (20, 40 and 60 µm s-1) and the use of 44Ca as
an internal standard (IS) for the qualitative image of 66Zn, 137Ba, 55Mn, 57Fe, 88Sr, 60Ni, and 26Mg
distribution in the sample were also evaluated. As results, the multivariate optimization revealed
positive interactions between the parameters evaluated (through Pareto’s chart at 95% confidence
level), i.e., the greater the laser intensity, LA frequency and spot diameter, the greater the analyte
signal and, thus the sensitivity. Therefore, 90% laser ablation intensity, 20 Hz repetition rate, and
100 µm spot diameter were selected. In the scan speed evaluation (Figure 1) was observed that by
increasing the scan speed, the pixel size increase, worsening the image resolution in general. In
higher scan speed seems to be an overlap of the data in the spatial distribution and the images
obtained with 40 and 20 µm s-1 were very similar for all isotopes. The results also shows that the use
of 44Ca as IS did not impact the resolution of the images, but in environmental analysis, the use of
44
Ca can provide important information about the speleothem formation and paleoclimate changes4.
Figure 1. 137Ba distribution on the stalagmite sample surface according to each scan speed evaluated without 44Ca as
IS.
1Bernal JP et al. Earth Planetary Science Letters, 2016, 450, 186-196.

2Francischini DS, Arruda MAZ, MIcrochemical Journal, 2021, 169, 106526.
3Novaes CA et al. Current Analytical Chemistry, 2016, 12(2), 94-101.
4Francischini DS, Amais RS, Strikis NM, Arruda MAZ. Brazilian Journal of Analytical Chemistry, 2022 (forthcoming).
[FAPESP, CNPq]

150
A SUSPECT SCREENING APPROACH FOR THE IDENTIFICATION OF
PHTHALATES IN WATER SAMPLES
Clara V. Miranda (UG)a*, Nicole H. Lazzari (PG)a, Tânia M. Pizzolato (R)a
a Universidade Federal do Rio Grande do Sul, Instituto de Química/Departamento de Inorgânica, Porto Alegre, RS,
Brazil, 90650-001
*e-mail: claravianamiranda@gmail.com
Phthalate is the name commonly given to a group of diesters derived from the reaction between
phthalic acid, or phthalic anhydride, and alcohols. They have a variety of applications, such as
plasticizers, solvents, and additives in polymers, therefore, they are widely used in innumerous
different products and are considered omnipresent environmental contaminants1,2. When used as
plasticizers, the phthalate molecules do not covalently bond to the polymer molecules, causing them
to leach from the plastic packaging into foods and beverages, they can also be expelled into the
environment during industrial processes, and eventually be absorbed by the body due to ingestion
of contaminated food and drink, and/or contact with contaminated bodies of water 3. There are more
than 20 different phthalates, but the most commonly used are : Bis(2-ethylhexyl) phthalate (DEHP),
Benzyl butyl phthalate (BBP), Dibutyl phthalate (DBP) e Diethyl phthalate (DEP)2, 3, 4, 5 and they are
considered high-priority emerging contaminants by many countries due to the environmental and
health risks they present, since phthalates are endocrine disruptors6,7, seen as many studies have
found links between high exposure to phthalates, especially DEHP, and fertility, thyroid and obesity
issues, in both men and women and a higher frequency of endometriosis in women3, 8. Because of
this, many countries have legislations to regulate the use of phthalates and consequently, limit the
contamination they may cause. The federal regulation in Brazil9, whereas the regulation established
by the World Health Organization10 limit the maximum concentration of DEHP in drinking water as 8
μg/L, whereas the European Union limits the maximum concentration of DEHP in surface waters as
1.3 μg/L11 and the United States Environmental Protection Agency (US EPA) establishes the
maximum concentration for DEHP in drinking water as 6 μg/L12. The objective of this work is to be
able to qualitatively study the spatial distribution and presence of phthalates in the water of the
Guaíba Lake, as well as calculate the risk coefficient for the contaminants in this work. The first step
involves a pre-selection of the phthalates of interest to this work, said selection criteria is based on
phthalates studied in pre-existing literature, and from there the second step is to use post-data
samples from the Guaíba Lake which have been processed via UHPLC-Q-TOF-MS to construct the
suspect screening table that will serve as a database. The suspect screening methodology was then
applied to drinking water and river water samples, collected from the Guaíba Lake, that will be
analyzed via HS-SMPE-GC/MS.
1 Serodio, P., Nogueira, J., Water Research, 40, 2006, 13.
2 Zia, A. I., Rahman, M. S. A., Mukhopadhyay, S. C., Yu, P.-L., Al-Bahadly, I. H., Gooneratne, C. P., Kosel, J., & Liao, T.-
S., Journal of Food Engineering, 116, 2013, 2.
3 Heudorf, U., Mersch-Sundermann, V., & Angerer, J., International Journal of Hygiene and Environmental Health, 210,
2007, 5.
4 Bodzek, M., Dudziak, M., & Luks-Betlej, K., Desalination, 162, 2004, 3.
5 Ouchi, Y., Yanagisawa, H., & Fujimaki, S., Polymers, 11, 2019, 4.
6 Bila, D. M., & Dezotti, M., Química Nova, 30, 2007, 3.
7 Pallotti, F., Pelloni, M., Gianfrilli, D., Lenzi, A., Lombardo, F., & Paoli, D., Journal of Clinical Medicine, 9, 2020, 2.
8 Rowdhwal, S. S. S., & Chen, J., BioMed Research International, 2018, 2018
9 Ministério da Saúde. Gabinete do Ministro. Portaria nº 888, de 4 de maio de 2021. Brasília, 2021.
10 World Health Organization. Guidelines for drinking-water quality: Fourth edition, 21 de maio de 2022. Geneva 2022
11 European Parliament and Council. SWD/2020/251. Bruxelas, 14 de outubro de 2020
12 United States Environmental Protection Agency. National Primary Drinking Water Regulations, May 2009.
[FAPERGS, CAPES, CNPq, UFRGS, IQ/UFRGS]

151
CHEMICAL PROFILE OF AROEIRA HONEY FROM THE JEQUITINHONHA
VALLEY
Adriana Nori de Macedo (R)a*, Mayra Conceição Meira Silva (PG)b, Olívia Brito de Oliveira
Moreira (PG)c, Marcone Augusto Leal de Oliveira (R)c, André Rodrigo Rech (R)b
a Universidade Federal de Minas Gerais, Departament of Chemistry, Belo Horizonte, MG, Brazil, 31270-901
b Universidade Federal dos Vales do Jequitinhonha e Mucuri, Faculdade Interdisciplinar de Humanidades, Diamantina,
MG, Brazil, 39100-000
c Universidade Federal de Juiz de Fora, Departament of Chemistry, Juiz de Fora, MG, Brazil, 36036-900
*e-mail: adrianamacedo@qui.ufmg.br
Aroeira honey is a monofloral honey produced by Apis mellifera from the Aroeira tree (Myracrodrum
urundeuva), found in the Brazilian Dry Forest.1 This type of honey has gained attention over the last
years mainly due to its antibacterial and anti-inflammatory activities, which have been studied in
samples from the North region of Minas Gerais.1,2 As a result, the Aroeira honey Geographical
Indication has been recognized in the North of Minas Gerais, bringing commercial notoriety to the
product.2 Although not included in the Geographical Indication, the Jequitinhonha Valley is a nearby
region with similar environmental conditions, where Aroeira honey is also produced by many family
beekeepers. In this context, the objective of this study was to compare the chemical profile of Aroeira
honey produced in the Jequitinhonha Valley with the one produced in the North region of Minas
Gerais, using a metabolomics approach, in order to evaluate their similarities in terms of chemical
profile. The Aroeira honey samples included in this study were obtained from cooperatives of
beekeepers from the Jequitinhonha Valley (AJV, n = 13) and from the North of Minas Gerais (ANM,
n = 20), as well as from some non-cooperated beekeepers from the Jequitinhonha Valley (AIJV, n =
11). In addition, a few samples of multifloral honey (mel silvestre) from the Jequitinhonha Valley
(MJV, n = 3) were also included in the study as a negative control. The honey samples were weighted
(0,2 g) in microtubes and extracted with 500 µL of 1:1 (v/v) methanol:acetonitrile. After vortexing for
10 s, the mixture was sonicated for 20 min on an ice bath and centrifuged at 7000 rpm for 5 min.
The supernatant was then transferred to a chromatographic vial and 4-fold diluted with ultrapure
water. Data acquisition was performed on a 1250 Infinity Series liquid chromatographer coupled to
a 6530 Accurate-Mass QTOF mass spectrometer (Agilent Technologies), using a Kinetex F5 column
(150 x 4,6 mm, 5 µm, Phenomenex) and a mobile phase composed of methanol and water with 0,5%
formic acid. Data processing was performed on XCMS Online, with parameters previously optimized
on the R-package IPO (Isotopologue Parameter Optimization). Uni- and multivariate statistical
analyses were performed on MetaboAnalyst 5.0. The principal component analysis (PCA) scores
plot revealed a clear overlay between the three groups of Aroeira honey (ANM, AJV and AIJV),
whereas the multifloral honey (MJV) formed a separate group. Similarly, the hierarchical cluster
analysis (HCA) was able to distinguish the multifloral honey from the Aroeira honeys, whereas no
identifiable clusters were formed among samples from ANM, AJV and AIJV. In the univariate
analysis, only 5 and 16 molecular features were found to be different in the two-by-two comparisons
of ANM x AJV and ANM x AIJV, respectively, from a total of about 4500 molecular features
evaluated. These results indicate that Aroeira honey from the North of Minas Gerais and from the
Jequitinhonha Valley are mainly indistinguishable from each other based on their chemical profiles,
which helps to support the possible expansion of the Aroeira honey Geographical Indication, in order
to include the Jequitinhonha Valley within the region already recognized.
1 Viana FR, Carmo LS, Bastos EMAF, Acta Scientiarum – Biological Sciences, 40, 2018, e36766.
2 Demier ADM, Doces matas do Norte de Minas: Atores, instituições e a obtenção do registro de Indicação Geográfica do
mel de Aroeira, Dissertação (Mestrado), UFMG, Unimontes, 2018.
[UFMG, UFVJM, COOAPIVAJE, COOPEMAPI, MCTI/FINEP, FAPEMIG, CNPq, CAPES]

152
CHEMICAL PROFILING OF PROPOLIS FROM THE AMAZON REGION
Brícia Marques Parreiras (UG)a*, André Rodrigo Rech (R)b, Adriana Nori de Macedo (R)a
a Universidade Federal de Minas Gerais, Department of Chemistry, Belo Horizonte, MG, Brazil, 31270-901
b Universidade Federal dos Vales do Jequitinhonha e Mucuri, Faculdade Interdisciplinar de Humanidades, Diamantina,
MG, Brazil, 39100-000
*e-mail: briciamp@hotmail.com
Propolis, also known as bee glue, is a natural resin produced by bees, which has been used for
thousands of years in folk medicine, due to its antioxidant, anti-inflammatory and antibacterial
properties. Propolis fulfills an important function in the beehive, where it is used for protection,
temperature control and promotion of a sterile environment1. These biological properties depend on
the chemical composition, which is complex and varied, depending on the region, bee species, as
well as the local flora. Due to this variability in composition and properties, the propolis are classified
into different types. However, there are only a few studies on the physicochemical characteristics of
the propolis produced in the Amazon region. The present work intended to study the chemical profile
of propolis samples from the Amazon, comparing it with other type of propolis from the southeastern
region of Brazil, named green propolis. Four samples of Amazonian propolis and five samples of
green propolis were analyzed. The extracts were prepared by weighting 50 mg of each propolis
sample and adding 2 mL of HPLC grade methanol in a screw capped test tube, followed by
sonication for 30 min and overnight incubation at -20 oC. The supernatant was transferred to a
microtube and centrifuged at 7000 rpm for 10 min. The supernatant produced in this step was
transferred to another microtube and kept at -20 oC until the analysis. A pooled sample was prepared
by mixing equal volumes of the 9 samples. Mass spectra were obtained by paper spray ionization
mass spectrometry (PSI-MS), using an ion trap analyzer, model LCQ Fleet (Thermo Scientific) in the
m/z range of 100 - 2000 in positive and negative ion modes. The chromatographic paper used in the
technique was cut in the shape of isosceles triangles, with a base of 1.0 cm and a height of 1.5 cm
and attached to an alligator clip at 5 mm from the mass spectrometry entrance. For analysis, 10 µL
of the sample and 30 µL of HPLC grade methanol were transferred directly to the paper. Samples
were analyzed in triplicate, in random order, interspersed with a blank (solvent only) in order to
monitor the occurrence of possible carry-over during the analyses. A data matrix was obtained by
computing the average relative intensity per m/z for each sample and removing values with relative
intensity below 10%, in order to minimize the contribution of background noise. Missing values were
replaced by one-fifth of the minimum detected value. The acquired data were analyzed using
multivariate statistics after mean-centering. Principal components analysis (PCA) and a dendrogram
were obtained using MetaboAnalyst 2.0 and Jupyter Notebook. As a result, the analysis showed a
good separation between Amazonian propolis and green propolis for the negative mode, which was
not observed for positive mode. This indicates that Amazonian propolis has a distinct composition
from green propolis, especially when considering the components preferentially ionized in negative
mode. Some compounds were tentatively identified by consulting the literature2 and will be further
studied using fragmentation mode in mass spectrometer. The results of this study will serve as a
basis for future studies about the biological applications of this scarcely studied type of propolis,
providing more information to Amazonian communities involved in the production and
commercialization of local propolis.
1 Saleh K, Zhang T, Fearnley J, Watson DG, Current Metabolomics, 2015, 3.
2 Costa A, Yoshida N, Garcez W, Perdomo R, Matos M, Garcez F, Journal of Natural Products, 2020, 83.
[UFMG, CNPq, CAPES, FAPEMIG]

153
QUANTIFICATION OF THE AMPHENICOL AND FLORFENICOL AMINE
METABOLITE IN MATRICES OF ANIMAL ORIGIN BY LC-MS/MS
Diana A. de Sousa (R)a*, Fabiano Barreto (R)a, Daniel Hillesheim (R)a, Caroline Tomaszewski
(R)a
a Ministry of Agriculture, Livestock and Food Supply, Federal Laboratory of Animal and Plant Health and Inspection,
Porto Alegre, Rio Grande do Sul, Brazil, 91780-580.
*diana.sousa@apoio.agro.gov.br
A quick and simple method for the determination of amphenicols and the florfenicol amine metabolite
(FFA) in products of animal origin1 is being developed for concentration levels in accordance with
the Maximum Residue Limits (MRL) established in NORMATIVE INSTRUCTION N° 51, DE
DECEMBER 19, 2019 (ANVISA). FFA is the main metabolite of florfenicol (FF) in poultry, swine and
cattle and is also considered a marker residue for the determination of FF in products of origin of
these species.2,3 Liquid chromatography coupled with mass spectroscopy detection (LC-MS/MS)
was used to detect and quantify the analytes. The extraction was performed with ethyl acetate4 and
reconstituted in hexane with solvent (water and methanol) in the respective proportion 85:15. The
extracts were analyzed in a chromatographic method in reverse phase with the Synergi Polar-RP
column. Analytical limits were set to 0.5ng/ml (limit of quantitation) to TAP, FF and FFA. CAP
presented 0.075ng/ml (limit of quantitation). The best results were obtained with an equilibration time
of 9 min and a flow of 300ml/min. For the bovine milk matrix, the coefficient of correlation (R) is
greater than 0.9792762 (FFA). FFA was the only compound that showed a low recovery of 12.81%.
The other analytes obtained 62.84% (TAP), 89.88% (CAP) and 102.49% (FF). Other merit values
such as precision, veracity, decision limit (CCα), method capability (CCβ), matrix effects, stability
and measurement uncertainty will be evaluated in the analytical validation process for other matrices
as well.
1 Hoff RB, Molognoni L, Deolindo CTP, Vargas MO, Kleemann CR, Daguer H, Journal of Chromatography B, 1152, 2020
2 Imran M, Habib F, Majeed S, Tawab A, Rauf W, Rahman MU, Iqbal M, Food Additives & Contaminants: Part A, 2018.
3 Jung YS, Kim Dan-Bi, Nam TG, Seo D, Yoo M. Food Chemistry, 382, 2022.
4 Barreto F, Ribeiro C, Hoff RB, Dalla C T, Journal of Chromatography A, 1449, 2016.
[MAPA, CNPq]

154
TOWARDS SINGLE CELL ICP-MS NORMALIZED QUANTITATIVE
EXPERIMENTS USING CERTIFIED SELENIZED YEAST
Juliana S. F. Pereira (R)a*, Roberto A. F. García (PG)b, M. Corte-Rodríguez (PG)b, A. Mateca
(R)c, J. Bettmer(R)b, K. L. LeBlanc (PG)d, Z. Mester (R)d, M. Montes-Bayón (R)b
a Universidade Federal do Rio Grande do Sul, Instituto de Química, Porto Alegre, RS, Brazil, 91501-970.
b University of Oviedo, Dept. of Physical and Analytical Chemistry and Instituto de Investigación Sanitaria del Principado
de Asturias (ISPA), Oviedo, Spain, E-33006.
c University of Oviedo, Faculty of Biology, Dept. Functional Biology and Instituto de Investigación Sanitaria del Principado
de Asturias (ISPA), Oviedo, Spain, E-33006.

d National Research Council Canada, Ottawa, Ontario, Canada, K1A OR6.
*e-mail: julianasfp@ufrgs.br
Selenium enriched yeast or selenized yeast has been widely used as a nutritional supplement in the
case of Se deficiency and also due to its potential nutraceutical product against some types of
cancer.1 The evaluation of the selenium species produced in yeast cells has been approached
through different analytical methods that tried to achieve a quantitative mass balance with respect
to the total incorporated Se.2 In fact, certified reference materials (CRM) have been brought to the
market with the aim of validating analytical procedures for the quantitative characterization of the
total Se content in selenized yeast. SELM-1, produced by the National Research Council Canada
(NRC), is certified for total Se as well as Se-methionine, the most abundant Se metabolite, content.3
However, little is known regarding the characterization of the yeast material at the individual cell
level. Nowadays, due to the widespread use of the single cell analytical techniques, this study can
be feasible to achieve a full characterization of the selenized yeast material. In order to obtain a
normalized procedure for single cell inductively coupled plasma mass spectrometry (SC-ICP-MS)
measurements, the quantitative characterization of the intra- and extracellular Se fractions of a CRM
of Se-enriched yeast (SELM-1) has been carried out. Different sample preparation procedures have
been studied and best results were observed with a sequential washing procedure to isolate the cells
from the extracellular Se content through which two fractions were obtained and analyzed by ICP-
MS (washing solution and washed/dried cells). After proving the cell integrity throughout the washing
and drying processes by confocal microscopy, the Se intracellular fraction was determined by SC-
ICP-MS as well as by acid digestion using microwave digestion. The extracellular Se concentration,
has been directly determined in the washing solutions. Regarding quantitative results, the sequential
procedure involving cell washing followed by the freeze drying of the washed material and
intracellular Se quantification using SC-ICP-MS provided best results instead of directly analysis of
suspended cells by dilution in water. In this case, intracellular Se accounted for 1304 ± 48 mg kg -1
(corresponding to 64% of the certified Se content). The average mass of Se per yeast cell was 41.6
fg Se with a dispersion of 1.6 to 279 fg Se/cell. In the isolated extracellular Se fraction, the Se
concentration accounted for 412 ± 48 mg kg-1 (about 21% of the total Se). Thus, the sequential
procedure provided a total Se recovery of about 85% with respect to the certified value. The direct
dilution and simultaneous measurement of intra and extracellular Se by SC-ICP-MS provided results
of 1024 ± 42 mg kg-1 for intracellular and 316 ± 30 mg kg-1 for extracellular Se representing a total
recovery of about 66%. These results clearly demonstrated that with appropriate sample preparation,
SC-ICP-MS is a unique tool, which is capable of providing quantitative information about intracellular
and extracellular Se. In addition, SELM-1 seems the ideal tool to enable data normalization at the
single cell level to replicate, benchmark, and improve new SC-ICP-MS studies by using the same
material for data validation.
1 Rayman MP, Br. J. Nutr. 92, 2004, 557.
2 LeBlanc KL, Mester Z, Metallomics. 13, 2021, https://doi.org/10.1093/mtomcs/mfab031.
3 Mester Z, Willie S, Yang L, Sturgeon R, Caruso JA, Fernández ML, Fodor P, Goldschmidt RJ, Goenaga-Infante H,
Lobinski R, Maxwell P, McSheehy S, Polatajko A, Sadi B.B.M., Sanz-Medel A, Scriver C, Szpunar J, Wahlen R, Wolf R.,
Anal. Bioanal. Chem. 385, 2006, 168.
[CAPES, ISPA, UNIOVI]

155
TRACKING THE IONIC AND NANOPARTICULATED TITANIUM RELEASED IN-
VIVO FROM METAL DENTAL IMPLANT USING (SINGLE-PARTICLE)-ICP-MS
Diogo Pompéu de Moraes (R)a*, Sara Gonzáles-Morales (PG)b, Jorge Toledano-Serrabona
(PG)c, M. Ángeles Sánches-Garcés (R)c, Mario Corte-Rodríguez (PG)b, Jörg Bettmer (R)b,
María Montes-Bayón (R)b
a UniversidadeFederal do Rio Grande do Sul (UFRGS), Institute of Chemistry, Porto Alegre, RS, Brazil, 91501-970
b Universityof Oviedo, Department of Physical and Analytical Chemistry, Oviedo, Spain, 33006
c University of Barcelona, Department of Oral Surgery and Implantology, Barcelona, Spain, 08907
*e-mail: diogo.moraes@ufrgs.br
Despite their wide use and tolerability, titanium-based prosthetic implants of all kinds may develop
some problems leading to implant rejection. In particular, dental implants are especially prone to
bacterial growth and infection causing several implant-related diseases, such as peri-implantitis. The
peri-implant bone-loss that may result from these diseases or the surgical treatments to overcome
them, can cause the implant surface to be exposed to the oral environment with an increased risk of
bacterial growth, infection and eventually implant rejection. Implantoplasty consists of polishing and
smoothing the dental implant exposed surface, reducing bacterial adhesion, and facilitating hygiene
by the patient. However, implantoplasty produces a great amount of metal debris, part of them
macroscopic that can be easily removed from the surrounding tissues, but part of them in the micro-
and nanometer scales, which are impossible to completely eliminate from the tissues and will stay
there after the treatment, affecting the surrounding tissues and potentially producing systemic
effects.1 In this animal study, ICP-MS was applied to study the release and biodistribution of Ti, Al
and V from implant debris implanted in rats, by acid-digesting the blood, liver, spleen, lung and brain
of the animals. For this aim, sample preparation for total Ti, Al, and V determination was carefully
optimized using microsampling inserts to minimize dilution during the acid attack of the lyophilized
tissues. The presence of a small percentage (0.005%) of hydrofluoric acid revealed to be mandatory
for quantitative recovery (> 85%) of the ionic Ti in the spiked samples analyzed using ICP-TQ-MS.
Additionally, since implantoplasty may also produce nanoparticulate debris, single-particle-ICP-MS
(sp-ICP-MS) and other complementary techniques were used to search and characterize titanium
nanoparticles in the different tissues that may have been transported from the implant site. In these
organs, the possible presence of Ti-containing nanoparticles mobilized from the dental implants was
tested using enzymatic digestions. Applying proteinase K in the presence of sodium dodecyl sulfate
(SDS) as solubilization media resulted in better reproducibility and highest recoveries. The presence
of Ti-containing nanoparticles was observed in all the analyzed tissues, however, differences on the
Ti mass per particle were found between the blanks and the enzymatic digested tissue and between
control and experimental animals in some organs. Although the significant content of
nanoparticulated Ti in control rats from environmental sources hampered the correct determination
of the nanoparticles with an origin in the implanted debris, the mass of Ti in the obtained events
seemed to point out the presence of different Ti nanoparticles in brain and spleen samples
distinguishable to those in the control counterparts. The removal of this kind of particles from the
solutions used for the sample preparation represents the next challenge to achieve such kind of
studies. For Al and V concentrations in the different organs no significant differences were observed
between control and experimental groups with a confidence level of 95% (t-test, p<0.05), with
average concentration of 1000 and 10 ng g-1, respectively.
1 Barrak FN, Li S, Muntane AM, Jones JR, Int. J. Implant Dent. 6, 2020, 50.
[CAPES/Print, CNPq]

156
NAPHTHENIC ACIDS CHEMICAL SPECIATION BY ORBITRAP-HRMS
Thamara Andrade Barra (PG)a*, Gleicielle Tozzi Wurzler (R)a, Vinícius Barreto Pereira
(PG)a*,Gabriela Vanini (R) a, Alexandre de Oliveira Gomes (R)b, Débora de Almeida Azevedo
(R)a
a Federal University of Rio de Janeiro, Institute of Chemistry, Ladetec, Rio de Janeiro, RJ, Brazil, 21941-598
b Petrobras-CENPES, Ilha do Fundão, Rio de Janeiro, RJ, Brazil, 21941-909
*e-mail: thamara.barra@iq.ufrj.br
The study of naphthenic acids (NAs) has been receiving particular attention from refining technology
point of view since it belongs to petroleum, a natural complex matrix1 of great economic interest, and
the cost-benefit ratio is a factor that directly impacts the oil industry. Besides that, regarding analytical
issues, NAs are relevant due to their very low concentrations in petroleum. The characterization of
NAs in petroleum samples represents an analytical challenge and industry demand for some specific
crude oils, mainly because the matrix’s complexity. Data obtained by Orbitrap high-resolution mass
spectrometry (Orbitrap-HRMS) technique is very interesting to the possibility of drawing the
relationship with TAN (total acid number) value and NAs, contributing to increasing knowledge at the
molecular level of complex crude oil samples. Thus, this study aims to characterize NAs in real
Brazilian crude oils by Orbitrap-HRMS, by assessing non-volatile polar compounds. Four Brazilian
crude oils were provided by CENPES/Petrobras SA and studied: 1) P1 - 22.2 °API, TAN = 1.25
mgKOH g-1; 2) P2 - 19.8 °API, TAN = 2.39 mgKOH g-1; 3) P3 - 29.9 °API, TAN = 0.22 mgKOH g-1;
and 4) P4 - 29.8 °API, TAN = 0.24 mgKOH g-1. The crude oils were weighed (ca. 1.0 mg) and diluted
with 1 mL of a mixture of deuterated carboxylic acid standards at 50 ng mL -1 for ESI(-)-Orbitrap
HRMS analyses. The ESI(-)-Orbitrap HRMS analyses were performed similarly as already described
elsewhere2. This study investigated the carboxylic acids that contribute to the acidity of the four crude
oils. The data obtained via Orbitrap-HRMS expresses that a higher variability of molecular formulas
with a large range of double bond equivalents (DBE) detected for P1 and P2, while the P3 and P4
oils showed a high percentage of linear carboxylic acids. Linear and monocyclic acid compounds
were identified for the four crude oils, and tricyclic acid compounds were identified for the lower °API,
P1 and P2 crude oils. In this study, monocyclic and linear acid molecules in P1 and P2 crude oils
with high TAN values indicated to be more related with TAN than linear acid molecules for the P3
and P4 crude oils with lower TAN values. The TAN value is defined as the mass of KOH (mg)
required to neutralize 1 g of crude oil. In the present case study and set of samples, the total O2
class concentration (mg goil−1) obtained by ESI(−)−Orbitrap−HRMS was converted to the TAN
equivalent value (TANequiv). The TANequiv value was obtained by converting the O 2 class
molecular weight relative concentration to the amount of KOH (mg) equivalent in the acid−base
neutralization reaction. The results present a good correlation with the TAN values (R2 = 0.9501,
y=1.0946x−0.0235), and the slope close to 1 express that the TAN and TANequiv values vary in
very similar proportions to each other. The ESI(-)-Orbitrap-HRMS results exhibited an O2
heteroatomic species predominance pattern in the crude oils studied. There is a great relationship
between TAN and TANequiv obtained from ESI(-)-Orbitrap-HRMS results considering the four oils
studied.
1 Tissot BP, Welt DH. Petroleum formation and occurrence. Second ed. Berlin: Springer-Verlag, 1984.
2 Vanini G, Barra TA, Souza LM, Madeira NCL, Gomes AO, Romão W, Azevedo DA. Fuel, 282, 118790, 2020.
[CNPq, FAPERJ, CAPES, Petrobras]

157
A FAST-GC×GC-TOFMS METHOD DEVELOPMENT FOR ENVIRONMENTAL
GEOCHEMISTRY STUDIES
Raquel V.S. Silva (R)*, Rafael S. Xavier (UG), Vinicius B. Pereira (PG), Ananda S. Antonio (R),
Débora A. Azevedo (R)
Federal University of Rio de Janeiro, Institute of chemistry, Rio de Janeiro, RJ, Brazil,21941-598
*e-mail: raquelvieira@iq.ufrj.br
Despite the innumerous advantages of comprehensive two-dimensional gas chromatography

coupled to time-of-flight mass spectrometry in the deconvolution of complex matrices, analyses by
GC×GC-TOFMS are time-consuming, which causes expenses related to consumables usage and
data archiving. However, fast-GC×GC has been a promising alternative to achieve chemical
characterization of environmental matrices1. Furthermore, the use of lower ionization energies
enables the identification of trace compounds, typically analyzed in environmental matrices, by
promoting lower fragmentation and therefore increasing molecular ions intensity2. This work focuses
on the comparation between a conventional method with a 1D column of 30 m and the application
of a 2² factorial design with central point in a set of fast methods with a 1D column of 10 m for the
analysis of a sedimentary organic extract from the Garça lagoon localized in National Park of
Restinga of Jurubatiba (RJ, Brazil) for environmental geochemistry purposes. Moreover, different
ionization energies (50, 60 and 70 eV) and acquisition rates (100, 200 and 300 Hz) were evaluated
aiming to increase molecular ion identification of biomarkers. Statistical tests were applied evaluating
as response variables the relation signal/noise of diagnostic and molecular ions, as well as the
concentration obtained for each class in their respective assays. Analyses were performed in a
Pegasus 4D (Leco, USA). The analyzes performed by the conventional method allowed obtaining
95% confidence interval values for the total concentration of each one of the classes, and relative
standard deviations of up to 10%. As expected, chromatographic resolution decreased using fast-
GC×GC method, but the selectivity remained practically constant and above 1, not generating
difficulties for the identification of compounds with very close retention times. Thus, the efficiency of
the proposed chromatographic method was satisfactory for most of the analytes, providing a shorter
analysis time and good chromatographic resolution values. The factorial experiment showed that the
lower ionization energies applied were not sufficient to promote significant increases in molecular
ion intensity, but promising results showed increased ratios compressing molecular ions and
diagnostic ions. The comparison between fast methods by Pareto charts using the S/N ratio as the
response indicated that only the voltage variable was significant for obtaining better S/N ratios of
diagnostic ions for the class of steroids, which was not observed as much for the other classes, and
for the molecular ion S/N ratio. For most of the analyzed classes, the trend surface method indicated
that the most optimized experimental condition for the S/N ratios of diagnostic and molecular ions
corresponds to 70 eV and 300 Hz. Quantification of compounds was compared using analysis of
variance (p<0.05), and the methods using 70 and 50 eV are statistically equivalent to the
conventional method for all analyzed classes. The results showed that the change from a
conventional method to a fast one resulted in a decrease in the analysis time from 60 to 20 min,
keeping the chromatographic resolution at values suitable for an efficient separation and
identification. In addition, the mass acquisition optimization revealed that the fast method using 70
eV and 300 Hz was the most adequate and statistically equivalent to the conventional one.
1 Schena T, Bjerk, TR, von Mühlen, C, Camarão EB, Talanta, 219, 2020, 121-136.
2 Dubois, LM, Perrault KA, Stefanuto PH, Koschinski S, Edwards M, Mcgregor L, Focant JF, Journal of Chromatography
A, 1501, 2017, 117-127.
[CNPq, FAPERJ]

158
ELEMENTAL DETERMINATION IN E-LIQUID SAMPLES USING ICP-MS
Guilherme V. Espinosa (UG)*, Tatiana D. Saint’Pierre (R)
Pontifícia Universidade Católica do Rio de Janeiro, Departamento de Química, Rio de Janeiro, RJ, Brazil, 22451-900
*e-mail: guiespin@aluno.puc-rio.br
Tobacco is one of the global main causes of premature death and smokers lose at least a decade
of life expectancy, compared to those who never smoked.1 Electronic Smoking Devices (ESDs), also
called e-cigarettes, which usually consist of a battery, heating element and a cartridge for the e-
liquid,2 an aqueous solution containing glycerol, propylene glycol, nicotine and flavors,3 is
considered a safer alternative to regular smoking, specially due to the absence of a combustion
reaction, to which most of the risks of smoking is associated. However, this assessment is
controversial, since the ESDs’ long-term effects are still insufficiently studied.4 Among other health
risks, e-cigarettes are sources of trace elements, potentially carcinogenic,5 derived from both the
device and the e-liquid.6 In this context, the present work investigates the presence and
concentration of trace elements in e-liquid, using concentration limit guidelines for inhaled drug
substances and excipients7 to evaluate potential health risks. While the concentrations determined
are smaller than such limits, the trace elements contained in the e-liquid can’t be considered
harmless to the health of the user, since the concentration increases in the aerosol, coming from the
device.8 It also aims to study the accuracy of the results obtained using the sample dilution in
ultrapure water, without the use of HNO3 as it’s usually prepared, to avoid the possible generation
of dangerous compounds such as propylene glycol dinitrate (PGDN) and nitroglycerin. 9 After an
initial semiquantitative analysis, the analytical method to determine Al, Ba, Co, Cu, Li, Mn, Ni, Pb,
Rb, Sb, Ti, Tl, V, and Zn by inductively coupled plasma mass spectrometry (ICP-MS) was developed,
by using five samples of e-liquid, of brands originated from the USA, Brazil and Paraguay, simply
diluted in ultrapure water. To correct non-spectral interferences, 103Rh was used as internal standard,
and external and standard addition calibration methods were used to evaluate such correction by
comparing the slopes using Student t-test. Each analysis was made in triplicate. Recovery studies
were performed to evaluate the method’s accuracy and the figures of merit were calculated using
the appropriate calibration method. The comparison of the calibration methods showed that the non-
spectral interferences for Ni, Ti and Zn were not significant, while for the other elements standard
addition calibration should be used. Concentrations above the method’s limit of quantification were
obtained, except for Pb, for which a smaller dilution should be used. The results stayed in the range
of 0.01 µg L-1 for Tl to 200 µg L-1 for Zn. The recovery experiments results were between 80 % to
110 %, ratifying the accuracy of the analytical method. Even though for most analytes the external
calibration was considered inappropriate, it generates accurate enough results, compared with the
standard addition calibration, to be used to estimate the real concentrations. Since the sale of ESDs
is forbidden in Brazil, the samples lack quality control and compositions vary even among products
from the same batch. Therefore, such estimated values are appropriate to evaluate the health risks
originated from the e-liquid and, since it’s responsible for only a fraction of the trace element
exposure, works quantifying such increase and reproducing the impurities effects in human body are
necessary to correctly protect users and potential users.
1 Jha P, Ramasundarahettige C, Landsman V, Rostron B, Thun M, Anderson RN, McAfee T, Peto R, New England Journal
of Medicine 2013, 368, 341.
2 Young SE, Henderson CA, Couperus KS, Current sports medicine reports 2020, 19, 146.
3 Margham J, McAdam K, Forster M, Liu C, Wright C, Mariner D, Proctor C, Chemical Research in Toxicology 2016, 29,
1662.
4 Marques P, Piqueras L, Sanz MJ, Respiratory Research 2021, 22, 1.
5 Hess CA, Olmedo P, Navas-Acien A, Goessler W, Cohen JE, Rule AM, Environmental Research 2017, 152, 221.
6 Ward AM, Yaman R, Ebbert JO, PLoS ONE 2020, 15, 1.
7 Kamilari E, Farsalinos K, Poulas K, Kontoyannis CG, Orkoula MG, Food and Chemical Toxicology 2018, 116, 233.
8 Williams M, Villarreal A, Bozhilov K, Lin S, Talbot P, PLoS ONE 2013, 8, 1.
9 Fettaka H, Lefebvre MH, Central European Journal of Energetic Materials 2016, 13, 199.

159
DEVELOPMENT OF A CHEMOTYPING METHOD BASED ON PHYTOCHEMICAL
MARKERS OF BRAZILIAN WOOD SPECIES
Thays V. C. Monteiro (R)a*, Antônio Unias (R)a, Ana Baddini (R)b, Alexandre Gontijo (R)c,
Werickson F. Rocha (R)a, João Latorraca (R)d, Álex A. Rosini (R)e, Salvador Sánches (R)e,
Andréia Porcari (R)e, Maíra Fasciotti (R)a
a Instituto Nacional de Metrologia, Qualidade e Tecnologia – INMETRO, Duque de Caxias, RJ, Brazil, 25250-020
b Instituto Federal do Rio de Janeiro – IFRJ, Rio de Janeiro, RJ, Brazil, 24315-375
c Laboratório de produtos florestais – LPF, Brasília, DF, Brazil, 70818-900
d Universidade Federal Rural do Rio de Janeiro – UFRRJ, Seropédica, RJ, Brazil, 23897-045
e Universidade de São Francisco – USF, Bragança Paulista, SP, Brazil, 12916-900
*e-mail: tvmonteiro@inmetro.gov.br
Illegal logging is a dramatic problem in Brazil, as the country concentrates some of the greatest
biodiversity and rich biomes in the world, such as the Amazon biome. Indiscriminate deforestation
has encouraged the government to create federal laws listing endangered wood which logging is
prohibited. These species, although protected by law, are still the target of illegal trade because they
are considered “noble” and commercially coveted. Despite efforts to fight illegal logging based on
laws, in practice, the systems fail to effectively be used against the marketing of wood products made
from illegal timber. As wood identification is technically difficult, this project aims to develop a method
at INMETRO to establish a methods to differentiate the wood species. A powerful tool is the use of
chemical profiles based on secondary metabolites, analysis called chemotyping, which are basically
molecular fingerprints according to genus and species. In this study, 61 different wood species and
their replicates were used, totaling 187 samples. The samples were extracted with MeOH/H2O 3:1
with the addition of 0.05 mol L-1 sodium bicarbonate aqueous solution and directly infused into the
ESI ion source of a Synapt HDMS QTOF mass spectrometer. The method was qualitatively
validated, evaluating parameters such as: repeatability, intermediate precision and selectivity.
Comparing the spectra, very characteristic profiles were observed for different samples and great
similarity between replicates of the same species, however there were some inconsistencies. Due
to the large amount of data, different statistical treatments were tested. An exploratory data analysis
was performed by PCA. Later, the construction of PLS-DA models for classifying samples of
Caryocar spp., Copaifera spp. and Eucaliptus spp., and when performing the external validation, the
models showed 100% selectivity. Some distance calculation models were evaluated, with the help
of an application developed for this work, and the angular distance model showed the best results.
Another approach used was the comparison of spectra by spectral similarity, obtaining similarities
greater than 88% for samples of the same species. In this way, it was possible to prove that the
method is capable of classifying and identifying different species of wood, which demonstrates that
the method is promissing to be employed for the control of illegal logging.

160
INSTRUMENTATION AND
AUTOMATION
161
3D-PRINTED ELECTROCHEMICAL PLATFORM WITH MULTI-PURPOSE
CARBON BLACK-POLYLACTIC ACID ELECTRODES
Habdias A. Silva-Neto (PG)a,*, Anderson A. Dias (R)a and Wendell K. T. Coltro (R)a,b
a UFG, Instituto de Química, Goiânia, GO, Brazil, 74690-900
b Instituto Nacional de Ciência e Tecnologia de Bioanalítica, Campinas, SP, Brazil, 13084-971
*e-mail: habdiasaraujo@gmail.com.br
In the last five years, different authors have explored 3D printing to create electrochemical platform
integrated with commercial or 3D printed electrodes, using filaments composed of acrylonitrile-
butadiene-styrene (ABS), polylactic acid (PLA) and PLA doped with carbon black (CB/PLA) or
graphene.1,2 However, the current state-of-the-art related to 3D printed electrochemical sensors
does not include analytical systems with multiple integrated electrodes. This report describes the 3D
printing of a complete system comprising a batch injection analysis (BIA) cell and an electrochemical
platform with eight sensing electrodes. Both BIA and electrochemical cells were printed within 3.4h
using a multi-material printer equipped with insulating, flexible and conductive filaments at cost of
ca. ~ U$ 1.2 per unit (Figure 1A). Printed electrodes were exposed to electrochemical/Fenton pre-
treatments to improve the sensitivity. Scanning electron microscopy and electrochemical impedance
spectroscopy measurements upon printed materials revealed high-fidelity 3D features (90 to 98%)
and fast heterogeneous rate constants ((1.5 ± 0.1)  10-3 cm s-1). The best analytical performance
was achieved using a dispensing rate of 9.0 µL s-1 and injection volume of 2.0 µL. The proof-of-
concept of the printed device for bioanalytical applications was evaluated using adrenaline (ADR) as
target analyte (Figure 1B) and its redox activities were carefully evaluated through different
voltametric techniques upon multiples 3D printed electrodes. The coupling of BIA system with
amperometric detection ensured fast responses with well-defined peak width related to the oxidation
of ADR applying a potential of 0.4 V vs Ag. The fully 3D printed system provided suitable analytical
performance in terms of repeatability and reproducibility (RSD ≤ 6%), linear concentration range (5
to 40 µmol L-1; R2 = 0.99), limit of detection (0.61 µmol L-1) and high analytical frequency (494 ± 13
h-1). Lastly, artificial urine samples were spiked with ADR solutions at three different concentration
levels and the obtained recovery values ranged from 87 to 118%, thus demonstrating potentiality for
biological fluid analysis. Based on the analytical performance, the complete device fully printed
through additive manufacturing technology emerges as powerful, inexpensive, and portable tool for
electroanalytical applications involving biologically relevant compounds.
(A) (B)
Figure 1. (A) Illustration process of fabrication and assembling of electrodes. (B) Schematic representation of
electrochemical oxidation of ADR.
1 Muñoz J, Pumera M, TrAC - Trends in Analytical Chemistry, 128, 2020

2 Richter EM, Rocha DP, Cardoso RM, et al, Analytical Chemistry 91, 2019, 12844
[UFG, CNPq, CAPES, LabMic, INCTBio]
–––––––––––––––––––– Instrumentation and automation ––––––––––––––––––––

162
A FLOW-BASED PROCEDURE FOR EXTRACTION AND DETERMINATION OF
TOTAL PHENOLIC COMPOUNDS IN SOLID SAMPLES
Kethlin F Curiel (PG)a*, Wanessa R. Melchert (R)b
a Center for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil.
b College of Agriculture “Luiz de Queiroz”, University of São Paulo, Piracicaba, SP, Brazil.
*e-mail: kethlin.curiel@usp.br
Multicommutation allows increasing versatility of the flow systems by means of discrete commuting
devices, such as solenoid micro-pumps and valves. The flow system was designed with four solenoid
micro-pumps (20 µL / pulse) in order to improve mixing conditions and minimize reagent
consumption as well as waste generation and quantification of total phenolic compounds (CPT) were
based on the spectrophotometric measurement by the Folin-Ciocalteu method. The extraction
module was constructed with a 1000 µL pipette tip, a frit and cotton used as a filter (Figure 1).
The fluidized bed column was under constant stirring
using a vibratory shaker for a better interaction between
the extraction solution (ethanol 5% (v/v)) and the solid
samples (0.05 g). The extraction of CPT was started by
pumping extraction solution through P1 (150 pulses). The
analytical cycle was started by pumping extraction
solution and reagents through P1 (4 pulses) and P3
(Folin-Ciocateu 20% (v/v); 1 pulses) and P4 (2% sodium
carbonate (w/v); 1 pulses) pumps, respectively. These
aliquots underwent fast mixing at the interfaces
establishing the first sampling cycle. The sequence was
repeated 7 times to form the sample zone and was
carried out towards detection at 740 nm by actuation of
P2. The analytical signal was based on peak height and
measurements were taken in triplicate. Under optimized
experimental conditions, a linear response was observed
Figure 1. Flow diagram of the system for CPT -1
extraction (A) and quantification (B). P1-P4: from 1 to 20 mg L gallic acid, as described by the
solenoid micro-pumps; VB: vibratory shaker; Col: equation Absorbance = 0,02966 + 0,03631 C; (r = 0,997).
column; SE: extraction solution; Ca: carrier The detection limit (99.7% confidence level), the
stream; R1-R2: reagents; R: 170-cm coiled reacto; coefficient of variation (n=11; 10 mg L-1), and the
D: spectrophotometer (740 nm); V: solenoid
valve; x: Perspex joint point and W: waste vessel.
sampling rate were estimated at 33 µg L-1, 0.4% and 120
determinations h-1, respectively. Reagent consumption
per determination was estimated at 8.36 µL of Folin-Ciocateu and 3.35 mg of sodium carbonate,
generating 4 mL of waste. Fermented grape residue samples were employed, then analyzed using
the optimal operational conditions selected for the proposed procedure and 28.62 ± 2.26 mg of gallic
acid per 100 g of sample was quantified. The accuracy of the measurements was assessed by
comparing the results using a reference method based on the spectrophotometric method. Typical
characteristics of systems using solenoid micro-pumps, as portability, low cost and low energy
consumption, improved mixing and reproducibility for handling micro-volumes of solutions were also
aggregated.
1 Silva ABP, Souza CSG, Paim SPA, Lavorante FA, J. Chil. Chem. Soc., 63, 2, 2018.
2 Rocha PD, Cardoso MR, Mendonça MHD, Richter ME, Silva GS, Batista DA, Munoz AAR, Electroanalysis 2018, 30,
180.
[CAPES, FEALQ]

163
A FULLY AUTOMATED, MINIATURIZED, AND LOW-COST IOT ANALYTICAL
PLATFORM FOR THE MONITORING OF NITROGEN DIOXIDE IN THE
ATMOSPHERE
Vanderli G. Leal (PG)*, João Flávio S. Petruci (R)a
a Federal University of Uberlândia, Institute of Chemistry, Uberlândia, Minas Gerais, Brazil, 38400-902
*e-mail: vanderlileal@gmail.com
In the troposphere, nitrogen dioxide (NO2) is mainly produced by the burning of biomass and fossil
fuels1. NO2 plays an important role in the formation of tropospheric ozone. increasing air pollution
and toxicity in plants1.NO2 is an urban pollutant that generates particulate matter, causing people to
breathe air with high levels of insalubrity. Also, NO2 is associated with respiratory diseases,
especially asthma, increasing hospital admissions in emergencies, indicating significant damage
even at low concentrations2. Despite its high pollutant behavior, only a very few cities in Brazil
monitor the NO2 levels. Therefore, portable and low-cost analytical platforms must be developed to
contribute to data acquisition regarding the concentration levels of air pollutants in the troposphere
of urban cities. In this work, an automated, miniaturized, and portable 3D printed IoT analytical
platform was developed for the determination of NO2 in atmospheric air. The platform was designed
using CAD software (Autodesk Inventor) and fabricated using a 3D printer (GTMax A2 Core). The
determination of NO2 was based on the bubbling of atmospheric air into a vial containing 800 µL of
the Griess-Saltzman reagent. A LED emitting at 525 nm and a digital light sensor (TSL2591)
positioned at 180º were employed to measure the absorbance of the solution. All the components
were attached to an Arduino Uno board and the written code enabled the acquisition of the analytical
signal and data transmission via Bluetooth to smartphones. Initially, the performance of the detection
device was evaluated using sodium nitrite and the results indicated that the relation between
absorbance and nitrite was linear (r2=0.9974) in the range of 5 – 100 μmol/L. Next, gaseous standard
solutions were prepared using a NO2 permeation tube (81,25 ng min-1). A solution of 50 ppb of NO2
was used to evaluate the sampling airflow and time, in which an airflow of 100 mL min-1 and sampling
time of 60 minutes was considered the optimum conditions. Also, the repeatability of three
measurements of 50 ppb of NO2 was evaluated, resulting in a relative standard deviation (RSD) of
2.6%. A linear relation was obtained in the range of 31 to 520 ppbv of NO2, with a limit of detection
of 2 ppbv. The platform was used to monitor the hourly NO2 levels of the atmosphere of the city of
Uberlândia-MG. Therefore, a miniaturized, automated, and low-cost portable platform was
developed for the monitoring of NO2 in the urban atmosphere with suitable sensitivity, repeatability
and linearity
1 BAIRD, COLIN; CANN, MICHAEL. Química Ambiental. 4. ed. Porto Alegre: Bookman, 2011.
2 CETESB – Companhia Ambiental do Estado de São Paulo. Emissões veiculares no estado de São Paulo 2017, São
Paulo: CETESB, 2018.
[UFU, CAPES, CNPq]

164
A LOW-COST COLORIMETER BASED ON A RGB LED−LDR APPARATUS
Alessandro Munhoz Taccani a, Alexandre Antônio Sabadin a, Isabela Mariá Giatti
Leopoldino a, Claudemir Adriano Borgo (R)a, Marcos Canto Machado (R)a
a FIEC - Fundação Indaiatubana de Educação e Cultura
Analytical techniques based on UV-VIS absorption spectroscopy are widely used in the analysis in
chemical laboratories in the most diverse areas of knowledge (e.g., chemistry, environment, biology,
etc.), but due to their value, the presence of equipment that performs these analyses is reduced or
non-existent in school laboratories for didactic purposes. In this context, several initiatives 1 seek to
develop projects to build simple and low-cost analytical instruments with the participation of students
as an academic activity, however many of these initiatives comprise detachable and fragile
instruments that operate on a single wavelength or that need to be exchanged for the cuvette/sample
for reading. The objective of this work consisted of the assembly of a robust colorimeter based on
the absorbance of light in the wavelength range of UV-VIS consisting of a system of RGB LEDs
operated by remote control to change the wavelength as a source of light, a 12 V source, a light
sensor (Light Dependent Resistor - LDR), cuvette/sample containers and a digital multimeter. For
the construction of the instrument's structure (base, lid, cuvette holder, rail, LED, and LDR protection)
high-impact black polyester sheets were used, manually cut (Figure 1a), and glued with super glue
(Figure 1b). Once the instrument structure was assembled, the electrical components were inserted
and their functionality verified (Figure 1c). The premise for the use of this equipment starts from the
measurement of the light transmitted by the LDR generating an electrical resistance signal (R) that
is measured by the multimeter and later converted to an absorbance signal using the relationship:
abs = log (Rsample/Rwhite). For this, cobalt nitrate solutions were used at concentrations of 0.02 mol/L;
0.04 mol/L; 0.06 mol/L; 0.08 mol/L, and 0.1 mol/L. The results were compared with a
spectrophotometer (QUIMIS, model Q898U) operating at 510 nm. Initially, the measurement of the
absorption spectrum of the samples was carried out, and later the construction of an analytical curve
to verify the linearity of the responses presenting equations equal to y = 4.017+0.010 (r 2 = 0.9912)
and y = 4.311+0.002 (r2 = 0.002 9995) for the built instrument and the commercial instrument,
respectively. Finally, it was demonstrated that the instrument constructed presented similar
sensitivity to commercial equipment, showing that it can be used for academic purposes as a low-
cost alternative at an approximate cost of R$ 120.
1 Kovarik ML, Clapis JR, Romano-Pringle A. Journal of Chemical Education 97, 2020.

165
A NOVEL APPROACH FOR LACTATE DETERMINATION IN MILK EXPLOITING
KINETIC MONITORING BY DIGITAL VIDEOS
Gabriel M. Fernandes (R)*, Fábio R. P. Rocha (R)
Center for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil, 13416-000
*e-mail: gabrielf@usp.br
Milk acidity is an indicator of spoilage and microorganism’s proliferation. Milk is characterized by pH

within 6.6-6.8, but pH levels of 4.0-5.0 may be reached because bacteria grow produce lactic acid1.
The official method for lactic acid quantification in milk is titratable acidity, which is time-consuming,
laborious, and may yield false-negative results due to fraud by the addition of neutralizers (e.g.
carbonate salts and sodium hydroxide). To overcome these drawbacks, alternative methods have
been developed for lactate quantification, typically involving high pressure liquid chromatography
(HPLC)2, which are also time-consuming, laborious, and expensive. In this work, a simple and cost-
effective method is proposed for lactate determination in milk based on colorimetric measurements
using a smartphone as a detector. It is based on the photochemical reduction of Fe(III)-lactate
complex under UV irradiation3, yielding Fe(II), detected as o-phenanthroline complexes (maximum
absorption at 512 nm) by digital-image colorimetry. Fe(III) reduction depends on the lactate
concentration and kinetic monitoring was carried out by digital videos taken with a smartphone
camera, under irradiation by an 11-W low-pressure mercury vapor lamp (Osram, TUV PL-S). Videos
were recorded for 10 min and analytical information (RGB color system, channel B) was extracted
using ImageJ® software from frames taken each 30 s. By the fixing time method (1 min of reaction),
a linear response was observed from 0.2 – 1.0 mmol L-1 (18 – 90 mg L-1), r > 0.996, with limits of
detection and quantification of 0.06 and 0.18 mmol L-1, respectively, and coefficient of variation of
3.2% (n = 10). These analytical features are suitable for monitoring milk samples, in which lactic acid
concentration higher than 20 mg L-1 indicates spoilage. Interference by an oxidant species (probably
hydrogen peroxide, illegally added to milk to inhibit microbial growth)4, observed in the analysis of
milk samples, was successfully circumvented by the addition of a fixed amount of Fe(II) to sample
and standards. Slopes of analytical curves obtained with and without addition of whole and semi-
skimmed milk samples were in agreement (differences lower than 8%) indicating the absence of
matrix effects. Therefore, the proposed method is a simple and cost-effective approach for the
determination of lactic acid and evaluation of conformity of milk.
1 Lu M, Shiau Y, Wong J, et al. Food Nutr Sci, 4, 2013, 113.
2 Aiello A, Pizzolongo F, Manzo N, Romano R. Food Anal Methods, 12, 2019, 2555.
3 Justyna G, Koncki R, Strzelak K. Anal. Chim. Acta 2022, 1210.
4 Nascimento CF, Santos PM, Pereira-Filho ER, Rocha FRP. Food Chem, 221, 2017, 1232.
[CNPq, CAPES, FAPESP, INCTAA]

166
A PORTABLE OPTICAL SENSOR FOR MONITORING ACETONE LEVELS IN
EXHALED BREATH USING MINIATURIZED GAS CELLS AND DEEP-UV LED AT
280 NM
Diandra N. Barreto (PG)a*, João Flávio da Silveira Petruci (R)a
a Federal University of Uberlandia, Institute of Chemistry. Av. João N. de Ávila, 2121, 13400-970, Uberlândia, MG, Brazil.
*e-mail: diandra.nbarreto@gmail.com
The quantification of volatile organic compounds (VOCs) present in the exhaled breath (EB) of
humans is a potential tool for diagnosing abnormal physiological conditions, prognosis diseases, and
monitoring responses to pharmacological interventions1,2. EB analysis is a non-invasive alternative
and has been performed by gold-standard techniques capable of detecting VOCs at concentrations
in the range of ppmv to pptv. However, such techniques usually require complex and highly cost
instrumentation as well as sample collection and storage. These facts make them unfeasible for
point of care applications3,4. Among the variety of species present in the exhaled breath, acetone
has been identified as a specific biomarker of diabetes mellitus, a disease that affects approximately
346 million people worldwide. Acetone levels in healthy and unhealthy patients are usually found in
the range of 0.1 to 2 ppm. In this study, we present a low-cost and portable optical sensor for
monitoring acetone in exhaled breath in real time based on absorbance measurements at the
ultraviolet range. The analytical platform combines a 280 nm light-emitting diode (LED), a
miniaturized gas cell (substrate-integrated hollow waveguide – iHWG), and a photodiode as a
detector. All components were aligned and mounted using 3D printed holders. According to the
Lambert-Beer law, the optical pathlength (OPL) is directly proportional to the absorbance of the
analyte, therefore, aluminium-coated iHWGs with OPLs of 15 and 7.5 cm were evaluated. The
sensor’s calibration was performed by bubbling an airflow of 200 mL min-1 into solutions containing
acetone in different mixture ratios. The vapor mixtures were directly injected into the iHWG enabling
the absorbance measurement. The optical sensor with the iHWG of 15 cm provided a maximum
response at 40 s and the relation of absorbance with acetone concentration was linear in the range
of 0.5 to 5.0 ppmv. Moreover, the limit of detection (LOD) of 150 ppbv and repeatability (n = 3) of
2.0% (RSD) were obtained. Further strategies to improve the sensor’s sensitivity have been
developed, such as iHWGs with longer optical paths and the usage of solid sorbents for the acetone
preconcentration. Finally, the evolution of optoelectronic devices, such as LEDs, photodiodes, and
electronic boards has contributed to the production of low-cost portable sensors for monitoring
gaseous samples, including exhaled breath, and consequently, enabling diagnostics and pro-
diagnosis at the point of care in near-real time.5
1 Spectrometry, F. I. M.; Hagemann, L. T.; Repp, S.; Mizaiko, B. Sensing for Exhaled Breath Analysis. 2019.
2 Fortes, P. R.; da Silveira Petruci, J. F.; Raimundo, I. M.; Fortes, P. R.; da Silveira Petruci, J. F.; Raimundo, I. M. Optical
Gas Sensors for Exhaled Breath Analysis; 2017.
3 Wang, Z.; Wang, C. Is Breath Acetone a Biomarker of Diabetes? A Historical Review on Breath Acetone
Measurements. J. Breath Res. 2013, 7 (3).
4 Perez-Guaita, D.; Kokoric, V.; Wilk, A.; Garrigues, S.; Mizaikoff, B. J. Breath Res. 2014, 8 (2).
5 Barreto, D. N.; Leal, V. G.; Conrado, J. A. M.; Fernandes, G. M.; MacHado, C. C. S.; Batista, A. D.; Da Silva, S. G.;
João, J. F. Quim. Nova, 44, 2021, 1184.
[IQ-UFU, CAPES, CNPq]

167
AIRQUALITY-LAB-ON-A-DRONE: A SELF-POWERED, AUTOMATED AND
LOW-COST 3D PRINTED ANALYTICAL IoT PLATFORM FOR MOBILE AIR
POLLUTION MONITORING
Vanderli G. Leal (PG)a, Habdias A. Silva-Neto (PG)b, Wendell K. T. Coltro (R)b, João Flávio S.
Petruci (R)a*
a Universidade Federal Uberlândia, Instituto de Química, Uberlândia, Minas Gerais, Brazil, 38400-902
*e-mail: jfpetruci@gmail.com
Measuring gaseous compounds in the atmosphere is a multi-challenging task due to the low
concentration range, long- and latitudinal concentration variations and the presence of multiple
sample interferents1. Moreover, air monitoring is a logistical challenge especially when considering
areas that have poor accessibility or the analysis requires vertical sampling. In this study, we present
a quadcopter drone deployed with a fully integrated and miniaturized 3D printed analytical laboratory
for gaseous H2S monitoring capable of sampling, analyzing and transmitting the results in real-time.
The analytical method is based on the selective reaction of FMA with H2S that leads to fluorescence
quenching at maximum emission peak of 525 nm2. A 5V micro-pump with constant airflow of 50 mL
min-1 was employed to deliver constant air sampling into a flask containing 1 mL of the reagent. The
detection device is comprised of a light-emitting diode (LED) and a miniaturized light digital detector
positioned at 90º to provide the fluorescence intensity of the reagent. A humidity and temperature
sensors were integrated to the platform to provide additional information of the environmental
conditions. All the devices were programmed and controlled using an Arduino powered by a portable
power bank (5V, 1100 mAh) and the results were transmitted to a smartphone via Bluetooth. The
final device resulted in weigh of 300g and overall costs less than 100 USD. The analytical device
enabled detection of H2S in the range of 20 to 150 ppbv with reproducibility of 5% in 10 minutes of
sampling time. The platform was employed to monitor the concentration of H2S in different intervals
next to a lake at ground and vertical levels. The ability to perform all analytical steps in the same
device, the low-energy requirements, low weight and attachment of data transmission modules
enable new possibilities for drone-based analytical systems for air pollution monitoring.
1 S.K. Pandey, K.-H. Kim, K.-T. Tang, TrAC-Trends Anal. Chem. 32 (2012) 87.
2 J.F.S. Petruci, A.A. Cardoso, Anal. Chem. 88 (2016) 11714.
[UFU, UFU, PETROBRAS, CNPq, CAPES, INCTBio]

168
ANALYSIS OF MONOETHYLENE GLYCOL IN PETROCHEMICAL SAMPLES BY
MICELLAR ELECTROKINETIC CHROMATOGRAPHY ON MICROCHIPS
Mauricio M. L. Pereira (UG)a*, Kariolanda C. A. Rezende (PG)a, Wendell K. T. Coltro (R)a
a Universidade Federal de Goiás, instituto de Química, Goiânia, GO, 74690-900
*e-mail: mauricio.matheus@discente.ufg.br
Monoethylene glycol (MEG) is widely used in the petrochemical industry as a thermodynamic

inhibitor of hydrate formation, preventing the obstruction of ducts due to the formation of gas
hyrates.1 This process is not only effective but also economically interesting, since MEG can be
reused in the extraction process after undergoing a regeneration step. In this regeneration, MEG
must be extracted to be reinjected for other extraction of petrochemical samples. On the other hand,
gas condensate samples should no longer have traces of this inhibitor. 2 Based on this, the present
study describes the development of an analytical methodology using electrophoresis microchips
(ME) devices integrated with capacitively coupled contactless conductivity detection (C 4D) for MEG
detection. In this study, it was used a commercial instrumentation supplied by eDAQ including glass
ME devices containing microchannels (100 µm wide; 10 µm deep) arranged in a double-T format
and two pairs of integrated sensing electrodes (200 µm wide × 500 µm long × 200 nm thick spaced
by 250 µm) made of Pt/Ta. The best sensitivity was obtained by applying a 1200- kHz sinusoidal
wave with 100% excitation voltage. The separation of MEG was performed through micellar
electrokinetic chromatography due to its ability to promote the separation of neutral analytes. 3 The
separation was performed using 50 mmol L-1 phosphate buffer (pH = 9) containing 30 mmol L -
1 sodium dodecyl sulfate (SDS). For sample injection, a potential of 600 V was applied in the injection
channel during 10 s. Afterwards, a separation voltage of 700 V was then applied to the separation
channel and the resukting signal was monitored in real time. As it can be seen in Fig. 1, MEG was
successfully detected within 60 s. Under these conditions it was possible to identify MEG in analyzes
of approximately 100 s. The separation through MEKC mode on microchips provided satisfactory
repeatability values. For 12 consecutive injections, the RSD values calculated for the peak area was
4.2%. The proposed strategy revealed a linear behavior in the concentration from 200 to 500 µmol
L-1 and limit of detection of 3 µmol L -1. Based on the achieved results, MEKC on chip emerges as
simple, inexpensive and powerful tool for routine and in-field analysis.
70
A) B)
1 mV 60
MEG
Signal C4D (mV)
50
40
30
20 Y=0,175x-27,33
R²= 0,996
10
0
0 20 40 60 80 100 120 140 160 180 200 200 250 300 350 400 450 500
Time (s) MEG (µmol/L)

Figure 1. MEG analysis in condensate gas supplied by Petrobras (A). Analytical curve obtained from the electrophoretic
detection of monoethylene glycol in range from 250 µmol L -1 to 500 µmol L -1 (B).
1 ALEF, Khalid et al., Fuel, 222, 2018, 638.

2 ALHAROONI, Khalifa et al., Energy & Fuels, 31, 2017, 12914.
3 REZENDE, Kariolanda CA et al., Food chemistry, 329, 2020, 127175.
[UFG, PETROBRAS, CNPq, CAPES]

169
ANALYTICAL PROCEDURE TO DETERMINE ANTIOXIDANT IN RED WINES
USING MULTICOMMUTED FLOW ANALYSIS AND CHEMILUMINESCENCE
DETECTION
Rafael A. Alexandre (PG)a, Boaventura F. Reis (R)a*
a University of Sao Paulo, Center for Nuclear Energy in Agriculture, Piracicaba, Sao Paulo, Brazil, Zip code 13416 000
*e-mail: resis@cena.usp.br
This work involves the development of instrumentation and automatic analytical procedure for
antioxidant determination in red wines, using multicommuted flow analysis process and
chemiluminescence as detection technique. The instrumental designed for this proposal consisted
the following devices: a syringe pump designed for this work, driven by a stepper motor; a
luminometer assembled with two photodiodes, which were equipped with a hollow coin-shaped flow
cell, and an analog interface for signal transduction (light to electrical potential difference). The
operations of the flow analysis module and luminometer were controlled using an Arduino Due
microcontroller, furnished with interfaces and software developed for this project. The hardware of
the flow system module, and as well as the control software were designed to provide facility to
change the concentration range of the analyte to be determined. Luminol solution in alkaline medium
(pH ≈11) and sodium hypochlorite solution were used for the chemiluminescence reaction. Gallic
acid standard solutions were used as a reference for antioxidants (polyphenols). The main
parameters that could affect the proposed method were evaluated, which included the pumping flow
rates, the sample and reagent aliquot volumes and the concentrations of the reagent solutions. The
performance of the proposed system was evaluated, using it for the determination of antioxidant in
red wines. The following parameters were observed: linear response ranging from 1.0 to 10.0 gL-1
gallic acid (r2 = 998); a 0.93 mg luminol consumption per determination; a sampling rate of 65
determinations per hour; a coefficient of variation of 3.5%; and a waste generation of 2.4 mL per
determination. The red wine samples were analyzed without prior dilution, which is an additional
advantage compared to the previously proposed methods.
[INCTAA, CAPES, CNPq]

170
AUTOMATIC METHOD FOR PHOTOMETRIC DETERMINATION OF
ALUMINUM IN HEMODIALYSIS SOLUTIONS USING THE MULTICOMMUTED
PROCESS IN FLOW ANALYSIS
Eliel G. Brandão (PG)a,b, Boaventura F. Reis (R)c*
a Federal University of Amazonas, Institute of Nature and Culture, Amazonas, Brazil, 69630-000.
b Federal University of Sao Carlos, São Carlos, São Paulo, Brazil, 13565-905
c University of São Paulo, Center for Nuclear Energy in Agriculture, Piracicaba, São Paulo, Brazil, 13416-000
*e-mail: reis@cena.usp.br
This work comprises the development of an automatic analytical method for aluminium determination
in solutions for hemodialysis, combining the processes of multicommuted flow analysis and flow-
batch for solutions handling, and photometry as detection technique. The flow analysis module
consisting of mini solenoid pumps and mini peristaltic pumps, and a flow-batch chamber, was
designed to comprise a compact structure of reduced dimensions, which allows the use of large
volumes of sample and small volumes of reagents solutions, thus minimizing sample dispersion, and
thus favoring the sensitivity. The photometer used a photodiode as detector and a light emitting diode
(LED) as a radiation source. It was equipped with a flow cell with a 50 mm optical path. Croazurol S
solution was used as a chromogenic reagent, which forms a complex with aluminium ion (Al 3+). In
medium with the surfactant cetylpyrinium chlride (pH ≈ 6), presented a maximum absorption around
612 nm, allowing the use as radiation source a LED with maximum emission at 627 nm. The control
of the flow analysis module operation, and as well as the data acquisition were performed using an
Arduino Due microcontroller, equipped with electronic interfaces and software developed for this
work. At the current stage of development, the following parameters were established: linear
response ranging from 5 to 75 µg L-1 Al3+ (r2 = 0.995); limit of detection of 7.3 µg L-1 Al3+; coefficient
of variation of 2.5%; consumption of chromogenic reagent and waste generation of 47 µg and 1.9
mL per determinations, respectively; and sampling rate of 35 determination per hour. These data
show that the overall performance of the proposal is very good, so there is a synergy between the
instrumentation control, data acquisition software, and the reaction medium.
1 Bouzid B, Macdonald AMG, Analytica Chimica Acta, 207, 1998, 337.
2 Rodrigues JL, Magalhães CS, Luccas PO, Journal of Pharmaceutical and Biomedical Analysis, 36, 2005, 1119.
3 Sampson B, Fleck A, The Analyst, 109, 1984, 369.
[INCTAA, CAPES, CNPq]

171
DEVELOPMENT OF A 3D PRINTED LOW-COST FLAME EMISSION
PHOTOMETER FOR USE IN CLASSROOM EXPERIMENTS
Matheus F. Pedrotti (R)a*, Francisco C. Rosa (R)a, Lucas G. O. Aguiar (UG)a
a Instituto Federal de Educação, Ciência e Tecnologia do Rio Grande do Sul Feliz, RS, Brazil, 95770-000
*e-mail: matheus.pedrotti@feliz.ifrs.edu.br
During the teaching process of the fundamentals of spectrometry for undergraduate and high school
students, the visualization of the atomic emission process in a real analytical equipment makes the
learning process more effective and motivates the students to understand this phenomenon.1
However, due to the limited budget of the educational institutions, the acquisition and maintenance
of a real spectrometer is not viable due to the associated high costs. In this sense, the purpose of
this work was to develop a low-cost prototype of a flame emission photometer to be used for
classroom experiments. Most part of the components of the prototype were 3D printed (Core A1,
GTMax, Brasil), such as the nebulization system, composed by a 50 mm nebulization chamber and
a cross-flow pneumatic nebulizer, consisted of 1/8” polymeric tubing place at 90°. The nebulization
chamber was connected to a ½” stainless steel tube (100 mm of length), with a flattened tip, that
was used as the burner. A propane/butane (bottled gas) and compressed air flame was used. The
fuel was injected in the nebulization chamber to enable the complete mixture with compressed air,
which was introduced through the cross-flow nebulizer at a constant pressure of 20 psi. The
stoichiometry of the flame could be adjusted by a needle valve connected to the fuel inlet. As ambient
light could interfere with the measurements, the flame was placed inside a metal tube (100 mm in
diameter), allowing the flame to be seen through a side opening. A digital camera was used to
measure the light emission intensity. This camera was coupled to the system by a 3D printed
adapter. During the classroom experiments, pictures of the flame were captured and processed with
ImageJ software (U. S. National Institutes of Health, Maryland, USA). The picture was split to RGB
components and the mean gray value was measured in the desired position of the picture. The
nebulization flow rate was about 5 mL/min, which is similar to commercial equipment. The flame
intensity remained relatively stable during the experiments and no more than 5% of deviation was
observed even after 2 h of operation. Calibration curves for Na, Li, K and Ca could be obtained in
the range of 0 to 100 mg L-1. The instrumental limits of detection were lower than 1.20 mg L-1. These
results could be successfully used to teach the students about the calibration process and,
eventually, to measure a real sample. It is important to mention that only one element could be
measured at a time as the spectra of the emitted light was not measured. In this sense, further
improvements will be performed to decompose the emission spectra by positioning a diffraction
grating and an adjustable slit in the optical part of the system. One of the advantages of this system
is that it can be replicated by other people interested in teaching general chemistry and/or
instrumental analytical chemistry, since the printing files can be shared and additional parts are
easily found in the market.
1 OKUMURA, F et al. Química Nova, 27, 2004, 5.
[IFRS]

172
DEVELOPMENT OF A COMPREHENSIVE TWO-DIMENSIONAL GAS
CHROMATOGRAPHY (GCXGC/Q-TOFMS) METHOD FOR AMINO ACIDS
ANALYSIS IN CELLS SAMPLES
Samantha C H Rodrigues(PG)a*, Helvécio C Menezes(R)a and Zenilda L Cardeal (R)a
aUniversity Federal of Minas Gerais, Chemistry Departament/Icex, Belo Horizonte, Minas Gerais, Brazil, 31270-901
*e-mail: samantharodriguess@gmail.com
Four types of column sets were used, with some modifications to the oven temperature to improve the
method of analyzing a standard containing 18 amino acids. The best set of columns was used for analysis
of human cell samples. Amino acids (AAs) are essential in cellular metabolism and therefore can be used
in biochemical insights for disease diagnosis and repair mechanisms. Thus, the analysis of AAs often
requires techniques capable of broad screening of sample content and greater separation efficiency, such
as comprehensive two-dimensional gas chromatography (GCXGC). This technique allows a detailed
profile of known compounds. In addition, two-dimensional (2D) chromatograms are highly accurate and
allow the acquisition of a unique fingerprint of a given sample. The aim of the present study was to develop
an amino acid analysis method in the GCxGC and apply it to a real sample of fibroblast. The
comprehensive two-dimensional gas chromatography coupled with quadrupole time of flight mass
spectrometry (GCxGC/Q-TOFMS/MS) analysis was performed on an Agilent 7890B GC coupled to an
Agilent 7250 Q-TOFMS. The system was equipped with a ZX2 thermal modulator Zoex. The first
dimension (1D) was comprised of a nonpolar HP-5MS capillary column (30 m × 250 µm × 0.25 µm). The
second dimension (2D) consisted of a Stabilwax high polarity capillary column (2 m x 150 μm x 0,15 μm).
The generated data were processed using GC Image Version 2.9R3 (Zoex) incorporated with the NIST
Mass Spectral Library (Version 2.3). The oven temperature was ramped up from 120 ºC to 150°C at a
rate of 3°C min−1, then to 160°C at 1 ºC min−1 and finally to 260°C at 5°C min−1, and held for 3 min. A 1,0
µL sample was injected in the split mode (1:50) at 280ºC using ultra high-purity helium as the carrier gas
at a constant flow rate of 1.2 mL min−1. The modulation period was set at 8 s. The derivatization method
was based on the study of Batista 20202. Sample preparation is described in Fig 1. Fig 2 shows that it
was possible to determine amino acids derivatized with propyl chloroformate, extracted from fibroblast
cells. These results indicate that the sample preparation method with analysis by GCxGC/Q-TOFMS were
effective for AAs analysis on dermal fibroblasts. In addition, these results are a path to more complex
studies such as metabolomics studies with a fingerprint of all compounds that involve the cells
metabolites. The developed method was able to identify 18 amino acids with high separation, and to
identify these analytes in real cell samples. Besides, the results indicate that this method is a gateway to
several types of studies involving amino acids, biochemistry and metabolomics.
Figure 1. Sample preparation steps for the analysis of Figure 2. Color diagram of AAs (1-Alanine; 2-Glycine; 3-
AAs derived from human dermal fibroblasts (HDFa) Valine; 4-Norvaline; 5-Leucine; 6-Isoleucine; 7-Proline; 8-
Threonine 9-Aminobutyric acid; 10-Methionine; 11-Acid
aspartic; 12-Serine; 13-Phenylalanine, 14-Glutamic acid,
15-Lysine; 16-Histidine, 17-Tyrosine and 18-Tryptophan)
extracted from HDFa.
[CNPq, Cemig]

173
DEVELOPMENT OF A FIELD TRIAL FOR ANALYSIS OF ACYLGLYCEROLS IN
BIODIESEL
Breno A. Britoa (PG), Cristiane G. de Souzab (R), Rafael C. dos Santosb (PG), Renan de O.
Munizb (PG), Débora F. de Andradea* (R), Luiz A. d’Avilab(R)
a Federal University of Rio de Janeiro (UFRJ), Department of Analytical Chemistry/Institute of Chemistry, Rio de Janeiro,
RJ, Brazil, 21941-909
b Federal University of Rio de Janeiro (UFRJ), Department of Organic Processes/School of Chemistry, Rio de Janeiro,
RJ, Brazil, 21941-909

*e-mail: debora.franca.andrade@gmail.com
The acylglycerols (mono-, di- and triacylglycerols) are intermediate compounds of transesterification that
affect some characteristics of the final product, causing negative impacts on the quality of biodiesel1. ANP
determines the maximum value of 0.23% (weight) of combined glycerin (sum of the content of mono-, di-
and triacylglycerols) in pure biodiesel (B100). The reference methodology used to determine combined
glycerin content in biodiesel is based on gas chromatography with flame ionization detector (GC-FID)
according to ASTM D6584:2014. However, this method requires a sample derivatization step through
derivatizing agents and needs internal standards for each class, which increases its cost. In addition, it
is a test that must be possible in laboratories, on expensive equipment and trained technicians. In this
context, the main objective of this work was to develop a simple, fast and low cost test, capable of being
carried out in field, based on previously obtained results2, including object of patent filing in France3 and
Brazil4. It is based on the processing of digital images from colorimetric reactions, using the smartphone
app PhotometrixPRO® for quantitative determination of combined glycerin in biodiesel. The linearity of
the method was determined in the range of 0.006 to 0.056% (weight) of glycerol trom both green and
blue channels. The results Cochran test showed C values calculated for both green and blue channels,
respectively, 0.6446 and 0.5805, using Microsoft software Excel. As the calculated C value for the blue
channel was lower than the tabulated C value (0.6161), the analytical curve was considered
homoscedastic. The coefficients of determination (R²) of analytical curves for both green and blue
channels were 0.9353 and 0.9298, respectively. The developed method presented detection limit for both
green and blue channels, respectively, 6,99x10-3% (weight) and 7,29x10-3% (weight). The method also
presented a limit of quantification for both green and blue channels, respectively, 2,31x10 -2% (weight)
and 2,41x10-2% (weight). The precision (repeatability) was determined through the relative standard
deviation (RSD%) under repeatability conditions and the results obtained ranged from 0.57% to 1.37%
for the green channel and from 0.86% to 1.60% for the blue channel. The accuracy was determined
through recovery and the results obtained ranged from 94.02% to 114.00%, demonstrating the validation
of this parameter. The results obtained for the determination of combined glycerin, in 6 samples of
biodiesel (B100), by both reference and proposed methods, were compared and considered statistically
similar, with 95% confidence, using the paired t test (both green and blue channels presented values of
calculated t, respectively, 0.5595 and 0.8867, smaller than the tabulated t value 2.5705). The developed
method has the potential to be used for the analysis of glycerin combined in biodiesel, in field, making
the monitoring of biodiesel more accessible and, therefore, improving its quality. An analysis kit for
determining acylglycerols is already being tested by the company Macofren Tecnologias, as a result of a
technical cooperation agreement with UFRJ and licensing of the technology, with the prospect of a brief
launch in the fuel market.
1. Lobo IP, Ferreira SLC, Cruz RSD, Química Nova, 32, 2009, 6
2. Muniz, R. O, Martins, S. B, Honório, G. G, Cunha, J. N, Souza, C. G, Andrade, D. F, Pradelle, R. N. C, Turkovics, F,
Neto, R. S, d’Ávila, L. A, D'Elia, E. Analytical Methods, 11 (6), 2019, 767-773.
3. Patent: innovation privilege. Registration number: FR1872032, títle: "Procede d’analyse du glycerol libre dans un
echantillon de biodiesel par voie enzymatique" , Registration institution: Institut National de la Propriete Industrielle.
Deposit: 11-21-2018
4. Patent: innovation privilege. Registration number: BR1020190021764, títle:“ANALYSIS METHOD OF A BIOFUEL
SAMPLE AND USE OF IT”, Registration institution: Instituto Nacional da Propriedade Industrial. Deposit: 11-03-2019
[CAPES]

174
DEVELOPMENT OF A PORTABLE LED UV-Vis SPECTROPHOTOMETER:
ORGANIC MATTER DETERMINATION IN NATURAL WATERS
Ismael F. Christmannª (PG), Maikel L. Kollingb (R), Isadora C. M. Silvac (UG), Roberta O.
Santosb (R), Jocelene Soaresa (PG), Demis P. Faquia (PG), Adilson B. da Costaª,b (R)
aUniv. de Santa Cruz do Sul, Prog. de Pós-Graduação em Tec. Ambiental, Santa Cruz do Sul/RS, Brazil, 96815-900
bUniv. de Santa Cruz do Sul, Prog. de Pós-Graduação em Sist. e Proc. Ind., Santa Cruz do Sul/RS, Brazil, 96815-900
cUniv. de Santa Cruz do Sul, Curso de Engenharia Química, Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96815-900
*e-mail: ichristmann@mx2.unisc.br
Traditional methods to detect organic matter (OM) in underground and superficial water are very
used all over the world. Consulting the textbooks (Standard Methods, 2017)¹, it is easily found
quantifying methods of OM based in different analytical principles, such as: Chemical Oxygen
Demand (COD), Biochemical Oxygen Demand (BOD), Total Organic Carbon (TOC), Oxygen
Demand (OD) by permanganate method, which are the most known traditional methods.
Nevertheless, these methodologies typically involve the high consume of chemical reagents, energy
and the generation of dangerous waste. In this sense, the present work has as a main purpose to
develop a portable device and a methodology based in the Green Analytical Chemistry (GAC),
Gaÿuszka (2013)², to determine the organic matter in natural water samples. The portable equipment
(Fig. 1) has a case printed in a 3D printer, a Wemos D1 R33 prototype board (processing board),
provided with an ESP 32 Chip (microcontroller), its development IDE (Integrated Development
Environment) is Arduino (prototype platform) and the software
in language C# (programming language) in back-end (the back
part of applications and business rules). The software
developed in front-end (application that interacts with the user)
used as an interface Javascript, HTML (Hypertext Markup
Language) and CSS (Cascade Styling Sheets), programming
languages. This device is equipped with two sets of detectors
and radiation emitters (LED, Light Emitting Diodes) arranged
orthogonally. One set (S1) is applied to the analysis in the
visible spectrum region and another (S2) in the ultraviolet 50 mm
spectrum region. The S1 uses an LDR (Light Dependent
Figure 1. Portable equipment
Resistor) detector and an RGB (red, green, blue) LED, while the
S2 uses Ultraviolet Sensor Uvm-30 and LED UV (Ultraviolet
Radiation - spectral range 250nm/256nm). This way, this device allows the development of different
molecular spectrophotometry methodologies. In this work it was applied to suppress the stage of the
method’s titration of oxygen determination consumed by the potassium permanganate method
(NBR1079)³ (S1). The S2 system allows the quantification of organic matter in natural waters through
the ultraviolet absorption method (STANDARD METHODS, 2017 – 5910 B)¹. A device software
allows the development of univariate calibration models, correlating the concentration values of the
reference samples and the electrical resistance values provided by the detectors. The same software
manages the analysis samples and results expression. The results obtained from this prototype
allowed the determination of organic matter in the concentration range between 0,10 and 2,95 mgL-
1, showing no significant difference (p>0.05) when compared to the results obtained by the reference
methods. This equipment, called Specket, allows the development of spectrophotometric methods
in a simple, reliable and fast way. Also meeting several GAC principles, such as the application of
direct analytical techniques, in situ measurements, reduction in the use of reagents and energy, and
reduction of waste generation.
1 Standard Methods for the examination of water and wastewater, American Public Health Association, 23, 2017, USA.
2 Galuszka, A; Migaszewski, Z; Namiesnik, J, Trends in Analytical chemistry, 50, 78-84, 2013.
3 Water - Determination of oxygen consumed, Potassium permanganate method, NBR10739, ABNT, 1989.
[FAPERGS, CAPES, CNPq]

175
DEVELOPMENT OF A WALL COATED OPEN TUBULAR COLUMN FOR NANO
LIQUID CHROMATOGRAPHY COUPLED TO MASS SPECTROMETRY
Natalia G P dos Santos (PG)a*, Fernando M Lanças (R)b

aUniversity of São Paulo, Institute of Chemistry of Sao Carlos, Sao Carlos, Sao Paulo, Brazil, 13566-590
bUniversity of São Paulo, Institute of Chemistry of Sao Carlos, Sao Carlos, Sao Paulo, Brazil, 13566-590
*e-mail: nataliagabrielly@usp.br
Liquid chromatography (LC) is one of the most versatile separation techniques being used in
numerous relevant areas of scientific knowledge. Its miniaturization, understood here as the
decrease in the internal diameter of the columns, followed the introduction of capillary columns in
gas chromatography by Golay [1]. Therefore, the current interest is focused on the development of
new columns for LC, aiming to increase the efficiency of the technique and the implementation of
the various benefits inherent to it. These mainly comprise the miniaturization of the system and its
implications, for example, the economy of solvents and samples, and the coupling with detection
systems commonly used in gas chromatography, such as mass spectrometry [2]. Among the
capillary columns that can be applied to LC, we would like to pay attention to the open tubular
columns [3], characterized by a hollow tube with the wall coated by the stationary phase. Therefore,
the objective of this work was to develop an open tubular column coated with a liquid stationary
phase, which is called wall coated open tubular column (WCOT) for nano liquid chromatography
coupled to electrospray mass spectrometry. For this, the authors prepared a 2 m x 50 µm column
(length x i.d.) coated with the OV-210 phase (50% trifluoropropyl 50% polydimethylsiloxane) to be
used in the nanoLC-ESI-MS/MS system. Three pesticides of different classes were used as
molecular probes: clomazone, atrazine and metallochlor. The column efficiency was calculated by
the number of theoretical plates, which proved to have N = 43.868, characterizing a high efficiency
column compared to a packed column. In addition, a small separation (gradient, flow of
300 nL min-1, 500 nL of injection volume, and analytes at a concentration of 500 µg L-1) was obtained
between the analytes atrazine (tR = 19.94 min), clomazone (tR = 20.50 min) and metallochlorine
(tR = 21.58 min). In conclusion, WCOT columns have high potential for application in nano LC, given
the efficiency obtained with this column. However, several challenges still need to be overcome,
such as finding the best condition between column length, i.d. and film thickness that are suitable for
achieving high resolution separation. Such factors have been studied by the authors in their works.
1 Ettre LS, Vieira LP, Journal of High Resolution Separation,10,1987, 221.

2 Mejía-Carmona K, Burato JSS, Borsatto JVB, Toffoli AL, Lanças FM, Trends in Analytical Chemistry,122,2020,1.
3 Medina DAV, Santos NGP, Burato JSS, Borsatto JVB, Lanças FM, Journal of Chromatography A,1641,2021,1.

176
DEVELOPMENT OF DATA SCIENCE FOR VOLTAMMETRIC ANALYSIS
USING ARDUINO PLATFORM
Gabriel A. Zaia André Olean-Oliveira Marcos F. S. Teixeira*
Faculty of Science and Technology, São Paulo State University (UNESP), Pres. Prudente, SP, BR.
Arduino is an open-source electronic board with inputs and outputs for connecting sensors and
computational devices for use and data analysis.1 This platform has practicality, with data
processing, less error security, a wide range of sensors, microcontrollers, and modules. These
characteristics make it a widely used platform for the development of equipment and prototypes.1
Arduino is applied for data analysis using electrochemical techniques such as cyclic voltammetry,
differential pulse voltammetry, and linear voltammetry2,3. Data Science is a promising computing
platform in data storage and sharing. This work reports the development of a computational server
to perform the processing of data obtained by the electrochemical cell.
Figure 1. Integrated circuit model between electrochemical cell, sensors, Arduino, server.
The electrochemical cell will be composed of 3 electrodes: counter electrode, work, and reference.
Keeping it as a conventional electrochemical configuration, between the work analysis and the
constant potential, between the work and the current counter-electrode. So that there is a connection
between electrochemical sensors and Arduino, potential and current will be used to make an
Arduino-server connection.2 The server is ready and some tests have already been done with graphic
voltammetric data from other electroanalytical studies. Data processing takes place by comparing
each Cartesian point, that is, after the Arduino indicates the results to the server, the server will
analyze the results from point to point, comparing them with the graphs already obtained previously.
The comparisons in the tests already carried out were satisfactory, due to the percentage points
correlated between the graph and the database. A graph with 100 points (x,y) and hundreds of dated
voltammetric graphs was used. The server compared and returned the satisfactory result of some
voltammograms that have the same electrochemical behavior. The work is currently in the
development of the connection between the electrochemical cell and sensors, later, it will end with
tests of the complete equipment.
1 DRYDEN, M. D. M.; Fobel, R.; Fobel, C.; Wheeler, A. R. Analytical Chemistry, v. 89, n. 8, p. 4330–4338, 18 abr. 2017.
2 Scholz, F. ChemTexts v.1, n.17, 2015.
3 RACE, A. M.; Palmer, A. D.; Dexter, A.; Steven, R. T.; Styles, I. B.; Bunch, J. Analytical Chemistry, v. 88, n. 19, p. 9451–
9458, 4 out. 2016.

177
DROPLET ANALYSIS USING A ROBUST AND PORTABLE ELECTROCHEMICAL
CELL PRODUCED BY 3D PRINTING
David L. O. Ramosa, Lucas V. de Faria (R)a, Diego A. C. Alvesb, Rodrigo A. A. Muñoza,
Wallans T.P. dos Santosc, Eduardo M. Richtera
aUniversidadeFederal de Uberlândia, Instituto de Química, Uberlândia, MG, Brazil, 38.408-100
bUniversidadeFederal de Uberlândia, Faculdade de Engenharia Mecânica, Uberlândia, MG, Brazil, 38.408-100
Universidade Federal do Vale do Jequitinhonha e Mucuri, Dep. de Farmácia, Diamantina, MG 39100-000, Brazil
*e-mail: daviluizor@ufu.br
Additive manufacturing or 3D printing is a very promising method for the production and prototyping of
complete electrochemical systems1. In this work we propose a simple and practical electrochemical
system, completely printed with thermoplastic material (ABS or PLA or PETG) that allows the analysis in
small volumes (from 50 to 1000 µL). The cell only consists of two parts printed by 3D-FDM that are joined
using 3 screws and nuts (Figure 1A, B, and C). The solution vessel was designed for easy and precise
positioning of both counter and pseudo-reference electrodes printed by 3D printing (Figure 1A). The
bottom of the microcell was designed for easy coupling of different conductive materials over a rubber
ring (prevents leakage and defines the geometric area of the working electrode; Fig. 1B). In short, all
square plate-shaped (1 x 1 cm) materials with thickness less than 5 mm can be easily used as disposable
or non-disposable working electrodes. Materials such as boron doped diamond (BDD), glassy carbon
plates, commercial graphite sheets2, laser-induced graphene sheets3 and conductive plates printed with
conductive filaments4 have been successfully used as working electrodes. As a proof of concept,
paracetamol was used as model analyte and linear calibration curves (r > 0.99) with similar slopes (0.29
± 0.01 µA uM-1; RSD = 3.4 %) were obtained using different volumes of standard solutions (50, 100, and
200 µL). So, we can conclude that the system here proposed works similarly to the well-known “screen-
printed electrodes (SPEs)”, that is, allows drop analysis and different materials can be easily used as
working electrodes (disposable sensors). However, as an advantage, the large-scale production of the
system here proposed is possible using low cost FDM 3D printers, while for SPEs production, more
unusual and expensive screen-printer machines need to be used. Finally, the coupling of robust materials
such as BDD electrodes (wider useful electrochemical window) is also possible and analysis at potentials
higher than +1.8 V (vs Ag/AgCl) can be performed5.
Figure1. Scheme of the 3D-printed cell: (A) upper view, bottom view of (B) solution vessel and, (C) base.
1 Whittingham M.J. et al., Talanta Open, 4, 2021, 100051

2 Silva L.A.J. et al., Journal of Electroanalytical Chemistry, 833, 2019, 560.
3 Costa W.R.P. et al., Microchimica Acta 189, 2022,185.
4 Richter E.M. et al., Analytical Chemistry, 91, 2019, 12844
5 Freitas J.M. et al., Sensors and Actuators B: Chemical 243, 2017, 557.
[UFU, FEMEC CNPq, FAPEMIG, CAPES, Polícia Federal]

178
EVALUATION OF AN INEXPENSIVE 10-CHANNEL COLOR SENSOR TO
QUANTITATIVE DETERMINATIONS
Caio C. S. Machado*a (PG), Sidnei G. da Silvaa (R)

a Institute of Chemistry, Federal University of Uberlândia, Uberlândia, MG, Brazil, 38.408-100
*e-mail: cesarcaio12@gmail.com
The AS7341 sensor is a spectrometer sensor that was evaluated for quantitative determinations.
The visible spectrum detection is based-on eight channels (415, 445, 480, 515, 555, 590, 630 and
680 nm) which given simultaneously light intensity for each channel. A white high brightness LED is
incorporated at the sensor, which emits light and illuminates the aimed surface. The light reflected
by that surface is measured by the detector. The LED, which is based on nitride (InGaN), has a
viewing angle of 120°, with maximum illumination in the desired spectrum ranges (400-700 nm). A
3D Printed cuvette white holder was manufactured in two parts: a holder for a standard cuvette, with
a hole to allocate the sensor and lid to cover the cuvette, using a 1-cm conventional cuvette for liquid
solutions. The sensor shows adjustable gain and integration time, in order to increase the sensitivity.
The measurements are based on the reflectance of the light, at specific wavelengths, that passes
through an analyte reflected to the detector. Aiming to investigate and relate the desired analytical
parameters to the color sensor, in order to obtain quantitative information, the determination of
Sunset Yellow (SY) and Tartrazine Yellow (TY) dyes were evaluated. The colored dyes TY and SY
show a maximum absorption wavelength close to 415 and 515 nm, respectively. For quantitative
determination of yellow dyes in beverage samples TY measures were performed at 415 nm, with
128x gain and 80 mA current, and a linear range of 1.5 – 15.0 µmol.L-1 was observed, (y = 0.0039x
+ 0.0048, R = 0.999) with a LOD and RSD estimated as 1.1 and 0.53%, respectively. SY
measurements were performed at 515 nm, with 32x gain and 50 mA current, and a linear range of 0
– 15.0 µmol.L-1 was observed (y = 0.018x + 0,0051, R = 0.998) with a LOD and RSD estimated as
0.9 and 0.46%, respectively. To demonstrate the applicability of the spectrometer sensor, the
Tartrazine and Sunset yellow dyes were determined in beverages samples (Juice and isotonic).
These samples and analytes were chosen to demonstrate the sensor ability to separate the light into
distinct wavelength regions (415 and 515 nm) and distinguish region from each other. The maximum
concentration established of total SY and TY in beverage samples, according the Agência Nacional
de Vigilância Sanitária (ANVISA) is 0.01 g in 100 g/mL [1]. The found concentrations ranged from
1.0 to 6.0 µg for SY in 100 mL of samples. The accuracy was estimated comparing the results
obtained with the proposed procedure with those obtained with a spectrophotometer and the results
agreed with those obtained by spectrophotometry at a 95% confidence level. Tartrazine
determination in presence of Sunset Yellow was not possible due the SY absorption in 415 nm. The
multi-channel spectrometer sensor approach, using a 1-cm conventional cuvette for liquid solutions,
shows potential to detect low concentrations (µmol L-1) of yellow dyes. Additionally, that device allows
the development of versatile and miniaturized system for the point-of-care determinations,
contributing to achieve the requirements to generate an Internet of Analytical Things dispositive [2].
1 BRASIL. Agência Nacional de Vigilância Sanitária. Resolução nº 382 a 388, de 09 de agosto de 1999. Diário Oficial [da]
Republica Federativa do Brasil. Poder Executivo, Brasília, DF, 09 ago, 1999.
2 Cadeado A, Machado C, Oliveira G, e Silva D, Muñoz R, Silva S. Internet of Things as a Tool for Sustainable Analytical
Chemistry: A Review. J Braz Chem Soc. 2022;33(7):681-692. doi:10.21577/0103-5053.20220048
[CNPq, FAPEMIG, CAPES]

179
INVESTIGATING OPTICAL PROPERTIES OF 3D-PRINTED SUBSTRATES
FABRICATED BY COMMERCIALLY AVAILABLE RESINS
Fernando Henrique M. Costa (PG)a*, Brenda M. de C. Costa (PG)a, Dhaniella C. de B. Oliveira
(PG)a, Daniele G. Rodrigues (PG)a, Ivo M. R. Junior (R)a, José Alberto F. da Silva (R)a,b
a State University Campinas, Analytical/IQ, Campinas, SP, Brazil, 13083-970
b National Institute of Science and Technology of Bioanalytics, INCTBio, Campinas, SP, Brazil.
*e-mail: f231964@dac.unicamp.br
3D printing technology has been an attractive alternative to the fabrication of microfluidic devices
due to its advantages such as the possibility to fabricate complex in a single step 1,2. Despite
significant advances, barriers related to material properties, such as optical transparency, there are
still challenges to be overcome in the device fabrication, aiming the integration of optical detectors
to the microfluidic devices3,4. In this direction, we report the absorption spectra of substrates
manufactured using different commercial resins available in Digital Light Processing (DLP) 3D
printing. Thus, this study aims to assist the development of devices with suitable optical properties.
Substrate samples were printed using commercial resins from Anycubic clear, Phrozen, Formlabs
clear, Power Sega, MicroSLA, Asiga RED and Solus. Shimadzu model 2450 UV-vis
spectrophotometer was employed to obtain absorbance and diffuse reflectance spectra in a spectral
range from 200 to 800nm. Optical transparency is a fundamental property of microfluidic devices.
Most cell analysis and cell culture on chip require the visualization of the inner channels and
chambers in order to follow cell dynamics. Moreover, when optical detectors are integrated, such as
UV-vis absorption and fluorescence detectors, is of utmost importance to know the behavior of the
microchip substrate. We have studied the absorbance spectra of a variety of substrates prepared by
DLP 3D. We observe an intense region of absorption for the resins ANY, Formlabs, Phrozen and
SOLUS for wavelengths below 410 nm. For these resins, there is no significant absorption was
observed to the resin POWER above 420 nm. Resin MicroSLA also present high absorption for short
wavelengths and transparency above 470 nm. Conversely, the Asiga RED resin present an
additional absorption band centered at 525 nm due to red dye included in the resin formulation. This
resin is transparent above 650 nm. The diffuse reflectance spectra are complementary to the
absorption spectra, and it is very advantageous to verify singularities caused by the absorption
process when solid samples are using in the measurements5. It is important to know the absorption
behavior of commercial resins prior applications in microfluidics. It is necessary to avoid the
wavelengths of absorption of the substrate since the incident or emitted beam will be completely or
partially blocked in absorbance or fluorescence applications, respectively. This work is useful for the
selection of a suitable wavelength range when it is desired to obtain optical data from target
compounds using microfluidic devices manufactured with the commercial resins used.
1 Barone, S. et al., Procedia Structural Integrity 2018, 12, 113-121.
2 Tiboni, M. et al., International Journal of Pharmaceutics 2021, 599, 120464.
3 Gonzalez, G. et al., Additive Manufacturing 2022, 55, 102867.
4 Krzesniak, A. et al., Optics and Lasers in Engineering 2022, 158, 107173.
5 Solé, J.; Bausá, L.; Jaque, D. An Introduction to the Optical Spectroscopy of Inorganic Solids. England: John Wiley &
Sons, 2005.
[CAPES, FAPESP, CNPq, INCTBio, INCTAA]

180
LOW-COST AND MODULATED 3D-PRINTED FLUOROMETER/PHOTOMETER
FOR THE DETERMINATION OF CAFFEINE AND FE(II) IN MEDICINE SAMPLES
Rafaela Silva Lamarca (R)a, João Pedro da Silva (R)a, Paulo C. F. de Lima Gomes (R)a*
a National
Institute for Alternative Technologies of Detection, Toxicological Evaluation and Removal of Micropollutants
and Radioactives (INCT-DATREM), Institute of Chemistry, São Paulo State University (UNESP), Araraquara, São Paulo,
Brazil, 14800-060
*e-mail: paulo.clairmont@unesp.br
In the last decades, the search for fast, portable and low-cost devices has been the object of study
in several areas of analytical chemistry.1 In this context, advances in the production of new detection
platforms using 3D printing have been highlighted. In this study, a device was developed to
determine caffeine and Fe(II) in medicine samples using a portable and modulated device which
could be used as a fluorometer or a photometer. This device presents dimension of 12 x 9 cm, it
contains LED used as light source and photodetector. The device was printed in black polylactic acid
polymer. The absorbance and fluorescence intensity was obtained by a digital multimeter monitoring
through the electrical resistance obtained by the photodetector. For the quenching of fluorescence,
a LED with maximum emission in 460 nm, and a reaction involving caffeine and the trisodium salt of
8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) at 50.0 µmol L-1 in the presence of phosphate buffer
(0.1 mol L-1, pH 6.3) was applied.2 For the absorbance studies, a LED with maximum emission in
530 nm and reaction between Fe(II), hydroxylamine (5.0 % m/V), sodium acetate (10 % m/V) and
1,10-phenanthroline (reaction performed at pH = 4.5 - 5.5) was evaluated.3 As a fluorometer, this
device presented limit of detection (LOD) and quantification (LOQ) of 25.0 mg L -1 and 50.0 mg L-1,
respectively. The method recovery obtained was 99.5 % at concentration level of 250 mg L-1, the
analytical curve was represented by F = 3.00 x 10-4 [caffeine]+ 1.00 and R2 = 0.993 in the
concentration range of 50.0 - 300 mg L-1. The inter and intra-day precision were 1.28 % and 1.02 %,
respectively. The medicine sample presented 211 ± 0.0671 mg in accordance with the labelled
concentration (210 mg). As a photometer, the device presented the following analytical curve A =
0.112 [Fe(II)] + 0.0126 and R2 = 0.998 in the concentration range of 1.00 - 5.00 mg L-1. The obtained
LOQ and LOD were 1.00 mg L-1 and 0.500 mg L-1, respectively. The medicine sample containing
Fe(II) presented a concentration of 39.1 ± 1.57 mg, in accordance with the sample label (40.0 mg).
The both methods were compared using a t-test at 95% confidence with a reference method using
a spectrofluorometer and spectrophotometer, respectively, and no significant differences between
both methods were observed. The printed device presented an average cost of U$25.00, both
methods required only 1 min to obtain the analytical signal with precision and accuracy. Moreover,
the device can be easily used as a fluorometer or a photometer, only switching the photodetector
position. Moreover, the LED can also be switched in order to adapt for other types of analysis.
Therefore, the printed-device is a promising alternative, especially in remote locations with few
laboratory resources.
1 SinghH, et al, Sensors and Actuators B: Chemical, 322, 2020, 128622.
2 Rochat S, et al, Chemical Communications Journal, 47, 2011, 10584–10586.
3 Sandell, E. B. The Journal of Physical Chemistry, 49, 3, 1945, 263–264.
[FAPESP, IQ-UNESP, CAPES, CNPq, INCT-DATREM]

181
NEW ADVANCE TOWARD PRINTING MULTI-MATERIALS BY RESIN 3D
PRINTERS
Reverson F. Quero 1, Dosil P. de Jesus 1, José A. F. da Silva 1
1 UNICAMP - Universidade Estadual de Campinas
Additive manufacturing, also known as 3D printing, in which objects with complex geometry are
constructed from drawings made on graphic software, emerged more than 30 years ago. However,
only in the last few years, with the availability of printers with high resolution and affordable cost,
they allowed the creation of devices in micro and nanometric scales to study concepts that were not
possible or difficult before.1 Although it is currently possible to create complex 3D microdevices using
a high-resolution technique, such as DLP (Digital Light Processing), which uses light-curing resins,
the next even more complex step is using multi-materials. This advance can allow using materials
with different compositions and properties in a single printing process, allowing the construction of
complete devices, with base materials and integrated conductivity parts, for example2. The present
work proposes an unprecedented strategy for 3D printing of multi-materials using a holder adapted
to a 3D resin printer that automatically performs the exchange of resins. For this, simulations were
performed using computational fluid dynamics to assess the best conditions for exchanging these
materials. The initial tests were conducted by adapting the holder to a commercial DLP 3D printing,
with two resin inlets and an outlet to remove the waste. Devices using two resins of different
colors and reagents dissolved in transparent resin were manufactured, demonstrating the feasibility
of multi-material printing. In the near future, the potential of multi-material 3D printing will be
revolutionary, allowing devices with specific functionalities for advanced applications. The 3D
printing development proposed in this work will allow exploring the benefits of combining different
properties of the materials, opening a wide range of structural applications, such as the integration
of electrodes and membranes, and improving biocompatibility into a device 3D printed with high
resolution and in a single step.
1 Mao M, He J, Li X, Zhang BL, Lei QH, Liu YD. Micromachines, 8, 113, 2017. 2Shaukat U, Rossegger E,
Schlögl S. Polymers, 14, 2449, 2022.

182
PORTABLE DEVICE AND MULTIVARIATE ANALYSIS: AN INNOVATIVE
METHOD FOR BASIC NITROGEN DETERMINATION IN DIESEL OIL
Alice P. Holkem(PG)a, Gabrielle D. Iop(PG)a, Edson I. Muller(R)a, Juliano S. Barin(R)b,
Paola A. Mello(R)a*
aUniversidade Federal de Santa Maria (UFSM), Departamento de Química, Santa Maria, RS, Brazil, 97105-900
bUniversidade Federal de Santa Maria (UFSM), Departamento de Tecnologia e Ciência dos Alimentos, Santa Maria, RS,
Brazil, 97105-900.
Basic nitrogen compounds, as pyredine, quinoline, and aniline are the most common in crude oil and can
be transferred to fractions during distillation. The presence of these compounds in diesel oil strongly
deactivates the catalysts that are used in the fuel-refining processes, such as hydrodesulfurization (HDS),
hydrodearomatization (HDA), hydrocracking, and reforming, even at very low concentrations1. The UOP
method 269-10 is the standard protocol to measure basic nitrogen in fuels. It is based on a nonaqueous
titration using diluted perchloric acid as the titrant (the final point can be identified using potentiometry or
with a colour indicator).2 However, several disadvantages have been reported, such as the use of high
amounts of reagents and waste generation, mainly acid residues, which contrasts with the
recommendation of green analytical chemistry. Alternatives methods can be applied for this purpose,
mainly focusing on miniaturization to circumvent or minimize those problems. The colorimetric analysis
using PhotoMetrix app can be an approach easily performed with low-cost, portable option, and with high
throughput. Thus, the aim of this work was to develop a simple protocol to provide basic nitrogen
determination directly in diesel oil samples, by using the digital images for colorimetric quantification. For
that, the following parameters for the reaction and detection were evaluated: i) the volume of violet crystal
indicator (30, 60, 90 or 120 µL), ii) the volume of HClO4 (20, 40, 60 or 90 µL), and iii) the concentration
of HClO4 (0.01, 0.02, 0.05, 0.075 or 0.1 mol L-1). The volume inside the microtube (1120 µL), the lighting
(80 lx), the focal distance (2.19 cm), the region of interest (64 x 64), and the time to carry out the
measurement (20 - 30 s after the indicator addition) were kept constant in all experiments. To obtain the
digital images, an USB camera was placed in an open-source 3D-printed chamber illuminated by a white
light-emitting diode (LED) with light intensity controlled. Sample and reagents (perchloric acid and the
indicator) were added to an Eppendorf vessel, which was placed inside the chamber. Thus, the digital
images were obtained and converted into red, green, and blue (RGB) histograms, and the partial least
squares (PLS) models were obtained within the app. Figure 1 shows an experimental overview.
Figure 1. The three steps for N determination using the PhotoMetrix app in a portable device: 1) reaction, 2)
image (directly from the microtube using the endoscopic camera), and 3) the calibration curve using
PLS model (obtained directly from the app).
The PLS regression results were evaluated in terms of the number of samples and factors, the coefficient
of determination (R2), the root mean squared error of calibration (RMSEC), the root mean squared error
of cross validation (RMSECV), and the root mean squared error of prediction (RMSEP). The calibration
curve (from 1 to 6 mg kg-1 of basic nitrogen) was obtained using a PLS model with a R2 higher than 0.99
using 60 µL of violet crystal and 20 µL of 0.02 mol L-1 HClO4. No significant differences (p < 0.05) between
the measured and predicted values were identified. The RMSEC, RMSECV and RMSEP of PLS model
were 0.06, 0.583 and 0.283 mg kg-1, respectively. The developed method was simple, minimized reagents
consumption and residues generation and can be an alternative for fast and accurate determination of
basic nitrogen content in diesel oil.
1 Poppi RJ et. al., Fuel, 160, 2015, 274-281.
2 UOP LLC, UOP269-10, 2010.
[CNPq, CAPES, FAPERGS, UFSM]

183
PREDICTING SOIL CLAY USING DIGITAL IMAGES BASED ON SMARTPHONE
José G. L. Abich a (IC*) Isadora C. M. Silvaa (UG), Roberta O. Santosb (PG), Jocelene
Soaresc(PG), Gilson A. Helfera (R), Adilson B. da Costab,c(R)
aUniv.de Santa Cruz do Sul, Depto de Eng., Arq. e Comp., Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96815-900
bUniv.de Santa Cruz do Sul, Prog de Pós-Graduação em Sist. e Proc. Industriais, Santa Cruz do Sul, Rio Grande do
Sul, Brazil, 96815-900
cUniv. de Santa Cruz do Sul, Prog de Pós-Graduação em Tec Ambiental, Santa Cruz do Sul, Rio Grande do Sul, Brazil,
96815-900
*email: joseabich@mx2.unisc.br
The demand for soil analysis has increased, mainly for clay determination, due to soil quality and nutrient
and carbon storage. The traditional method used by most soil analysis laboratories is considered time
consuming, uses dangerous regents and generates a a large amount of waste.
In view of this, the aim of this work was to develop a methodology to clay determination in soils, using a
smartphone coupled to a digital image acquisition system. Preliminary tests were carried out to choose
the operating parameters such as the type of camera, distance from the sample, luminosity and the size
of the region of interest (ROI). The conditions that provided the best result and the lowest error consisted
of using the smartphone's own camera (Pixel Phone by Google), with indirect lighting through a white
LED light strip (4.6 W/m, 12 V, LED line M-2835, 8 mm) with 13 lamps (513.16 LUX) in a box with with
dimensions (width x depth x height) measuring 950 mm x 155 mm x 125 mm. The walls were covered
with white paper and the bottom with green paper to improve light dispersion and not interfere with the
colouring of the sample. The camera was positioned at a
height of 14 cm away from the sample, and the size of the
ROI of the image was 128X128 pixels (Fig. 1) Through the
PhotoMetrix PRO® application, two calibration models were
built. The first was a univariate analysis an analysis of the
RGB Vector using a set of 45 soil samples, and the second
was multivariate analysis where the same set of soil samples
was used and Partial Least Squares Regression (PLS) was
performed. Prediction errors (root mean-square error of the
prediction - RMSEP) were obtained for the 10 test samples
(Table 1). All samples had the percentage of clay determined
by the traditional method (Calderano, 2011).
Table 1. Results of the univariate and multivariate models.
Clay, %
Univariate Multivariate
R2 cal 0,900 0,863
RMSEC 9,83 5,36 Figure 1. System for capturing images from
RMSEP 2,95 6,29 smartphone
Range 10 - 60 10 - 60
PhotoMetrix can be considered as an alternative for the determination of clay in soil samples in
miniaturized system with low cost, operational ease and speed in obtaining and processing digital images.
1 Gonzatti, CA, de Souza, ARR., Faqui, DP., Molz, RF, Santos, RO, Tischer, B, da Costa, AB. Journal of Food Process
Engineering, e14099.
2 Helfer, GA, Barbosa, JLV, Alves, D, da Costa, AB, Beko, M, Leithardt, VRQ, (2021). Journal of Sensor and Actuator
Networks, 3, 2021,40
3 Donagema, GK., de Campos, DB, CALDERANO, SB, TEIXEIRA, WG, & VIANA, JH, Manual de métodos de análise de
solo. Embrapa Solos-Documentos (INFOTECA-E). 2011.
[FAPERGS, CNPq, CAPES]

184
RAPID MOLECULAR DIAGNOSTICS OF COVID-19 BY RT-LAMP IN A
CENTRIFUGAL POLYSTYRENE-TONER BASED MICRODEVICE WITH END-
POINT VISUAL DETECTION
Kézia G. de Oliveira (PG),a* Paulo F. N. Estrela (PG),a Geovana de M. Mendes (PG),a Carlos A.
dos Santos (PG),b Elisângela de P. Silveira-Lacerda (R) b and Gabriela R. M. Duarte (R) a
a Instituto de Química, Universidade Federal de Goiás, Goiânia, GO, Brazil, 74690-900
b Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiânia, GO, Brazil, 74690- 900
*e-mail: oliveirakg2@outlook.com
Infection caused by the new coronavirus (SARS-CoV-2) has become a serious worldwide public
health problem, and one of the most important strategies for its control is mass testing.1 Real-time
reverse transcription PCR (RT-qPCR) is the gold standard for molecular diagnosis of COVID-19.2
However, RT-qPCR is costly and time consuming, and it requires highly trained personnel and
expensive laboratory equipment.3 Currently, isothermal techniques of nucleic acid amplification have
emerged to overcome the limitations of PCR, providing faster and lower cost molecular diagnostics,
which can be especially useful for developing countries or in the current situation where the lack of
PCR reagents has become an important issue.3 Loop-mediated isothermal amplification (LAMP) has
emerged as an important alternative to simplify the diagnostics of infectious diseases. 4 In addition,
an advantage of LAMP is that it allows for easy reading of the final result through visual detection.
However, this step must be performed with caution to avoid contamination and false-positive results.
LAMP performed on microfluidic platforms can minimize false-positive results, in addition to having
potential for point-of-care applications. Here, we describe a polystyrene-toner (PS-T) centrifugal
microfluidic device manually controlled by a fidget spinner for molecular diagnosis of COVID-19 by
RT-LAMP, with integrated and automated colorimetric detection. The amplification was carried out
in a microchamber with 5 µL capacity, and the reaction was thermally controlled with a thermoblock
at 72 °C for 10 min. At the end of the incubation time, the detection of amplified RT-LAMP fragments
was performed directly on the chip by automated visual detection (Figure 1). Our results demonstrate
that it is possible to detect COVID-19 in reactions initiated with approximately 10−3 copies of SARS-
CoV-2 RNA. Clinical samples were tested using our RT-LAMP protocol as well as by conventional
RT-qPCR, demonstrating comparable performance to the CDC SARS-CoV-2 RT-qPCR assay. The
methodology described in this study represents a simple, rapid, and accurate method for rapid
molecular diagnostics of COVID-19 in a disposable microdevice, ideal for point-of-care testing
(POCT) systems.
Figure 1. Schematic illustration of the visual detection LAMP in centrifugal PS-T microdevices.
1 WHO, World Heal. Organ, 2019, 2020, 2633.

2 H. Zhu, Z. Fohlerová, J. Pekárek, E. Basova and P. Neužil, Biosens. Bioelectron., 2020, 153, 112041.
3 B. Udugama, P. Kadhiresan, H. N. Kozlowski, A. Malekjahani, M. Osborne, V. Y. C. Li, H. Chen, S. Mubareka, J. Gubbay
and W. C. W. Chan, ACS Nano, 2020, 14, 3822.
4 T. Notomi, H. Okayama, H. Masubuchi, T. Yonekawa, K. Watanabe, N. Amino and T. Hase, Nucleic Acids Res., 2000,
28, e63
[CAPES/FAPEG, CNPq, MPT]

185
SIMULTANEOUS DETERMINATION OF CARBON AND NITROGEN IN SOIL BY
NEAR INFRARED SPECTROSCOPY
Jocelene Soaresa*(PG), Camila A. Gonzattib (PG), José G. Abichc (UG), Gilson A. Helferd (R),
Roberta O. Santosd (R), Adilson B. da Costaa,d (R)
a Universidade de Santa Cruz do Sul, Programa de Pós-Graduação em Tecnologia Ambiental, Santa Cruz do Sul, RS,
Brazil, 96815-900.
b Instituto de Química, Universidade Federal do Rio Grande do Sul, Programa de Pós-Graduação em Química, Porto
Alegre, RS, Brazil, 90650-001.

c Universidade de Santa Cruz do Sul, Departamento de Engenharias, Arquitetura e Computação, Santa Cruz do Sul, RS,
Brazil, 96815-900.
d Universidade de Santa Cruz do Sul, Programa de Pós-Graduação em Sistemas e Processos Industriais, Santa Cruz do
Sul, RS, Brazil, 96815-900.

*e-mail: s.jocelene@gmail.com
Near infrared spectroscopy (NIRS) associated with chemometric methods is a clean methodology,
due to advantages such as high analytical frequency, non-use of chemical reagents and does not
generate waste and, in addition, it stands out for its miniaturization and ease in the development of
portable devices1. The aim of this study was to develop multivariate calibration models using NIRS
for the determination of carbon and nitrogen in soil samples. The samples were dried and ground to
a thickness of 1 mm for determination carbon and nitrogen concentrations were measured for
reference method2, using an elemental analyzer (Flash EA1112, Thermo Electron Corporation,
Milan, Italy). A conventional infrared spectrometer (Spectrum 400, Perkin Elmer) was used with
accessory of near infrared reflectance (NIRA), in region 1250 - 2470 nm, obtained in absorbance
mode, with 16 scans for sample. As a pre-processing method was applied standard normal variate
(SNV) followed by first derivative and mean center. The partial least squares regression (PLS)
calibration model was carried out for simultaneous determination of carbon and nitrogen, using
SOLO+MIA software (Eigenvector Research, Inc), with 40 samples measured in duplicate, and
applied to the determination of carbon and nitrogen in 27 samples also in duplicate. The using 7
latent variables, presenting values of coefficient of determination to calibration (R2Cal) > 0.92, and
square root of the average error of prediction (RMSEP) values of 0,03% and 0,73% to nitrogen and
carbon, respectively. Others analytical parameters, such as coefficient of determination to cross-
validation (R2 CV) and prediction (R2Pred), and square root of the average error of calibration
(RMSEC) and cross-validation (RMSECV) are presented in Table 1. The methodology developed in
this project, still in evolution, intends to consolidate the use of NIRS as a viable alternative for the
analysis of soil fertility, also expanding its application to other variables, such as clay, silt and sand,
minimizing the generation of residues in agronomic laboratories.
Table 1. Results of the PLS simultaneous regression models.
Parameter Nitrogen Carbon
R2 Cal 0.928 0.950
R2 CV 0.870 0.903
R2 Pred 0.740 0.665
RMSEC, % 0.03 0.31
RMSECV, % 0.04 0.43
RMSEP, % 0.05 0.73
Range, % 0.03-0.5 0.2-5.2
1 BARRA I. HAEFELE S. M., SAKRABANI R., KEBEDE F. Soil spectroscopy with the use of chemometrics, machine
learning and pre-processing techniques in soil diagnosis: Recent advances–A review, Trends in Analytical Chemistry,135,
2021.
2 TEIXEIRA, P. C.; DONAGEMMA, G. K.; FONTANA, A.; TEIXEIRA, W. G. Manual de Métodos de Análises de Solos,
Empresa brasileira de pesquisa agropecuária – Embrapa - Solos, Brasília, DF, 2017.
[CNPq, CAPES, FAPERGS]

186
STENCIL-PRINTING OF PAPER-BASED ANALYTICAL DEVICES FOR
COLORIMETRIC SENSING
Jordana C. Jaimea,b* (UG), Habdias A. Silva-Netob (PG) Danielly S. Rochab (PG), Wendell K.
T. Coltrob,c (R)
aUniversidade Federal de Goiás, Faculdade de Farmácia, Goiânia, Goiás, Brazil, 74605-170
bUniversidade Federal de Goiás, Instituto de Química, Goiânia, Goiás, Brazil, 74690-900
eInstituto Nacional de Ciência e Tecnologia de Bioanalítica, Campinas, São Paulo, Brazil
*e-mail: jordanacj20.06@gmail.com
Paper-based devices including spot test, microfluidic and distance-based analytical devices (PAD and
µPAD) have been successfully employed in the analytical chemistry field. The popularity of the above
sensors can be associated to the electrochemical and colorimetric detectors that are easily assembled
on paper substrates. In this way, some manufacturing techniques as stamping, pencil drawing, laser
scribing, screen-printing and 3D printing have been reported for this purpose.1 However, the combination
of stencil-printing method and alternative binder does not have been satisfactory explored for fabricating
PAD and µPAD. Here, we development three different devices, spot test, PAD and μPAD exploring the
stencil printing approach and alkyd resin as hydrophobic chemical agent. As proof-of-concept, the
colorimetric assay of copper (Cu) was realized in authentic and adulted alcoholic beverages. For
manufacturing the different devices (Figure 1), the desired layout was modeled through a cameo studio
software. Next, three sheets composed by vinyl adhesive/chromatographic paper/thermoplastic were
sandwiched and cut by using a desktop cut plotter. Finally, the liquid hydrophobic agent was deposited
on stencil mask via spatula tool. The proposed device was characterized in terms of morphological, fidelity
and chemical resistance experiments. Considering to the colorimetric assay of Cu, reaction zones were
sequentially spotted with four different aliquots 1.5 μL bathocuproine solution (50 mg mL-1) prepared in
chloroform solvent, 1.0 mol L-1 acetate buffer (pH = 4.5), 1 mg L-1 hydroxylamine and the target samples
or standard solution of Cu. After finishing the colorimetric reaction, colorimetric zones were digitalized by
desktop scanner and the pixel intensity (gray channel) analyzed by using Corel Photo software. Through
scanning electron microscopy (SEM) measurements, it was possible visualize three important
morphological characteristic upon the manufactured devices: (i) solid binder was well-incorporated on
cellulose fibers (Figure 1B); (ii) satisfactory definition between native fibers and barrier and (iii) that the
theoretical values of microfluidic channels (ranged from 400 to 5000 µm) is in agreement with the
calculated values (R2 ≥ 0.99). Also, the proposed hydrophobic barrier when exposed to different chemical
solutions composed of acid-basic, organic solvent and surfactants (Figure 1C) exhibited excellent
chemical resistance. The analytical applicability for detection of Cu exhibited linear range (0.1 and 4.0
mg L-1; R2= 0.99; Figure 1D), limit of detection (0.05 mg L-1) and measured level of Cu in authentic
beverages samples ranged from 0.12 to 0.16 mg L-1. Future studies will be dedicated to detect Cu in
adulterated beverage samples. We believe that the proposed fabrication based on stencil-printing of
analytical devices on paper emerge as novel, simple, inexpensive and high-fidelity microfluidic channel
and portable analytical approach for colorimetric sensing.
Figure 1. Examples of PAD and µPAD; (B) SEM image of the barrier and (C) the chemical resistivity; (D) analytical
curve of Cu.
1. NISHAT, Sumaira et al. Sensors and Actuators B: Chemical, 336, 129681
[ INCTBio, CAPES, CNPq, LabMIC, UFG, GME]

187
SULFUR SPECIATION IN PETROLEUM ASPHALTS BASED ON REDOX
PROCESSES, LIQUID CHROMATOGRAPHY AND ED-XRF DETERMINATION
Patrícia Schmidt (PG)a*, Milena Pendeza (PG)a, Arthur Jacques (UG)a, Kezia de Souza (UG)a,
Jhovana Maciel (UG)a, Juliana Bortoluzzi (UG)a, Luiza Garcez (UG)a, Luiza Franzin (UG)a,
Gabriela Strieder (UG)a, Ana Amaral (UG)a, Leandro Carvalho (R)a
aFederal University of Santa Maria (UFSM), Department of Chemistry, Santa Maria, RS, Brazil, 97105-900
*e-mail: milenapendeza@gmail.com
Sulfur is the most abundant heteroatom present in

Figure 1. Distribution (%, w/w) of sulfur
petroleum asphalts, and these compounds are known
compounds over aromatic/saturated,
as precursors of asphalt oxidation reactions1. In this
aliphatic sulfides, and thiophenes.
context, specific classes of sulfur compounds were
investigated in four different Brazilian petroleum
asphaltic binders after two fractionation methods2,3.
Regarding the fractionation in thiophenes, aliphatic
sulfides and aromatic/ saturated sulfur fractions, most of
sulfur was found in aromatic/saturated fractions, a less
reactive form of sulfur. The results are demonstrated in
Figure 1. On the other hand, the separation in sulfur into
total acids, bases and neutral compounds allow the
creation of a reactivity index that classify the samples
regarding their aging susceptibility1. These results
indicated that sample C is the less reactive, and sample
D the most reactive. The speciation in the Brazilian
binder samples investigated in this work
showed a significantly higher amount of less reactive compounds (neutral compounds) compared to acid
and basic ones, as can be seen in Figure 2. It is worth to note that the results over the asphaltenic fraction
speciation present a higher amount of basic (negatively charged) compounds, what may indicate the
higher reactivity of this fraction. The sulfur speciation into acidic, basic and neutral compounds in the
asphaltenic fraction is demonstrated in Figure 3. The results permitted a comparative study of the
chemical processes taking place in 4 asphaltic binders based on the sulfur speciation, what can be useful
to predict the aging susceptibility of asphalts used for paving.
Figure 2. Distribution (%, w/w) of sulfur Figure 3. Distribution (%, w/w) of acid, basic and
compounds into acidic, basic, and neutral neutral compounds in the asphaltenic fraction.
compounds in the whole asphalt.
1 Carvalho LM, Nascimento PC, Bohrer D, Claussen LE, Ferraz L, Grassmann C, Braibante HTS, Dias D, Cravo M, Leite
LFM, Energy and Fuels, 29, 2015, 3.
2 Green JB, Hoff J, Woodward PW, Stevens LL, Fuel, 63, 1984, 9.
3 Payzant JD, Mojelsky TW, Strausz OP, Energy and Fuels, 3, 1989, 4.
[CAPES, CNPq, Petrobras]

188
UNDERWATER DETERMINATION OF METAL IONS BY LASER-INDUCED
BREAKDOWN SPECTROSCOPY (LIBS)
Nilvan A. Silva (PG)*, Ivo. M. Raimundo Jr. (R)

State University of Campinas - UNICAMP, Department of Analytical Chemistry - Institute of Chemistry, Campinas, São
*e-mail: nilvan_alves@hotmail.com
The deposition of inorganic ions in pipelines of marine platforms for oil exploration is one of the main
causes of the productivity reduction of the oil industry. The chemical composition and the
combination of waters used in oil production are factors responsible for scale formation1. Therefore,
the determination of scaling ions, such as Ca(II) and Sr(II), in produced waters can be used to
indicate whether these waters have a scaling potential. The purpose of the present work is the direct
determination of Ca(II) and Sr(II) in liquid samples by underwater LIBS measurements. A Q-switched
Nd:YAG laser (Quantel, Brio) operating at the fundamental wavelength (1064 nm), with a pulse
duration of 5.5 ns was employed (Figure 1). The laser beam was expanded by a concave lens (L1,
f = -25 mm, Ø = 12,5 mm, Thorlabs) and a convex lens (L2, f = 150 mm, Ø = 25 mm, Thorlabs) to
achieve a large focusing angle for better stabilization of plasma in water. The laser was reflected by
a dichroic mirror (Ealing Electron-Optics) and an achromatic doublet (L3, f = 60 mm, Ø = 25 mm,
Thorlabs) and a meniscus lens (L3, f = 100 mm, Ø = 25 mm, Thorlabs) were employed to minimize
the spherical aberration. Then, the laser beam was focused into a cuvette filled with 25 mL solution
containing 200 mg L-1 of Ca(II) and Sr(II). A 5-mm diameter lens was employed to collect the radiation
emitted by the plasma and focus it on the tip of an optical fiber (L4) with a diameter of 105 µm. Signal
acquisition was performed by an echelle polychromator and an ICCD detector (iStar DH 734, Andor
Technology). Parameters such as laser pulse energy (19 - 50 mJ), gate delay (200 - 1500 ns), gate
width (0.5 - 5.0 µs), number of pulses (20 - 100 pulses), repetition rate (2 - 20 Hz) and solution
salinity (0 - 5.0% (m/v) NaCl) were studied. A laser pulse energy of 31 mJ, a gate delay of 300 ns
and a gate width of 5 µs were chosen based on a univariate approach. A total of 100 laser pulses
and a 10 Hz repetition rate provided the highest signal intensity. For Sr(II), an increase in RSD(%)
occurs when the repetition rate increases from 10 to 20 Hz, due to the increase of bubbles produced
in the solution that can deviate the laser pulses, affecting the repeatability of the measurements.
Emission intensities increased around 20% for Ca (422.7 nm) and 27% for Sr (460.7 nm) when NaCl
concentration was varied from 0 to 0.2% (m/v). However, the intensity for Ca and Sr decreased
around 57% when the salinity varied from 0.2 to 5% (m/v). These results indicate that the underwater
LIBS measurements have potential application in the quantification of scaling ions and can be used
as a tool to help solve problems of industrial interest.
Figure 1. Schematic diagram of the LIBS setup.
1 Chen M, Shafer-Peltier K, Veisi M, Randtke S, Peltier, E. Separation and Purification Technology, 222, 2019, 1
[INCTAA, CNPq, FAPESP]

189
UNMANNED AERIAL VEHICLE ADAPTED FOR SAMPLING OF
ENVIRONMENTAL WATERS AND IN SITU VOLTAMMETRIC ANALYSIS OF
POLLUTANTS
Vinícius A Carvalho (UG)a, Vagner B dos Santos (R)a,*, Rodrigo A Avelino (PG)a
aFederal University of Pernambuco/ Department of Fundamental Chemistry, Recife, Pernambuco, Brazil, 50740-560
*e-mail: vagner.bsantos@ufpe.br
Water is an essential resource for the maintenance of life and for the development of society. It is
important to emphasize that the water is intended for several applications, such as irrigation,
domestic, energy generation, and industrial uses. In Brazil, there is a growing demand for this
resource according to Conjuncture of Water Resources in Brazil 2021 produced by National Agency
for Water and Basic Sanitation (ANA)1. According to this report, in the last two decades there has
been a significant growth in water withdrawals for irrigation, which went from 640 to 965 cubic meters
per second. In addition, there is a prospect of growth of about 42% of water withdrawals until 20401.
These data corroborate with the importance of preservation of waters. In this context we proposed
an automatized water sampling system based on an unmanned aerial vehicle (UAV) with in situ
electrochemical analysis of pollutants. The automatized method was developed to be a useful tool
compared to the traditional processes of water sampling in which it requires human intervention, it
can lead to a contamination of the ecosystem and it exposes the analyst to toxic substances that
may be contained in contaminated water samples. Moreover, the adapted UAV could be used to
access remote and inaccessible areas2. The UAV adapted is an autonomous system for surface
water sampling employing a miniaturized pump and solenoid valve controlled by an open source
microcontroller. The water sampling system is powered by a 9 V battery and a booster electronic
circuit. In this project, a quadcopter Mavic Air (DJI) which has a maximum flight time of 21 min was
used, Fig. 1. All automated water sampling system coupled to the UAV is controlled by an algorithm
developed in C language. This algorithm was developed to optimize the battery life to perform the
displacement of the UAV for sequential sampling without any cross contamination. For this, all water
sampling system is switched OFF during the displacement of the aircraft to collect points. A portable
potentiostat, EmStat Pico (PalmSens) is used to perform the in situ voltammetric analysis. For this
purposes an electrochemical cell with 3 electrodes such as boron-doped diamond (BDD) as work
electrode (WE), Ag/AgCl as reference electrode (RE), and platinum as auxiliary electrode (AE) was
used. Moreover, a screen-printed electrode (SPE) based on graphite as WE, Ag as RE and Pt (AE)
was used. To evaluate the performance of this compact and portable system, a cyclic voltammetry
was performed to a solution containing 1.0 mM of K3[Fe(CN)6] with 0.1 M KCl as supporting
electrolyte and the voltammogram obtained using BDD is presented in Fig. 1.
A B C D
Figure 1. The UAV adapted with a water sampling system (A) with highlights the frontal face of the UAV and a 1.5 mL
sampling flask supported by a customized 3D piece (B). A open source microcontroller to control the microchip potentiostat
coupled to a SPE (C). A voltammogram of 1.0 mM K3[Fe(CN)6] with 0.1 M KCl at a scan rate of 100 mV/s with n= 9 cycles
(D).
1. Agência Nacional de Águas e Saneamento Básico. Conjuntura dos Recursos Hídricos no Brasil 2021. Disponível em:
https://relatorio-conjuntura-ana-2021.webflow.io/capitulos/usos-da-agua. Available: 06/07/2022. 2. Terada A, Morita Y,
Hashimoto T, Mori T, Ohba T, Yaguchi, M, Kanda W, Earth, Planets Space, 70, 2018, 64.
[CNPq, FACEPE]

190
UNMANNED AERIAL VEHICLE FOR ENVIRONMENTAL MONITORING
Vagner B dos Santos (R)a,*, Vinícius A Carvalho (UG)a, Rodrigo A Avelino (PG)a
aFederal University of Pernambuco/ Department of Fundamental Chemistry, Recife, Pernambuco, Brazil, 50740-560
*e-mail: vagner.bsantos@ufpe.br
An unmanned aerial vehicle (UAV) is an aerial vehicle that is remotely controlled or programmed to
perform a task autonomously. UAVs are rotary wing aircraft type quadcopter (four rotors) or hexacopter
(six rotors), highly foldable, compact, and they have sensors to avoid collisions with flight time on average
20-30 min and traveling from 2 to 8 km for the most common models1. Previously restricted to the military
area, UAV has been used for agriculture, support for humanitarian actions and environmental
preservation1. In this work, the UAV was adapted to perform water sampling after an environmental
disaster such as the spill of oil and to monitor residues of pesticides in water, Fig. 1. For this, a solenoid
micropump and a peristaltic pump controlled by an open source microcontroller were embedded on an
UAV. For this, an algorithm in C language was developed to perform two samples sequentially at a flow
rate of 30 mL/min. This time is enough to fill a sample flask of 100 mL for a time lower than 5 min. By
performing this setup, the flow line is cleaned and no cross-contamination is observed. This sample
procedure is important because inexpensive UAV has a limitation of flight time, and the battery is a
limitation mainly for great distances. Moreover, this method is quick and safe, and only one professional
is needed compared to the traditional way [1]. After the liquid-liquid microextraction was performed using
5 mL of each collected sample and 5 mL of the hexane this aliquot was preconcentrated to 500 μL and 1
μL was injected in the GC-MS equipment for analysis of BTEX (benzene, toluene, ethylbenzene, and o,
m, p-xylenes) for a concentration range from 1.00 μg L-1 to 100 μg L-1. The column temperature was 50
°C for 1 min, 60 °C at 2 °C min-1 and afterwards to 200 °C at 50 °C min-1 and then held at 200 °C for 2
min. The analysis was carried out in the selected ion monitoring mode using electronic impact ionization
of 70 eV keep at 250 °C to detect m/z 78 (benzene), 92 (toluene), 106 (ethylbenzene and xylenes), and
106 (benzaldehyde), respectively. The BTEX concentration ranged from 0.3 to 5.0 μg L-1 in the samples
with a recovery from 83.9 to 118% with a relative standard deviation lower than 7.0%. In the second
application we advance for in situ voltammetric analysis of 2,4-dichlorophenoxyacetic acid (2,4-D) in
environmental water samples, using a electrochemical cell made in 3D printer using a biodegradable
polylactic acid (PLA) filament. A boron-doped diamond electrode, Pt and Ag/AgCl (KCl 3M) were used as
work, auxiliary and reference electrodes, respectively. After optimizing the supporting electrolyte, pH and
parameters of the square wave voltammetry a linear analytical curve for 2,4-D in 0.5 mol L-1 Na2SO4 (pH
2.0 regulated using 0.5 mol L-1 H2SO4) in a concentration range from 100 nmol L-1 to 911 nmol L-1 with
34 nmol L-1 as the limit of detection was obtained. The same samples were sent to the laboratory for GC-
MS analysis and no statistical difference at a 95% confidence level was obtained. This is the first time
that an adapted UAV was used for these applications being a tool to alert the presence of environmental
hazard compounds or to evaluate the environmental water quality with response in real time.
a b c d
Figure 1. Mavic 2 Pro adapted for water sampling (a). UAV used to collect mangrove water (b). Mavic air adapted for
water sampling (c), a 3D customized piece coupled to a 1.5 mL flask (d).
1 Terada A, Morita Y, Hashimoto T, Mori T, Ohba T, Yaguchi, M, Kanda W, Earth, Planets Space, 70, 2018, 64.
[CNPq, FACEPE]

191
USING HIGH-RESOLUTION 3D PRINTING TO MANUFACTURE
MICRONEEDLES FOR CHEMICAL ANALYSIS
Reverson F. Quero (PG)*a, William R. de Araujo (R)a, Dosil P. de Jesus (R)a
aUniversidade Estadual de Campinas, Instituto de Química, Campinas, São Paulo, Brasil, CEP 13083-970
*e-mail: reverson.quero@iqm.unicamp.br
In recent years, additive production technology has attracted the attention of the scientific community
in several areas. The various 3D printing techniques have allowed a paradigm shift in designing and
manufacturing customized devices according to individual needs1. The increase in print resolution,
equipment precision, and the accessibility of low-cost and biocompatible printing materials have
allowed 3D printing to be used to manufacture microneedles (MN).2 Despite the fact that current 3D
fabrication techniques of MN can produce an acceptable and cost-effective resolution, it still requires
complex steps and high proficiency in microfabrication, which strongly delays exploration in research
and new applications.The advantages of MNs are increasing recently with the possibility of
manufacturing a variety of structures, including hollow, solid, coated, and dissolvable MN, with
various sizes and features for various applications. In this work, we used an affordable high-
resolution DLP 3D printer (Phrozen Sonic Mighty 8K) with resin from Siraya Tech to perform the
fabrication of microneedles. This fabrication started from a 3D BIM project (Figure 1A), and the initial
prototypes (Figure 1B) were printed and optimized until reaching the best resolution and the smallest
diameter possible. The projected MN measure 200 µm at their base and 800 µm in height and have
perforations throughout their structure (Figure 1C). A microscopy image confirmed the possibility of
high-resolution printing of these complex microneedles. (Figure 1D).t b
e writtArial font, size 11, single spa justified paragraph
.
Figure 1. (A) 3D computational microneedle design. (B) 3D printed device. (C) Detail of the holes in the
microneedle structure. (D) Microscopy image of the printed needles.
The development of these 3D-printed MN can revolutionize the drug delivery systems, extraction of
biofluids from tissues (human and plants), and chemical and point-of-care analysis, benefiting
several areas such as medicine and agriculture.
1 Gross B, Lockwood SY, Spence DM Analytical Chemistry, 89, 2017.
2 Economidou SN, Douroumis DB, Advanced Drug Delivery Reviews, 173, 2021.
[CAPES, FAPESP, FAEPEX, CNPq,INCTBio]

192
SEPARATION METHODS
193
AN INTEGRATED MULTIVARIATE APPROACH FOR THE DEVELOPMENT OF
A COMPREHENSIVE CHROMATOGRAPHIC METHOD FOR THE ANALYSIS OF
BIOACTIVE COMPOUNDS IN CITRUS SPP.
Igor M. Santanaa (PG), Maurício A. Rostagnob (R), Márcia C. Breitkreitza (R)*
aUniversity of Campinas, institute of Chemistry, Campinas, SP, Brazil, 13083-970
b University of Campinas, School of Applied Sciences (FCA), Limeira, SP, Brazil, 13484-350
*e-mail: marciacb@unicamp.br
Chromatographic methods are usually developed by changing one factor at a time (univariate
approach). However, the academic and industry environments are drifting their practices towards a
multivariate risk-based approach [1]. In this context, the relationship between one or more responses
(e.g., resolution between critical pairs) and the critical method parameters (such as stationary and
mobile phases composition, temperature, gradient time, etc.) are linked through design of
experiments (DoE). This work focused on the development of a method using an ultra-high
performance liquid chromatography for comprehensive analysis of bioactive compounds found in
the Citrus genus (coumarins, furocoumarins, flavones, and flavanones). This plant genus was
chosen because of the increasing interest in natural products and the high availability of these
compounds from the wastes due to the citrus processing industry activities [2]. The experimental
designs were created by the software Fusion QbD (S-Matrix, USA), which was integrated with
Empower (Waters, Milford, USA) to perform chromatographic runs. Then, the data were statistically
treated in the same software that built visualisation graphics and statistical reports as decision tools.
A first step involved the screening of four stationary phases (YMC Triart C18, YMC Triart Phenyl,
YMC Triart Pentafluorophenyl, and BEH C8, fully porous particles with size of sub-2μm) and two
strong solvents (methanol and acetonitrile) in different gradient times and temperatures using an
optimal design based on the A and G criteria. A second step was the optimization of the gradient
time, temperature and flow rate in the column YMC Triart C18, and the mobile phase was composed
by a weak solvent (acetic acid 0.20 % v/v) and a strong solvent (acetonitrile:methanol 3:1 v/v). This
optimization step employed a rotatable central composite design with five replicates in the center
point. The responses retention of the last peak, overall resolution, peak capacity, and resolution
between critical pairs were simultaneously evaluated by a desirability criterion. Monte Carlo
simulation was carried out to predict the robustness of the models by establishing the method
operable design region (MODR). The final conditions for the stationary and mobile phase
abovementioned were 0.68 mL.min-1, 33.3 °C and gradient time of 18 min (strong solvent from 10 to
80 %). Finally, the quantitative performance was evaluated for selected compounds: hesperidin
(polar flavanone), nobiletin (nonpolar flavone), bergapten (furocoumarin), citropten and herniarin
(coumarins). The validation was based on the tolerance interval which combines systematic (bias)
and random (intermediate precision) errors to obtain an accuracy profile as unique decision tool [3].
Limits of detection were as low as 0.05 μg.mL-1, lower limits of quantification were 0.10 μg.mL-1, and
upper limits of quantification were 100 μg.mL-1. Based on this approach, it is possible to state that a
proportion of 95 % of the future samples will fall within an acceptance limit of ± 15 % in the range of
0.10 – 100.00 μg.mL-1 with a confidence level of 95 %. Also, the developed method allowed the
distinction between Citrus species based on the bioactive compounds (oranges, tangerines,
grapefruit, limes, and lemons). Therefore, this method can be applied for qualitative and quantitative
performance regardless the Citrus fruit to be analyzed.
1. R. Deidda et al., Journal of Pharmaceutical and Biomedical Analysis, 161, 2018, 110
2. E. Gómez-Mejía et al., Food Chemistry, 295, 2019, 289
3. P. Hubert et al., Journal of Pharmaceutical and Biomedical Analysis, 36, 2004, 579
[CNPq, CAPES, Waters Corporation and S-Matrix]
––––––––––––––––––––– Separation methods –––––––––––––––––––––

194
APPLICATIONS OF ARTIFICIAL INTELLIGENCE CONCEPTS IN
COMPREHENSIVE TWO-DIMENSIONAL GAS CHROMATOGRAPHY COUPLED
TO HIGH RESOLUTION MASS SPECTROMETRY
Leandro Wang Hantao (R)
University of Campinas, Institute of Chemistry, Campinas, São Paulo, Brazil, 13083-862
*e-mail: wang@unicamp.br
Comprehensive two-dimensional gas chromatography takes advantage of the sharp peaks that are
distributed in a retention plane to improve the overall peak capacity of the composite system [1]. The
sharp peaks generated in the GC×GC experiments determine the hardware settings required to
produce reliable chromatographic data. Hence, data acquisition is vital for proper peak
reconstruction in the GC×GC chromatograms. In addition to the well-known and described
transmission quadrupoles (QMS) and time of flight mass analyzers (TOFMS), the electrostatic ion
trap mass analyzer (FT-Orbitrap) uses an optimized ion management system that requires careful
optimization for GC×GC experiments. The orbital ion trap decomposes the ion trajectories of a well-
defined ion package across the fusiform central electrode for m/z measurements assisted by Fourier
transform. The acquisition time of the induced current image is highly dependent on the desired
mass resolving power. So, the ion management is assisted by the curved flatpole (i.e., C-trap) and
a charge collector. The high ion transmission is attained by carefully alternating the events in the
mass spectrometer. As the image current is being acquired and deconvoluted to produce the mass
spectrum at the desired mass resolution, the C-trap stores the incoming ions. The size of the ion
package is defined by the ion injection time (IT) and the ion current reaching the C-trap. In order to
find the most acceptable settings for GC×GC experiments, we have carefully evaluated the
interaction of IT and desired mass resolving power, as well as non-conventional hardware settings
to measure (or estimate) the ion current. For instance, instead of using the charge collector, the ion
current can be estimated using the one (or two) previous spectra) to determine the IT value [2].
GC×GC investigations are well known to generate a substantial amount of information-rich and
structurally complex data, requiring advanced data processing strategies like chemometrics. Many
workflows are available for data handling and processing, such as the peak-table and pixel-based
approaches. The goal of the first case study was to present a solution based on method development
to solve the missing pixel problem that may be encountered in experiments performed GC×GC-
HRMS. Data input is vital for pixel-based chemometric analyses (e.g., using MATLAB), as some
post-processing solutions may lead to significant loss of chemical information in the data set. Hence,
a key requisite is that the chemical information is consistently indexed in the data arrays for proper
pixel-based data handling and analysis [2,3]. The second set of case studies highlights some
interesting opportunities for the use of chemometrics in GC×GC, which include partial least squares
regression (PLS) [4] and artificial neural networks [5] for quantitation of physical-chemical properties
of petroleum derivatives, and data-dependent soft independent modelling of class analogy for one-
class classification [6] to monitor a quality parameter of waste water produced by the petroleum
industry, total oil and grease content.
[1] Pollo, BJ; Alexandrino, GL; Augusto, F; Hantao, LW; Trends in Analytical Chemistry 105 (2018) 202-217.
[2] Oliveira, AM; Teixeira, CA; Hantao, LW; Analytical Methods 14 (2022) 1646-1654.
[3] Hantao, LW; Teixeira, CA; Ferreira, VHC, (2022) https://doi.org/10.7910/DVN/WMTEMF
[4] Berrier, KL; Freye, CE; Billingsley, MC; Synovec, RE; Energy Fuels 34 (2020) 4084-4094.
[5] Marini, F; Bucci, R; Magri, AD; Microchemical Journal 88 (2008) 178-185.
[6] Ferreira, VHC; Poppi, RJ; Hantao, LW; Talanta 234 (2021) 122616.
[Petrobras, Nova Analítica, ThermoFisher]

195
ASSESSMENT OF AS AND CD CONTAMINATION IN MARINE FISH BY
COPRECIPITATION-MICROWAVE INDUCTIVELY OPTICAL EMISSION
SPECTROMETRY ARRANGEMENT
Ívero Pita Sá (PG)a*, Julymar M. Higuera (UG)b, Carlos Alberto Silva (R)c, Ana Rita A
Nogueira (R)a
aEmbrapa Pecuária Sudeste, São Carlos, São Paulo State, 13560-970, Brazil
bGroup of Applied Instrumental Analysis, Department of Chemistry, Federal University of São Carlos, São Carlos, São
Paulo State, 13565-905, Brazil
c Embrapa Coastal Tablelands, Aracaju, SE, 49025040, Brazil
*e-mail iveropita@gmail.com
Coprecipitation is a remarkable separation/preconcentration method, still not very well explored by

combining Microwave-Induced Plasma Optical Emission Spectrometry (MIP OES). It has
advantages such as simplicity, low consumption of reagents, and high preconcentration factor, which
are essential in the determination of trace elements1,2. Therefore, a procedure for As and Cd
determination in fish samples is described. It combines MIP OES with silver nitrate and potassium
chromate coprecipitation. The procedure is based on the separation and preconcentration of the
analytes from the addition of silver nitrate and potassium chromate to a 20 mL solution containing
the analytes. In this way, the analytes are coprecipitated in the precipitate formed (Ag 2CrO4),
separated by centrifugation, diluted in 700 µL of acid, and determined by MIP OES. Factors that
directly affect the coprecipitation procedure, such as silver and chromate concentrations and solvent
volume, were optimized using the Box-Behnken design. The optimized conditions were 5.46x10-4
mol L-1 [CrO42-] and 1.16x10-3 mol L-1 [Ag+], without need for solvent. The procedure provided
preconcentration factors of 25 and 11 for As and Cd, respectively, and LOD adequate to the
international legislation. The trueness was assessed from the analysis of certified reference
materials (CRM) of dogfish liver (DOLT-5) and lobster hepatopancreas (TORT-3) and compared with
Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Relative standard deviations < 9% and
3.5% and recoveries > 91% and 88% were obtained for As and Cd, respectively. The method was
applied to local market fish samples, and it showed suitable for As and Cd determination by MIP
OES.
1 I.P. Sá, L.N. Santos, G.P. Erik, D.D.C. Lima, F. Alan, C. Amorim, Pré-Concentração Baseada na Coprecipitação usando
Cromato de Prata como Carreador para Determinação de Cobre por FAAS, Quim. Nova. 42 (2019) 10–16.
2 D.A. Skoog, D.M. West, F.J. Holler, S.R Crouch, Fundamentals of Analytical Chemistry, 9a Ed., Cengage Learning,
2014.
[FAPESP. CNPq, CAPES, INCTAA]

196
AUTOMATED FILTERING SCRIPTS BASED ON ACCURATE MASS APPLIED TO
GC×GC-HRMS FOR GROUP-TYPE ANALYSIS OF NAPHTHENIC ACIDS
Naiara M. F. M. Sampaioa*, Juliana Crucelloa, Iris Medeiros Jr.b, Rogerio M. Carvalhob,
Leandro W. Hantaoa
aInstitute of Chemistry, University of Campinas, 13083-862 Campinas, SP, Brazil
bLeopoldo Américo Miguez de Mello Research and Development Center, Petrobras, Rio de Janeiro 20031-912, RJ,
Brazilc
*e-mail: naiaramfms@gmail.com
Produced water (PW) is the primary effluent generated by the oil and gas industry, having a
diversified composition including apolar and polar compounds. Naphthenic acids (NAs) have gained
significant awareness among these analytes since they can cause operational and environmental
issues[1]. NAs are naturally occurring compounds found in crude oil with structures comprising one
or more carboxylic groups bonded to a naphthenic moiety substituted with linear or branched alkyl
side chains[2]. The complexity of NAs mixtures typically requires high peak capacity techniques for
analyzing diastereomers. In this context, GC×GC-MS is an interesting alternative to monitoring these
compounds in complex PW samples. Nevertheless, analyte identification remains one of the major
challenges in untargeted approaches[3], especially for compounds that underwent a derivatization
step, such as NAs. The contemporary petroleomics approach avoids using custom databases, which
requires authentic standards and the analytical methods' standardization[4]. However, the
petroleomics protocol may produce false-positives for GC-based methods due to the high
fragmentation of analytes caused by the ionization process, requiring double-checking every entry
of the peak table, which is very time-consuming since GC×GC generates overwhelmingly large and
complex sets of data[5]. In this work, we have conducted a detailed evaluation of filtering scripts for
GC×GC-HRMS to perform group-type analysis of NAs in complex mixtures, like produced water
(PW) samples. These automated scripts were based on high-resolution mass spectral information
acquired by using electron ionization (EI) and chemical ionization in the positive mode (PCI). The
most common derivatization methods were carefully evaluated, namely silylation with N-methyl-N-
tert-butyldimethylsilyl trifluoroacetamide (MTBSTFA) or N,O-bis(trimethylsilyl)trifluoroacetamide
(BSTFA) and methylation using a Lewis acid catalyst (BF3) in methanol. NAs were extracted from
reference and unknown PW samples by liquid-liquid extraction with dichloromethane. After
derivatization, the analytes were separated in a GC×GC-FT-Orbitrap MS system with a non-polar ×
mid-polar column set and flow modulation using the reverse fill/flush configuration. GC Image 2020r2
software was used to create the Computer Language for Identify Chemicals scripts. The best
performing approach for qualitative analysis was attained by combining the MTBSTFA derivatization
agent with EI, which resulted in reproducible fragmentation patterns and high-intensity even-electron
ions ([M-57]+). These data enabled the development of selective scripts, exhibiting high rates of true
positives above 98 % and 88 % for the reference and unknown PW samples, respectively. The
retention pattern of GC×GC was used to discriminate ambiguous identification, highlighting the
synergy between GC×GC and HRMS. The proposed scripts are an alternative to profile 154 classes
of MTBSTFA-derived NAs, considerably reducing the time spent in manual data curation and
annotation. Therefore, this was an important step to aid in the development of standard methods for
studying and regulating water-soluble compounds found in PW samples.
[1] J. Crucello, R. M. Carvalho, L. W. Hantao, LC GC N. Am. 2021, 39, 478–485.
[2] J. S. Clemente, P. M. Fedorak, Chemosphere 2005, 60, 585–600.
[3] I. Blaženović, T. Kind, J. Ji, O. Fiehn, Metabolites 2018, 8, 1–23.
[4] J. Lee, T. Kind, D. J. Tantillo, L. P. Wang, O. Fiehn, Metabolites 2022, 12, 1–19.
[5] S. E. Prebihalo, K. L. Berrier, C. E. Freye, H. D. Bahaghighat, N. R. Moore, D. K. Pinkerton, R. E. Synovec, Anal.
Chem. 2018, 90, 505–532.
[PETROBRAS, FAPESP, CNPq,,CAPES, Nova Analítica, ThermoFisher Scientific]

197
CAPILLARY ELECTROPHORESIS WITH UV DETECTION APPLIED TO
ANALYSIS OF URINARY ORGANIC ACIDS FOR TARGETED METABOLOMICS
Marcone Augusto Leal de Oliveiraa,c, Jéssica Cordeiro Queiroz de Souzaa, João Marcos
Beraldo Candidoa, Maria Patrícia do Nascimentoa, Olívia Brito de Oliveira Moreiraa, Paula
Rocha Chellinia,b.
aGroup of Analytical Chemistry and Chemometrics - GQAQ, Chemistry Department, Institute of Exact Sciences, Federal
University of Juiz de Fora (UFJF), 36026-900, Juiz de Fora-MG, Brazil.
bFaculdade de Farmácia, Federal University of Juiz de Fora (UFJF), 36026-900, Juiz de Fora-MG, Brazil.
cNational Institute of Science and Technology for Bioanalytics – INCTBio, Institute of Chemistry, University of Campinas
(UNICAMP), 13083-970, Campinas, SP, Brazil

e-mail: marcone.oliveira@ufjf.br
Capillary electrophoresis (CE) is an electromigration technique that has become a part of the
outstanding high-throughput platform for bioanalysis. By using the selectivity associated to CE-UV-
based methods, the aim of our work was to define an organic acid profile for individually collected
human urinary samples. Organic Acids (OAs) is present in urine as products of a wide range of
metabolic pathways and could indicate a series of health-related abnormalities, consequently, the
resulting OAs screening from the CE-UV analysis could be used as entrance data for bioinformatic
and machine learning protocols for discriminative statistics of any human anomaly under study.1
A total of 100 human urine samples were individually collected by the volunteers and prepared
considering protein extraction with minimum handling and dilution. The supernatant was directly
injected in triplicates under the optimized CE method. An adaptation of a CZE with indirect UV
detection method previously developed at our group2 designed originally for phytochemical analysis
was made considering this specific biological matrix, for better visualization of peak profile in urine,
we enhanced the analysis by adjusting the pH of
the electrolyte (Tris/Phthalic Acid) at 3.8 rather
than the original pH 3.2. Additionally, we used a
20-cm longer TSP capillary and optimized the
conditioning between runs and the system clean-
up for better reproducibility. Over the analysis of
all samples, a pool of urine was injected between
6 runs as quality control (QC). The EOF
migration time was used as precision parameter,
for the analysis of a hundred samples that lasted
approximately 80 hours, the EOF signals was
exhibit at 14.77 min average with 1.54 % RSD.
Fig 1. Shows the resolution of a standard mixture
of low chain dicarboxylic acids and examples of
three random urine with distinguishing OA
profiles. Considering the possible interferents,
the urine major components (urea and
creatinine) are invisible in this condition as the
major inorganic salts migrate faster than the low-
chain OA under study.
Figure 1. Examples of electropherograms of urinary specimens.
(1) tartrate; (2) malate; (3) lactate; (4) succinate,
*chloride and other unidentified fast anions.
1 Moreira OBO, Souza JCQ, Candido JMB, Nascimento MP, Penna EA, Chellini PR, Oliveira, MAL, Brazilian Journal of
Analytical Chemistry, (Forthcoming), 2022.
2 Vaz FAS, Silva PA, Passos LP, Heller M, Micke GA, Costa ACO, Oliveira MAL, Phytochemical Analysis, 23, 2012.
[CNPq, CAPES, Fapemig, INCTBio, Lemos Laboratórios]

198
DETERMINATION OF ABAMECTIN IN SOYBEAN ROOTS BY LIQUID
CHROMATOGRAPHY COUPLED TO TANDEM MASS SPECTROMETRY
Eliézer de Oliveira (PG)*, Carla B. G. Bottoli (R)
UNICAMP, Institute of Chemistry, Campinas, SP, Brazil, 13083-872
*e-mail: e266022@dac.unicamp.br
Abamectin is registered to control root-parasitic nematodes for use on soybean and other
crops. As a nematicide, abamectin has been applied as a seed treatment and its application by foliar
spray is limited since abamectin has low foliar translocation1. Therefore, some strategies have been
used to improve the uptake and translocation of pesticides by plants2. In this study, we have
evaluated the combined use of abamectin with copper complexes in the translocation of this
nematicide in soybean plants. For that, an HPLC-MS/MS method has been developed for analysis
of abamectin in soybean roots. Samples were washed with water and dried under a nitrogen stream.
One g of homogenized sample was weighed into a 50 mL Teflon centrifuge tube and mixed with 10
mL of acetonitrile in a Vortex mixer for 1 minute. For phase separation, 4 g anhydrous sodium sulfate
(MgSO4) and 1 g sodium chloride (NaCl) were added and mixed immediately for 1 min. After
centrifugation for 5 minutes at 5000 rpm, the supernatant was transferred to a clean tube containing
150 mg PSA sorbent plus 1 g anhydrous MgSO4 and vortexed for 30 s. Subsequently, the extract
was centrifuged for 5 minutes at 5000 rpm and 1 mL aliquot of the extract was filtered before injection
into the HPLC–MS/MS system. The HPLC-MS/MS system consisted of a Waters Alliance 2695
separation module coupled to a Waters Micromass Quattro Micro API tandem mass spectrometer
equipped with an electrospray ionization (ESI) source. HPLC-MS/MS analysis was performed on a
Nova-Pak C18 column (150 mm x 3.9 mm, 4.0 μm, Waters) that was operated at 40 ºC. The
separation was performed with an isocratic mode run and the mobile phase consisted of 15% water
(pH 4.0) (solvent A) and 85% methanol (solvent B), both modified with 10 mmol L−1 ammonium
formate. The injection volume was 10 μL, the flow rate was set at 1 mL min −1 and the total run time
was 10 minutes. The electrospray was operated in the positive mode and the source parameters
were: capillary voltage 3.5 kV; cone voltage 20 V; source temperature 150 ºC; and desolvation gas
temperature 450 ºC. The flow rates for desolvation gas and cone gas (N2) were set at 50 and 1000
L h−1, respectively. The detection was carried out in the selected reaction monitoring (SRM) mode.
The collision energies were 28 eV (m/z 890,57 → m/z 305,3) and 18 eV (m/z 890,57 → m/z 567,4).
By comparing the response of the analyte in fortified matrix extract and that in solvent, the matrix
effect was considered as significant (>20%). Consequently, the samples were quantified with matrix
matching. The limit of detection was 0.10 mg/kg and the limit of quantification was 0.25 mg/kg. Good
linearity of the analytical curve was obtained over the studied range of concentrations from 0.10 to
1.0 mg/kg, with a correlation coefficient of 0.994. The trueness and precision of the method were
evaluated by spiking the blank samples of soybean roots with abamectin at three levels of
concentration (0.1, 0.5, and 1.0 mg/kg). The method presented adequate trueness with recoveries
ranging from 96.1 to 105.6%. Intra-day RSD values in the 3.1 - 9.0% were obtained. Inter-day (3
days) RSD values were all within 5.9 - 7.2%. Finally, we compared the translocation of abamectin
and abamectin + Fulland® after foliar spray application in a greenhouse. Root samples were
collected at 4, 7, 14, 21, 28, 35, 42 and 49 days after treatment. All the samples analyzed were seen
to be free of the abamectin, at least above the detection limit.
1 Faske TR, Starr JL, Journal of Nematology, 39, 2007, 27.
2 Wang CJ, Liu ZQ, Pesticide Biochemistry and Physiology, 87, 2007, 1.
[CAPES, Fapesp, CNPq, INCTBio]

199
DETERMINATION OF NITROGEN-CONTAINING AROMATIC COMPOUNDS IN
AIR PARTICULATE MATTER FROM ASPHALT MIXING PLANT BY HPLC-DAD
Milena Pendeza (PG)a*, Leandro Carvalho (R)a
*e-mail: milenapendeza@gmail.com
Petroleum asphalt is the name given to the petroleum refining residue, which is a product widely
used in the road pavement1. The handling and application of the asphaltic binder on the pavement
is only possible with the use of high temperatures, which causes the matrix aging process through
the volatilization of compounds such as polycyclic aromatic hydrocarbons (PAHs) and their
nitrogenated derivates2. This process also results in atmospheric emissions of harmful carcinogenic
and mutagenic compounds3. Therefore, the present work investigated new liquid chromatographic
methods for the determination of nitrogen-containing aromatic compounds (NACs) in air particulate
matter from an asphalt mixing plant based on reverse-phase and ion-pair separation mechanisms
and diode array detection. The optimized methods were applied to the identification of NACs in the
air particulate collected in a mixing plant of asphalt located in Brazilian Northeast (Fortaleza, Ceará).
The air particulate matter (PM10) was collected by using a large volume sampling device (HiVol 3000)
and 10 m cellulose filter papers. The collected filters were extracted in dichloromethane4 and
resuspended in acetonitrile prior to HPLC analysis. In addition, a sample of asphalt binder used in
the mixing plant was analyzed for the selected NACs. Thus, 1-nitronaphthalene, 1-nitropyrene, and
N,N-dimethylaniline were identified in all the studied air particulate samples, being 1-
nitronaphthalene (8.17−28.30 µg per filter) and 1-nitropyrene (0.83−9.20 µg per filter) quantified in
the samples, as can be seen in figure 1. In addition, 9-nitroanthracene and 1-nitropyrene were
identified and quantified in a SRM1975 reference material (particulate diesel extract). 1-
Nitronaphthalene was identified and quantified (350.0 ± 22.4 mg/kg) in a virgin asphalt sample. The
presence of the NACs in the studied samples were confirmed by tandem mass spectrometry
(MS/MS) with atmospheric pressure photoionization (APPI). The methods were found to be useful
for studies involving petroleum asphalts and its atmospheric emissions.
Figure 1. HPLC-DAD chromatogram of an air particulate sample extracted in CH2Cl2 with increasing additions of 1-
nitronaphthalene and 1-nitropyrene using a Pursuit PAH column (Agilent®). Mobile phase: 0.5 mmol L-1 H3PO4/ACN
(gradient elution). UV detection: 245 nm. Column temperature: 30°C
1 Bernucci LB, Motta LD, Ceratti JAP, PETROBRÁS: ADEBA, 2010.

2 Lesueur D, Advances in colloid and interface science, 145, 2009, 42.
3 Schreiner CA, Regulatory Toxicology and Pharmacology, 59, 2011, 270.
4 Heikkilä P, Riala R, Hämeilä M, Nykyri E and Pfäffli P, Am. Ind. Hyg. Assoc. J., 63, 2002, 156.
[Petrobras, UFCE, CAPES

200
]
DEVELOPMENT OF A CHROMATOGRAPHIC METHOD FOR THE

DETERMINATION OF PESTICIDE RESIDUES IN TOMATOES
Gabriela Emídio Saud Pachá (UG)a*, Marcos Martins Gouvêa (R)b, Flávia Ferreira de
Carvalho Marques (R)c
aFederal Fluminense University, Department of Analytical Chemistry/Institute of Chemistry, Niterói, RJ, Brazil, 24020-140
*e-mail: gabrielapacha@id.uff.br
Pesticides have been widely used to control pests and to regulate the growth of plantations, with a
significant increase in the number of registrations in Brazil in recent years. 1 Active ingredients such
as chlorpyrifos and imidacloprid, which, due to their toxic effects, have already been banned by the
EU, and the first one also by the USA, still appear among the most commercialized in Brazil.2 Some
foods stand out due to the amount and diversity of residues detected, such as tomatoes.3 This work
consists in the development of a chromatographic method for the determination of residues of two
classic active ingredients (chlorpyrifos and imidacloprid) and two recently released ones
(flupyradifurone and dinotefuran) in tomato, using high performance liquid chromatography with
array detector diodes (HPLC-UV/DAD). Column C18 was employed with mobile phase A (ultrapure
water) and B (acetonitrile) in gradient mode (0 to 2 min: 30% B; 2 to 4 min: 30-90% B; 4 to 9 min:
90% of B) with a flow rate of 1.0 mL/min at 25°C and an injection volume of 20 µL. The linearity of
the method was verified through the analytical curves prepared in the range of 0.05 to 1 µg/L, which
values of the coefficients of determination were greater than 0.99. Sample preparation consisted in
extraction by an adapted QuEChERS method,4 without the use of the cleaning step, which was
compared with a Soxhlet extraction. The first method showed less interferences in the
chromatograms. In addition, the QuEChERS method showed better recovery percentages for the
determined concentration of 1 mg/L, ranging from 72% to 94%. The values for the limits of detection
(LOD) and quantification (LOQ) obtained for flupyradifurone (LQ = 0.10 mg/kg and LD = 0.03 mg/kg)
and chlorpyrifos (LQ = 0.10 mg/kg and LD = 0.03 mg/kg) allowed their determination in
concentrations up to 5 times lower than the maximum residue limit (MRL) permitted in Brazil for
these substances in tomatoes. However, it was not possible to determine dinotefuran in the matrix
at the lowest concentration of the analytical curve (0.05 mg/L) due to the presence of interferents in
this region. Therefore, it would be necessary to make adjustments in the extraction method for the
determination of this substance. Additional studies will also be done for imidacloprid that did not
show adequate accuracy at the 0.1 mg/kg level.
1 MAPA (2005-2021). Disponível em: https://www.gov.br/agricultura/pt-br/assuntos/insumos-agropecuarios/insumos-
agricolas/agrotoxicos/informacoes-tecnicas.
2 EFSA. Search Active substances, safeners and synergists. Disponível em:
https://ec.europa.eu/food/plant/pesticides/eu-pesticides-database/active-substances/?event=search.as
3 PARA. Disponível em: https://www.gov.br/anvisa/pt-br/assuntos/agrotoxicos/programa-de-analise-de-residuos-em-
alimentos.
4 Frenich AG, Vidal JLM, Montoro E, González R. Anal Bioanal Chem, 390, 947-959, 2008.

201
DEVELOPMENT OF A GC METHODOLOGY FOR QUANTIFYING GLYCEROL
ESTERS FROM THE ESTERIFICATION OF GLYCEROL WITH LEVULINIC ACID
Evellyn P. S. da Silvaa (PG), Lucas N. de Meloa (PG), Janaína H. Bortoluzzia (R), Simoni M. P.
Meneghettia* (R)
Federal University of Alagoas, Institute of Chemistry and Biotechnology, Maceió-AL, Brazil, 57072-900
*e-mail: simoni.plentz@gmail.com
Glycerol (GLY) and levulinic acid (LA) are important building blocks which can be obtained as a co-
product of biodiesel and from carbohydrate conversion, respectively. In the context of biorefineries, the
esterification reaction is an important route for the production of chemical inputs of industrial interest and
high added value. Therefore, the union of these two molecules in the synthesis of new products has
attracted attention for promising applications.1 The quantification of reactants and formed products is a
key step of this process, which brings the necessity of developing efficient methodologies capable of
simultaneously determining GLY, LA and the acylglycerols formed. In this work, the results and
discussions of the development and optimization of a gas chromatography (GC) methodology for that
purpose are reported. A chromatograph equipped with a flame ionization detector (FID) at 300 °C and a
nonpolar 5%-diphenyl-95%-dimethylpolysiloxane column (30 m × 0.25 mm i.d. × 0.25 µm) was used.
Based on previous works by our research group, the oven temperature was initially programmed to 150
°C, heating to 160 °C at a 5 °C min-1 rate, to 250 °C at a 120 °C min-1 rate with a 2 min hold, and to 300
°C at a 50 °C min-1 rate with a 2.25 min hold, with a total run time of 8 min. The split/splitless injection
system was maintained at 240 °C, split ratio of 30:1. Benzyl benzoate was used as internal standard. The
ascending elution order of all compounds was: GLY, LA, monoacylglycerol (MA), benzyl benzoate (IS),
diacylglycerol (DA) and triacylglycerol (TA), as confirmed by analysis of standards and GC-MS
determinations. The method was optimized by means of variation of temperature programming and
carrier gas (H2) flow rate in order to minimize analysis time while retaining appropriate resolution for
quantification purposes especially between the peaks of GLY and LA. As it can be seen in Figure 1,
increasing H2 flow rate from 0.50 to 2.25 mL min-1 resulted in a reduction of analysis time from 6.85 to
5.30 min. However, 2.00 mL min-1 was chosen for quantitative purposes, maintaining resolution values ≥
1.25.2 Moreover, changes in temperature programming made possible reducing analysis time to 4.40 min
with a significant gain in resolution (> 3.00) between GLY and LA (Fig. 2). The developed methodology
proved to be faster than the reports available in the literature for identification and quantification of the
products of this reaction.3-5
Figure 1. Chromatograms of sample containing GLY, LA Figure 2. Reduction of analysis time (A) and increase
and products formed after esterification. A: Flow rate of resolution between GLY and LA (B): bottom, 150 –
variation (bottom to top: 0.50, 0.75, 1.00, 1.25, 1.50, 1.75, 160 °C (5 °C min-1), 160 – 250 °C (120 °C min-1,
2.00, 2.25 mL min-1). B: Reduction in resolution from 1.40 2.00 min hold), 250 – 300 °C (50 °C min-1, 2.25 min
to 1.20 after increasing flow rate. hold), t = 5.40 min; top, 140 (2.00 min hold) – 300 °C
(120 °C min-1, 2.00 min hold), t = 4.40 min.
1 Silva EPS, Meneghetti SMP, Molecular Catalysis, 528, 2022, 112499.

2 Collins H, Gilberto L, Bonato SP, Fundamentos de Cromatografia, Editora da UNICAMP, 2006, Brasil.
3 Mukesh C, Nikjoo D, Mikkola JP, ACS Omega, 5, 10, 2020, 4828.
4 Gonzalez-Arellano C, De S, Luque R, Catalysis Science & Technology, 4, 12, 2014, 4242.
5 Umrigar V, Chakraborty M, Parikh P, International Journal of Chemical Kinetics, 51, 4, 2019, 299.
[CNPq, CAPES, FINEP, FAPEAL, INCT-Catálise]

202
DEVELOPMENT OF A RESPONSE FACTOR BASED FAST GC METHODOLOGY
FOR QUANTIFYING TOTAL FATTY ACID METHYL ESTERS FROM DIFFERENT
SOURCES
Filipe L. Silvaa (PG), Lucas N. de Meloa (PG), Simoni M. P. Meneghettia (R), Janaína H.
Bortoluzzia* (R)
aUniversidade Federal de Alagoas, Instituto de Química e Biotecnologia, Maceió, AL, Brazil, 57072-900
*e-mail: janaina.bortoluzzi@iqb.ufal.br
In this work, the results and discussions of the development and validation of a Fast GC methodology
based on response factors (Fast GCRF) are reported. A conventional chromatograph equipped with
a flame ionization detector (FID) and a nonpolar dimethylpolysiloxane column (10 m × 0.1 mm i.d. ×
0.1 µm film thickness) was used. The method was validated and applied to the quantification of total
fatty acid methyl esters (FAME) produced from various raw materials (soybean oil, cottonseed oil,
canola oil, coconut oil, palm oil, sesame oil, sunflower oil, castor oil, corn oil, macauba oil, almond
oil, peanut oil, chia seed oil, residual frying oil, and chicken fat) in five different ranges of FAME
concentration obtained by varying transesterification reaction parameters.1 The developed
methodology proved to be selective regardless of the type of raw material and robust when subjected
to changes in sample preparation. The Fast GCRF method presented maximum relative standard
deviation (RSD) values for instrumental precision, repeatability and intermediate precision of 2.3%,
3.6%, and 2.8%, respectively. The method proved to be accurate by the means of recovery tests
with values between 86 and 119% and after showing a high degree of agreement with two reference
methodologies [r2 = 0.9962 (GCSCRF) and r2 = 0.9953 (EN14103)] and a total analysis time of 5
min, which is up to 4 times shorter than the methods described in EN14103 and GCSCRF (Figure
1).2,3
Figure 1. Fast GCRF versus the reference method GCSCRF (A) and Fast GCRF versus the reference method EN14103
(B).
1 Meneghetti SMP, Bortoluzzi JH, Silva FL, Freitas JAS, Salgueiro B, Melo LN, Silva WWL, Nascimento JR, Parâmetros
físico-químicos para a reação de transesterificação: estudos envolvendo biodiesel metílico e etílico de soja, in: Suarez
PAZ, Meneghetti SMP (Eds.), Parâmetros Físico-Químicos Para os Processos de Produção de Biodiesel, MCTI/ RBTB,
2015, Brasil.
2 EN 14103. EUROPEAN STANDARD; Fat and oil derivatives – fatty acid methyl esters (FAME) – determination of ester
and linolenic acid methyl ester contents. European Committee for Standardization. Brussels. 2011.
3 Silva FL, Melo LN, Wolf CR, Meneghetti SMP, Bortoluzzi JH, Journal of the Brazilian Chemical Society, 29, 2018, 1336.
[CNPq, CAPES, FINEP, FAPEAL, INCT-Catalysis, LASSOP/UFAL, NPRMN/UFAL]

203
DEVELOPMENT OF METHODS WITH METROLOGICAL TRACEABILITY FOR
MEASUREMENT OF MASS FRACTION OF TIN AND ORGANOTIN IN LEATHER
Marcelle Figueiredo de Souzaa, Marcia Silva da Rochaa, Thiago de Oliveira Araujoa*
a Instituto Nacional de Metrologia, Qualidade e Tecnologia (Inmetro), DIMCI/DIMQT/LABIN, Duque de Caxias, RJ, Brazil,
25250-020
*e-mail: toaraujo@inmetro.gov.br
This work presents the development of methods for the determination of tin (Sn) using ICP-MS and
organotin compounds (OTCs, especially tributyltin TBT) using the HPLC-ICP-MS coupling in leather
samples. At first, studies were carried out on the behavior of the calibration curve used to quantify
Sn with the ICP-MS technique and thus be able to quantify them with reliable and traceable results.
The study of the calibration curve (Sn: ICP-MS) was carried out in the range of 0.3 µg kg-1 to 1.8
µg kg-1 (n=7) by diluting the NIST SRM 3161a standard in HNO3 2%, with R2 of 0.999. The adequacy
of the calibration curve model was evaluated by visual inspection of residuals, outliers, normality of
residuals, homoscedasticity, regression significance and zero intercept. The fortification study or
recovery of the amount of Sn added to the samples showed 100% recovery, proving that there were
no losses or contamination in the samples. The limits of detection and quantification were
determined: instrumental: 3 ng kg-1 and 9 ng kg-1 respectively and in the sample: 292 µg kg-1 and
877 µg kg-1. The instrumental repeatability ranged from 0.54% to 0.77% for n=7 depending on the
levels of the calibration curve. Chromatographic separation was carried out by reverse phase
chromatography (C18) and mobile phase composed of MeOH: 30% CH3COOH/ 0.5% NH4OH/ 70%
H2O in the proportion of (60:40). The study of the calibration curve (TBT: HPLC-ICP-MS) was carried
out in the range of 1.5 mg kg-1 and 3.5 mg kg-1 (n=5) by diluting the MRC GBW NIM standard 08710
(1547 ± 47) mg kg-1 with R2 of 0.999. The adequacy of the curve model was evaluated using the
same statistical tests used for Sn by ICP-MS. The study of extraction efficiency was investigated and
the highest extraction efficiency obtained was 90.87% using shaker and MeOH (25%) /
CH3COOH(75%) as TBT extracting agent, during 16 h of extraction (overnight). The detection and
quantification limits were determined: instrumental: 12.5 µg kg-1 and 35.5 µg kg-1, respectively, and
in the sample: 70 µg kg-1 and 210 µg kg-1, respectively. Instrumental repeatability ranged from 1.23%
to 1.92% for n=5 depending on the levels of the calibration curve.
[PRONAMETRO]

204
ELECTROPHORESIS MICROCHIPS FOR SCOPOLAMINE AND
BUTYSCOPOLAMINE ANALYSIS
Hellen I. Santos (PG)a*, Kemilly M. P. Pinheiro (R)a, Eduardo M. Richter (R) b and Wendell K.
T. Coltro (R)a,c
aUniversidade Federal de Goiás, Instituto de Química, Goiânia, GO, 74690-900
bUniversidade Federal de Uberlândia, Instituto de Química, Uberlândia, MG, 13400-970
c Instituto Nacional de Ciência e Tecnologia de Bioanalítica, Campinas, SP, Brasil, 13084971
*e-mail: hellendossantos16@gmail.com
The number of cases of drug-facilitated sexual assault (DFSA) have increased drastically in the last
decades, and there are several drugs that can be used for this purpose, such as scopolamine (SCO)
and butylscopolamine (BSCO)1. Although SCO can only be acquired with a prescription, there are
reports in the literature in which SCO can be produced from BSCO (non-prescription drug) by simple
heating, which increases the risk of contamination or facilitates access to scopolamine by criminals1.
Thus, different analytical techniques have been used to determine BSCO and SCO in different types
of matrices, so, among these techniques, the use of microchip electrophoresis systems (ME) coupled
with contactless conductivity detection (C4D) is advantageous, due to its miniaturization; portability;
simple, fast and low cost analysis2. Therefore, this study demonstrates the development of an
analytical method using ME-C4D for the identification, separation and quantification of SCO and
BSCO in beverages and pharmaceutical formulations. First, standard solutions of SCO and BSCO
were analysed and so far, the best separation condition was obtained using a running buffer
composed of 40 mmol L-1 of butyric acid and 25 mmol L-1 of sodium hydroxide (Figure 1 - B). For
C4D measurements, the highest sensitivity was obtained by applying a sinusoidal wave with 1200
kHz frequency with an excitation voltage of 20 Vpp. After this analysis, the developed and validated
method will be applied in the study of real samples to identify and quantify these two compounds.
Therefore, with the use of ME-C4D it is expected to achieve satisfactory results, enriching the
scientific-technological field, as well as forensics and analytics.
Figure 1. Electropherograms showing the optimization of the buffer (A) with the pH variation from 4.0 to 5.5
and (B) maintaining pH 5.0 varying the proportion of the buffer (a) 10mM C₄H₈O₂ and 3mM NaOH; (b) 20mM
C₄H₈O₂ and 13mM NaOH; (c) 30mM C₄H₈O₂ and 19mM NaOH; (d) 40mM C₄H₈O₂ and 25mM NaOH.
1 Ribeiro, M. MAC; Barreto, D. N.; Petruci, J. F. S.; Richter, E. M. Microchemical Journal, 172, 2022, 106985.
2 Saylor, R. A.; Lunte, S. M. Electrophoresis, 39 (3), 2018, 462-469.
[CAPES, INCTBio, UFG]

205
EXTRACTION OF VOLATILE COMPOUNDS-VCs FROM MIXED FRUIT AND
VEGETABLES REFRESHMENT BY HS-SPME/GC-MS
Felipe S. da Silvaa (PG), Sheyla M. B. Amarala (PG), Francisca V. Pintob (UG), Vitor P. de A. Silvac (R), Crisiana de
A. Nobrec (R), Maria A. L. Milhome*a (R)
a Federal Institute of Education Science and Technology of Ceará- IFCE, Postgraduate in Food Technology- PGTA,
Limoeiro do Norte, CE, Brazil, CEP 62.930-000

bFederal Institute of Education Science and Technology of Ceará- IFCE, Undergraduate in Nutrition, Limoeiro do Norte,
CE, Brazil, CEP 62.930-000

c Nucleus of Technology and Industrial Quality of Ceará-NUTEC, Instrumental Chemistry Laboratory-LQI, Fortaleza, CE,
Brazil, CEP 60.440-552

*e-mail: maria.milhome@ifce.edu.br
Food industry industries have been advancing in the use of chromatographic techniques, through
the qualitative monitoring of volatile-CV's and bioactive compounds, which are related to product
quality, in addition to the sensory aspect1.Thus, the present study aims to optimize the sample
preparation technique using Solid Phase Microextraction, headspace mode (HS-SPME) and
detection by Gas Chromatograph Coupled to Mass Spectrometry- GC-MS to evaluate the volatile
composition of Mixed Refreshment consisting of fruit and vegetables. Pineapple pulp, kale, ginger,
lime and drinking water were used to prepare the drink, which was then filtered (0.2 mesh) and stored
under refrigeration at 10°C ². Three SPME fibers were evaluated in this study: polyacrylate (PA, 85
µm), polydimethylsiloxane-carboxen (PDMS-CAR, 80 µm) and polydimethylsiloxane-divinylbenzene
(PDMS-DVB, 100 µm). The fibers were exposed in headspace mode, in sealed vials containing 20
mL of the sample. The exposure time (15 and 30 min) and temperature (65 and 85 °C) ³ were
evaluated under different extraction conditions. After extraction, the samples were analyzed in the
GC-MS equipped with a quadrupole analyzer, using a DB-5MS column (5% phenylmethylsiloxane)
(30 m x 0.25 mm x and 0.25 μm), operated in Full Scan mode, 35-400 m/z, electron impact-EI
ionization with energy of 70 eV. Helium was used as carrier gas (1.0 mL/min) and splitless injection
mode. To obtain the volatile profile were considered the major components with area% ≥1% 4. The
results show that the PA fiber, 85 µm was able to extract the largest number of compounds (total of
90 compounds), followed by PDMS-CAR (80 µm) and PDMS -DVB (100 µm) with 85 and 82
compounds, respectively. In the first test, it was observed that the PA fiber (85 μm) was more efficient
in extracting the analytes from the sample, with expressive area% levels (≥1%), composing a volatile
profile with 17 major components. The extraction time and temperature were also optimized, and the
best conditions obtained for mixed beverage were: 65 °C/20 min using PA fiber (85 μm). The
qualitative analysis of the sample reveals that most of the compounds identified are used by the food
industry as natural additives. The HS-SPME method proved to be fast and satisfactory for the
extraction of volatile components in mixed drinks. The data suggest the importance of a more
detailed study to evaluate the relationship of CVs with the sensory characteristics and stability of the
final product.
1 Assunção DA, Pinto BG, Leal AC, Augusti R, Melo JOF, Ramos AS, Silva MR, Capobiango M. Research, Society and
Development, v. 9, 2020, e55491110054
2 Fouchy MV, Santos IC, Ahn CSv, Mendonça, CRB, Nogueira MB. Brazilian Journal of Technology, Curitiba, v. 2, 2019,
601.
3 Xu X, Bi S, Lao F, Chen F, Liao X, Wu J. Food Chemistry, v. 341, 2021, 128118.
4 Nascimento RF, Lima ACA, Barbosa PGA, Silva, VPA. Cromatografia Gasosa: aspectos teóricos e práticos. Fortaleza:
Imprensa Universitária, 2018.
[FUNCAP, CAPES, IFCE, NUTEC]

206
LARGE VOLUME INJECTION FOR THE DIRECT DETERMINATION OF
PARAQUAT USING HILIC SEPARATION WITH UV DETECTION
Ismael dos S. Belmonte (PG), Tania M. Pizzolato (R), Mariana R. Gama*(R)
Institute of Chemistry, Federal University of Rio Grande do Sul, Porto Alegre - RS, Brazil
*e-mail: mrgama@ufrgs.br
Paraquat (PQ) is a quaternary ammonium pesticide, widely used as an herbicide for several crops,
such as soy, cotton, and citrus. Due to its high toxicity, paraquat was banned in European Union,
China, and more recently, Brazil, but later allowed for rural producers to use the stocks already
purchased. In 2021, the Brazilian Ministry of Health has established 13 µg L-1 as the maximum limit
for PQ in drinking water samples1, considering that this pesticide can offer a risk to human health,
such as mutagenic effects and Parkinson's disease for long-term exposition2. Multiresidue methods
are routinely applied to analyze pesticide residues, generally carried out as reversed-phase liquid
chromatographic separations coupled to mass spectrometry. The high polarity of PQ (log P = -4,5)
hinders its determination by multiresidue separation approaches, thus requiring a substitute
instrumental method of analysis3 or adjusted chromatographic methods, such as those based on
ion-pairing or different column chemistries for the improved retention of PQ. In this sense, hydrophilic
interaction chromatography (HILIC) emerged as an option for the successful analysis of PQ,
providing effective interactions between the PQ structure and a high polar stationary phase. The low
concentration of PQ residue usually requires a highly sensitive detection technique, commonly
supported by a preconcentration strategy. Thus, the injection of large sample volumes represents a
suitable alternative to extraction techniques, since this approach requires little (or no) sample
preparation and reduces waste generation associated with the traditional off-line solid-phase
extraction (SPE) as pre-concentration step4. Generally, the large volume injection is successfully
applied for reversed-phase separation, with a few descriptions for HILIC mode. This work describes
the direct determination of PQ in water samples using large volumes of injected samples for avoiding
the requirement for SPE sample preparation. The analyte determination was performed using HILIC
separation mode with UV detection. Tap water samples were filtered in a PTFE 0.45 membrane.
The chromatographic separation employed a core shell zwitterionic HILIC column (100 x 4.6 mm
i.d., 4.0 µm particle diameter) and 65:35 (v/v) ACN/ammonium formate 50 mmol L–1 as mobile phase,
at a flow rate of 0.8 mL min-1. The injection volume was 50 µL and the UV detection was performed
at 257 nm. The LOD and LOQ values were estimated as 1.10 and 3.60 ng mL–1, respectively. The
analytical curve showed linearity in the range of 2.50 to 100 ng mL–1 (R² = 0.9998). The precision
(calculated for 50 ng mL–1 of PQ) obtained an RSD of 0.68%. Recoveries of 103.5, 95.0, and 100.4%
(RSD < 2.0%) were estimated for tap water samples fortified with 5.0, 25, and 100 ng mL–1 of PQ,
respectively. There was no matrix effect for analytical curves in the aqueous organic solvent,
ultrapure water, or water sample. The proposed method reached a LOQ about 4 times lower than
the maximum limit established for PQ, without the need for a pre-concentration step, enabling its
application for analyses with minimum sample manipulation. Additionally, the injection of large
amounts of the sample was greatly combined with LC-UV, with remarkable method performance and
not requiring sophisticated detection strategies, such as mass spectrometry.
1 BRASIL. Ministério da Saúde. Gabinete do Ministro. Portaria nº 888, de 4 maio de 2021. Brasília, 2021.
2 Arsac JN, Sedru M, Dartiguelongue M, Vulin J, Davoust N, Baron T, Mollereau B, International Journal of Molecular
Sciences, 27, 2021, 11613.
3 Belmonte IS, Pizzolato TM, Gama MR, Trends in Environmental Analytical Chemistry, 35, 2022, e00171.
4 Kutlucinar KG, Hann S, Electrophoresis, 42, 2021, 490.
[CAPES]

207
METHOD FOR THE ANALYSIS OF BIOGENIC AMINES BY MICELLAR
ELECTROKINETIC CHROMATOGRAPHY USING BENZOYL CHLORIDE AS A
DERIVATIZING AGENT
Jéssica O. F. Mantoanelli (PG)a, Elisabete A. Pereira (R)b*
a University of São Paulo, Chemistry Department/Chemistry institute, São Paulo, SP, Brazil, 05508-900
b Federal University of São Carlos, DFQM department, Sorocaba, São Paulo, Brazil18052-780
*e-mail: ealves@ufscar.br
Biogenic amines (BAs) occur naturally in a wide variety of foods, especially those that are
rich in proteins being formed by the degradation of amino acids by bacteria [1]. Although its presence
in foods and beverages is recognized as undesirable due to the toxicity of these compounds,
Brazilian legislation establishes maximum limits only for histamine in some species of fresh fish and
derivatives [2]. The development of analytical methods for determining biogenic amines in foods is
essential because of their toxicological effect on consumers and because they are considered quality
indicators. The present work describes the new method using benzoyl chloride (Bnz-Cl) as a
derivatizing reagent for the separation and analysis of biogenic amines by micellar electrokinetic
chromatography (MEKC). The biogenic amines (putrescine, cadaverine, tyramine, histamine, and
tryptamine) were reacted with Bnz-Cl reagent to form adducts with maximum absorbance at 206 nm.
Amines derivatization was performed by mixing 2 mL de amines standard mixture, 2 mL Bnz-Cl 6 g
L-1, and 0.8 mL of NaOH 6 mol L-1. The organic phase was dried by nitrogen flux, and the residue
was dissolved in 2 mL acetonitrile/water (10:90 % v/v). Different analytical and instrumental
parameters were evaluated. The optimized conditions were performed using a CE system (Agilent
Technologies, model 7100 A) with a diode array detector set to 206 nm and temperature control
maintained at 25C. Fused silica capillaries (48.5 cm total length) were used. The mixture consisting
of 15 mmol L-1 borate, 40 mmol L-1sodium dodecyl sulfate (SDS), and 20 % acetonitrile (CAN), pH 9
was used as a running buffer. The system was operated in normal polarity and constant voltage of
17 kV. Samples were injected hydrodynamically, 40 mbar during 5 s (Figure 1).
Legend. Standard amines Bnz-Cl (20 mg L-1 each amine). Electrolyte: 40 mmol L-1 SDS + 15 mmol L-1 borate + 20 % v/v ACN. Run
conditions: 25˚C; 5s x 40 mBar; 17 kV; 206 nm. Peaks: R. reagent; *. unknown; 1. histamine; 2. tyramine; 3. putrescine; 4. Cadaverine
and 5. Tryptamine.
Figure 1. Optimized separation of amines-Bnz-Cl.
A new methodology was successfully developed for biogenic amines determination. The proposed
method will be validated and applied to beverage /food samples.
1. Abdurrahman, M.M.; Badri, Haji, B. H.; J ANAL METHODS CHEM.2020, 2020, DOI: 10.1155/2020/5814389.
2. Ruiz- Capillas, C.; Herrero, A. M.; Foods2019, 8, 62
[FAPESP, CAPES]

208
MULTIDIMENSIONAL GAS CHROMATOGRAPHY DATA AS AN AUXILIARY
TOOL TO PREDICT SENSORY FEATURES OF BEERS
Isabela C. M. Cunha and Fabio Augusto *
Institute of Chemistry and National Institute of Science and Technology in Bioanalytics (INCTBio), University of
Campinas (Unicamp) – CP 6154, 13083-970 Campinas SP, BRAZIL
*e-mail: faugusto@unicamp.br
The assessment of sensory features is a key step both on the development of new products
and for the quality control of industrial products. This task is usually performed using several different
strategies based on the use of panel where people (not always specifically trained for this task) taste
the products and attribute qualitative or semi-quantitative scores related to sensorial aspects relevant
to the quality of the sample. The use of sensory panels as primary tool to assess food flavour and
aroma properties is unvaluable and it is irreplaceable; however, chemometric proccessing of data
generated by analytical tools such as Comprehensive Multidimensional Gas Chromatography
(GC×GC) can provide a valuable alternative to replicate these processes, allowing faster
qualification of sensory parameters of samples using a potentially robust tool. In this work, we
present the use chemometric models obtained from GC×GC-MS chromatographic fingerprints of the
volatile fraction of lager beers previously isolated by Headspace Solid Phase Microextraction (HS-
SPME) as auxiliary tool for sensory assessment of these samples. Commercial lager beers available
in the local market were used as calibration and validation samples (22 different brands); their key
sensory features such as hop flavour, bitterness and barley flavour were obtained by Quantitative
Descriptive Sensory Analysis (ADQ) performed by a trained pannel of users: each panellist was
coached to assign values on an hedonic scale from 0 to 9 to each tasted parameter. The fingerprint
of the volatile organic compounds for each sample was assessed by a previously optimized HS-
SPME+GC×GC-MS procedure. The correlation between the chromatographic data and sensory
features for the sample set was modelled through Partial Least Squares Determinant Analysis (PLS-
DA). Results showed that it is possible to use GC×GC data to screen beer samples according to
each sensory feature, qualifying each beer as being “high” or “low” on that particular sensory
attribute. Further examination of the models also allow to pinpoint specific volatile compounds
associable to each sensory attribute; although in most cases this certainly are non-causal
correlations, the compounds are potential markers for the that particular attribute. Althogh the results
here presented are preliminary, they point out that GC×GC when combined to use of data provided
by panels can be a powerfull tool on sensory analysis.

209
OPTIMIZATION OF A CHIRAL LIQUID CROMATOGRAPHY METHOD FOR
ANALYSIS OF THE PESTICIDES ACEPHATE AND METAMIDOPHOS
Leandro O. Duarte*,a(US), Maria C. H. Grili (UG)a, Anderson R. M. de Oliveira (R)a
aUniversidade de São Paulo, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Departamento de Química,
Ribeirão Preto, SP, Brazil, 14040-900
*e-mail: leandro.oka@usp.br
Acephate and metamidophos are pesticides that acts as acetylcholinesterase inhibitors and their
high uses have induced health issues and environmental pollution1. Due to their chiral centers,
enantioselectivity had raised important questions about their unpredictable toxic effects on nontarget
organisms. Nonetheless, in order to investigate its enantiomeric behaviour, efficient analytical
methods for the differentiation of enantiomers have become essential. In chiral liquid
chromatography, the recognition ability of chiral stationary phases (CSPs) depends not only on the
structure and substituents of the chiral polymers but also of the mobile phase composition2. Since it
is hardly possible to predict enantiomeric resolution, separation strategy includes screening and
optimizing a set of CSPs and organic modifiers to detect a suitable enantioselectivity and improve
the resolution obtained. Therefore, in this proposed work, the desired retention time (< 15 min) and
resolution (Rs ≥ 1.5) were assessed by evaluating eight chiral columns (Table 1) under normal, polar
organic, and reversed modes of elution. In polar organic mode, acetonitrile, methanol, or ethanol
were used. In reversed-mode, mobile-phases were prepared by mixing acetonitrile or methanol with
water at 30/70 or 70:30 (v/v) ratios. Finally, in normal-mode, two hexane-based mobile phases,
hexane/isopropanol or hexane/ethanol, both a 90:10 v/v, were applied. If necessary, formic acid or
ammonium hydroxide (both at 0.1% v/v) was added in mobile phase. The column temperature was
maintained at 30 ºC and the mobile phase flow rates were 0.5 mL/min at polar organic conditions
and 0.8 mL/min at both normal and reversed conditions. Detection wavelength was set at 235 nm.
The results showed the enantiomers of at least one racemic pesticide mixture were resolved from
each other employing aprotic organic modifiers in Chiralpak AD, Chiralcel OJ, and/or Chiralpak AS
columns. After optimization, the best condition was accomplished on a Chiralpak AS-H column (150
x 4.6 mm i.d., 5 μm) under acetonitrile/water/formic acid (98:2:0.1, v/v/v) or hexane/ethanol/methanol
(85:14:1, v/v/v) mobile phases.
Table 1. Chiral Stationary Phases (CSPs) used in screening step
Column Stationary phase Dimension

Chiralpak AD Amylose tris-(3,5-dimethylphenylcarbamate) A
Chiralpak AS Amylose tris-[(S)-α-methylbenzylcarbamate] A
Chiralcel OD Cellulose tris-(3,5-dimethylphenylcarbamate) A
Chiralcel OJ Cellulose tris-(4-methylbenzoate) A
Lux Cellulose-2 Cellulose tris-(3-chloro-4-methylphenylcarbamate) B
Lux Amylose-2 Amylose tris-(5-chloro-2- methylphenylcarbamate) B
Chirobiotic V Vancomycin B
Chirobiotic T Teicoplanin B
A: 250 x 4.6 mm, 10 µm and B: 150 x 4.6 mm, 5 µm
1 Lin, Z, Pang, S., Zhang, W, Mishra, S., Bhatt, P., Chen, S. Frontiers in Microbiology, 11, 2020.
2 Tarafder A, Miller L, Journal of Chromatography A, 1638, 2021.
[FAPESP, CNPq, INCT-DATREM, CAPES]

210
SIMULTANEOUS DETERMINATION OF CRUDE OIL QUALITY PARAMETERS
VIA GC×GC-HRMS AND MULTIVARIATE DATA ANALYSIS
Amilton Moreira de Oliveira*, Carlos Alberto Teixeira, Leandro Wang Hantao
University of Campinas, Institute of Chemistry, Campinas, São Paulo, Brazil, 13083-862
*e-mail: amilton.moliveira@gmail.com
Characterization of crude oil and its fractions is fundamental to addressing exploration and
processing challenges in an optimized supply chain. For instance, the oil density and sulfur content
significantly affect its price and refining prospects1. Thus, the petroleum industry has employed both
classical and instrumental methods to determine properties of interest in feedstock and distillate
products. However, the individual assessment of several properties is laborious, demands
equipment diversity, and consumes a high sample volume. In this context, a method for the
simultaneous determination of oil parameters is necessary to enhance the analytical workflow. The
present study evaluated an approach using comprehensive two-dimensional gas chromatography
coupled with high-resolution mass spectrometry (GC×GC-HRMS) and multivariate data processing.
The gain in selectivity enabled the investigation of constituents related to the modeled properties
(Figure 1). The experiment was designed with eight crude oils from various origins provided by the
Brazilian Petroleum Company research center (CENPES – Petrobras). The oils had been previously
characterized regarding different properties by reference methods. The sample set was expanded
by combination according to a mixture design, which covered a wider composition range. GC×GC
improved isomers separation while the mass resolving power enabled to distinguish critical ion pairs
(0.004 Da mas split). The sample complexity and high throughput of the technique produced a large
dataset. Thus, data-driven pixel-based analysis2,3 was performed on MATLAB®. Raw
chromatograms were concatenated to obtain an augmented data tensor. The individual arrays were
aligned and submitted to preprocessing steps to mitigate instrumental artifacts and reduce
discrepancies in signal intensity. Variable selection tools were studied to eliminate information
unrelated to the goal of the experiment. Finally, partial least square regression was used as
multivariate data analysis in order to predict different oil properties.
4
(a) TIC
Retention time / s
4
(b) EIC
7 Retention time / min 74
Figure 1. GC×GC-HRMS contour plots of (a) crude oil and (b) a class of sulfur-containing compounds (dibenzothiophenes).
The combined high resolution/accurate mass measurement and structured chromatogram improve compound
identification. TIC: total ion chromatogram, EIC: extracted ion chromatogram.
1 Han Y, Zhang Y, Xu C, Hsu CS, Fuel, 221, 2017, 144.

2 Oliveira AM, Teixeira CA, Hantao LW, Analytical Methods, 14, 2022, 1646.
3 Ferreira VHC, Hntao LW, Poppi RJ, Talanta, 234, 2021, 122616.
[Petrobras, Nova Analítica, ThermoFisher]

211
SMALL, HIGHLY CHARGED, BRIGHT, AND PHOTOSTABLE FLUORESCENT
LABELS FOR SUCCESSFUL APPLICATIONS IN TOROIDAL CAPILLARY
ELECTROPHORESIS
Bruna Nitzke Minuzzi (UG)a, Jean G. Novello Gregolon (UG)b, Filssen N. Souza Schereiber
(UG)b, Melissa M. Rodrigues (R)c, Klester S. Souza (R)a, and Tarso B. Ledur Kist (R)b
a Instituteof Chemistry, UFRGS, Porto Alegre, RS, Brazil, 91.501-970
b Institute
of Biosciences, UFRGS, Porto Alegre, RS, Brazil, 91.501-970
c Univ. of Caxias do Sul, UCS, Caxias do Sul, RS, Brazil, 95.070-560
*E-mail: tarso@ufrgs.br or tarso.kist@gmail.com
Labelling analytes with fluorescent dyes prior to separation and analysis in HPLC, CE, and microchip
electrophoresis is considered a time demanding task and a potential source of errors in the analytical
practice. Consequently, such dyes are not used as often as they could be despite the improvements in
selectivity and detection limits they promote. Recently optimized reactive moieties and automation of the
derivatization procedure have the potential to revive the popularity of these methods. Moreover, small
and highly charged labels have the additional benefit of increasing both separation efficiencies and time
efficiencies1. Toroidal capillary electrophoresis1 exhibits extremely high separation efficiencies with the
number of theoretical plates crossing the 2 million landmark. However, as the analyte bands repeatedly
cross through the detection point (with electroosmosis2 or without electroosmosis3) in this method, the
photobleaching phenomenon continuously decreases peak area and peak hight over time and
compromises quantitative analysis. Therefore, fluorescent dyes with high photostability (or
photorobustness) are required. In order to overcome these shortcomings, the photophysical properties
of over seven-hundred fluorescent dyes were collected from the literature (some have to be measured)
and examined to facilitate the selection of the best fluorescent labels which exhibit the following desired
properties: 1) High net charge and small size (small friction coefficient). This is important to optimize both
separation efficiency and time efficiency (both concepts are explained in chapter five of reference 1). 2)
High molecular brightness (which is defined as the product of molar absorption coefficient and
fluorescence quantum yield). Knowledge of this photophysical parameter facilitates the optimization of
detection limits. 3) High photostability. This is important as it assures reproducibility in the quantitative
analysis (peak height and peak area repeatability and reproducibility) in toroidal capillary electrophoresis.
4) Finally, conjugation ability (so that it can be conjugated as many functional groups as possible, such
as primary and secondary amines, thiols, and others). Among all dyes studied, only a group of few dozens
were found to exhibit the desired properties and they belong to the following classes: rhodamines,
pyronines, borondipyrromethenes, trianguleniums, squaraines, cyanines, and pyrenes (a few of these
are shown in Figure 1).
Figure 1. Molecular structures at neutral pH of some fluorescent dyes to be used as labels to maximize separation
efficiency and time efficiency1 in toroidal capillary electrophoresis. They exhibit small sizes, high net charge, high
brightness, and high photostability. Parameters in blue are λ max, Stokes shift, and brightness expressed in mM-1cm-1.
DAOTA 4 is triangulenium derivative4.
1 Kist, T.B.L. Open and Toroidal Electrophoresis: Ultra-High Separation Efficiencies in Capillaries, Microchips, and Slabs.
First edition. John Wiley & Sons Ltd, Chichester, 2020. ISBN: 978-1-119-53940-7.
2 Video: https://youtu.be/X7ol2g-7BxE
3 Video: https://youtu.be/OOn5ma12bpc
4 Bisballe, N. and Laursen, B.W. (2020). Chem. Eur. J. 26: 15969-15976.

212
SYNTHESIS OF HYBRID MONOLITHIC STATIONARY PHASES USING A CLICK
CHEMISTRY REACTION FOR APPLICATION IN CAPILLARY LIQUID
CHROMATOGRAPHY
Marcella E. P. Schmidt (PG), Carla B. G. Bottoli (R)
aUNICAMP , Institute of Chemistry, Campinas, SP, Brazil, 13083-970
*e-mail: carlab@unicamp.com.br
Hybrid monoliths share characteristics common to both silica and organic monoliths, such as
good mechanical properties, broad pH tolerance, high permeability, stability and reduced shrinkage1.
Usual methods for producing such materials use alkoxysilanes and organic monomers through sol-
gel processes and click chemistry reactions. In this work, the authors developed hybrid monolith
column based on tetramethoxysilane (TMOS) and vinyltrimethoxysilane (VTMS) as precursors for
the sol-gel process. The hybrid stationary phase was functionalized with dodecanethiol through a
click thiol-ene reaction. The synthesis of stationary phases were performed in a pre-treated silica
capillary with an internal diameter of 100 µm. Initially, PEG 10000 (porogenic agent), urea (mesopore
formation inducer), VTMS (silica precursor), TMOS (silica precursor) and 0.01 mol/L acetic acid
(catalyst and solvent) were mixed. This mixture was stirred in an ice bath (0 °C.h -1). The mixture
was introduced into the capillary with a syringe pump. After filling, the capillary was incubated in a
water bath for 12 h at 55 °C, then heated at 120 °C for 2 h in an oven. After this process, the capillary
was treated with a solution of vinyldimethylethoxysilane (VDMES)/methanol/triethylamine (TEA) for
24 h at 80ºC, to increasing the availability of vinyl groups. The capillary was subjected to thiol
functionalization, then filled with a mixture containing 2,2'-azobisisobutyronitrile (AIBN, radical
initiator), dodecanethiol and ethyl ether. Finally, the capillary was heated at 70 °C for 12 h. Monolithic
capillary columns were subjected to chromatographic evaluation on a Thermo Scientific Dionex
UltiMate3000 chromatograph with diode array detection. The test mixture was composed of
alkylbenzenes (ethylbenzene, propylbenzene, butylbenzene and pentylbenzene) and the mobile
phase was composed of water:acetonitrile 55:45 (v/v), flow rate of 1.2 μLmin-1, detection at 215 nm
with cell of 3 nL and 0.08 μL injection volume. Other two columns were synthesized to evaluate the
differences between hybrid thiol-eno column: (1) a dodecyl silane bonded-phase and (2) low content
of vinyl groups on the stationary phase. The stability of the thiol-ene columns was assessed over
time, and the effect of different methods was investigated in detail by scanning electron microscopy
(SEM), scanning capacitively coupled non-contact conductometric detection (sC4D), Fourier
transform infrared (FTIR), and retention behavior in cLC. The best separation results used hybrid
monolithic columns with a vinylization step, in order to increase the rates of available ene groups
and favor the thiol-ene reaction.
1GAMA, Mariana R.; ROCHA, Fábio RP; BOTTOLI, Carla BG. TrAC Trends in Analytical Chemistry,115, 2019
[FAPESP, CNPq, CAPES, INCTBionallítica]

213
SAMPLE PREPARATION
214
SUPRAMOLECULAR SOLVENT-BASED MICROEXTRACTION METHOD OF
SEVEN PESTICIDES AND DETERMINATION BY HPLC-DAD
Luciane Effting (PG)*a, Ismael R. Amador (R)a, César R. T. Tarley (R)a,b
a State University of Londrina (UEL), Department of Chemistry, Londrina, PR, Brazil, CEP: 86050-482
b State University of Campinas (UNICAMP), National Institute of Science and Technology in Bioanalysis (INCTBio),
Institute of Chemistry, Department of Analytical Chemistry, Campinas, SP, Brazil, CEP: 13083-861
*e-mail: luciane.effting@uel.br
Pesticides are widely used in agriculture to eliminate or control various agricultural pests that can
damage crops and livestock and reduce farm productivity. Due to their widespread use, pesticides
have been released to different environmental matrices, including freshwater sources, therefore it is
essential to develop convenient, sensitive, and rapid methods for pesticides determination in natural
waters1. Sample preparation is the most crucial part of the analytical process, due to the complexity
of matrices and low analyte concentrations. SUPRASs can used during micro-extraction, provide a
number of different interactions with pesticide molecules and these results in an increase in the
extraction efficiency1. In this work, seven pesticides of different classes (acetamiprid, azoxystrobin,
bifenthrin, carbendazim, chlorpyrifos, imidacloprid and tebuconazole) were pretreatment with
supramolecular solvents and quantified by HPLC-DAD. To select the significant factors that play an
essential role on supramolecular solvent-based microextraction, the optimization was performed in
two steps: 25 fractional factorial and a Doehlert design was used to obtain the optimal conditions for
the extraction system. Under optimized conditions and a preconcentration volume of 40.0 mL, the
analytical performance of the method was evaluated and applied in lake and river water samples.
The results were analyzed from the Pareto chart (95 % confidence level), indicating that all factors
investigated within the experimental domain were appreciably important. The pH value and the
volume of decanol proved to be the most important variables individually and in their interactions
(DECxTHF, pHxDEC, THFxVOR). Because the results obtained through the factorial design
indicated that the isolated variables pH, volume of decanol and interaction between volume of
decanol and pH, volume of decanol and volume of THF strongly influenced the responses, these
factors were investigated in greater detail using the Doehlert design method. From the partial
derivative model, the critical values were obtained (for this, the volume THF was fixed at the central
point value (500 µL) due to the fact observed in the response surfaces): 113.0 µL of the volume of
decanol and pH of 6.3. Under optimum conditions and using a preconcentration volume of 40.0 mL,
analysis performance of the SUPRAs method for preconcentration of seven pesticides with HPLC-
DAD quantification was assessed. The linearity values demonstrated a wide working range for all
pesticides and all linear equation exhibit a greater correlation coefficient than 0.993. The LOQ values
obtained were lower than the MRL value in water for all the pesticides established by ANVISA
(Brazil). RSD values lower than 4.07 % were obtained at both spiking levels, in repeatability and
inter-day studies, confirming the precision of the method. To study the applicability of the proposed
method in real water analysis, and no residues of the seven pesticides were detected in blank water
samples. The accuracy of the suggested method was verified by means of addition and recovery
tests and recovery values ranged from 80.52 to 106.51 % were obtained, indicating the absence of
matrix effect, no interference, and the method feasibility to determinate simultaneously those
pesticides at low levels from several water types.
1 Musarurwa, H. & Tavengwa, N. T. Talanta 223, 2021, 121507.
[UEL, INCTBio, CAPES, CNPq]
––––––––––––––––––––– Sample preparation –––––––––––––––––––––

215
OPTIMIZATION AND VALIDATION OF A SCALE REDUCED QuEChERS METHOD
FOR EXTRACTION OF PESTICIDE RESIDUES IN FROG ADIPOSE TISSUE
Igor M. Limaa, Allyson L.R. Santosa, Andressa T. Vieiraa, Giselle O. Carvalhoa,
Patrícia M. Gondimb, Paulo Casconb, Anizio M. Fariaa,*
a Federal University of Uberlândia, Institute of Exact and Natural Sciences of Pontal, Ituiutaba, MG, Brazil, 38304-402
b Federal University of Ceará, Departament of Biology, Fortaleza, CE, Brazil, 60440-900
*e-mail: anizio@ufu.br
The expansion of monocultures to regions close to conservation areas has put biodiversity in
Brazilian biomes at risk, mainly due to the intense use of pesticides. Anuran species are highly
susceptible to pesticides and have shown a strong decline worldwide.1 Pesticide residues in frog
tissues may be a key factor in confirming the relationship between frog decline and its polluted
habitat. As part of this work, a modified QuEChERS method2 was developed for the extraction of
atrazine, chlorpyrifos, α- and -endosulfan, α-, -, - and -cypermethrin into frog adipose tissues.
The method was optimized for a scale according to the size of the samples, using chicken adipose
tissue as a model organism. In addition, the extracting solvent and different cleaning sorbents were
optimized, achieving better recoveries (85-113%) with 1.5 mL of ethyl acetate and without the
cleanup step, Figure 1. Extract analyzes were performed by high-performance liquid
chromatography with photodiode array detection, with pesticide separation in 15 min, Figure 2. The
scale reduced QuEChERS method (mini-QuEChERS) was validated following the European
Community guidelines, document No. SANTE/2019/12682.3 The method showed a moderate matrix
effect for some pesticides that were corrected using the matrix-matched calibration. The limits of
quantification for the pesticide residues in adipose tissues varied from 0.6 to 4.5 mg kg-1. Pesticide
recovery rates were 73.8 to 114.8%, and repeatability and laboratory reproducibility showed relative
standard deviations below 10.9% that met acceptance criteria.3 The mini-QuEChERS method was
used to extract pesticides from the adipose tissues of two species of frogs, Leptodactylus
macrosternum and Scinax x-signatus, harvested in the Baixo Jaguaribe/Ceará area. Some samples
tested positive for cypermethrin and atrazine. The results confirm the efficacy of the QuEChERS
method developed and indicate the risk of anuran species as agricultural production expands in the
study area.
1
120
Atrazine blank adipose tissue
Chlorpyriphos spiked adipose tissue
100
Endosulfan (a+)
Cypermethrin (a+++)
Recovery (%)
80
60
3
40 * 4
6
2 5 7 8
20
*
0
PSA Si(PDAS) Si(PGPMS) C18 Si(PDMS) none
0 5 10 15 20
Sorbents Time (min)
Figure 1. Evaluation of pesticide recovery from adipose tissue samples, with and without cleaning step.
Figure 2. Chromatogram of separation of atrazine (1), endosulfan (2 and 4), chlorpyrifos (3), and cypermethrin (5, 6, 7,
and 8) in adipose tissue extracts fortified at 0.1 mg L -1, using the proposed method.
1 Billet LS, Hoverman JT, Ecotoxicology, 29, 2020, 1476.

2 Musarurwa H, Chimuka L, Pakade VE, Tavengwa NT, Journal of Food Composition and Analysis, 84, 2019, 103314.
3 Golge O, Food Analytical Methods, 14, 2021, 1432.
[CNPq, FINEP]
––––––––––––––––––––– Sample preparation –––––––––––––––––––––

216
SENSITIVE DETERMINATION OF TRIAZINE PESTICIDES IN SURFACE WATER
BY DLLME-LC-MS/MS
Renata P. Gabardo (PG)*a,b, Bianca Amaral (R)b, Gilcélia A. Cordeiro (R)c,
Patrício G. Peralta-Zamora (R)a
a Federal University of Paraná, Department of Chemistry, Curitiba/PR, Brazil, 81530-001
b Itaipu Technological Park, Territorial Intelligence and Management, IT.DT, Foz do Iguaçu/PR, Brazil, 85867-900
c Federal University for Latin American Integration, Foz do Iguaçu/PR, Brazil, 85867-900
*e-mail: renata.p.gabardo@gmail.com
Brazil is one of the world’s largest grain producers and, consequently, one of the largest consumers
of pesticides, representing a latent risk of environmental contamination, especially in the water
environment. Thus, the need for environmental monitoring is more than evident. Preparation
methods that match the analytes concentration and the matrix complexity with the available
analytical technique are still challenging. Recently, miniaturized extraction techniques, such as
dispersive liquid-liquid microextraction (DLLME), have been used to determine pesticides in aqueous
matrices1–5. Therefore, this work aimed to optimize the DLLME system to enable the simultaneous
determination of 10 triazines in natural water, using liquid chromatography coupled to mass
spectrometry (LC-MS/MS). Different extracting solvents (DCM, CLF e C2Cl4) and dispersing solvents
(can, MeOH, acetone) were initially evaluated. The dichloroethane-acetone solvent pair showed the
highest recovery rates (>70% for most triazines). Then, the effect of ionic strength, extractor solvent
volume, and dispersor solvent volume was evaluated by a factorial design (2³ with central point).
According to the observed results, all variables have a positive effect on extraction efficiency, having
been selected as conditions for application of the method: 10% ionic strength, 600 µL of extractor
solvent (DCM) and 1000 µL of dispersor solvent (acetone). Seeking to know the limits of
quantification (LOQ) of the method, an analytical curve was prepared to evaluate the working range
from 0.05 to 0.50 µg L-1, under optimized conditions. LOQ of 0.05 µg L-1 was obtained, and the
determination coefficients were satisfactory for the concentration range evaluated (R²>0.9961).
Therefore, the linearity was got for all analytes in ultra-pure water. Univariate and multivariate
strategies were successfully used to optimize a method based on dispersive liquid-liquid
microextraction (DLLME) to10 triazines by LC-MS/MS. The method allows efficient extraction of the
species of interest and enables quantification limits of the order of ng L -1, which is compatible with
the analysis of natural waters.
1 Caldas SS, Gonçalves FF, Primel EG, Prestes OD, Martins ML, Zanella R. Quim. Nova 2011, 34, 1604.
2 Da Silva AS, Dores EFGC, Villa RD. Rev. Virtual Quim. 2017, 9, 550.
3 Martins ML, Primel EG, Caldas SS, Prestes OD, Adaime MB, Zanella R. Sci. Chromatogr. 2012, 4, 35.
4 Nagaraju D, Huang S. J. Chromatogr. A 2007, 1161, 89.
5 Primel EG, Caldas SS, Marube LC, Escarrone ALV. Trends Environ. Anal. Chem. 2017, 14, 1.
[Itaipu Binational, UNILA, PTI]
Sample Preparation 217

PESTICIDE RESIDUES DETERMINATION IN BLACK BEANS BY BUFFERED
QuEChERS METHOD AND UHPLC–MS/MS
Karina Ues (PG), Cleusa F. Zanchin (PG), Leonardo R. Amaral (PG), Martha B. Adaime (R),
Renato Zanella (R), Osmar D. Prestes (R)*
Laboratory of Pesticides Residue Analysis (LARP), Chemistry Department, Federal University of Santa Maria,
Santa Maria, Rio Grande do Sul, Brazil, 97105-900
*e-mail: osmar.prestes@ufsm.br
Brazil is the world’s third largest common bean (Phaseolus vulgaris L.) producer, and 60% of its
population consumes this legume.1 Pesticides are used globally to improve crop productivity and
quality by protecting crops from pests such as insects, microorganisms, rodents, and weeds.
However, owing to their adverse effects on human health, there are growing concerns about the
health risks associated with exposure to residual pesticides in foods. Consequently, many countries
around the world have established maximum residue limits (MRLs) for pesticides in foods. 2
Analytical monitoring is often conducted to determine if residues are present for food safety,
regulatory, product liability, quality, research, and/or food labeling purposes. 3 The aim of this work
was to evaluate the simple and rapid multiresidue method for the determination of more the 100
pesticides in black beans using ultra-high performance liquid chromatography coupled with tandem
mass spectrometry (UHPLC-MS/MS). The extraction step was optimized testing the unbuffered
(original) and buffered (acetate and citrate) versions of the QuEChERS method. The final extraction
step was performed with 10 g of sample (slurry) and 10 mL of acetonitrile, with the addition of the
citrate QuEChERS salts. The performance of different sorbents for the cleanup step were evaluated,
such as C18, secondary primary amine (PSA) and graphite carbon black (GCB). The pesticide
residue analysis was performed with Acquity UPLCTM liquid chromatography coupled to a Xevo TQ®
MS/MS triple quadrupole mass spectrometer (Waters Corporation, USA) with positive and negative
electrospray ionization interface in multiple selected reaction monitoring (SRM) mode. The
chromatographic separation was performed using an BEH C18 column (50 mm x 2.1 mm, 1.7 µm
particle size). The mobile phase used consisted of (A) water:methanol (98:2, v/v) and (B)
methanol:water (98:2, v/v), both containing 5 mmol L-1 of ammonium formate and 0.1% (v/v) formic
acid, with a flow rate of 0.225 mL min-1. The injection volume was 10 μL and total run time was 10
min. The results of the accuracy and precision tests, evaluated at levels of 10, 25 and 50 µg kg-1 (n=
6), proved to be adequate, since they showed recoveries between 70 and 120%, with RSD ≤ 20%
for more than 115 compounds. The method LOQ values were between 10 and 50 µg kg -1. The
present results suggest that the developed method is suitable for the regulatory purpose analyses
of pesticides in black beans.
1Barreto NMB. et al. Foods, 10, 2021, 900.
2Saito-Shida S, Nemoto S, Akiyam H. Analytical Methods, 13, 2021, 894.
3Koesukwiwatj U. et al. Journal of Agricultural and Food Chemistry, 58, 2010, 5950–5958.
[UFSM, CAPES, CNPq]

MINIATURIZED REVERSED-PHASE DISPERSIVE LIQUID-LIQUID
MICROEXTRACTION IN MICROTUBES FOR FURTHER LITHIUM
DETERMINATION IN LUBRICATING GREASES
Ana C. C. Kusbiak (UG)a, Murilo O. Beato (UG)b, Mariana C. G. Fileres (G)c,
Paulo R. S. Bittencourt (R)a,c, Oldair D. Leite (R)a,c, Eder L. M. Flores (R)a,c*.
a Universidade Tecnológica Federal do Paraná – Campus Medianeira, Programa de Mestrado em Tecnologias
Ambientais, Medianeira, Paraná, Brazil, 85884-000
b Universidade Tecnológica Federal do Paraná – Campus Curitiba, Programa de
Pós-graduação em Química, Curitiba, Paraná, Brazil, 85884-000

c Universidade Tecnológica Federal do Paraná – Campus Medianeira, Departamento de Química,
Medianeira, Paraná, Brazil, 85884-000

*e-mail: eder.utfpr@gmail.com
Lithium is a metal that has been used in several branches of technology, given its multiple
applicability. In this way, lithium hydroxide is widely used as an additive in automotive lubricating
greases. Quality control of this grease is an important step to ensure its functionality. In the present
work, the use of a Reversed-Phase Liquid-Liquid Microextraction (RP-DLLME) in the sample
preparation procedure was evaluated, using miniaturization with microtubes for the determination of
lithium in different brands of commercial automotive lubricating greases. In this work, a miniaturized
RP-DLLME procedure was used in accordance with the principles of Green Analytical Chemistry for
the analysis of automotive lubricating grease, in contrast to the Official method, based on dry-ashing
Initially, the volume of sample mass sufficient for recovery, the solvents n-propanol and isopropanol
as dispersants, the concentration of the extractant solvent HNO3 (0.0, 0.25; 0.50; 1.00; 2.00; 3,00,
4.00 mol L-1) and the proportion of dispersant/extractant solvents (50:50; 60:40; 70:30 and 80:20
(v/v)), as well as the volume of the dispersing/extracting solvent solution used (250; 500 and 750
uL). The temperature in a water bath (60, 65, 70 and 75 ºC) and the centrifugation time (5, 10 and
15 min) were also tested. With the RP-DLLME conditions adjusted, Polypropylene Eppendorf
microtubes with a capacity of 1.5 mL were used. It was used 0.25 g of sample, diluted with 500 µL
of xylene and heated to 65 ºC, a mixture of isopropanol and HNO3 3 mol L-1 in the proportion of 300
uL / 200 uL, shaking with Vortex for 30 s and centrifugation for 15 min at 6000 rpm. The determination
of Lithium was performed by flame atomic emission spectrometry. This study showed that the
miniaturized RP-DLLME procedure is viable with adequate accuracy showed by analyte recovery
tests (89 to 101%) and comparison with official method. The evaluation of how green the proposed
RP-DLLME method was performed using the AGREE® software to establish Green Chemistry
metrics. The proposed RP-DLLME was applied for the first time in samples of lubricating greases,
showing advantages as simple operation, low cost and low consumption of reagents.
1ASTM. D7303-17 – Standard Test Method for Determination os Metals in Lubricating Greases by Inductively Coupled
Plasma Atomic Emission Spectrometry.
2Lourenço EC; Eyng E; Bittencourt PRS; Duarte FA; Picoloto RS; Flores ELM. Talanta, 199, 2019,
3Pena-Pereira F; Wojnowski W; Tobiszewski M; Analytical Chemistry, 92, 2020, 10076.
[CEANMED, UTFPR - Campus Medianeira, CAPES]

]

THYMOL-BASED HYDROPHOBIC DEEP EUTECTIC SOLVENTS AS AN ECO
FRIENDLY APPROACH FOR SCREENING NITROSAMINES IN SARTANS MEDICINES
Thiago S. Novotnya,b, Mychelle A. Monteirob, Patrícia C. de Limab, Soraya M. Ochsb,
Ricardo L. N. Maranhob, Fernando A. Simas Vaza, Flávia F. de Carvalho Marquesa*
aUniversidade Federal Fluminense, Analytical Chemistry Departament/Chemistry Institute, Niterói, RJ, Brazil, 24020-141
b Instituto Nacional de Controle de Qualidade em Saúde, Chemistry Departament, Rio de Janeiro, RJ, Brazil, 21.040-900
*e-mail: flaviamarques@id.uff.br
N-nitrosamines (NA) are carcinogenic substances that should not be present in medicines. In 2018,
Regulatory Agencies detected for the first time the presence of NA in medicines, bringing great
concern worldwide as medicines should not pose additional risk to patients.1 Hence, the agencies
instructed manufacturers to monitor the batches produced, posing considerable costs and
challenges to the pharmaceutical industry and regulators as well.2 Therefore, fast, sensitive, and
cost-effective alternative methods for screening NA in medicines are highly desirable. In recent
years, hydrophobic deep eutectic solvents (HDES) have emerged as a novel “green” alternative to
solvents commonly used in microextraction techniques as they are biocompatible, chemically stable,
biodegradable, low volatile, and non-expensive.3 In the present study, a simple and rapid sample
preparation method designated ultrasound-assisted dispersive liquid-liquid microextraction with
thymol based HDES followed by high performance liquid chromatography with diode array detection
(UADLLME-HPLC-DAD) was developed for the extraction and determination of polar NA from
candesartan, irbesartan, losartan, olmesartan, telmisartan and valsartan drug substances, and from
losartan medicines. The chosen analytes were N-Nitrosodimethylamine (NDMA) and N-Nitroso-N-
methylamino butyric acid (NMBA), in accordance with Brazilian regulations4,5 for active
pharmaceutical ingredients (API), belonging to the class of angiotensin II receptor blockers. The
HDES were prepared by combining thymol (hydrogen bond donor) with menthol, octanoic acid,
tetraheptylammonium bromide, polyethylene glycol 400 and benzyl alcohol as hydrogen bond
acceptors in different proportions. The influence of HDES type, HDES:sample ratio, salt addition and
sample pH was investigated. Best extraction efficiencies were achieved with the combination
thymol:benzyl alcohol (1:1), with enrichment factors ranging from 7.7 to10 times (Figure 1). Under
optimal conditions, linearities ranging from 15 to 1000 ng mL-1 for both NDMA and NMBA (R² > 0.99)
were obtained. Limits of detection in each sartan drug and medicine were 1.1-2.2 ng mL−1 for NDMA
and 1.0-2.9 ng mL-1 for NMBA. Recoveries from 82 to 104% and precision (repeatability and
intermediate precision) < 14% were obtained. Finally, the proposed method was successfully applied
to the determination of polar NA in spiked sartans standards and in losartan potassium tablets
collected in the local market.
Figure 1. Comparison between

chromatograms of 1 mg L-1 NDMA
and NMBA solution (a) before and (b)
after extraction with HDES. Injection
volume = 25 uL.
1WHO. "Update on Nitrosamine Impurities", World health organization, n. November, p. 1–4, 2019.
2EMA. "Assessment report: Nitrosamine impurities in human medicinal products", v. 5, n. June, p. 1–90, 2020.
3Zainal-Abidin MH, Hayyan M, Wong WF. Journal of Industrial and Engineering Chemistry, v. 97, p. 142–162, 2021.
4ANVISA. Resolução RDC N° 283 de 17 de maio de 2019.
5ANVISA. Resolução RDC N° 677 de 28 de abril de 2022.
[FAPERJ-E 26/202.707/2018, INCQS/FIOCRUZ, ANVISA]

EFFICIENT EXTRACTION OF PROTEIN FROM THE BRAZIL NUT
(Bertholletia excelsa) USING DIFFERENT BUFFERS
Carla M. da S. Medeiros*a, Mikael K. de A. Mendesa, Christian B. dos S. Oliveiraa,
Cícero A. L. Júniora, Edivan Carvalho Vieiraa.
aUniversidade Federal do Piauí, Departamento de Química, Teresina, Piauí, Brazil, CEP: 64049-550
*e-mail: Carlamarianadasilvamedeiros@gmail.com
The investigation of proteins from natural sources has shown a high potential for developing
functional foods with beneficial effects on health. Since proteins are macromolecules responsible for
almost all the processes in the cell, exercising is an essential function in health1. Nuts, in general,
have a high protein content. The Brazil nut (Bertholletia excelsa) is widely consumed due to its
pleasant flavor and protein content. However, protein is sensitive to several factors, and the
extraction and sample preparation must be done carefully because the solubility of the protein in the
extractant is a crucial factor in the analysis, so the selection of a buffer is an essential component in
the extraction2. Therefore, this work aimed to investigate the effect of different buffers in obtaining
protein concentrates from Brazil nuts. Initially, the sample was submitted to cold mechanical
pressing, and then the residual lipid content was extracted in a soxhlet system using the binary
combination hexane:ethanol (1:4) and diethyl ether:petroleum ether (1:1), for 5 g of sample in each
combination. After this procedure, the sample was collected, and the solvent was dried until a
constant mass was obtained. The extractions with ethereal solvents resulted in a better extraction
due to the low polarity extracting 21.55%, while the combination hexane:ethanol results in increased
polarity caused by the increased concentration of OH groups, extracting 19.18%. With the defatted
flour obtained, the protein content was extracted using two methods: tris-buffer (pH 7.4, 10 mmol)
and phosphate-buffer (pH 7.4, 10 mmol), at a ratio of 1.5 mL of the extractant solution to 15 mg of
sample. Then the mixture was stirred for 5 minutes in a vortex. Subsequently, separation of the
extract and supernatant was done in a refrigerated centrifuge at 4°C for 30 minutes, 13000 rpm. The
quantification of protein content was performed in a 96-well microplate reader using the Bradford
reagent. The results in Figure 1 indicate that extraction with Tris buffer results in better interaction
with Bertholletia excelsa protein since extraction with this buffer showed almost double the increase
in protein content compared to phosphate buffer. Thus, it is suggested that the best solution for
extracting the total content of soluble proteins for this matrix is Tris buffer. This is due to the limited
ability of phosphate buffer to extract proteins related to its low ability to solubilize weakly hydrophobic
biomolecules. In contrast, Tris has amino and hydroxyl groups, which enable the dissolution of
protein species through interaction with the side chain groups.
Figure 1: Protein concentration of the Brazil nut (Bertholletia excelsa) by extraction with Phosphate Buffer and Tris Buffer.
1Alves-Santos AM, Fernandes DC, Naves MMV. Journal NFS, 24, 2021, 26-36.
2Zbacnik TJ, Holcomb RR, Katayama DS, Murphy BM, Payne RW, Coccaro RC, Evans GJ, Matsuura JE, Henry CS,
Manning MC. Journal of Pharmaceutical Sciences, 2016
[CNPq, CAPES, UFPI]

ACID DECOMPOSITION OF DENIM SAMPLES FOR THE DETERMINATION OF
METALS BY ICP OES
Amanda L. Barbosa (PG), Briyitte S. S. Torres (PG), Ana P. S. Paim (R)*
Universidade Federal de Pernambuco, Departamento de Química Fundamental, Recife, PE, Brazil, 50740-560
*e-mail: ana.paim@ufpe.br
Several operations are important in the manufacturing of clothing articles: from raw materials to
consumer goods, these operations define durability, performance, design, and quality of clothes.
However, some of these operations make use of chemicals and add toxic elements on the surface
of textiles. These elements migrate from textile to skin during daily use [1-3]. Considering the global
demand for cotton for textile purposes, denim garments have one of the largest markets and are
very popular among everyday clothes. In this sense, for the safety of customers, it is essential the
development of methods for the determination of harmful elements in denim clothes. Therefore, it
was developed a sample preparation method, aimed at the determination of multiple elements by
ICP OES. The samples were prepared in a digester block with small volumes of 98% (w/w) H2SO4
and 30% (v/v) H2O2 as reagents. The sample digestion was studied to reach smaller values of
Residual Acidity (RA) and Residual Carbon Content (RCC) using of tools from Design of
Experiments (DOE). For the DOE, it was chosen a sample of difficult digestion: denim with high
polyester content (82% cotton; 16% polyester; 2% elastane). The variables selected were, as
follows: ratio of H2SO4 and H2O2, Step 1 (A); the volume of H2O2, Step 2 (B); heating time (C). A full
factorial design with a central point was applied and all the experiments proceeded in triplicate. The
chosen condition successfully digested 27 of the 29 denim samples, with values of RCC and RA
lower than 2% (w/w) and 10% (v/v), respectively. The particle size distribution was determined by
Dynamic Light Scattering (DLS) analysis and was fewer than 1 μm, independent of the denim sample
composition [4]. The method was validated through addition and recovery tests. Values between 75
and 125% were observed for the following elements: Al, Co, Cu, Mg, Mn, Ni, and Zn [5]. The
proposed method was compared with a reference method [1] and there was no significant difference
at a 95% confidence level according to paired t-tests. Also, the comparison between the determined
values and the certified ones, from a certified reference material (CRM), did not demonstrate
statistical differences. The elements Al (78.7 mg.kg-1), Mg (322.2 mg.kg-1) and Mn (69.0 mg.kg-1)
showed the highest average concentrations, while Co (0.34 mg.kg-1) and Ni (0.67 mg.kg-1) showed
the lowest ones. Taking into account international regulations and standards for toxic elements in
textiles, there is evidence that the element copper might be, in some samples, in higher
concentrations than the established limit [6,7]. The developed method proved to be effective for the
digestion of denim garments, with different compositions, and for the subsequent determination of
seven different metals, namely: Al, Co, Cu, Mg, Mn, Ni, and Zn. This study allows further investigation
of the elementary content of denim clothes, and the health risks concerning the daily use of these
articles.
1Herrero M; Rovira J; Nadal M; Domingo JL. Environ. Res. 172 (2019) 310.
2Matoso E. e Cadore S. Talanta 88 (2012) 496–501.
3Rovira J; Nadal M; Schuhmacher M; Domingo JL. Environ. Res. 140 (2015) 308.
4Costa WKOC, Silva CS, Figueiredo JFD; Paim APSJ. Pharm. Biomed. Anal. 155 (2018) 247.
5US EPA. Method 6010D (SW-846): ICP AES (Update VI, Revision 5). (2018).
6International Association for Research and Testing in the Field of Textile and Leather Ecology. Oeko-Tex®. (2021).
7Global Organic Textile Standard International Working Group, GOTS IWG (2014).
[FACEPE/NUQAAPE, CAPES, CNPq/INCTAA]

NEW SELECTIVE AND EFFICIENT MAGNETIC ION-IMPRINTED POLYMER FOR
PRECONCENTRATION OF TRACE Ni2+ IN WATER, TEA, AND COCOA SAMPLES
Paula M. dos Santosa*, Marcela Z. Corazzaa, César R. T. Tarleya,b
a StateUniversity of Londrina, Department of Chemistry, Londrina, PR, Brazil.
b State University of Campinas, National Institute of Science and Technology (INCT) of Bioanalytical, Campinas, SP,
Brazil.
*e-mail: paulamantovani@live.com
The use of magnetite (Fe3O4) has gained prominence in sample preparation methods due to its
magnetic properties, which are used to separate the extracting phase from the analytical matrix
without the use of filtration and/or centrifugation1. However, the MNPs has some deficiencies in the
process of adsorption of metal ions, such as the lack of selectivity and the possibility of agglomeration
and oxidation under severe conditions of analysis. To overcome these shortcomings, different types
of coatings and/or decorations of MNPs have been proposed, including coatings based on ionically
imprinted polymers (IIPs) technology, which has been noted for its high selectivity, in addition to its
synthesis process being considered simple, fast, low cost, presenting possibilities for different
synthetic routes and applications. Therefore, the main objective of the present study is the synthesis
of a magnetic ionically imprinted poly(allylthiourea) loaded with 1-(2-pyridylaz)-2-naphthol (PAN)
ligand (m@IIP-PAN-Ni2+) and its use in the development of Dispersive Magnetic Solid Phase
Extraction (DMSPE) for sensitive and selective preconcentration of Ni2+ ions and FAAS detection.
The choice of poly(allylthiourea) is due to its ability to form metallic complexes with cation ions, while
azo compounds, such as PAN, can form stable and colored complexes with metals.2,3 In order to
verify the selectivity of m@IIP-PAN-Ni2+ and the influence of PAN on the extraction of Ni2+ ions,
polymers without chemical imprinted as well without PAN were also synthesized. The
characterization of the polymers was performed by Fourier-transform infrared spectroscopy (FTIR),
textural analysis, scanning electron microscopy (SEM), transmission electronic microscopy (TEM),
thermogravimetric analysis (TGA), and vibrating sample magnetometry (VSM). The morphological
characteristics observed in the SEM and TEM analysis of the m@IIP-PAN-Ni2+ demonstrated a
complex reticular structure compared to m@NIP-PAN, due to a higher proportion of polymeric
network in the material, favoring the Ni2+ ions adsorption. The selectivity of m@IIP-PAN-Ni2+ was
assessed from competitive adsorption of binary mixtures, such as Ni2+/Co2+, Ni2+/Cd2+, Ni2+/Cu2+,
Ni2+/Pb2+, and Ni2+/Zn2. Under optimized conditions derived from a 24 factorial design, a 45 mL aliquot
of the sample was preconcentrated onto 50 mg of nanocomposite for 4 min using vortex agitation.
The nickel ions adsorbed on the nanocomposite were eluted using 2 mol L-1 HCl with 1% of thiourea
for 4 min. The proposed method provided a preconcentration factor of 76.70 times, yielding limits of
detection (LOD) and quantification (LOQ) of 0.25 and 0.88 µg L-1, respectively, and a linear range
between 0.88 and 100.00 µg L-1. The method was compared to other methods of extraction and
preconcentration of Ni2+ ions reported in the literature and proved to be advantageous and promising.
The accuracy and precision of the method were investigated using two certified reference materials
(SRM – 1573a and BCR – 191) and good recoveries (104% and 107%) with low RSDs were
achieved. Finally, the optimized method was successfully applied for the determination of nickel in
different brands of mineral water, three types of teas, and samples of powdered chocolate with
different cocoa contents.
1Silva AK, Sobieski E, Viana LH, Lanças FM, Nazario, CED, Scientia Chromatographica, 8, 2016, 239.
2Suquila FAC, Tarley CRT, Talanta, 202, 2019, 460.
3Bai, X, Li, Y, Gu, H, Hua, Z, RSC Advances, 2015, 77217.
[UEL, LADEMA, CAPES, CNPq, INCT]

MAGHEMITE NANOPARTICLES (γ-Fe2O3) FOR THE PRECONCENTRATION
AND DETERMINATION OF As IN EYESHADOW SAMPLES BY FIA-HG-AAS
Marina Araujo João Lopes da Costa (PG)a*, Ana Paula Benevides (PG)b,
Deborah Vargas Cesar (R)a,band Jefferson Santos de Gois (R)a,b
aGraduate Program in Chemical Engineering, Rio de Janeiro State University, Rio de Janeiro, RJ, Brazil, 20550013
bDepartment of Analytical Chemistry, Rio de Janeiro State University, Rio de Janeiro, RJ, Brazil, 20550013
*e-mail: marina.ajlc@yahoo.com.br
Arsenic determination in eyeshadow is challenging due to its low concentration and the complex
composition of the matrix, mostly composed of other elements in a high concentration such as
alumina, silica, titanium oxide, and mica1. In general, dissolution or decomposition using inorganic
acids is required for As determination using hydride generation atomic absorption spectrometry (HG-
AAS) and may jeopardize the detection limit of the analytical method due to successive dilutions
required. To overcome this problem, a preconcentration step can be added to the sample
preparation, therefore, the aim of this work is to present a new method for arsenic preconcentration
in eyeshadow samples using maghemite nanoparticles (γ-Fe2O3) and the determination by flow
injection analysis coupled to HG-AAS (FIA-HG-AAS). Maghemite nanoparticles were synthesized
based on a co-precipitation method2 and characterized using X-ray diffraction (XRD), Fourier-
transform infrared spectroscopy (FTIR), and N2 adsorption measurements at -196 °C. The XRD
results were consistent with a γ-Fe2O3 structure, with no indication of crystalline phases other than
maghemite. The FT-IR spectrum showed peaks at 628, 580 e 447 cm-1 that correspond to Fe–O
stretching and bending vibrational mode, characteristics of γ-Fe-O bond2. The adsorption and
desorption isotherms of N2 presented a type IV behavior, typical of mesoporous materials. The
material presented a pore diameter equal to 101.8 Å, a crystalline size of 9.1 nm, and a surface area
of 106 m2 g−1. The experimental conditions for the hydride generation system and the
preconcentration process were optimized using a central composite design (CCD). For the hydride
generation, the optimal conditions for the concentration of HCl, NaBH4, and the carrier gas flow were
4 mol L−1, 1.1 % (m/ v), and 145 mL min−1, respectively. The refined quadratic model presented an
R-squared of 0.96 and an adjusted R-squared of 0.95. The residues of the models were proven to
follow a normal distribution according to the Shapiro-Wilk test (p=0.05). The major inorganic
composition of the sample was determined by inductively coupled plasma optical emission
spectrometry after microwave-assisted digestion using 120 mg of the sample, 4.60 mL of HNO3 65
% (w/w), and 1.40 mL of HF 48 % (w/w), follow by a complexation step with H3BO3 5% (w/v). The
results showed an average content of 24, 4, 7, 11, 3, and 110 mg g−1 for Al, Ca, Fe, K, Ti, and Si,
respectively in the samples The optimized condition for the adsorption process was achieved at pH
= 5.5, the mass of γ-Fe2O3 = 180 mg, and a stirring time of 40 min in the presence of the sample
matrix. The optimization of the adsorption process simulating the matrix allowed establishing
experimental conditions in which the sample matrix did not affect the adsorption process. Desorption
of As was carried out using 400 μL of a solution of 0.10 mol L−1 EDTA with 5% w/v NaOH and stirring
for 20 min. The enrichment of 50 of achieved, while the limit of detection (LOD) obtained was 0.9 μg
g−1. The accuracy of the method was accessed through recovery tests, which ranged from 75% to
99%. The analytical method proved to be efficient for the determination of As in eyeshadow samples
after microwave-assisted digestion, providing a low LOD that met the requirements for As monitoring
in makeup samples.
1Bocca B, Pino A, Alimonti A, Forte G, Regulatory Toxicology and Pharmacology, 68, 2014, 447.
2Ferreira FN, Benevides AP, Cesar DV, Luna AS, De Gois JS, Microchemical Journal, 157, 2020, 104947
[CNPq, FAPERJ, UERJ]

MICROWAVE-ASSISTED DIGESTION FOR MINERAL DETERMINATION IN BEANS,
BASIL, AND MINT BY ICP OES AND FAES: AN ECO-FRIENDLY ALTERNATIVE
Iago J. S. da Silva (R)a,*, Maria C. da Rocha (UG)a, Micherlane M. da Silva (UG)b, Elvis J. de
França (R)c, Maria J. F. Gomes (R)b, Kenia K. B. da Silva (R)d, Mario T. Kato (R)a
aUniversidade Federal de Pernambuco (UFPE), Depto. de Engenharia Civil e Ambiental, Laboratório de Saneamento
Ambiental (LSA), Recife, Pernambuco, Brazil, 50740-530
bUniversidade Federal Rural de Pernambuco (UFRPE), Depto. de Química, Recife, Pernambuco, Brazil, 52171-900
cCentro Regional de Ciências Nucleares do Nordeste (CRCN-NE), Recife, Pernambuco, Brazil, 50740-545
dUniversidade Federal de Pernambuco (UFPE), Centro Acadêmico do Agreste,
Laboratório de Engenharia Ambiental (LEA), Caruaru, Pernambuco, Brazil, 55002-970

*e-mail: iago.jose@ufpe.br
Beans (Phaseolus vulgaris), basil (Ocimum basilicum), and mint (Mentha sp.) are vegetables of great
relevance in the human diet and agricultural production because of the composition of proteins and
minerals (beans), which act on the human immune system, and essential oils of high added value
(basil and mint)1-3. For consumption, it is important to determine the mineral profile in order to assess
the intake of essential elements and toxic metals contained in those vegetables. Therefore, a simple,
sensitive, precise, and accurate method was developed following the Principles of Green Chemistry.
Calcium, Cd, Co, Cu, Mg, Mn, Mo, Pb, S, V, and Zn were determined in beans, basil, and mint by
inductively coupled plasma optical emission spectrometry (ICP OES, Model 5100, Agilent) and Na
and K, by flame photometry (FAES, DM-61, Digimed). Initially, microwave-assisted acid digestion
was performed for leaf, stem and root of basil and mint and seed of cowpea bean. The used
analytical conditions were those indicated by the microwave oven manufacturer (Ethos Easy,
Milestone): 0.5-g sample plus 8-mL HNO3 14.4 mol L-1. The sample decomposition followed a heating
program consisting of: (i) 20 min to reach 180 °C, (ii) 20 min hold at 180 °C, and (iii) 15 min for
cooling. The resulting residual carbon content (RCC) ranged from 7.1 to 17.8% for mint stem and
leaf, respectively. These relatively low values demonstrate the good potential and viability of using
faster decomposition procedures and diluted HNO3. Further, a factorial design 23 using mint leaf and
stem samples was used to verify the effects of the following variables on the RCC: (i) digestion time
(5, 12.5, and 20 min), (ii) HNO3 concentration (1.0, 2.5, and 4.0 mol L-1), and (iii) volume of 30%
H2O2 (0.5, 1.5, and 2.5 mL). The RCC values (Figure 1) indicated the decomposition conditions of
experiment E6 as the most efficient, i.e., digestion time of 20 min, concentration of 1.0 mol L−1 HNO3,
and 2.5 mL of H2O2. Therefore, the use of only 1.0 mol L-1 HNO3 represents: (i) a saving of reagent
by a factor of 14 times compared to the p.a. solution (14.4 mol L-1), (ii) a considerable decrease in
the concentration of acid residues, and (iii) higher working safety for the analyst, by handling less
corrosive solutions. The analytical method was validated, and satisfactory results were obtained in
terms of linearity (r>0.999), limits of quantification ranging from 0.031 (Mn) to 60 mg kg-1 (Na and K),
and relative standard deviation of up to 11% in recovery tests. The accuracy of the methodology was
evaluated using three reference materials (Spinach Leaves NIST 1570a; Hay Powder IAEA-V-10;
Marandu USP/Embrapa Agro E1001a), with the agreement between the determined and certified
concentration values. The computations for the factorial design were performed using the Statistica
10 (StatSoft) software. All variables
had significant effects on the RCC Figure 1. Cubic diagrams for interpreting factorial design results using
(p<0.05), emphasizing the volume of mint leaf (right) and stem (left), with RCC as the dependent variable.
H2O2, with a mean value of 28%.
Most of the bean, basil, and mint
digestion methods available in the
literature employ concentrated
acids, making the proposed method
a greener, cheaper, and safer
alternative.
1Santos WPC et al., Microchemical Journal,
91, 2009, 153.
2Burducea M et al., Scientia Horticulturae,
249, 2019, 407.
3Rubio C et al., Journal of Pharmaceutical and Biomedical Analysis, 71, 2012, 11.

MICROWAVE-ASSISTED EXTRACTION INDUCED BY EMULSION BREAKING BASED
ON NADES FOR ELEMENTAL DETERMINATION IN EDIBLE OILS BY ICP OES
Floriatan S. Costa (PG)a, Letícia L. Ludovico (UG)a, Luana S. Moreira (PG)a,
Jaqueline Volpe (PG)a, Mayara P. Santos (PG)a, Erik G.P. Silva (R)b, Fábio A.C. Amorim (R)b,
Marco T. Grassi (R)a, Mario H. Gonzalez (R)c, Clarice D.B. Amaral (R)a*
a Federal University of Paraná, Department of Chemistry, Curitiba, PR, Brazil, 81531-980.
b Santa Cruz State University, Department of Exact and Technological Sciences, Ilhéus, BA, Brazil, 45662-900.
c São Paulo State University, Department of Chemistry and Environmental Science,
São José do Rio Preto, SP, Brazil, 15054-000.

*e-mail: clariceamaral@ufpr.br
Natural Deep Eutectic Solvents (NADES) are low-cost new customizable safe solvents with high
extractability.1 The application of NADES in chemical analysis provides promising results, especially
in the development of Green Analytical Chemistry (GAC) methods. In this work, a new Microwave-
assisted Extraction Induced by Emulsion Breaking (MW-EIEB) based on Natural Deep Eutectic
Solvent was developed aiming at the elemental determination by inductively coupled plasma optical
emission spectrometry (ICP OES). NADES composed of carboxylic acids and choline chloride were
prepared (microwave heating)2 and evaluated for extractability, in addition to molar ratio and water
addition. In this approach, the oil samples were emulsified with a surfactant/extractant solution to
form a stable water-in-oil emulsion.3,4 Further, the emulsion breaking was promoted by microwave
heating and the aqueous phase was used in the measurements by ICP OES. Satisfactory results
were obtained in the proposed method applied to edible oil samples. The highest recovery range
(86–90%) was obtained using the NADES composed of choline chloride, oxalic acid, and water
(1:1:4 molar ratio). Also, surfactant type was evaluated, and Triton X-100 (5% v/v) provided
recoveries >93% for all analytes (Al, Ca, Cr, Cu, K, Mg, Mn, and Zn). A mixture design was applied
to optimize sample and solvent volumes and the optimal condition was provided when using 5.0 mL
of sample, 1.7 mL of NADES, and 1.3 mL of Triton X-100. The vortex time, extraction, and MW
assisted-emulsion break were optimized by the Box-Behnken design. Optimal results were obtained
with 36 s vortexing, 5 min of extraction, and 10 s emulsion breaking. The combination of NADES
and MW-assisted EIEB provided high recovery rates, reaching interesting experimental speed and
simplicity when compared to conventional methods (e.g., acid digestion). Also, the GAC metrics
were complied3 with and the results obtained support the application of NADES as an efficient
extractor in methods based on the formation of emulsions. As far as we know, this study is the first
work proposing the application of NADES combined with MW radiation in EIEB methods prior to
elemental determination by ICP OES.
1Liu Y. et al., J. Nat. Prod. 2018, 81, 679.

2Costa FS. et al., J. Food Compos. Anal. 2022, 109, 104510.
3Valasques GS. et al., Appl. Spectrosc. Rev. 2017, 52, 729.
4van Osch DJGP et al., Phys. Chem. Chem. Phys. 2020, 22, 2181.
[CAPES, CNPq, INCT-DATREM, INCTAA, PETROBRAS, LAMIR, FAPESP, PRPPG, UFPR]

INNOVATIVE FIBER-IN-TUBE SPME-MS/MS METHOD TO DETERMINE
β-AMYLOID PEPTIDES IN CEREBROSPINAL FLUID SAMPLES
Israel D. Souzaa*, Jared L. Andersonb and Maria Eugênia Costa Queiroza
a FFCLRP-Universidade de São Paulo, Departamento de Química, Ribeirão Preto, SP, Brazil, CEP 14040-901
b Iowa State University, Department of Chemistry, Ames, IA, U.S.A, CEP 50011
*e-mail: israeldsz@usp.br
Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by progressive impairment

of cognitive skills, behavioral changes, and loss of functional autonomy. Pathologically, AD is
characterized by accumulation of extracellular β-amyloid (Aβ) plaques and intracellular
neurofibrillary tangles in the brain. Aβ peptides in cerebrospinal fluids (CSF) have been evaluated
as biomarkers for early AD diagnosis 1. Quantitative analyses of Aβ peptides in CSF have relied
almost exclusively on immunoassay-based procedures. However, these assays are time-consuming
and expensive, require highly specific antibodies and reagents, present poor dynamic ranges, can
present cross-reactivity, and are very sensitive to matrix interferences. In this context, quantification
methods based on tandem mass spectrometry (MS/MS) can overcome many of the problems
associated with immunoassay-based methods. Recently, microextraction devices directly coupled
to MS/MS have redefined the analytical workflow by providing faster screening and quantitative
methods for complex matrixes 2. In this context, this study presents direct coupling of fiber-in-tube
SPME to MS/MS as a high-throughput approach to determine Aβ peptides in CSF samples. The
biocompatible fiber-in-tube SPME capillary was developed by using a PEEK tube segment
longitudinally packed with fine fibers (nitinol wires coated with zwitterionic polymeric ionic liquid (Zw-
PIL)) as the selective extraction medium 3. The fiber-in-tube SPME-MS/MS method efficiently
integrated analyte extraction/enrichment and sample clean-up (elimination of the matrix effects) in
less than 10 min. In addition, the analytical method presented lower limit of quantification (0.2 ng mL-
1
for Aβ1-38 and 0.1 ng mL-1 for Aβ1-40 and Aβ1-42), as well as precision (coefficient of variation values,
CV) and accuracy (relative standard deviation, RSD) lower than 7.3% and 11.6%, respectively. This
method was successfully applied to determine Aβ in CSF samples from AD patients (n = 8) and
controls (healthy volunteers, n = 10). The basal concentrations (mean ± S.D.) of Aβ in the CSF
samples were 1674 ± 365 (Aβ1-38), 7023 ± 1614 (Aβ1-40), and 942 ± 167 pg mL-1 (Aβ1-42) and
1693±1006 (Aβ1-38), 7449±2685 (Aβ1-40), and 558±231 pg mL-1 (Aβ1-42) for the control and AD
patients, respectively. Results showed that Aβ1-42 levels in the CSF from AD patients were
significantly decreased compared to healthy individuals (p < 0.05). Additionally, the Aβ1-42 results
obtained by using the fiber-in-tube SPME-MS/MS method exhibited strong positive linear correlation
(r = 0.95, Pearson) with the results obtained by ELISA immunoassay. Although there were no
substantial changes in the CSF Aβ1-38 and Aβ1-40 levels, the AD group presented significantly lower
levels when the peptide levels were compared by the CSF Aβ1-42/Aβ1-40 ratio. On the basis of ROC
analysis, the Aβ1-42/Aβ1-40 ratio (AUC = 0.950, p < 0.01; 95%), distinguished the AD group from
healthy groups better than Aβ1-42 (AUC = 0.913, p < 0.01; 95%). Moreover, the ratiometric analysis
(CSF Aβ1-42/Aβ1-40) was more reliable because it compensates the intraindividual fluctuations among
the patients. In conclusion, the innovative advances described in this study are related not only to
the clinical study of Aβ peptides as AD biomarkers, but also to the direct online coupling of fiber-in-
tube to MS/MS for high-throughput bioanalysis.
1Lee JC, Kim SJ, Hong S, Kim Y, Exp. Mol. Med. 51, 2019, 1–10.
2Gionfriddo E, Gómez‑Ríos GA, J. Mass Spectrom. 56, 2021, e4665.
3Souza ID, Anderson JL, Queiroz MEC, Anal. Chim. Acta. 1193, 2022, 3393942.
[FAPESP, Instituto Nacional de Ciência e Tecnologia Translacional em Medicina]

DEEP EUTECTIC SOLVENT FOR DISSOLUTION OF DISCARDED
LITHIUM-ION BATTERIES
Adriane T. da Silva (PG)a, Jamille C. Souza (PG)a, Leonardo S. P. Alves (UG)a,
Victor S. Teixeira (UG)a, Jossemyller F. Damascena (UG)a, Kaíque A. M. L. Cruz (PG)a,
Helena I. S. Passos (R)b, Nicolas Schaeffer (R)b, Maria C. Hespanhol (R)a*
aGrupo de Análises e Educação para a Sustentabilidade (GAES), Departamento de Química, Centro de Ciências Exatas
e Tecnológicas, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil, 36570-900
bCICECO - Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, Aveiro, Portugal, 3810-193
Lithium-ion batteries (LBIs) are being used increasingly due to the high consumption of portable
electronic equipment and electric vehicles. However, as their life cycle is short, many LBIs are
discarded, creating waste and contaminating the environment. LIBs are valuable because 10 to 55
% (w/w) cobalt can be found at the cathode.1 As a result, efforts are being made to obtain efficient
processes for reusing metals.1 One of the most used processes is hydrometallurgy, in which the
cathode is dissolved, followed by liquid-liquid extraction, and the pure metal is obtained.1 In this work,
special attention was given to the dissolution step to replace acids, which are typically used and are
harmful to health and the environment, with deep eutectic solvent (DES). DES has similar properties
to ionic liquids, such as low volatility and adjustable polarity/acidity, but has lower toxicity and cost
and is considered the future solvent. Thus, the feasibility of using DES for cathode dissolution of
discarded LIBs was evaluated by investigating the influence of the presence of water and molar ratio
of DES, dissolution time, type of DES (choline chloride (ChCl):acetic acid (AcA),
ChCl:methanesulfonic acid (MSA) and ChCl:citric acid (CiA), dissolution temperature and solid/liquid
ratio. The sample of cathodes was calcined at 550 oC for 5 h, homogenized, and subjected to
different dissolution conditions. The dissolution with aqua regia (1HNO3:3HCl) yielded the total
concentration of Co, Li, Mn, and Ni equal to (33 ± 4), (3.7 ± 0.4), (7.2 ± 0.7), and (4.3 ± 0.2) % (m/m),
respectively. ChCl:AcA containing 30 % H2O provided higher dissolution efficiency compared to
ChCl:AcApuro (Figure 1a). The ChCl:AcApuro has a higher viscosity making it more difficult to dissolve
the metals. On the other hand, the molar ratio ChCl:AcA influences the dissolution to a lesser degree,
with higher leaching for 1ChCl:2AcA, i.e., when a larger amount of acid is employed (Figure 1a). The
dissolution of metals was not influenced by time, except for Co, where there was no increase in the
amount of Co leached after 30 min. The DES 1ChCl:2MSA provided higher Co leaching efficiency
when compared to DES 1ChCl:2AcA and 1ChCl:2CiA (Figure 1b). The dissolution of Co first occurs
by reducing CoIII to CoII due to the presence of ChCl, followed by the interaction between MSA and
Co, forming the soluble (2CH3O3S)2Co compound in the medium. By increasing the dissolution
temperature from 40 to 80 oC (Figure 1c) and the solid:liquid ratio from 1:10 to 1:25 (Figure 1d), a
significant increase in the amount of leached metal occurs where Co, Li, Mn, and Ni is 89, 98, 83
and 95 % leached, respectively. The high capacity of DES 1ChCl:2MSA to promote the almost
complete dissolution of metals is a promising result. Since the DES medium is conducive to
performing electrodeposition of Co. Thus, metallic cobalt could be obtained without needing a liquid-
liquid
8
extraction step. 15 25 30
ChCl:MSA (1:2)30 % water Co
Co ChCl:AcA Co Co
metal concentration / % (m/m)
ChCl:MSA (1:2)30 % water

Li Li Li Li
Mn Mn 20 Mn 25 Mn
12 Ni
6 Ni Ni Ni
20
9 15
4 15
6 10
10
2
3 5
5
0 0 0 0
(1:2) (1:2) (2:1) (1:1) 1ChCl:2MSA 1ChCl:2AcA 1ChCl:2CitA 40 80 1:10 1:25
puro o
30 % water 30 % water temperature / C solid:liquid ratio
(a) (b) (c) (d)
1 Mansur MB, Guimarães AS, Petraniková M, Miner. Process. Extr. Metall. Rev., 43, 2022, 489.
[INCTAA/CNPq, CAPES/FCT, FAPEMIG, CNPq, CAPES]

MICROWAVE-ASSISTED UV DIGESTION OF ORGANIC SAMPLES WITH
DILUTED HNO3 EMPLOYING O2 AND H2O2 AS AUXILIARY REAGENTS
Vanessa R. do Nascimento (PG)*, Karen Giacobe (PG)a, Débora P. de Almeida (PG)a,
Cezar A. Bizzi (R)a
*e-mail: vanessa.nascimento@acad.ufsm.br
Wet decomposition has several advantages over the combustion technique such as the use of lower
temperatures, which reduce the loss of analytes but may commit the digestion efficiency. In this
context, microwave heating can be used as a tool to improve digestion efficiency.1 Microwave
heating combined with closed vessels allows the use of higher temperatures and pressures, which
allows reducing the amount of required reagents without impacting the digestion efficiency, even
when dilute HNO3 is used. In addition, the digestion efficiency can be further improved by the use of
ultraviolet (UV) radiation combined with microwave-assisted digestion.2 Based on this aspect, this
work aims to evaluate the microwave assisted UV digestion using diluted HNO3 with the aid of
auxiliary reagents such as O2, and H2O2, for the digestion of organic samples (milk powder, potato
starch, and imipramine hydrochloride) for further metals determination. The samples were digested
in a microwave oven (Multiwave 3000®, Anton Paar) with an electrodeless lamp with cadmium vapor
(1 mg), filled with low pressure of argon (4 mbar). The optimization of sample digestion was
performed using 15 mL of 0,093 mol L-1 HNO3. Up to 100 mg of sample were digested i) with, and ii)
without the UV-lamp. For each condition, 20 bar of O2, or H2O2 were evaluated as auxiliary reagent.
Besides that, instrumental parameters were also evaluated, such as heating time (30 min up to 60
min), and external cooling (FAN mode 1, 2 and 3). For Fe recovery 100 µL of HCl was used. The
digestion without acid was also evaluated. After selecting the most efficient digestion condition, the
metals (Ca,Cd, Co, Cu, Fe, K, Mg, Mn, Mo, Na, Pb and Zn) determination step by ICP OES (Spectro
Analytical Instrumental, Spectro Ciros CCD) was performed. The evaluation of the digestion
efficiency was performed by dissolved organic carbon (DOC, mg L-1) determination, and by residual
carbon content (RCC, %) calculation, based on the total carbon content (TOC, %) originally present
in the sample. The value of RCC obtained for milk powder after digestion with H2O2 or with 20 bar
O2, without the lamp was 6.5 ± 0.5 and 7.2 ± 0.2%, respectively. When only 20 bar of O2 was used,
the digestion was not efficient (RCC 55.9 ± 5.7%), which was not improved by the use of UV-lamp.
However, a better condition was obtained when UV-lamp was used with the aid of H2O2 and O2 (RCC
1.2 ± 0.1%). This condition, the one that presented the better RCC results for milk powder, was
applied to imipramine hydrochloride sample (RCC 3.8 ± 0.5%). For potato starch, all the conditions
showed an RCC of less than 1% except when O2 was used alone, without UV-lamp. The optimal
operational parameters were i) 60 min of digestion (RCC de 0.4 ± 0.1, 0.3 ± 0.1 e 1.6 ± 0.1% to
samples of potato starch, milk powder, and imipramine hydrochloride); ii) microwave irradiation with
external cooling at FAN 3, which improve the digestion efficiency (RCC lower than 0.9 ± 0.2% for all
the samples). The use of HCl combined with HNO3 improved the digestion efficiency (RCC of 1.5 ±
0.2, 1.5 ± 0.2 e 5.4 ± 0.4% potato starch, milk powder, and imipramine hydrochloride, respectively).
Finally, it was evaluated the decomposition with UV radiation with no acid, just H2O2 and O2 for 30
min with the external cooling at FAN 1 mode, and the value of RCC 12.2 ± 0.6, 0.4 ± 0.1 e 4.9 ±
0.8% potato starch, milk powder, and imipramine hydrochloride, respectively, were obtained. For the
elemental determination step, the accuracy of the proposed method was evaluated using CRMs of
milk powder (BCR 151), and muscle bovine (NIST 8414), which were in agreement with the certified
values.
1Krug, FJ Métodos de Preparo de amostras para análise elementar, EditSBQ – Sociedade Brasileira de Química, 2,
2019, Brasil.
2Cerveira C, Hermann PRS, Pereira JSF, Pozebon D, Mesko MF, Moraes DP, Evaluation of microwave-assisted
ultraviolet digestion method for rice and wheat for subsequent spectrometric determination of As, Cd, Hg and Pb, 92,
2020, 103585.
[FAPERGS, CAPES, CNPq]

SYNTHESIS AND APPLICATION OF MAGNETIC POLY(ALLYLTHIOUREA-MAA) IN
THE PRECONCENTRATION OF Cd(II) IONS AND DETERMINATION BY FAAS
Liriana M. Roveda (PG)a*, Magno A. G. Trindade (R)a,b, Marcela Z. Corazza (R)c
aFederal University of Grande Dourados, FACET, Dourados, MS, Brazil, CEP 79.804-970.
bUnesp, National Institute for Alternative Technologies of Detection, Toxicological Evaluation and Removal of
Micropollutants and Radioactives (INCT-DATREM), Institute of Chemistry, Araraquara, SP, Brazil, CEP 14.800-900
cState University of Londrina, Department of Chemistry, Londrina, PR, Brazil, CEP 86.057-970
*e-mail: lirianaroveda@ufgd.edu.br
In recent years, SPE has become the ultimate choice in the separation and preconcentration of
heavy metals at trace levels, especially from the synergistic integration of adsorption technology with
nanotechnology1. This is because nanoscale materials have unique characteristics resulting from
their finite size, such as high specific surface area, adjustable pore size and surface reactivity, large
number of active sites on the surface and high adsorption capacity2. Thus, in this study, a new
magnetic polymeric sorbent with double monomer was synthesized, characterized and applied in
the Cd(II) separation and preconcentration, once in addition to the desirable characteristics of
nanomaterials, the magnetic propertied provides speed and simplicity to SPE. For the synthesis of
the adsorbent, the magnetite was coated with silica and vinyl functionalized with VTES. Allylthiourea
and methacrylic acid were chosen as monomers in order to insert functional groups that would direct
the selectivity of the sorbent towards Cd(II) ions. The sorbent was successfully characterized by
FTIR, BET, XRD, SEM and TEM. Kinetic studies were performed in batch using 30 mg of polymer
and 30 mL of 1.0 mg L-1 of Cd(II) solution under optimized conditions (pH 6.0). The equilibrium time
for Cd(II) adsorption was 20 min and the experimental data showed an excellent fit for the pseudo-
second order model. Adsorption isotherm experiments were performed with solutions in the
concentration range of 0.4 – 6.5 mg L-1 of Cd(II) and the Langmuir model was the one fitted to the
experimental data of the polymer with a maximum adsorption capacity of 2.56 mg g-1. The optimized
variables (sorbent mass = 30 mg; extraction time = 2 min; eluent type = HNO 3 and elution time = 2
min) were obtained through a complete factorial design 24 and FAAS was used as a detection
technique. A preconcentration factor of 37 times and detection and quantification limits of 0.12 and
0.43 µg L-1 were achieved, respectively. The retention of Cd(II) ions in the sorbent phase was not
affected in the presence of foreign ions, studied in different proportions. The proposed method was
satisfactorily applied to the determination of Cd(II) in different water samples, obtaining recovery
percentages ranging from 91.2 – 97.0% and relative standard deviations lower than 2.5%.
1Li Z, Yu B, Cong H, Yuan H, Peng Q, Rev. Adv. Mater. Sci. 48,2017, 87

2Li R, Zhang L, Wang P, Nanoscale, 7, 2015, 17167
[UFGD, UEL, CNPq, INCT-DATREM]

PREPARATION AND CHARACTERIZATION OF A POLY(TMPTA-CO-EDMA)
MONOLITHIC SORBENT FOR DISPOSABLE PIPETTE EXTRACTION
Allyson L.R. Santos (PG)a*, Anizio M. Faria (R)a
*e-mail: allyson@ufu.br
The search for quick, eco-friendly, and portable sample preparation methods for chemical analysis
has been increasingly incisive. From this perspective, one method that has stood out is disposable
pipette extraction (DPX). DPX is considered a form of solid phase extraction (SPE), in which the
particulate sorbent is dispersed inside a micropipette tip and retained by Teflon filters. 1 Due to the
reduced scale of the sorbent mass and the sample and eluent volumes, DPX has also considered
an SPE miniaturization. However, for complete portability of the DPX, two factors still require further
studies (i) the few availabilities of sorbents, limiting the selectivity of the method, and (ii) the reduction
of the pressure when suctioning and dispensing of liquids due to the presence of microporous Teflon
filters inside the tip. We propose to prepare and functionalize macroporous polymer monoliths inside
the micropipette tip for use in DPX. Monoliths are single macroporous particles made up of long
channels that allow the flow of liquids with less resistance (low pressure) which does not require
filters to retain them in the tip.2 Organic polymer monoliths are easy to prepare and functional, as
well as they are chemically stable across a wide pH range. In this work, a synthesis of organic
monoliths was developed and optimized from the in situ thermal polymerization of trimethylolpropane
triacrylate (TMPTA) and ethylene glycol dimethacrylate (EDMA), using 2,2-azobisisobutyronitrile
(AIBN) as a radical initiator and polyethylene glycol (PEG) and isopropanol as porogenic at 60 °C
for 20 h. The functionalization of the monolith occurred by molecularly imprinted polymer (MIP) of
Congo red (CR) simultaneously with the polymerization step to evaluate the extraction selectivity.
To optimize the monolithic sorbent, the molar ratios TMPTA:EDMA, monomer:porogen and
CR:TMPTA, and certain porogenic solvents were evaluated. The flow of liquids through the produced
monoliths was monitored in the optimization. The monoliths were characterized morphologically and
physicochemically. CR adsorption on MIP monolithic sorbent was measured and compared with
non-imprinted polymer (NIP) monolithic. The monoliths were efficiently produced (Figure 1) and
showed 60% of macroporosity, as can be seen by scanning electron microscopy, Figure 2. The
specific surface area ranged from 12 to 82 m2 g-1 with total pore volume from 0.82 to 0.95 g cm-3.
Typical stretches in the infrared region confirmed the polymerization of TMPTA-co-EDMA and the
imprinting of CR on the monolithic MIP. The CR retention in the MIP was higher than in the NIP,
validating the molecular imprint of the CR. The liquid flows in the MIP monolithic were 1,24 mL
min-1, confirming the high permeability of the fluids through the monoliths using a manual
micropipette.
Fig. 1. Photo of poly(TMPTA-co-EDMA) monolith. Fig. 2. SEM of poly(TMPTA-co-EDMA) monolith at 5 kX magnification.
1Bordin DCM, Alves MNR, Campos EG, De Martinis BS, Journal of Separation Science, 39, 2016, 1168.
2Barberán MV, Correa EJC, García MJL, Alfonso EFS, Martínez JMH, Analytica Chimica Acta, 1084, 2019, 1.
[CNPq, FINEP]

A NEW METHOD BASED ON THE COAGULATION PROCESS FOR THE
DETERMINATION OF SILVER NANOPARTICLES IN WATER BY GFAAS
Priscila L. S. Estevão*a, Patricio Peralta-Zamoraa, Frederico L. F. Soaresa, Noemi Nagataa
a Universidade Federal do Paraná, Chemistry Department, Curitiba, PR, BR
*e-mail: priscilalagner@hotmail.com
The growing global use of silver-based nanomaterials (nano-Ag), has resulted in inevitable
environmental contamination. However, the monitoring of nano-Ag in low concentrations is still a
challenge for analytical chemistry, especially because of the problems in differentiating the metal
nanoparticles from their ions and the low instrumental sensitivity, which makes the use of pre-
concentration techniques essential. Several studies have demonstrated the efficiency of the
coagulation process in removing engineered nanomaterials, such as nano-Ag, during the treatment
of natural and wastewater.1,2,3 In this context, this work aims to investigate the potential use of the
organic coagulating agent liquid Tanfloc SG® (Tanac S.A), for the extraction and pre-concentration
of nano-Ag from drinking and natural waters followed by GFAAS analysis. Commercial citrate (50
nm, Sigma Aldrich) and synthesized polyvinylpyrrolidone (PVP, 40 kDa) coated nano-Ag standards
were used as models for the method optimization and validation. The samples were fortified with Ag+
(TraceCert®, Sigma Aldrich), Ag-PVP, and Ag-Citrate, separately, to evaluate the extraction
efficiency for different silver species, and the method performance was assessed by Graphite
Furnace Atomic Absorption Spectroscopy (GFAAS). At the optimized conditions, the extraction was
carried out with 45.0 mL of aqueous samples added with 75.0 µL of Tanfloc SG (33.3 % w/w aqueous
solution). The pH was adjusted to 6 with tetramethylammonium hydroxide solution (1.67% w/v in
water, Sigma-Aldrich) to assist the coagulation process. The samples were manually stirred and then
centrifuged to accelerate phase separation and allow the recovery of the colloidal precipitate. The
supernatant was decanted and the extracts obtained were digested with H2O2 in a medium of HNO3
by microwave heating using a domestic microwave (Electrolux ME850). The extracts were stored at
room temperature until the GFAAS analysis. High extraction efficiency values (>76%) were observed
for all silver species in the digested extracts obtained in the absence of interferences, primarily
indicating the potential of the method in determining the total silver content in the samples. Adequate
linearity (r>0.9999) and low limits of detection (10.1 ng L-1) and quantification (33.7 ng L-1) were
achieved by the matrix-matched calibration curves, reaching enrichment factors close to 46 times.
Nevertheless, a matrix effect that significantly reduced Ag+ extraction efficiency was observed in real
samples, with recovery rates < 20 % while adequate values were observed for nano-Ag (86-118%).
Preliminary tests indicated that the presence of humic acids can stabilize Ag + in solution, thus
reducing the extraction efficiency. Furthermore, the presence of Ca2+, which competes for humic
acid with Ag+, makes it possible to increase the extraction of silver ions. Future assays will be
conducted to evaluate the calcium and humic acid contents to activate/deactivate the matrix effect
and guarantee the desired differentiation between Ag+ and nano-Ag, which is not possible to be
achieved only by the instrumental technique (GFAAS) under conditions used in this work. Thereby,
besides presenting high sensitivity, accuracy, and low cost, the proposed method still has great
potential in ion/nanoparticle differentiation for metal-based nanomaterials.
1Sun Q. et al. Journal of Hazardous Materials, 261, 2013, 414.
2Weinberg,H.; Galyean A.; Leopold M. TrAC Trends in Analytical Chemistry, 30, 2011, 72.
3Choi S. et al. Science of The Total Environment, 590–591, 2017, 809.
[CAPES, Dr. Marco T. Grassi, Dr. Andrea P. Oliveira.]

EVALUATION OF EXTRACTION PARAMETERS FOR THE ANALYSIS OF LIPID
CLASSES IN PLANTS
Taynara S. Matosa*, Emerson A. F. dos Santosa, Mariana S. Marquesa, Thales F. D. Pereiraa,
Cleber J. N. Chavesb, Flávia S. Zandonadia, Alessandra Sussulini (R)a
a Laboratory of Bioanalytics and Integrated Omics, Institute of Chemistry, University of Campinas,
Campinas, SP, Brazil, CEP 13083-970
b Laboratory of Molecular Ecology and Evolutionary Genomics of Plants, Institute of Biology, University of Campinas,

*e-mail: t262827@dac.unicamp.br
Sample preparation is a crucial step to select the class of metabolites to be analyzed, reduce sample
complexity, and define parameters of the chosen analytical method. Lipids are plant components
that participate in many processes as in cell signaling, biotic and abiotic stress tolerance, besides
energy storage properties. Plant lipidomic studies have been employing different extraction methods
including different solvents and proportion of them1. The processes used in this study were based
on the method developed by Hummel et al. (2011) for plant lipid analysis using UHPLC-HRMS. In
this method, the sample was separated in 3 phases2: organic, aqueous and protein phases. The
parameters evaluated in our study were different solvent proportion (1:2, 1:3 or 1:4 v/v) and time
under ultrasonic in ice-cold bath (10, 20 or 30 min). The extraction solvents were methanol (MeOH)
and methyl tert-butyl ether (MTBE). For each condition, experiments were prepared in triplicate
resulting in 27 samples, as described in Table 1. The aim of this study was preliminarily evaluating
the lipid profile from the leaves of Pitcairnia flammea followed by the lipid class enriched analysis
based on changes in the extraction parameters. In order to achieve this goal, an UHPLC-MS method
and the MSDial platform were used to perform data preprocessing and statistical analysis. Samples
extracted in MeOH:MTBE (1:2 v/v) for 10 min and 20 min ultrasonic bath did not separate in three
phases. Then, their organic phase chromatographic analyses were not performed. 42
chromatograms were obtained from the LC-ESI-(+)-MS and LC-ESI-(-)-MS analyses. Quantitatively,
the combination of LC-ESI-(-)-MS analysis and MeOH:MTBE 1:3 v/v extraction consisted in the most
efficient method according to the total ion chromatograms. Statistical analysis of LC-ESI-(-/+)-MS
data allowed visualization of a trend distribution according to the extraction time. Besides, it was
possible to observe that solvent proportion has more influence to separation between the groups in
the data samples of 30 min of extraction than 10 and 20 min. Moreover, the lipid concentrations were
directly proportional to the extraction time. This result is more pronounced in the LC-ESI-(+)-MS
data. Besides, fatty acids, glycerophosphoinositols and glycerophosphates were mostly observed in
the negative mode data while in the positive mode data, phosphatidylcholines and diacylglycerols
were the lipid classes more relevant.
Table 1. Experimental design of extraction with proportion of solvents and time under
ultrasound in ice bath
Proportion/Time A_10 min B_20 min C_30 min
1_1:2 A1.1, A1.2, A1.3 B1.1, B1.2, B1.3 C1.1, C1.2, C1.3
2_1:3 A2.1, A2.2, A2.3 B2.1, B2.2, B2.3 C2.1, C2.2, C2.3
3_1:4 A3.1, A3.2, A3.3 B3.1, B3.2, B3.3 C3.1, C3.2, C3.3
1Kehelpannata C, Rupasinghe T, Hennessy T, Bradley D, Ebert B, Roessner U, Molecular Omics, 17, 2021, 894.
2Hummel J, Segu S, Li Y, Irgang S, Jueppner J, Giavalisco P, Frontiers in Plant Science, 2, 2011, 1.
[CAPES, FAPESP, FAEPEX, INCTBio]

NICKEL-MODIFIED PAPER-BASED DEVICE FOR CYANIDE QUANTIFICATION
USING HEADSPACE SINGLE-DROP MICROEXTRACTION
Josiele A. M. Conrado (PG)a*, Diele A. G. Araújo (R)b, João Flávio S. Petruci (R)a
aFederal University of Uberlândia, Institute of Chemistry, Uberlândia, MG, Brazil, 38408-902.
b University of São Paulo, Institute of Chemistry, Department of Fundamental Chemistry, São Paulo, SP, Brazil, 05508-
900
*e-mail: josikpela@hotmail.com
Cyanide (CN-) is a highly toxic compound due to its combination with human red blood cells that
leads to asphyxia and can be lethal in a short time. It is used by some plants as a mechanism of
defense against herbivores, furthermore, it can be found in low quantities in foodstuffs such as
apples, mangoes, cassava, germinated potatoes, bitter almonds and rapeseeds [1]. Besides, the
presence of CN- in natural waters is regulated by most of the global legislation. Therefore, the
quantification of CN- in a variety of matrices is essential. Most of the conventional analytical methods
are time-consuming, high-cost, and require bulky instruments. In this study, we present a sensitive,
selective, low cost and portable analytical system for the quantification of cyanide anion in water.
The analytical procedure is based on the formation of gaseous HCN and its collection using single-
drop microextraction followed by a colorimetric reaction on a paper surface. 5 mL of the sample or a
cyanide standard solution, 500 µL of NaCl 3% and 1000 µL of H3PO4 2 mol L-1 were added into a
glass vial and immediatelly sealed. The generated HCN was collected using a single-drop of a
palladium dimethylglyoximate solution (Pd(DMG)22-) prepared in KOH 3 mol L-1 (extraction solution)
fixed in the flask headspace using a syringe. In strong basic solutions, Pd(DMG)22- reacts selectively
with cyanide leading to the desmasking of dimethylglyoxime (Equation 1 and 2).
𝑃𝑑(𝐷𝑀𝐺𝐻)2 + 2𝑂𝐻 − ⇌ 𝑃𝑑(𝐷𝑀𝐺)2− 2 + 2𝐻2 𝑂 (1)

𝑃𝑑(𝐷𝑀𝐺)2−
2 + 4𝐶𝑁 −
⇌ 𝑃𝑑(𝐶𝑁4 )2−
+ 2𝐷𝑀𝐺 2−
(2)
After the sampling time, the extraction solution was added to a paper-based analytical device
previously impregnated with Nickel (II). The red color originated by the reaction of Ni2+ and DMG was
used as indicative of cyanide in the sample (Equation 3).
𝑁𝑖 2+ + 2𝐷𝑀𝐺𝐻 ⇌ 𝑁𝑖(𝐷𝑀𝐺)2 + 2𝐻 + (3)
Then, the paper device was dried and the digital image was obtained using a flatbed scanner (HP
Deskjet G4050) with resolution of 300 dpi. The color information was extracted using ImageJ® as
RGB parameters and the difference between the green values of blank and sample (∆G) was
considered as the analytical signal. A drying time of 5 min, a paper device with 10 mm of diameter,
3 µL of the extraction solution, and 15 minutes of extraction time were chosen as the optimum
conditions. This analytical system enabled a linear range of 1.0 to 11.0 µmol L-1 of CN- described by
the equation: ∆G = 2,266 [CN-] + 15,627 (r2 > 0.99), with a limit of detection of 0,40 µmol L-1. This
approach enabled the development of an inexpensive, highly selective and sensitive analytical
method for the quantification of cyanide. In addition, the proposed device shows high degree of
portability for in-situ applications.
1 Randviir EP, Banks CE. TrAC - Trends Anal. Chem. 64 (2015) 75.
[UFU, CAPES, FAPEMIG, CNPq]

ANALYTICAL PERFORMANCE OF TOTAL LIPID EXTRACTION USING
TRIZOL® SOLUTION IN BIOLOGICAL SAMPLES
Ricardo A. Bernardoa, Charles I. O. Júnior b, Almir C. B. Juniora*, Carlos A. Sorgic,
Boniek G. Vaza, Andréa R. Chavesa,b
a
Instituto de Química, Universidade Federal de Goiás, 74690-900, Goiânia, GO, Brazil
b Departamento de Química, Universidade Federal de Jataí, 75804-020, Jataí, GO, Brazil
c Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto,
Universidade de São Paulo, 14015-130, Ribeirão Preto, SP, Brazil

*e-mail: almircustodio@discente.ufg.br
Metabolomics is often applied to the investigation of changes in metabolic pathways as lipid

metabolism and/or RNA transcription which leads to information about differential gene and
transcript expression patterns in healthy and disease cells/tissues1,2. The RNA extraction is based
on Liquid-Liquid Extraction (LLE) using TRIzol® solution (a mixture of phenol, water, and guanidine
isothiocyanate), and chloroform for phase separation3. The aqueous phase contains the RNA
portion, used for transcriptomics essays while the organic phase, rich in lipids, is discarded3. In this
study, a method was validated for total extraction of lipids in samples stored in TRIzol® solution
adapting the Bligh & Dyer4 methodology and Podechard3 methodology to total lipid extraction from
pig’s tongue samples. The extracted samples were analyzed by direct infusion mass spectrometry
(MS). A solution of pig’s tongue in TRIzol® was prepared in the ratio of 1000 µL of TRIzol® for each
100 mg of tissue and 200 µL of chloroform were added for each 1 mL of tongue solution in TRIzol®.
The resulting solution was homogenized by vortex for 1 min and centrifuged at 1500 G, 4 °C for 10
min, forming a three-phase system. An aliquot of 250 µL of the organic phase was taken adding 1,2-
diheptadecanoyl-sn-glycero-3-phosphocholine (PC 17:0/17:0) in a concentration range from LOQ to
2000 ng mL-1 and caffeine-(3-methyl-13C) was added as internal standard at a concentration of 800
ng mL-1. This organic phase was submitted to a Bligh & Dyer methodology adding 500 µL of
methanol, 500 µL of chloroform, and 250 µL of water, resulting again in a three-phase system. The
organic phase, containing lipids, was collected and the aqueous phase was reextracted, maintaining
the ratio of the solvents. Both organic phases were mixed and dried for posterior resuspension in
200 µL of methanol. The same extraction was performed using a pig’s tongue homogenized in
methanol for comparison. The extracted samples were analyzed by direct infusion MS performed on
a Bruker Daltonics micrOTOF III (Bremen, Germany) with an electrospray ion source in the positive
ion mode. The parameters used for the MS analysis were capillary voltage 4 kV, nebulizer 1.0 Bar,
dry temperature 200 °C, and end plate offset 500 V. The lipid extraction in TRIzol® solution was
linear in the range from 330 to 2000 ng mL-1, r² > 0.99, intra and inter-day precision and accuracy
ranging from -9.599 to 4.938%, and absolute recovery values ranging from 90.40 to 103.15%. In the
present study, a reliable, accurate, and precise methodology is reported for the simultaneous
extraction of RNA and lipids from the same sample. The transcriptomic analysis was not performed,
since they have been already reported by Podechard et al., 20183. The methodology presented here
was evaluated regarding analytical parameters often used for methods validation. Intra and inter-
day precision, accuracy and absolute recovery are in accordance with the Food and Drug
Administration regulation5, and the matrix effect was not significant in the analysis. The methodology
described here could enhance the data output using the same amount of samples that only a
lipidomics assay would require.
1 Wang L el al.. Oncotarget 2017.
2 Valdes-Mora F el al.. Frontiers in Immunology 2018, 9, Review.
3 Podechard N et al. Scientific Reports 2018.
4 Bligh EG; Dyer WJ. Canadian Journal of Biochemistry and Physiology 1959.
5 Swartz ME, Krull IS, CRC Press, 2018.
[LaCEM/IQ-UFG, CNPq, CAPE, FAPEG]

STUDY OF ACID DIGESTION PROTOCOLS IN POLY (ETHYLENE
TEREPHTHALATE) RESIN FOR ELEMENTARY DETERMINATION BY ICP OES
Abinoan S. Rodrigues (PG)a *, Licarion Pinto (R)b, Ana P. S. Paim (R)a
a Federal University of Pernambuco, Department of Fundamental Chemistry, Recife, Pernambuco, Brazil,50740-560
b Rio de Janeiro State University, Department of Analytical Chemistry, Rio de Janeiro, Rio de Janeiro, Brazil, 20550-013
*e-mail: Abinoan.rodrigues@ufpe.br
Poly (Ethylene terephthalate), PET, is a thermoplastic polymer with a wide application in food
packaging 1. Due to the incorporation of certain inorganic species as additives, the control of resin
impurities is essential to guarantee the quality and safety of the final product 2. Since PET resin is a
complex matrix, a methodology for its acid digestion was optimized with polar coordinates based on
an experimental design for subsequent quantification of Cd, Co and Pb in PET pellets marketed in
Pernambuco, Brazil, by ICP OES. The condition was defined by the response surface methodology,
which was built using the results of a process-mixture factorial design with polar coordinates
strategy 3. This experimental design used a polar coordinate strategy to joint mixture with process
parameters into a single central composite design (CCD). The ternary mixture (HNO3 + H2SO4 +H2O)
was converted to the parameters r and θ, which was evaluated jointly with the pellet mass and
heating time with a CCD. Sample digestion was performed using a microwave-assisted system
(Start-D, Milestone), equipped with Teflon tubes. The analysis using an ICP OES (Perkin Elmer,
Model Optima 7000 DV) was held in an axial configuration and argon with minimum purity of
99.999% (White Martins-Praxair, Brazil) and was used for plasma generation, nebulization and
auxiliary gas. The responses used during the optimization of statistical design were defined by the
lowest values for carbon and residual acidity in diluted digested corresponding to the use of 100 mg
of PET, a mixture composed of 54% of HNO3 + 6% of H2SO4 + 40% of H2O, during 20 minutes of
heating at 220 ºC. The resins analyzed present higher levels of Cd, Co and Pb than other works
(Table 1), but the sum of the maximum concentrations of Pb and Cd indicates that the pellets are
considered suitable even for the manufacture of food packaging, in accordance with Directive
94/62/EC (EU, 1994). This resolution establishes that the sum of Pb, Cd, Cr (VI) and (Hg) must
be below 100 mg kg-1.
Table 1- Concentration of elements, in µg g-1, reported in the literature for PET samples after
microwave-assisted digestion
Element This work Curtzwiler et al. (2011) Pereira et al. (2011)

Cd 0.00 – 0.20 (± 0.03) < 0.005 < 0.015
Co 0.12 (±0.03) – 85.10 (±0.61) 14.33 (± 2.50) < 0.015
Pb < 0.31 – 1.47 (±0.61) < 0.005 < 0.025
The experimental design used suggests a digestion protocol that enables efficient digestion. This
was confirmed visually with the transparent solution obtained and by the residual carbon and residual
acid analysis. The protocol was tested in 11 PET resins and enabled an efficient and repeatable
digestion in all cases, even without the use of fluoridated agents. The elements’ concentration of the
analyzed samples are in accordance with the established limits of Directive 94/62/EC 4. The highest
level of Co was expected in this work, as two of the digested resins were doped with this element
during industrial processing.
1 Curtzwiler G, Vorst K, Danes JE, Auras R, Singh J, J. Plast. Film Sheeting, 27, 2011, 65.
2 Pereira JSF, Knor CL, Pereira LSF, Moraes DP, Paniz J, Flores EMM, Knapp G, J. Anal. At. Spectrom., 26, 2011, 1849.
3 Teglia CM, García MDG, Galera MM, Goicoechea HC, J. Chromatogr. A, 1353, 2014, 40.
4 European Parliament and Council Directive 94/62/EC, Official Journal of the European Communities, 1994.
[NUQAAPE, FACEPE, CENA-USP, INCTAA, CNPq]

COMPARISON OF SAMPLE PREPARATION PROCEDURES FOR THE
DETERMINATION OF TOXIC METALS IN HAIR DYES BY ICP-MS
Jessica S. S. Lira (PG)a*, Tatiana D. Saint’Pierre (R)a
a Pontifícia Universidade Católica do Rio de Janeiro (PUC-Rio), Department of Chemistry,
Rio de Janeiro, RJ, Brazil, Zip code: 22451-900
*e-mail: lira.jessica@live.com
Hair dyes (natural or synthetic) are the most widely used cosmetics around the world, mainly by
women, to cover up grey hair or just to change the shade. Since this is a commonly used product,
it is important to guarantee that its components are not harmful to human health.1 The Agência
Nacional de Vigilância Sanitária (ANVISA)’s Resolution number 79 from August 28th, 2000
determines that the maximum metal impurities allowed for artificial organic dyes are 20 ppm (mg kg-
1
) for lead and 100 ppm (mg kg-1) for the sum of heavy metals other than lead.4 In the present work,
the concentrations of toxic and potentially toxic metals present in hair dyes were determined. Three
different brands of hair dyes in black color were purchased in the Rio de Janeiro market. The samples
were submitted to two different types of treatment. The first one was acid leaching with 8 mL of HNO3
65% and 2 mL of H2O2 30%, under heat for 2 h at 100 ºC on a hot plate. The second one was
decomposition in a digester block with 10 mL of HNO3 65% for 6 h in a temperature gradient up to
215 ºC. The solutions were diluted and analyzed by inductively coupled plasma mass spectrometry
(ICP-MS). Analytical curves were prepared using multi and monoelemental standards with 10%
HNO3. For the first treatment the concentrations of Al, Ba, Ca, Cd, Ce, Co, Cr, Cu, Fe, Ga, K, Li, Mn,
Mo, Nb, Ni, Pb, Rb, Sn, Sr, Ta, V, and Zn were above the limit of quantification, which varied from
0.4 µg kg-1 (Li) to 10.5 mg kg-1 (Ca) for all samples. The concentrations measured in the dyes ranged
from 1.0 ± 0.2 µg kg-1 (Ta) in all samples to 141.5 ± 20.6 mg kg-1 (K) in sample 2. For the second
treatment the concentrations of Ag, Al, Au, Ca, Cd, Co, Cr, Cu, Fe, Ga, K, Li, Mg, Mn, Mo, Ni, Pb,
Pt, Rb, Sn, Sr, V, Y, and Zn were above the limit of quantification, which varied from 0.5 µg kg-1 (Y)
to 27.5 mg kg-1 (Ca) for all the samples. The concentrations scaled up from 1.0 ± 0.5 µg kg-1 (Y) to
173.0 ± 4.9 mg kg -1 (K) in sample 2. For assessment of the accuracy of the method, samples were
fortified in two levels with analytical standards, before the treatments, and recoveries were within 80
and 110%. Treatment 2, using the digester block, was the most effective to extract metals, since the
concentrations obtained were higher, as in the aluminum case, which presented a concentration of
24.1 ± 1.9 mg kg-1 in sample 3 for the first treatment and 39.8 ± 7.7 mg kg-1 in sample 3 for the
second treatment. No sample presented a concentration of lead above 20 mg kg-1. The concentration
of aluminum in samples 2 and 3 were higher than in sample 1, which have metal packaging, different
from sample 1 which has plastic packaging. Considering ANVISA’s Resolution the hair dyes tested
are safe for human health.
1 Ozbek N, Akman S. Regulatory Toxicology and Pharmacology.Vol.79, 2016, Pag. 49-53

2 Iwegbue C et al, Toxicology Reports. Vol 3, 2016, Pag 796-803
3 Popwicz A, Santos-Tonial L, Revista Tecnológica. Vol. 25, 2021, Pag 59-72
4 ANVISA. Agência Nacional de Vigilância Sanitária. Diário Oficial da União de 31 de agosto de 2000. Resolução RDC
nº 79, de 28 de agosto de 2000.
[CAPES, CNPq, FAPERJ PUC-RJ]

DEVELOPMENT OF A METHOD FOR THE DETERMINATION OF TOXIC AND
ESSENTIAL ELEMENTS IN CANINE FUR BY ICP-MS
Rafael C.C. Rocha (PG)a*, Rachel A. Hauser-Davis (PG)b, Tatiana D. Saint’Pierre (PG)a
aPUC-Rio, Chemistry Department, Rio de Janeiro, RJ, Brazil, Zip Code: 22451-900
bFIOCRUZ, Oswaldo Cruz Institute, Rio de Janeiro, RJ, Brazil, Zip Code: 21040-360
*e-mail: rafaelccrocha@hotmail.com
Canines may be good environmental exposure bioindicators, as they share the same environment
as humans, in some cases, drinking the same water and ingesting the same food.1 Canine fur has
almost the same composition as human hair, varying only in its growth. As human hair can be used
to verify individual health status, we aimed to develop a method to determine essential and toxic
elements (Ag, Al, As, Au, B, Ba, Be, Bi, Cd, Cr, Co, Cu, Ga, Ge, Hg, Li, Mn, Mo, Pb, Pd, Sb, Se, Sn,
Sr, Ti, U, V, Zn, I, Ca, Fe K, Mg, Na, P and S) in dogs’ fur by inductively coupled plasma mass
spectrometry (ICP-MS) and investigate possible mineral imbalances or intoxication.2 Standard hair
washing procedures were optimized, determining a minimal sample mass to ensure homogeneity
and an adequate decomposition. A conventional method of decomposition was used, using
polypropylene tubes. For every 0.05 g of sample added, 0,5 mL of HNO 3 and 0.2 mL of H2O2 are
added. First, HNO3 is added and heated for 4 hours at 100ºC and then H2O2 is added and heated
for another 15 min at 100ºC. The best conditions were washing with water, others washing tests
showed removal or deposition effects, a sample mass between 150 and 300 mg (Figure 1), larger
or smaller masses showed high standard deviations, and decomposition employing both HNO3 and
H2O2, high standard deviations were obtained using only HNO3. All optimization steps were
accompanied by the analysis of two human hair CRMs to validate the results.
Figure 1. Mass test between 50 and 500 mg, where the best range was obtained between 150 and 300 mg.
The results were compared with those obtained by inductively coupled plasma optical emission
spectrometry (ICP OES), a well-established technique for determining essential and toxic elements
in biological samples. The method proved to be versatile due to its multi-element analytical capacity,
with adequate sensitivity and accuracy, displaying recovery values between 80 and 120 % compared
to the reference sample (NCSDC 73347a and ERM-DB001) and the washing step showed that there
is no need to use other solvents than water, making it less complex. A need for further studies to
establish reference mineralogram values for dogs is noted, as well to verify potential differences
between breeds.
1Kriya LD, Arleigh JR, Peter MB, Lawrence D. Science of the Total Environment, 2007, 385, 80–85.
2Miekeley N, Fortes LMC; Porto da Silveira CL, Lima MB, J. Trace Elem. Med. Biol., 2001, 15, 46-55.
[PUC-RIO, CAPES, LABIN – INMETRO, LabÁguas, LABSPECTRO, Fiocruz]

COMPARISON OF ORGANIC AND INORGANIC STANDARDS FOR THE
DETERMINATION OF Ni AND V IN PETROLEUM SAMPLES BY ICP-MS
Enrique R. Dionisio Calderon (R)a*, Rafael C.C. Rocha (PG)a, Eduardo R. Salazar (G),
Tatiana D. Saint’Pierre (R)a
aPontifícia Universidade Católica do Rio de Janeiro (PUC-Rio), Department of Chemistry,
Rio de Janeiro, RJ, Brazil, Zip Code: 22453-900
*e-mail: enriquedionisio@puc-rio.br
The characterization of crude oil presents difficulties mainly with regard to the separation and
identification of its components, due to the complexity of its chemical composition and its high
molecular weights.1,2 Although Ni and V are minor components in its composition, they act as
markers (V/Ni ratio) and allow each oil to have a unique fingerprint. 3,4 There are few reference
materials can be used to monitor the accuracy of the analytical methods for the analysis of oil
samples. In turn, it is convenient that the samples solutions have physio-chemical properties similar
to the analytical solutions.5 This work evaluates the determination of Ni and V concentrations
obtained by inductively coupled plasma mass spectrometry (ICP-MS) in reference materials by
comparing the analytical curves prepared with organic standards (CONOSTAN-S-21) in butanol and
xylene or inorganic standards (PE17 and PE29) in nitric acid solutions. The following reference
materials were analyzed: NIST 8505; 1084a; and 1634c. The samples for analysis were weighed
and diluted in 40 and 60 % by mass of butanol and xylene respectively. The quantification of the
elements was performed using two external calibration curves of, at least five points each one. The
concentration range used in the curve was from 0.8 ug.g-1 to 100 ug.g-1. The instrumental limit of
instrumental detection (LD) for Ni varies from 0.013 ug.g-1 to 0.033 ug.g-1 for the curve organic and
inorganic respectively. For vanadium, the LD was 0.005 ug.g-1 to 0.004 ug.g-1 for the curve organic
and inorganic respectively.Figure 1 shows the concentration values for the reference materials
tested using the organic and inorganic
analytical curves. The recoveries
obtained for Ni and V were slightly
better in the analytical curve prepared
with the organic standard than in the
inorganic. Values of 88.5 and 99.6 %
of recovery for Ni in the references
1084a and 1634c respectively were
obtained on the organic analytical
curve. For vanadium, the recovery
were 84.3, 104.7 and 107.7% in the
references NIST 8505, 1084a and
1634c on the organic analytical curve.
Among the reference materials
analyzed little variation was observed
in the recovery percentages with
organic and inorganic standards
except for the reference 1084a read in
the inorganic curve (74.3% recovery).
Figure 1. Ni and V recoveries
1 Mello P.A et al. Journal of Analytical Atomic Spectrometry 24, 2009, 911-916.
2 Souza RM, Microchemical Journal, 82, 2006, 137-141.
3 Al-Shahristani, MJ Al-Atyiauyonnet, Geochim. Cosmoquim., Acta 36, 1972, 929-938.
4 M. El-Gayar et al., Tecnologia 79, 2002, 13-28.
5 C. Duyck, Thesis, Department of Chemistry, PUC-Rio, 2001, 203 p.
[PETROBRAS, CNPq, FAPERJ, CAPES, PUC-RJ]

DEVELOPMENT OF CELLULOSE ACETATE-BASED POLYMERIC MONOLITHS
FOR ORGANIC CONTAMINANTS EXTRACTION
Poliana Ricci (PG)a*, Marcos V. Ferreira (PG)a, Anizio M. Faria (R)a,
Rosana M.N. Assunção (R)a
aFederal University of Uberlândia, institute of Exact and Natural Sciences of Pontal, Ituiutaba, MG, Brazil, 38304-402
*e-mail: ricci.poliana@gmail.com
Contamination by effluents from human activities is a severe problem due to the impacts caused by
environmental pollution. Substances such as organic dyes have received substantial attention due
to their harmful and antagonistic effects on the biosystem. Among them, Crystal Violet (CV) is a
cationic dye belonging to the triphenylmethanes group widely used as an acid-base indicator, in
fabric dyeing, leather, in the food and paint industry. Either the accumulation or the excess of CV in
the human body could lead to harmful effects on health.1 Thus, the development of efficient systems
for the removal of organic dyes is paramount for environmental remediation. Hierarchically porous
monolithic materials have interesting adsorbent properties that can fit the challenge of extracting
organic contaminants. Particularly promising are the polymer-based monoliths due to their porous
three-dimensional structure, good mechanical properties, and eligibility for chemical modification to
improve the adsorption process. Cellulose diacetate (CA) is a cellulose derivative that meets all
those requirements and can be easily processable in the form of a porous monolithic structure.
Furthermore, CA can be chemically modified to produce other cellulosic derivatives. In this work, CA
was used in the production of monoliths using dimethylformamide (DMF) as solvent and n-octanol,
as non-solvent, by applying the combined techniques NIPS/TIPS (non-solvent-induced phase
separation/temperature-induced phase separation), as seen in Fig. 1.
Figure 1. Process of the CA monolithic structure formation.
Three monoliths were developed to evaluate the influence of chemical modification on CV

adsorption: CA monolith (CAM); cellulose monolith (CM), produced by deacetylation of CA monolith;
and cellulose octanoate monolith (COM), synthesized by the chemical modification of CM. It was
observed from Fig. 2(a) that the CM was more efficient in removing the CV than the CAM or the
COM, for the same adsorbate concentration (CV).
Figure 2. (a) Adsorption and (b) extraction of the CV dye by the monoliths.
The results suggest that the amount of CV removed by the monoliths accompanies the increase
according to their hydrophilicity (CM > CAM > COM). However, the CV desorption (Fig. 2(b)) from
the used monoliths showed that less than 25% of CV was successfully desorpted. These ongoing
studies show the need for improvements in the efficiency of CV desorption.
1 Xin Y, Ma S, Chen G, Huang L, Xie Z, Journal of Water Process Engineering, 48, 2022, 102901.
2 Kumbhar P, Narale D, Bhosale R, Jambhale C, Kim JH, Kolekar S, Jornal de Engenharia Química Ambiental, 10, 2022,
107893.
[UFU, LAMEP, CAPES, ProPP, PROAP, FINEP]

CAFFEINE MOLECULARLY IMPRINTED POLYACRYLATE MONOLITH AS
DISPOSABLE PIPETTE EXTRACTION SORBENT
Laís M.B. Castilho (PG)a*, Allyson L.R. Santos (PG)a, Anizio M. Faria (R)a
aFederal University of Uberlândia, institute of Exact and Natural Sciences of Pontal, Ituiutaba, MG, Brazil, 38304-402
*e-mail: lais.castilho@ufu.br
In chemical analyzes, sample preparation is the fundamental step for reproducible, accurate, and
reliable determinations. In addition to these requirements, methods are currently required that meet
the sustainable development goals (SDGs) defined by the United Nations and that are simple and
portable, allowing for in loco sample preparation.1 Disposable pipette extraction (DPX) is presented
as an excellent opportunity to address all these requirements. DPX is considered miniaturization of
solid phase extraction (SPE), in which particulate sorbent is dispersed within the micropipette tips
retained by Teflon filters.2 However, the current limitations of DPX reside in the low selectivity of
sorbents and in the pressure generated by the filters in the tip, making the manual operation of the
extraction process difficult. We propose replacing the particulate with a monolithic organic sorbent
to overcome these limitations. The monolith is a continuous macroporous structure with long
integrated channels, allowing high permeability and which adheres to the wall of the tips, eliminating
the need for filters. Organic monoliths are easy to prepare and functionalize3 and, in this work, the
functionalization of the absorbent will be achieved by molecular imprinted polymer (MIP) of caffeine.
Thus, the monoliths were prepared by in situ thermal polymerization of trimethylolpropane ethoxylate
triacrylate (TMPTA) and ethyleneglycol dimethylmethacrylate (EDMA), using isopropanol as a
porogen and 2,2-azobisisobutyronitrile as a radical agent at 60 °C for 20 h. The MIP was carried out
with caffeine solutions in isopropanol. The monolith synthesis was optimized by evaluating the molar
ratios of TMPTA:EDMA, TMPTA:porogenic, and TMPTA:caffeine and different porogenic solvents.
The optimization of the monoliths was assessed by the degree of permeability and porosity. The
poly(TMPTA-co-EDMA)@caf MIP monoliths were characterized morphologically and structurally.
The optimized monoliths consisted of 10 % TMPTA, 20 % EDMA, and 70 % isopropanol with 0,01
mol L-1 of caffeine and presented permeabilities of 0,008 mL s-1. The monolithic structures showed
a macroporosity of 59,95%, a rigid structure, and high adhesion to the tips, Figure 1. The
poly(TMPTA-co-EDMA)@caf sorbent showed thermal stability up to 345 °C and characteristic
signals by infrared spectroscopy referring to the C=O and -COOR- groups, indicating the polymeric
formation with the two monomers used, as well as actives left by caffeine when compared to non-
imprinted polymer (NIP).
Figure 1. Images of poly(TMPTA-co-EDMA)@caf monoliths for use in DPX.
Studies of the adsorption capacity of caffeine in the poly(TMPTA-co-EDMA)@caf sorbent indicated

stronger retention than in the monolithic NIP sorbent, showing that caffeine was efficiently imprinted
and yielding the selectivity for the monolith.
1 Lorente AIL, Pereira FP, Bjergaard SP, Zuin VG, Ozkan SA, Psillakis E, Trends in Analytical Chemistry, 148, 2022,
116530.
2 Bordin DCM, Alves MNR, Campos EG, De Martinis BS, Journal of Separation Science, 39, 2016, 1168.
3 Xu L, Shi ZG, Feng YQ, Analytical and Bioanalytical Chemistry, 399, 2011, 3345.
[CAPES, ProPP/UFU, FINEP]

DEVELOPMENT OF A NEW METHOD FOR MELATONIN AND ITS METABOLITES
DETERMINATION IN BREAST MILK BY DPX-MIP-HPLC-MS/MS
Lucas S. Machado (PG)a*, Steffany S. Santos (UG)a, Naiara R. C. Lopes (PG)a, Flavia S.
Oliveira (PG)b, Rodolfo R. Silva (R)a, Jussara V. Roque (R)a, Flaviana V. M. Vieira (R)b,
Andrea R. Chaves (R)a
aUniversidade Federal de Goiás, Instituto de Química, Goiânia, Goiás, Brazil, CEP: 74690-900
b Universidade Federal de Goiás, Faculdade de Enfermagem, Goiânia, Goiás, Brazil, CEP: 74605-080
*e-mail: lucas.s.m@live.com
Melatonin is the main hormone synthesized in the pineal gland, being secreted chiefly at night, in
which it is distributed to different regions of the body1. In breast milk, it could be associated with colic
and sleep disturbs in the newborn, and its concentration in this biological fluid varies according to
the humour of the lactating2. Consequently, determining melatonin and its metabolites in breast milk
seems to be an essential tool for newborn and mothers' health. The High performance liquid
chromatography coupled to mass spectrometry detector (HPLC-MS) has been used for these
analyses; however, due to the complexity of the milk and the low concentration of melatonin, a
previous sample preparation step is rouged necessary. The disposable pipette extraction (DPX) is a
miniaturized sample preparation technique that requires less extraction time and solvent
consumption, however, the number of commercial available phases is still limited3. An alternative
sorbent has been the molecularly imprinted polymers (MIP) once it has high selectivity for a
compound or class of compounds. So, this study´s aim is the development of a new method for
melatonin and metabolites in breast milk determination by DPX-MIP-HPLC-MS/MS. The synthesis
of MIP was carried by mass polymerization in the proportion of 1 mmol of melatonin, 3 mmol of
functional monomer (methacrylic acid or 1-vinylmidazole or 4-vinylpyridine or 2-hydroxyethyl
methacrylate), 15 mmol of ethylene glycol dimethacrylate, 0.31 mmol of azobisisobutyronitrile, in 6
ml of solvent porogenic (acetonitrile: ethanol 5:1), the mixture was heated at 70ºC for 24 hours, after
that, the polymer was crushed and washed with methanol in soxhlet to remove the template. The
synthesized MIP was characterized by Fourier transform infrared spectroscopy (FTIR) from 400 cm-
1
to 4000 cm-1, thermogravimetry, nitrogen adsorption and desorption, and scanning electron
microscopy. For the DPX-MIP, 20 mg of MIP was added to the a 1 mL empty tip with an inferior filter,
the extraction process was optimized following univariate parameters (best functional monomer and
desorption solvent, and washing step of the phase) and an multivariate mode for the parameters:
desorption solvent volume, extraction equilibrium time, number of desorption and adsorption cycles,
and sample pH, all the analyses were assessed by HPLC-MS/MS. The characterization analyses
confirmed that the synthesis of the MIPs was effective, presenting characteristic bands for the
synthesized MIPs and typical SEM images, the synthesized polymers were thermostable up to
250ºC, and presented surface area of 220 m2/g, and volume pore of 0.90cm3/g. It was observed that
the conditioning and cleaning process of the phase, in addition to the functional monomer selected
in the MIP synthesis, did not significantly interfere in the extraction efficiency. Besides, the
granulometry of the crushed MIPs and the desorption solvent significantly interfered in the DPX
extraction efficiency. The multivariate optimization step demonstrated that the solvent volume and
sample pH presented significantly effect in the extraction efficiency. According to this, the synthesis
of MIP for the DPX-MIP extraction of melatonin and metabolites was successfully performed and
also was observed that for the MIP synthesized presented higher influence by the particles
granulometry, also by the volume and type of desorption solvent, as well as the sample pH value.
The next step the developed methodology will be analytical validated and applied to breast milk from
patients samples.
1 Sack RL, Lewy AJ, Hughes RJ, Ann. Med., 30(1), 1998, 115–121.
2 Engler AC, Hadash A, Shehadeh N, Pillar G, Eur. J. Pediatr., 171 (4), 2012, 729–732.
3 Machado LS, Soares FQ, Martins RO, Bernardo, RA, Cardoso AT, Ruggiero MA, Rabelo D, Souza PS., Chaves AR,
J. Chromatogr. A,1651, 2021, 462260. |
[FAPEG, CAPES, CNPq]

DEVELOPMENT OF A SD-DLLME SAMPLE PREPARATION FOR ANALYSIS OF
FUNGICIDES IN WINE BY UHPLC-MS/MS
Magali Kemmerich (R)a*, Gabrieli Bernardi (R)b, Osmar D. Prestes (R)c, Renato Zanella (R)c
aUniversidade Federal do Pampa, Itaqui, RS, Brazil, 97650-000
b Universidade Federal de Santa Catarina, Chemistry Department, Florianópolis, SC, Brazil, 88040-900
c Universidade Federal de Santa Maria, Chemistry Department, Santa Maria, RS, Brazil, 97105-900
*e-mail: magalikemmerich@unipampa.edu.br
Fungicides are applied from plant development to grape production, transport, and storage. Even
after fruit processing, residues of these compounds can be present in grape by-products such as
wine and juice. This fact raises concerns among consumers and producers and emphasizes the
importance of monitoring pesticides in wine.1 Regarding the analysis of fungicides in wine samples,
liquid chromatography tandem mass spectrometry (LC-MS/MS) is preferably used, with different
sample preparations, such as solid-phase extraction (SPE)2, QuEChERS method3 and dispersive
liquid-liquid microextraction (DLLME).4 Since 2006, several advances have been proposed for the
technique to overcome possible drawbacks, thus leading to different modifications in DLLME, like as
the solvent demulsification-dispersive liquid-liquid microextraction (SD-DLLME) proposed by
Zacharis et al. (2010)5. In this way, a fast and straightforward method based on SD-DLLME sample
preparation was developed, validated, and applied for the multiresidue determination of 30
fungicides in wine using ultra-high performance liquid chromatography tandem mass spectrometry
(UHPLC-MS/MS). Chromatographic analyses were performed using a Waters Acquity UPLC system
with Xevo TQ mass spectrometer equipped with electrospray source (USA), nitrogen generator
(Peak Scientific, Inchinnan, Scotland) and argon gas (White Martins, Brazil) as collision gas for the
MS/MS system. The evaluated SD-DLLME parameters were agitation types (equilibrium time,
vortex, ultrasound, and ultrasound with controlled temperature), extraction solvent type (1-heptanol,
1-octanol, and 1-decanol), dispersive solvent type (acetonitrile, methanol, and acetone), extraction
solvent volume (80, 100, and 120 μL), dispersive solvent volume (400, 500, and 600 μL), sample pH
(2.0, 4.2, and 6.0), and salt addition (ammonium sulfate, sodium chloride, and magnesium sulfate).
Established SD-DLLME procedure consists in transferring 10 mL of wine sample into a 10 mL glass
volumetric flask, and a mixture of 1-octanol (extraction solvent) and acetonitrile (dispersive solvent),
rapidly injected into the sample. The mixture must be vortexed for 1 min, and the emulsion must be
broken down by adding acetonitrile (500 μL). Then, 50 μL of the upper layer is withdrawn with a
microsyringe and transferred to a vial with acetonitrile (150 μL) and ultrapure water (50 μL) prior
UHPLC-MS/MS analysis. Validation was performed using spiked blank sample and satisfactory
results were achieved for accuracy, with recoveries ranging from 70 to 117%, and precision, in terms
of relative standard deviation ≤20%, except for three compounds at the lowest spiked level. For most
of the compounds, the practical method limits of detection and quantification were 0.03 and 0.1 μg
L-1, respectively. The method developed is environmentally friendly and does not require the use of
a centrifuge. Also, it was successfully applied in commercial wine samples in which several
pesticides were found at concentrations ranging from 0.109 to 10.6 μg L-1.
1 Čepo DV, Pelajić M, Vrček IV, Krivohlavek A, Žuntar I, Karoglan M, Food Chemistry, 246, 2018, 394.
2 Carpinteiro I, Ramil M, Rodríguez I, Cela R, Journal of Chromatography A, 1217, 2010, 7484.
3 Bernardi G, Kemmerich M, Adaime MB, Prestes OD, Zanella R, Analytical Methods, 12, 2020, 2682.
4 Bernardi G, Kemmerich M, Machado FF, Prestes OD, Adaime MB, Zanella R, Food Analytical Methods, 15, 2022,
2026.
5 Zacharis CK, Tzanavaras PD, Roubos K, Dhima K, Journal of Chromatography A, 1217, 2010, 5896.
[CNPq, CAPES]

MINIATURIZED METHODOLOGY FOR QUANTIFICATION OF DHA IN PACU
(Piaractus mesopotamicus)
Natalie P. Toyamaa,d*, Bianca do Amarala, Andre Luiz Watanabeb, Marcela Boroskic,
Adelmo Lowe Pletschd
a PTI, Itaipu Technological Park, Territorial Intelligence and Management, IT.DT, Foz do Iguaçu, PR, Brazil, 85867-900
b Itaipu Binacional, MARR.CD, Foz do Iguaçu, PR, Brazil, 85867-970
c Federal University of Latin American Integration, ILACVN, Foz do Iguaçu, PR, Brazil, 85867-970
d Federal University of Technology - Paraná, Academic Department of Chemistry and Biology,
Curitiba, PR, Brazil 81280-340

*e-mail: natalie.toyama@pti.org.br
Docosahexaenoic acid (DHA, C22:6 n-3) is a long-chain polyunsaturated fatty acid (PUFAs) of
extreme importance in disease prevention, and its intake is necessary since humans are not able to
synthesize it spontaneously1. In this sense, fish that have a high capacity for synthesis and/or
retention of DHA, such as pacu (Piaractus mesopotamicus), a commercial, native and abundant
specie from the Prata Basin2, have great potential as a toll/source for PUFAs human nutrition. In this
work, a protocol of extraction, identification and quantification of DHA in mechanically separated
meat (MSM) of Piaractus mesopotamicus was established. The procedure consisted in
miniaturization of the original method based on Bligh and Dyer (1959) for lipid extraction. An amount
of 0.275 g of fish CMS, 1.2 mL of water and 4.5 mL of chloroform:methanol solution (1: 2 v/v) in a
15 mL Falcon tube were set as the best conditions, followed by homogenizer for 20 s in a mixer type.
In the mixture1.5 mL of chloroform was added, homogenized for 20 s and added 1.5 mL of saturated
NaCl solution with vortex for 30 s in order to obtain a biphasic system. The solution was filtered on
quantitative filter paper into a 10 mL hypodermic syringe. The chloroform with the extracted lipids
was collected and dried at 50°C in nitrogen flow. After extraction, esterification was performed
following the method of Santos Junior (2014)3, using methyl tricosanoate (C23:0) as internal
standard. Samples were analyzed by gas chromatography with flame ionization detector (GC-FID).
The validation parameters selectivity, specificity, limit of detection (LOD), limit of quantification
(LOQ), accuracy and precision were evaluated according to INMETRO (2020)4.
The validation data of the miniaturized extraction methodology are in accordance with the
specifications established in INMETRO (2020) its application in real samples with the levels will be
evaluated later.
1 Subbaiah PV, Dammanahalli KJ, Yang P, Bi J, O’Donnell JM Biochim Biophys Acta, 1861, 2016, 723
2 Carneiro WF, Castro TFD, Reichel T, Uzeda OLC, Martínez-Palacios CA, Murgas LDS, Aquaculture, 574, 2022, 37402
3 Santos Junior OO, Montanher PF, Bonafé EG, do Prado IN, Maruyama SA, Matsushita M, Visentainer JV Journal of the
Brazilian Chemical Society, 25, 2014, 1712
4 INMETRO DOQ-CGCRE-008, 2020, revisão 09, Brasil
[Itaipu Binacional, Itaipu PTI, UTFPR, IFPR, UNILA]

DEVELOPMENT OF A MONOLITHIC CELLULOSE STIR BAR FOR THE
SORPTIVE EXTRACTION OF GLYCEROL IN BIODIESEL
Pablo H.S. Martinsa,*, Allyson L.R. Santosa, Anizio M. Fariaa
*e-mail: pablomartins@ufu.br
Biodiesel production is usually carried out through a transesterification reaction of short-chain alcohol
with vegetable oils or animal fat, obtaining a mixture of fatty acid esters and glycerol as a
contaminant. The maximum recommended glycerol content in biodiesel is 0.25% w/w, which can
cause irreversible damage to modern motors. The glycerol content in biodiesel is determined by the
methods ASTM D6584 and EN14105, using gas chromatography with flame ionization detection
(FID).1 However, these methods require a derivatizing agent for glycerol and high temperatures (380
°C) of the column oven and FID, requiring columns and detectors of high thermal resistance,
resulting in poor detectability and low-efficiency detection. A more efficient alternative may be the
extraction of glycerol from biodiesel and its subsequent quantification by high-performance liquid
chromatography with refractive index detection (HPLC-RID). As cellulose is efficient in the adsorption
of glycerol in biodiesel,2 this work sought to produce a cellulose-based adsorbent for glycerol
extraction. Monolithic cellulose was produced in the form of a magnetic bar using it as a sorbent in
stir bar sorptive extraction (SBSE). Cellulose acetate monoliths were produced using a 1 mL syringe
as a mold. Monoliths were obtained by dissolving commercial cellulose acetate in
dimethylformamide, using octanol as a non-solvent. During the phase separation process, a
neodymium magnet was included in the cellulose acetate solution. After the formation of the
monolithic structure, a solvent exchange was performed to remove the octanol. Subsequently, the
monolith was subjected to a deacetylation step using a NaOH solution at 0.25 mol L-1 to obtain the
cellulosic monolith (Figure 1). The efficiency of deacetylation was confirmed by infrared spectroscopy
from the disappearance of the signal at 1750 cm-1 referring to the carbonyl stretching (Figure 2). The
characterization of the cellulose monolith was also performed by scanning electron microscopy,
thermogravimetric analysis, surface area and porosity (ASAP), and x-ray diffraction. The cellulose
monolith micrographs exhibited a highly porous structure, confirmed by the specific surface area
obtained by ASAP above 70 m2 g-1. The efficiency of the SBSE extraction of glycerol, using the
cellulose monolith, was evaluated in simulated biodiesel constituted by methyl oleate fortified with
0.25 % w/w glycerol, obtaining a recovery close to 90 % by HPLC-RID. The magnetic bar of
monolithic cellulose has therefore proved to be an attractive alternative to the extraction and
determination of the glycerol content of biodiesel.
0,20
Cellulose monolith
Acetate cellulose monolith
1214
0,15 1742
Absorbance (a.u.)
1368
0,10
3350
0,05
4000 3200 2400 1600 800

Wavenumber (cm-1)
Figure 1. Image of monolithic cellulose stir bar. Figure 2. FTIR spectra of monolithic celluloses.
1 Dias AN, Kurz MHS, Fagundes CAM, Caldas SS, Clementin RM, D’Oca MGM, Primel EG, Journal of the Brazilian
Chemical Society, 25, 2014, 1161.
2 Reis AMS, Vieira AT, Santos ALR, Ferreira MV, Batista ACF, Assunção RMN, Rodrigues Filho G, Ribeiro EAM, Faria
AM, Journal of the Brazilian Chemical Society, 31, 2020, 1011.
[FINEP]

PRECONCENTRATION OF DIURON FROM NATURAL WATER SAMPLES USING
A BIFUNCTIONAL HYBRID MOLECULARLY IMPRINTED POLYMER
Daniel Morais Nanicuacua (PG)*a,b, César Ricardo Teixeira Tarley (R)a,c
aUniversidade Estadual de Londrina, Departamento de Química, Londrina, PR, Brazil, CEP 86050-482
b Universidade Púnguè, Faculdade de Ciências Exatas e Tecnológicas, Chimoio, Manica, Mozambique
cUniversidade Estadual de Campinas, Departamento de Química Analítica, Campinas, São Paulo, Brazil,CEP13083-970
*e-mail: nanicuacua@gmail.com
The use of diuron herbicide for control weeds that grows on croplands, such as sugarcane, cotton,
coffee and citrus fruits is recurrent in agriculture, as it guarantees the improvement of productivity
and quality of agricultural products. However, the increases use of diuron contributes for its releasing
in environment, contaminating sources of drinking water supply. Due to its persistent in environment
associated to toxic effects to human health, which can cause diseases such as cancer and endocrine
disruption1, European Union established 1.8 µg L-1 as the allowed maximum limit in natural water.
Many researches based on solid-phase extraction (SPE) have been developed aiming to detection
of diuron, in natural and drinking water. The use of adsorbents obtained from imprinting technology
in SPE become attractive in pesticides quantification from environmental samples, because they
present cavities with size, form and complementary groups to the target analyte, that allow selective
and specific binding, so called molecularly imprinted polymers (MIPs). Many researches of extraction
and quantification of diuron, involving use of MIPs are pointed in literature, however, the possibility
of swelling or shrinking when filled in SPE columns, low maximum adsorption capacity as well as the
poor reproducibility of binding sites responsible for the interaction with the analyte, make them
unsuitable for the analysis of diuron in environment samples2. Thus, in this study, a bifunctional
hybrid organic-inorganic molecularly imprinted polymer (BHMIP) was synthesized using methacrylic
acid and phenyltrimethoxysilane as organic and inorganic functional monomers, respectively
exploring the sol-gel process and free radical polymerization and applied for extraction and
quantification of diuron herbicide from natural water, being the analyses performed by HPLC-DAD.
The BHMIP was characterized by means by transmission electron microscopy (TEM), Scanning
electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron
spectroscopy (XPS) and thermogravimetric analysis (TGA). Equilibrium time of diuron adsorption
onto BHMIP was reached in 130 min, which was described by pseudo-first order model, while the
adsorption isotherm was described as a dual-site Langmuir-Freundlich model. The maximum
adsorption capacity, determined at pH 4, was found to be 140.1 mg g-1. Then, the BHMIP was used
as adsorbent in solid-phase extraction (MISPE) for development of a diuron pre-concentration
method. The optimized conditions of the MISPE were: 150.0 mg of BHMIP mass filled in the SPE
cartridge, sample pre-concentration flow rate (pH 4) of 14.0 mL min-1, pre-concentration volume of
50.0 mL, elution flow rate of 2.0 mL min-1 using 6.0 mL of methanol as elution solvent. The linear
range was from quantification limit (0.27 µg L-1) to 200.0 µg L-1 (R2= 0.999). The MISPE method
showed a high pre-concentration factor, evaluated at 173. The applicability of the method was
checked by means of stream water analysis. Samples of stream water spiked with the standard
yielded recovery percentages that ranged from 96-104%. The BHMIP was used for 23 consecutive
cycles of diuron adsorption-desorption without losing the quality recovery, attesting to the good
capacity of its reusability.
1 Alves, GF, Vinícius de Faria L, Lisboa TP, Matos MAC, Muñoz RAA, Matos RC, Journal of Food Composition and
Analysis, 106, 2022, 104284
2 Kalogiouri NP, Tsalbouris A, Kabir A, Furton KG, Samanidou VF, Microchemical Journal, 157, 2020, 104965.
[CNPq]

DEVELOPMENT OF A METHOD FOR DETERMINING PESTICIDES RESIDUES
IN FISH LIVER
Jéssica F B Wendta*, Natalie P Toyamaa, Bianca do Amarala
a Itaipu Technological Park, Territorial Intelligence and Management, IT.DT, Foz do Iguaçu, PR, Brazil, 85867-900
*e-mail: froes.jb@gmail.com
Fish can be used as bioindicators in assessing the quality of the aquatic ecosystem, as they ingest
contaminants directly from the water and through their diet1. The determination of pesticides requires
the development of advanced multi-residues analytical methods, with instrumental technologies of
high selectivity and sensitivity, such as gas chromatography coupled with tandem mass spectrometry
(GC-MS/MS)2. In addition, it is necessary to use extraction and cleaning protocols due to the
complexity of biological matrices. Among the methodologies used for this purpose, QuEChERS
stands out for monitoring pesticides in food and environmental matrices1,3. In this work, the
QuEChERS method was developed for 40 pesticides and metabolites extraction and pre-
concentration in the fish liver with subsequent determination by GC-MS/MS under optimized
conditions. A study of the three official methods (AOAC, and EN 15662) was conducted. The original
method showed recovery ranging from 12 to 238%, with the lowest recovery for nicotine and the
highest for pyraclostrobin. The method AOAC showed recoveries from 6 to 149% for nicotine and
fluxapyroxad, respectively, and the method EN 15662 showed recovery results ranging from 2 to
189% for nicotine and pyraclostrobin, respectively. The similarity in the recovery results between the
three methods led to the choice of the original method for further optimization. For the optimization,
the sorbents were analyzed: chitin, chitosan, PSA, and C18, through the Box-Behnken experimental
design. The most efficient recovery results were found with chitosan and PSA, without the influence
of C18 in the recovery of the analytes. Central Composite Design 2² was used to evaluate the
proportion of each sorbent related to the sample mass. The analysis of the effects and critical points
showed there was trend toward more significant recoveries and better sample cleanliness with the
use of a higher mass of chitosan (>300 mg) than PSA (>100 mg), generating chromatograms without
interferences, with stable baseline and regular peaks. Therefore, the proportion of 346.6 mg of
chitosan and 187.5 mg of PSA presented the most significant effect and was selected as the best
condition for most analytes. The reconstitution volume was also evaluated, with better recovery
results and lower relative standard deviation at 200 µL. The optimized procedure was validated using
a matrix-matched approach to obtain the highest efficiency at the lowest concentration levels. The
coefficients of determination (R2) were higher than 0.9904 for all analytes except for parathion-methyl
(0.9221), ethion (0.9471), pyraclostrobin (0.9609), profenofos (0.9666) and propachlor (0.9765). The
linearity range was within 4 to 170 µg kg-1. The mean recovery values were between 79% and 120%,
with relative standard deviation values from 0,08% (trifloxystrobin) to 28% (atrazine-desethyl). The
matrix effects (ME) were calculated based on the analytical signal area. The responses obtained
showed a low ME (ME<±20%) for 85% of the analytes, 12.5% of the analytes had a moderate ME
(±20%<ME<±50%), and only for pyraclostrobin showed a high ME (ME>±50%), ranging from 28%
for the lowest point of the curve to 75% of matrix effect for the highest point of the curve. The modified
version of the QuEChERS method with chitosan can be successfully applied to determine selected
pesticides in fish liver samples, which will be collected in streams of the Paraná 3 basin.
1 Kaczynski P, Lozowicka B, Perkowski M, Szabunko J, Ecotoxicology, and Environmental Safety,138, 2017, 179.
2 Villaverde JJ, Sevilla-Morán B, López-Goti C, Alonso-Prados, JL, Sandín-España P, TrAC Trends in Analytical
Chemistry, 80, 2016, 568.
3 Kim L, Lee D, Cho HK, Choi, SD, Trends in Environmental Analytical Chemistry, 22, 2019.
[Itaipu Binacional, UNILA, PTI]

FEASIBILITY OF EXTRA VIRGIN OLIVE OIL DIGESTION USING SINGLE
REACTION CHAMBER AND REE DETERMINATION BY USN-ICP-MS FOR
AUTHENTICITY ASSESSMENT
Bruno O. Poletto (PG)a, Stéfani S. Corrêa (UG)a, Aline R. Santos (UG)a, Cezar A. Bizzi (R)a,
Érico M. M. Flores (R)a, Jussiane S. Silva (R)a*
aUniversidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil, 97105-900
*e-mail: jussiane.silva@ufsm.br
The health benefits associated with the sensory characteristics of extra virgin olive oil (EVOO) are
some factors that add commercial value to this globally accepted product. There are some ways to
adulterate EVOO to sell it at a lower market cost, such as adding cheap oil with lower nutritional
value or producing olive oil with olives of unknown origin, which alters the authenticated and certified
properties of the product.1 The National Health Surveillance Agency (ANVISA) nº 270/2005 and
Codex Alimentarius, Italy, (2003) establish the ratings, regulations, and other information about
EVOO as a way of obtaining the olive oil. The determination of trace elements has been studied to
provide geographic traceability, which can help to verify the provenance of EVOO. In this sense, the
elemental profile of olive oil, among them the rare earth elements (REE), has been considered an
effective and valid marker of the geographical origin of the oil. However, the REE are in low
concentrations (level ng g-1) in the EVOO, requiring sufficient analytical methods for simultaneous
multielementar determination with low limits of quantification (LOQs). In this context, a sample
preparation method using single reaction chamber system (SRC-UltraWave™) was proposed for
EVOO digestion and further REE determination by inductively coupled plasma mass spectrometry
(ICP-MS) with ultrasonic nebulizer (USN). Taking into account the difficulty of determining REE due
to spectral interference and low concentration of analytes in EVOO, efficient digestion of high sample
mass is required. Thus, initially, the heating program was evaluated to allow the digestion of high
masses of EVOO (1.0 to 2.0 g) without the projection of the sample. The following heating program
was used: (i) 5 min of ramp to 115 ºC; (ii) 50 min of ramp to 180 ºC; (iii) 10 min ramp to 270 ºC, and
hold for 20 min. Digestion conditions such as the volume of HNO3 14.4 mol L-1 (6, 7, and 8 mL),
mixture of concentrated HNO3 14.4 mol L-1 with H2O2 10.3 mol L-1 (ratio 7:1 and 6:1 v/v), the
maximum temperature (220, 250, and 270 ºC) also were evaluated. The digestion efficiency was
evaluated based on the concentration of residual carbon (RC) and residual acidity (RA).2 The smaller
volume (6 mL) of 14.4 mol L-1 HNO3 resulted in higher RC and RA compared to RC and RA using 8
mL of 14.4 mol L-1 HNO3. This fact can be explained due to the formation of carboxylic acids from
the reaction between linoleic and linolenic acids from the matrix with HNO3.3 Digestion of 2 g of
sample was not efficient, high values of RC and RA were obtained, being 22628 ± 412 mg L-1 and
16,9%, respectively. Using the mixture of HNO3 and H2O2 resulted in the projection of the samples
out of the vessels. Digests obtained up to 1.0 g using 8 mL of 14.4 mol L-1 HNO3 and hold for 20 min
at 270 ºC presented values of RC and RA lower than 129 ± 11 mg L-1 and 7.2%, respectively. Under
these RC and RA conditions, the digest was introduced into the ICP-MS without previous dilution
and it was possible to determine the REE without matrix interference. REE concentrations in EVOO
were between 0.1 and 6.1 ng g-1. Accuracy was evaluated by standard addition (CCS-1 solution)
and the recovery obtained ranged from 99 to 118%. The proposed method enabled low limit of
detection (0.007 to 0.51 ng g-1). It was possible to determine low REE concentrations after high-mass
sample digestion efficiently, which can be used for EVOO authenticity studies.
1 Aceto M, Calà M, Musso Db, Regalli N, Oddone M. Food Chemistry, 298, 2019.
2 Flores, EMM. Microwave-assisted sample preparation for trace element determination. Newnes, 2014, p. 400.
3 Krug FJ, Rocha FRP. Métodos de preparo de amostras para análise elementar, 2 ed. São Paulo: SBQ, 2019, Brasil.
[UFSM, CAPES, CNPq, FAPERGS]

ANALYTICAL METHOD FOR DETERMINATION OF METALS IN KRAFT CELLULOSE
Miguel P. Soares (UR)a*, Charlie G. Gomes (PG)a, Daísa H. Bonemann (PG)a,
Jéssica R. Porto (PG)a, Anderson S. Ribeiro (R)a, Mariana A. Vieira (R)a
aFederal University of Pelotas, Center of Pharmaceutical and Food Chemical Sciences, Pelotas, RS, Brazil, 96160-000
*e-mail: sp.miguel@gmail.com
The paper is obtained from fibrous materials, from the pulping of virgin wood fibers of the coniferous
or hardwood group. Conifers produce long-type fibers, from woods such as pine, araucarias, among
others. Hardwood, on the other hand, produce short-type fibers, obtained mainly from eucalyptus.1,2
The acquisition of these fibers can take place through mechanical, thermal or chemical processes
of pulping, where the choice of this results in the quality and characteristics of the final product. The
pulp obtained from the Kraft process is used as raw material for various purposes, such as stationery,
personal hygiene materials, or even for the manufacture of packaging for the storage of food
products, fresh or long-lasting, for cooking food through the incidence of heat and also for filtration.3
Therefore, the objective of this work is to develop an analytical method for determination of Al, B,
Ba, Ca, Cd, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, V and Zn in Kraft pulp samples by Microwave Induced
Plasma Optical Emission Spectrometry (MIP OES) aiming at quality control in relation to the
elemental composition of Kraft cellulose pulp. The sample preparation was performed in a closed
low-pressure system, by acid decomposition in borosilicate tubes with screw caps, on forced
ventilation, being optimized through the use of a rotational central compound design (DCCR),
considering the following independent variables: proportion between the volume of HNO3 and H2O2
(from 1.2 to 1.8 mL of HNO3 and 0.8 to 0.2 mL of H2O2), mass sample (from 80.4 to 169.6 mg) and
decomposition time (from 144 to 216 min). The decomposition temperature parameter remained
fixed at 150 °C due to equipment limitations. The responses obtained through the experimental
results were analyzed by free software R®, allowing the analysis of the interactions between the
variables studied and their influence on the analytical response of interest. The optimal conditions
found: sample mass of 169.6 mg, 1.2 mL of HNO3 with 0,8 mL of H2O2 and decomposition time of
216 minutes. The final volume of 15 mL was filled with deionized water. Before analysis in MIP OES
spectrometer, the solutions were filtered. For accuracy evaluation, the certified reference materials
CRM-Agro FT_012026 and C1005a were employed and satisfactory values of recoveries were
obtained, ranged between 82 and 115%, being within the confidence interval of the certificate. Thus,
the method developed proved to be efficient and accurate for the determination of Al, B, Ba, Ca, Cd,
Cr, Cu, Fe, K, Mg, Mn, Na, Ni, V and Zn in Kraft pulp samples, being a simple, practical, safe and
easy to apply method in order to ensure greater food and environmental safety. Therefore, it is
important to highlight the study of the determination of metals in this type of sample due to its varied
applications in storage of food products, daily use, or even as a basis for adsorption materials in
other studies.
1 PUBLIO R. Aplicação de celulases no refino de fibras celulósicas Kraft branqueadas de eucalipto. 2012. Tese de
Doutorado. Universidade de São Paulo.
2 ALVES ED et al. Revista Liberato, v. 16, n. 26, p. 205-218, 2015.
3 SOARES NS ET al. Floresta, v. 40, n. 1, 2010.
[CNPq, CAPES, INCTAA]

ACID DECOMPOSITION IN A LOW PRESSURE CLOSED SYSTEM FOR THE
DETERMINATION OF METALS IN WHEY PROTEIN BY MIP OES
Jéssica da R. Porto (UG)a*, Yasmin R. Bloedorn (UG)a, Miguel P. Soares (UG)a,
Charlie G. Gomes (PG)a, Daisa H. Bonemann (PG)a, Anderson S. Ribeiro (R)a
aFederal University of Pelotas, Center for Chemical, Pharmaceutical and Food Sciences,
Capão do Leão, RS, Brazil, 96160-000.
*e-mail: jporto8.jp@gmail.com
Proteins are essential compounds in the human diet and are present in several processes that occur
in the body. The food supplements available on the market act in the complementation of protein
intake.1 Among the supplements, whey protein stands out, which is a by-product of the cheese
manufacturing process, with proteins of high nutritional value, low level of fat and carbohydrates, in
addition to containing macro and microminerals.2 Despite the many benefits they bring to the body,
protein supplements can contain, in addition to essential elements, toxic elements that can have
harmful effects on health. These elements come mainly from anthropogenic actions such as disposal
of industrial waste and excessive use of agrochemicals, which may be related to animal feed and,
consequently, present in milk. The consumption of these elements occurs through the food chain
causing serious health complications.3,4 Monitoring the concentration of toxic elements in food is of
extreme nutritional and toxicological importance.4 Therefore, this work aims to evaluate the total
concentration of Cu, Fe, Mn and Pb in whey protein samples by microwave induced plasma op-tical
emission spectrometry (MIP OES). For sample preparation, an acid decomposition method was used
in a low pressure closed system, as described by Miranda.5 In order to optimize the best experimental
conditions for sample preparation, a central compound factorial design was used, where the
following variables were investigated: sample mass, HNO3 volume, pre-digestion time and digestion
time. For sample preparation, 162.5 mg were weighed directly into the borosilicate reaction tube,
followed by the addition of 1.064 mL of HNO3 65% (v/v) and 0.96 mL of H2O2 30% (v/v). The tubes
were closed and after 30 minutes of pre-digestion, were placed in a digester block heated at 150 ºC
for 155 minutes. After cooling, the samples were transferred to polypropylene tubes and filled up to
15 mL with deionized water and refrigerated until the time of analysis. The limits of detection (LOD)
were 0.267; 0.236; 0.009 and 0.091 mg kg-1 for Cu, Fe, Mn and Pb, respectively. The accuracy of
the method was evaluated using bovine liver certificate reference material (NIST 1577c) with
recovery values between 81 and 108%. The results of the total concentration of samples A, B, C and
D were 0.92; 2.77; 4.61 and 3.51 mg kg-1 for Cu, 9.85; 28.02; 49.48 and 8.0 mg kg-1 for Fe, 2.78;
2.78; 4.61 and 0.18 mg kg-1 for Mn and 0.92; 0.71; 3.26 and 3.08 mg kg-1 for Pb, respectively.
Considering that the recommended daily intake value (DRI)6 for adults of Cu, Fe, Mn and Pb is 0.9;
14.0; 2.3 and 0.9 mg, respectively, and considering that one dose of each sample is equivalent to
30 g as indicated by the manufacturer, it can be concluded that the samples contribute to the
ingestion of these elements, but without exceeding the recommended value.
1 Lin D, Lu W, Kelly AL, Zhang L, Zheng B, Miao S, Trends in Food Science and Technology, 68, 2017, 130.
2 Aquino LFMC, Ribeiro ROR, Simoes JS, Mano SB, Mársico ET, Conte Junior CA, Journal of Food Composition and
Analysis, 59, 2017, 141.
3 Latif A, Bilal M, Waleed A, Azeem M, Ahmad MI, Abbas A, Ahmad MZ, Shahzad T, Journal of Environmental Analytical
Chemistry, 5, 2018, 1.
4 Kumar P, Dipti, Kumar S, Singh RP, Environmental Pollution, 301, 2022, 118959.
5 Miranda K, Pereira Filho ER, Neto JAGA, Journal of Analytical Atomic Spectrometry, 29, 2014, 825.
6 Brasil. Agência Nacional de Vigilância Sanitária. Resolução RDC nº 269, de 22 de setembro de 2005. Regulamento
técnico sobre ingestão diária recomendada (idr) para proteína, vitaminas e minerais.

A MULTI-PUMPING FLOW SYSTEM FOR IN-LINE EXTRACTION AND
SPECTROPHOTOMETRIC DETERMINATION OF FREE GLYCEROL IN BIODIESEL
Vivian Maringolo (PG)a*, Alexandre Z. Carvalho (R)a, Diogo L. Rocha (R)a
a Federal University of ABC, Center for Natural Sciences and Humanities, Santo André,
São Paulo, Brasil, CEP 09210-580
*e-mail: vivian.maringolo@ufabc.edu.br
Biodiesel is an important alternative fuel produced from animal fats or vegetable oils via
transesterification (1). According to National Petroleum Association (ANP), the threshold limit for free
glycerol in biodiesel is 200 mg kg-1(2). Exceeding content may yield high amounts of acrolein after
combustion. In this work, a multi-pumping flow system was proposed for in-line liquid-liquid extraction
of free glycerol in biodiesel followed by spectrophotometric determination. The analyte was
transferred to the aqueous phase using the manifold depicted in Figure 1A by mixing sample and
water. The sample zone was directed towards a filtration unit before chemical derivatization (Figure
1B), in which glycerol was oxidized prior to the reaction with acetylacetone to yield 3,5- diacetyl-1,4-
dihydrolutidine (MAX= 412 nm(2)).
Figure 1: Flow manifold for free glycerol extraction and Figure 2: Response surface considering acetylacetone
determination. P1–P6: solenoid micro-pumps; V: three-way and pH of a 0.6 mol L−1 ammonium acetate buffer.
solenoid valve; S: sample; C: H2O, P: reference solution, F: .
filtration system, R1: 50% ethanol, R2: 8 mmol L−1 NaIO4 in 0.6
mol L−1 ammonium acetate (pH 5.5), R3: 1 mol L−1
acetylacetone in 0.6 mol L−1 ammonium acetate (pH 5.5).
B1: 70-cm extraction coil, B2: 155-cm reaction coil; D: detection
system; W: waste vessels
Table 1: Evaluated parameters and selected values for

glycerol determination.
Parameters -1 0 1 Optimum values
NaIO4, mmol L -1 6 8 10 8
Acetylacetone, mmol L -1 400 800 1200 1000
pH 4 5.5 6.5 5.5
Flow interruption, s 30 45 60 45
The multivariate method was used for the optimization of glycerol determination aiming at maximum
sensitivity and determination frequency. The screening of variables was performed by fractional
factorial design 24-1. The refinement of the model was carried out by central composite designs with
validation by analysis of variance, which yielded a satisfactory F test value. The optimal conditions
and the obtained response surface are shown in Table 1 and Figure 2, respectively. Linear range
from 3.0 to 50 mg L-1 glycerol was observed. Detection limit, coefficient of variation and determination
frequency were estimated at 2.0 mg L-1 (99.7 confidence level), 4.2% (n = 20) and 16 h-1,
respectively. A preliminary study with in-line extraction was assessed by mixing 75 mg of sample
(3.7 mg kg-1 free glycerol) and 1500 µL of water. The process was performed during 30s. The
extraction efficiency, which was also affected by dispersion in B1, was estimated at 54%. After each
extraction, the filtration column (filled with 185 mg of glass microfiber) was washed with ethanol
solution prior to the next determination to avoid carry-over. The extraction will be optimized aiming
at best efficiency exploiting micropumps for the first time for in-line liquid-liquid extraction followed
by direct determination.
1 Bondioli P, Della Bella L, Eur. J. Lipid Sci. Technol. 107 (2005) 153–157. DOI 10.1002/ejlt.200401054.
2 Silva SG, Rocha, RPF, Talanta 83 (2010) 559–564. DOI:10.1016/j.talanta.2010.09.06
[CAPES]

DEVELOPMENT OF A PRESSURIZED INFRARED RADIATION ASSISTED
DIGESTION SYSTEM
Fábia P. R. Peixoto (PG)a,b, Dalton M. de Abreu (UG)a, Paulo G. R. Peixoto (R)b,
Wladiana O. Matos (R)a, Gisele S. Lopes (R)a*
a Laboratório de Estudos em Química Aplicada (LEQA), Departamento de Química Analítica e Físico-Química,
Universidade Federal do Ceará, Fortaleza - CE, Brazil, 60.440-900
b Instituto Federal do Ceará, Fortaleza – CE, Brazil, 60040-531
*e-mail: gslopes@ufc.br
Sample preparation methods have been the target of many researchers, aiming not only for fast
procedures, but also reduction in sources of errors, profitability and waste reduction 1,2. Pressurized
digestion systems, particularly microwave assisted, are very attractive as they allow faster and more
efficient digestions, lower risks of contamination and loss of volatile analytes, and lower consumption
of reagents, due to high temperatures that can be reached 3,4. Unfortunately, purchase of a
microwave assisted system typically presents high acquisition and maintenance costs. The use of
infrared radiation (IR) as a heating source for sample digestion has been shown to be a very
interesting alternative, promoting fast and efficient mineralization of organic samples 5. The main
objective of this work was to develop a prototype infrared radiation assisted system using
pressurized vessels fitted with lock and relief valves and temperature control for organic sample
digestion. Parameters such as infrared lamp position, design of the vessel cap, temperature control
and cooling system were investigated. Sample masses of 0.5 g of Reference Materials (fish tissue,
bovine liver and rice) were weighed and placed in the quartz vessels along with 2.0 mL of nitric acid
(65% w w-1) and 1.0 mL of H2O2 (30% v v-1). The vessels were closed and their temperature
controlled via a sensor during a 3-step program: ramp from 25 to 180 oC over 18 min, 180 oC hold
(rice) and 200 oC (fish and bovine liver) for 8 minutes and then cooling over 15 min (or until room
temperature was reached). Control of temperature variation in the system to ± 5 °C was achieved
by regulating voltage to the IR lamp (1000 W). Residual carbon content (RCC) was measured in the
digested samples to test efficiency. RCC results were compared to those achieved using a closed
microwave assisted digestion system for the same samples. Rice samples presented a decrease of
78.4 % RCC (22.4 mg L-1) in the digested solution using the proposed pressurized infrared assisted
system, in agreement with that achieved using conventional microwave assisted procedures. Both
systems showed higher RCC values for fish and bovine liver samples when compared to the
microwave system, likely because the temperature reached in the latter was 200 oC higher and due
to the lipids and protein content in these samples higher temperatures are required for better
digestion efficiency. The heating program of the IR assisted system remains to be optimized for the
digestion of samples that present higher protein and fat content. While the prototype used in this
work is still under study, it can be considered a useful alternative to microwave heating due to its
efficiency, with radiation acting directly on the reaction mixture. The proposed system also allows
the use of nitric acid digestion of organic samples in addition to the reduction of time compared to
conventional heating.
1 Casado N; Pérez-Quintanilla D.; Morante-Zarcero S.; Sierra I. Trends in Analytical Chemistry, 88, 2017, p. 167-184.
2 Fanali C; D'orazio G; Fanali S; Gentili A. Trends in Analytical Chemistry, 87, 2017, p. 82-97.
3 Krug F. Rocha F. Métodos de Preparo de Amostras para Análise Elementar. Ed.2, Piracicaba, 2019.
4 Vieira A. Dissertação de Mestrado. Instituto de Química de Araraquara. Universidade Estadual Paulista, 2016, p.80.
5 Oliveira J, Silva F; Monte R; Matos W; Lopes G. Food Chemistry, 224, 2017, p. 335-341.
[CNPq, CAPES, National Research CouncilCanada (NRCC), INCTAA, UFC, PGQUIM-UFC]

A MINIATURIZED LLE METHOD FOR Na, K, Ca, AND Mg DETERMINATION IN
CRUDE OIL FROM PRE-SAL BRAZIL’S RESERVOIRS BY FAAS
Rochele S. Picoloto (R)a*, Felipe A. Silva (PG)a, Beatriz R. Rigui (UG)a,
Cristian R. Andriolli (PG)a, Érico M. M. Flores (R)a, Paola A. Mello (R)a
*e-mail: rochele.picoloto@ufsm.br
Alkaline and alkaline-earth metals are usually found in the aqueous fraction of crude oil emulsions,
as inorganic salts.1 The main salts present are chloride, carbonate, and sulfate, in general,
associated with alkaline and alkaline earth metals (e.g. NaCl, KCl, MgCl2, and CaCl2). The presence
of these elements in crude oil can cause several problems, as corrosion of the distillation towers in
refineries, obstructions and clogging of pipes, valves, and pumps.1,2,3 In this sense, the miniaturized
liquid-liquid extraction (LLE) method was developed for further Na, K, Ca, and Mg determination by
flame atomic absorption spectrometry (FAAS) in crude oil from Brazilian pre-salt. The miniaturized
LLE method is based on the complete extraction and preconcentration of analytes by using a suitable
extraction solution. For the LLE procedure, a water bath (TI-H 10, Elmasonic Ultrasonic, Germany)
with capacity of 8.6 L and temperature control up to 80 °C was used. The microwave-assisted wet
digestion using concentrated nitric acid and analyte determination by FAAS were applied in order to
obtain the reference values. A microwave oven (SRC-UltraWaveTM, Milestone, Italy) equipped with
5 quartz vessels (40 mL) was used for reference method. Sodium, K, Ca, and Mg determination was
carried out using a flame atomic absorption spectrometry (model AAS vario 6, Analytik Jena,
Germany) after LLE and MAWD-SRC methods. Five natural emulsions of crude oil samples from
pre-salt Brazil’s reservoirs were used in this study. The following parameters were evaluated: type
of extraction solution (1000 µL of 0.5, 1.0, 2.0, and 4.0 mol L-1 HNO3), sample mass (2.5 and 5.0 g),
use of chemical demulsifier (50 mg L-1) in 500 µL toluene, temperature (25, 50, 60, 70, and 80 °C),
heating time (0, 2, 5, and 10 min), stirring time (0, 10, 20, 40, and 60 s), and centrifugation time (1,
5, and 10 min). Suitable results were obtained by using 2.5 g of crude oil; 1000 µL of 2 mol L-1 HNO3,
50 mg L-1 of chemical demulsifier in 500 µL of toluene, 10 min of heating at 80 °C; 60 s of stirring,
and 10 min of centrifugation. No statistical difference (t-test, confidence level 95%) was observed
between the reference values and those using the optimized conditions by proposed miniaturized
LLE method. Relative standard deviations were lower than 6%. The limits of quantification (LOQ)
were 1.2, 1.5, 5.0, and 0.5 µg g-1 for Na, K, Ca, and Mg, respectively. The proposed miniaturized
LLE method presents advantages, such as ease-of-use, high throughput, and uses a high sample
mass (up to 2.5 g). In addition, the use of a low and diluted extraction solutions, consequently
reduces the laboratory residue generation. The proposed LLE method can be considered an
alternative for sample preparation of crude oil for further Na, K, Ca, and Mg determination at low
concentration for routine analyses.
1 Speight JG, Handbook of Petroleum Analysis, John Wiley and Sons Inc, New Jersey, (2001) 519 pp.
2 Mello PA, Pereira JF., Mesko M, Barin JS, Flores EMM, Anal. Chim Acta Anton, 746 (2012) 15-36.
3 Huang KG, Zhu YA, Chemistry Solutions to Challenges in the Petroleum Industry, American Chemical Society, (2019)
311-326.
[CAPES, CNPq, FAPERGS, UFSM]

DETERMINATION OF CHLORIDE AFTER REVERSED-PHASE DISPERSIVE
LIQUID-LIQUID MICROEXTRACTION IN CRUDE OIL BY POTENTIOMETRY
Cristian R. Andriollia (PG), Ariadne G. L. P. Gonçalvesa (UG), Heitor M. Mendesa (UG),
Érico M. M. Floresa(R), Paola A. Melloa(R), Rochele S. Picolotoa*(R)
Crude oil is commonly produced as a water-in-oil emulsion with high stability. This effect is more
critical for heavy and extra-heavy crude oil due to the presence of resins and asphaltenes in high
concentration, which can increase the stability of emulsions.1,2 In addition, generally, emulsified
water contains high concentration of inorganic salts (mainly CaCl2, MgCl2, KCl, and NaCl).1 Salts are
impurities in crude oil emulsion and can be hydrolyzed to hydrochloric acid, causing corrosion on
refinery distillation towers, valves, pumps, and pipes.2 Therefore, the chloride determination in crude
oil is mandatory. For this purpose, the standard method proposed by American Society for Testing
and Materials (ASTM) D6470 is used.3,4 However, the ASTM D6470 method presents several
disadvantages, such as low throughput, use of high amount of reagents, and consequently high
waste generation, which are not in agreement with the Green Analytical Chemistry
recommendations. In order to overcome these problems, a reversed-phase dispersive liquid-liquid
microextraction (RP-DLLME) method was investigated for further chloride determination in heavy
and extra-heavy crude oil by potentiometry using an ion selective electrode (ISE). The RP-DLLME
method is based on the analyte extraction from the organic phase (oil sample) to the aqueous phase
using a mixture of dispersant (isopropyl alcohol) and extractant (diluted alkaline solution) solvents.
For the RP-DLLME procedure, a water bath (TI-H 10, Elmasonic Ultrasonic, Germany) with capacity
of 8.6 L and temperature control up to 80 °C was used. The determination of chloride in extracts
obtained by RP-DLLME was performed by using a potentiometer (Model 7.1 pH/Ion meter, Metrohm,
Switzerland) equipped with an ion-selective Ag+ electrode (Model 6.0726.100, Metrohm) and
reference electrode of Ag/AgCl (Model 6.0726.100, Metrohm). In order to obtain the reference
values, heavy and extra-heavy crude oil samples were digested by microwave-induced combustion
(MIC) using diluted NH4OH and chloride determination was performed by ion chromatograph (UG).
Analyte determination was carried out by using an IC (850 Professional IC, Metrohm, Switzerland).
Five samples of heavy and extra-heavy crude oil from Brazil’s reservoirs were used in this study.
The following experimental parameters were evaluated: extractant solvent concentration (50 to 400
mmol L-1 NH4OH), dispersant and extractant mixture proportion (50:50 to 85:15% v/v, isopropyl
alcohol:diluted NH4OH), dispersant and extractant mixture volume (0.5 to 1.5 mL), sample mass (2
to 10 g), solvent type (xylene and toluene), and solvent mass (1.0 to 7.0 g). No statistical difference
(t-test, 95%) was observed between the results obtained by the proposed method with those
obtained by the reference method. Suitable results for the RP-DLLME method were obtained by
using 5 g of crude oil, 5 g of toluene, sample heating of 15 min at 75 °C, 1 mL of a mixture of 65:35%
(v/v) dispersant (isopropyl alcohol):extractant (400 mmol L-1 NH4OH) solvents, 1 min of stirring, and
10 min of centrifugation. The limits of detection and quantification were 2.4 and 8.0 µg g-1 for Cl,
respectively. The proposed RP-DLLME method combined with potentiometry presents several
advantages for routine analysis, such as simplicity, high throughput, high sample mass, and use of
low amount of reagents.
1 Diehl LO, Moraes DP, Antes FG, Pereira JSF., Santos MFP., Guimarães RCL., Paniz JNG., Flores EMM. Separation
science and technology, 2011. 46(8): p. 1358-1364.
2 Mello PA., Pereira JFS., Mesko M., Barin JS, Flores EMM, Anal. Chim Acta Anton, 746 (2012) 15-36.
3 American Society for Testing Materials D3230–Standard test method for salts in crude oil (electrometric method). 2019.
4 American Society for Testing Materials D6470–Standard test method for salts in crude oil (potentiometric method). 2015.

INFLUENCE OF HEAT TREATMENT ON THE CONCENTRATION
OF Cu, Fe AND Zn IN SHEEP MEAT
Daisa H. Bonemann (PG)a*, Charlie G. Gomes (PG)a, Sabrina H. Scherdien (PG)a,
Eduardo G. Carvalho (UG)b, Solange Cadore (R)c and Anderson S. Ribeiro (R)a
aFederal University of Pelotas, Chemistry Postgraduate Program, Pelotas, Rio Grande do Sul, Brazil, 96010-900
b Federal University of Pelotas, Center for Chemical, Pharmaceutical and Food Sciences,
Pelotas, Rio Grande do Sul, Brazil, 96010-900

cInstitute of Chemistry, University of Campinas, CEP 6154, 13083-970, Campinas, SP, Brazil.
*e-mail: daisabonemann@gmail.com
Several types of meat are consumed around the world, resulting in a high demand for these products.
Among these, the sheep meat shows in your nutritional composition essential elements such as Ca,
Fe, K, Mg, P and Zn.1 Meat is generally cooked before consumption, to increase palatability,
digestibility and safety.1,2 However, the thermal processing of meat, such as grilling, frying, and
roasting, promotes physicochemical changes in its composition, affecting the final quality of the
product for several reasons, such as loss of vitamins, minerals, moisture and protein content.2,3
Therefore, it is essential to know the concentrations of the elements (Cu, Fe, Zn and others), as well
as to identify and quantify if there are losses during the cooking process. 2 Thus, this work aims to
evaluate the influence of cooking processes on the concentrations of Cu, Fe and Zn in sheep meat
samples. For this, three samples (A, B and C) of sheep meat from different breeders in the southern
region of Rio Grande do Sul were acquired and subjected to two cooking processes (conventional
oven and microwave oven).3 For the microwave oven cooking process, pieces of approximately 100
g were cut and baked in a glass container covered with plastic film at 650 W power for 6 min. Already
the conventional oven, pieces of approximately 100 g were also cut and baked in a glass container
covered with foil for 45 min at 180 °C in a preheated conventional oven. After cooking, the samples
were ground in a food processor and frozen at −18 °C. To determine the concentrations of the
elements, present in the raw and cooked samples, an acid decomposition method was developed in
a reflux system. The best conditions were optimized through a multivariate design that evaluated the
independent variables (decomposition time, digester block temperature and volume of H2O2).
Sample mass and volume of HNO3 were set at 1 g and 5 mL, respectively. The best decomposition
conditions obtained for the elements were 140 °C for temperature, 187 min for time and 1.8 mL of
H2O2, which were determined by MIP OES. The accuracy of the results was assessed using certified
meat reference materials (NIST 1546a and NIST 1577c) and recoveries ranged from 87 to 111% for
the CRMs. The samples A, B and C (without heat treatment) showed Cu concentrations of 1.61; 2.07
and 0.95 mg kg-1; Fe 67.4; 88.6 and 79.1 mg kg-1 and Zn 99.4; 70.4 and 79.4 mg kg-1, respectively,
demonstrating that the concentration of the elements varies according to the location, type of soil,
food and animal husbandry. The concentrations, for the samples prepared in the microwave oven,
were 0.71; 0.83 and 0.49 mg kg-1 for Cu; 35.8; 40.7 and 34.1 mg kg-1 for Fe and 87.6; 62.7 and 32.4
mg kg-1 for Zn and in a conventional oven were 0.58; 0.85 and 0.59 mg kg-1 for Cu; 25.9; 38.1 and
29.1 mg kg-1 for Fe and 28.5; 26.1 and 32.0 mg kg-1 for Zn in samples A, B and C, respectively. The
losses of the elements during the cooking processes varied between 20 and 70%. Zinc suffers a
great influence depending on the process, since greater losses were observed when subjected to
thermal processing using the conventional furnace. This may be related to the extended cooking
time when compared to the microwave oven in which the time was shorter. Therefore, it is evident
that during the thermal processing there are losses of elements, which are leached into the cooking
medium.
1 Ponnampalam EN, Holman BWB, Scollan ND, Encyclopedia of Food and Health, 1, 2016, 750.
2 Higuera JM, Santos HM, Oliveira, AF, Nogueira, A.R.A, Journal of the Brazilian Chemical Society, 32, 2021, 2111.
3 Menezes EA, Oliveira AF, França CJ, Souza GB, Nogueira ARAN, Food Chemistry, 240, 2018, 75.

A SIMPLE METHOD BASED ON ULTRASOUND-ASSISTED EXTRACTION FOR
FURTHER Fe, Ni, AND V DETERMINATION IN CRUDE OIL BY ICP-OES
Ariadne G. L. P. Gonçalves (UG)a, Bruna C. Padoin (UG)a, Cristian R. Andriolli (PG)a,
Paola A. Mello (R)a, Érico M. M. Flores (R)a, Rochele S. Picoloto (R)a*
Iron, Ni, V are usually found in the organic fraction of crude oil emulsions, as nonpolar porphyrin and
polar organic complexes.1 The presence of these elements in crude oil can cause problems, as
environmental pollution and catalyst poisoning during the crude oil refining process.1,2 In this way,
accurate methods for further Fe, Ni, and V determination in crude oil are required. However, some
drawbacks are observed for conventional methods for crude oil, such as the problems related to
contamination, use of high amount of concentrated acids, as well as the limited sample mass for
closed systems.2 In order to solve these problems, in the present study, the ultrasound assisted
extraction (UAE) method was investigated for further Fe, Ni, and V determination in crude oil by
inductively coupled plasma optical emission spectrometry (ICP-OES). For the proposed method,
samples were weight directly in the vessels, an acid solution was added and ultrasound was applied.
After extraction, the aqueous-phase, containing the analytes, was collected and analyte
determination was performed. For the UAE procedure, ultrasound water baths (Elmasonic,
Germany), with frequency operation of 37 or 80 kHz, capacity of 9 L, effective power of 330 W, and
temperature control (up to 80 °C) was used. Natural emulsions of crude oil sample from pre-salt
Brazil’s reservoirs were used in this study. The microwave-assisted wet digestion (MAWD) using
concentrated HNO3 and analytes determination by ICP-OES were applied in order to obtain the
reference values. A microwave oven (SRC-UltraWaveTM, Milestone, Italy) equipped with 5 quartz
vessels (internal volume of 40 mL) was used for the reference method. The determination was
carried out using an inductively coupled plasma optical emission spectrometer with axial view (Ciros
CCD, Spectro Analytical Instruments, Germany) after UAE and MAWD-SRC methods. The following
operational parameters were evaluated for the proposed method: sample mass (1 to 5 g), toluene
mass (0.5 to 5.0 g), US frequency (37 and 80 kHz), extraction time (2 to 15 min), chemical demulsifier
concentration (5 to 100 g L-1), chemical demulsifier volume (0.5 to 2.0 mL), extraction solution type
and concentration (3 to 7 mol L-1 HNO3 and 6 to 12 mol L-1 HCl), and the use of two extraction steps.
An Analytical GREEnness calculator was used to evaluate the proposed method greenness.3
Suitable results were obtained by using 2.0 g of crude oil, 2.0 g of toluene, 1.0 mL of 50 g L-1 chemical
demulsifier, ultrasound frequency of 37 kHz for 10 min, an extraction solution with 2.0 mL of 7 mol
L-1 HNO3 (first extraction step) and 1.0 mL of 8 mol L-1 HCl + 1.0 mL of 7 mol L-1 HNO3 (second
extraction step). No statistical difference (t-test, confidence level 95%) was observed between the
obtained results by proposed UAE method and the reference values obtained by MAWD-SRC and
determination by ICP-OES. Due to the use of a high sample mass (2.0 g), diluted extraction solutions
and the low blank values, the quantification limits were 0.05, 0.25, and 0.30 µg g-1 for Fe, Ni, and V,
respectively. The proposed UAE method can be considered a simple and fast tool for sample
preparation of crude oil for further Fe, Ni, and V determination at low concentration avoiding the use
of concentrated acids and satisfied equipment for the sample preparation step.
1 Speight JG, Handbook of Petroleum Analysis, John Wiley and Sons Inc, New Jersey, (2001) p. 519.
2 Mello PA, Pereira JFS, Mesko M, Barin JS, Flores EMM, Anal. Chim Acta Anton, 746 (2012) p. 15-36.
3 Pena-Pereira F, Wojnowski W, Tobiszewski M. Analytical Chemistry, v. 92, n. 14, (2020) p. 10076-10082.
[PRH-ANP, FAPERGS, UFSM]

A MICROWAVE-ASSISTED DIGESTION METHOD FOLLOWED BY MAGNETIC
SOLID PHASE MICRO EXTRACTION FOR TRACE-ELEMENT DETERMINATION
IN LIGNOCELLULOSIC BIOMASS BY ICP-OES
Mikaelle de Carvalho Gomes (UG)a,b, Camilla Matos Belmiro Preissler (PG)a,

Marcia Angelica Fernandes e Silva Neves (R)b and Jefferson Santos de Gois (R)a
a Department of Analytical Chemistry, Rio de Janeiro State University, Rio de Janeiro, RJ, Brazil, 20550013
b Federal Institute of Education, Science and techonology, Rio de Janeiro, RJ, Brazil, 26530-060
*e-mail: jeffersongois@gmail.com.br
Lignocellulosic biomass is composed , mainly, of cellulose, hemicellulose, and lignin. It can be used
as an eco-friendly material to produce second-generation fuels to replace fossil fuels and their
products hence it is essential to determine the chemical composition of lignocellulosic biomass
before its use1. Therefore, this work aims to develop a microwave-assisted digestion method (MAD)
using diluted HNO3 for the determination of minor elements and a magnetic solid phase micro
extraction with maghemite nanoparticles for trace-elements determination in biomass by inductively
coupled plasma optical emission spectrometry (ICP-OES). Eight lignocellulosic biomass samples
were obtained, dried and sieved to 200 mesh, these samples were kept in a light-protected flask until
the sample preparation procedure. The levels of the factors sample mass, the volume of HNO3, and
volume of H2O2 were optimized for the microwave-assisted acid digestion procedure using a central
composite design (CCD). The residues of the mathematical models were proven to follow a normal
distribution according to a Shapiro-Wilk test (p=0.05). The optimal conditions were achieved at 280
mg of sample mass, 2.0 mL of HNO3 (≈14 mol L-1), and 2.0 mL of H2O2 (≈ 9 mol L-1). The accuracy
of the MAD method was accessed by recovery tests and a certified reference material analysis. For
the trace-element determination the preconcentration was carried out using maghemite
nanoparticles as a sorbent for the dispersive magnetic solid-phase microextraction process. The
maghemite nanoparticles were synthetized and characterized by Fourier Transform Infrared
Spectroscopy and X-ray diffraction 3. The levels of the factors mass of maghemite, pH, and time
were optimized using a CCD, where the optimal conditions were achieved at 10 mg of maghemite,
100 min of stirring time, and pH 9,5. Two solutions were tested for the desorption process (1) 0,1
mol/L-1 de EDTA and 5% (w/v) NaOH,and (2) HNO3 5 % (v/v), where the solution EDTA + NaOH
exhibited better desorption efficiency. The accuracy of the method was accessed through recovery
tests.
1 Okolie, JA, Chemistry and specialty industrial applications of lignocellulosic biomass. Waste and Biomass Valorization,
Springer, 2021.
2 Bajpai P, Pretreatment of lignocellulosic biomass. In: Pretreatment of lignocellulosic biomass for Biofuel Production,
Springer, 2016.
3 Mendes, MSL, Síntese e caracterização de material nanoparticulado a base de γ-Fe2O3 E Fe3O4, Revista Eletrônica
Perspectivas da Ciência e Tecnologia, 2019.
[CNPq, FAPERJ, CAPES, UERJ]

DETERMINATION OF THE TOTAL CONCENTRATION
OF Al, Ba, Ca and Cu IN COCOA POWDER
Mariana M. Da Cunha (UG)a*, Sabrina H. Scherdien (PG)a, Daisa H. Bonemann (PG)a,
Charlie G.Gomes (PG)a, Anderson S. Ribeiro (R)a
aFederal University of Pelotas, Center for Chemical, Pharmaceutical and Food Sciences,
Capão do Leão, RS, Brazil, 96160-000
*e-mail: cunha.mariana2003@gmail.com
Cocoa beans are derived from the fermented and dried seeds of the cocoa tree (Teobromacacau),
which originates from the tropical portion of the Americas and is widely consumed by the Brazilian
population for having a significant importance in the production of chocolate. This grain is commonly
known for being the basis to produce different variations of chocolate in which its concentration
interferes from the color to the taste of the product1. Food contaminants, especially metals (Al, Ba,
Cd, Pb, among others), can come from industrial agricultural activities since industrial discharge can
accumulate in water or soil and subsequently transfer to foodstuffs. cocoa beans during vegetation
through the root system. Because of this, the development and validation of analytical methods for
the control of metals in cocoa powder samples becomes important, since it is widely consumed by
the population, especially by children mixed with milk. Furthermore, knowing its composition of
essential elements, such as Ca, Cu, Fe, Mg, among others, is also important, as the consumption of
cocoa can also lead to the contribution of the ingestion of these elements to the human body2.
Therefore, the respective work aims to determine the concentration of the elements Al, Ba, Ca and
Cu by microwave-induced plasma optical emission spectrometry (MIP OES) technique in 100%
cocoa samples. For this, an acid decomposition method was developed in a low pressure closed
system, as described by Miranda3, through a DCCR-type factorial design to optimize the best sample
preparation conditions. After evaluating all assays, the best optimized condition was 200 mg of
sample, weighed directly into borosilicate tubes with screw caps, addition of 1.375 mL of HNO3 and
0.625 mL of H2O2 and a pre-decomposition time of 45 min. Subsequently, the tubes were placed in
the block at a constant temperature of 150 °C for 2 hours and 30 minutes. The method was applied
to 4 samples (A, B, C and D). To evaluate the accuracy of the method, a comparative method 4 was
applied to sample A. The values found were compared using the t test with 95% confidence and it
was observed that there was no significant difference between the values of the developed method
and the comparative method. In addition, calibration curves were obtained with R² > 0.99 and
quantification limits of 0.203 mg kg-1 for Al; 0.023 mg kg-1 for Ba; 2.08 mg kg-1 for Ca and 0.149 mg
kg-1 for Cu. The concentrations of Al, Ba, Ca, Cu were determined in samples A, B, C and D of 100%
cocoa, which ranged from 136.1 to 225 mg kg-1 for Al; from 9.4 to 17.1 mg kg-1 for Ba; from 3462 to
2142 mg kg-1 for Ca and from 29.9 to 53.5 mg kg-1 for Cu, respectively. Considering the use of 200
mg of cocoa, the values for the elements Al and Ba are below the maximum values allowed by
legislation. Regarding Ca and Cu, it is possible to observe that cocoa contributes with small amounts
of intake of these elements to the human organism. With this, there is a need for control in cocoa
samples, because in addition to having potentially toxic elements if consumed in large quantities, it
is a food linked to children's daily lives.
1 Mohamed R, Zainudin BH, Yaakob AS, Food Chemistry,303, 2020, 125392.
2 Dico GML, Galvano F, Dugo G, D´ascenzi C, Macaluso A, Vela A, Giangrosso G, Camilleri G, Ferrantelli, 245, 2018,1163.
3 Miranda K, Pereira Filho ER, Neto JAGA, Journal of Analytical Atomic Spectrometry, 29, 2014, 825.
4 Vanderschueren R, Montalvo D, Ketelaere B, Delcour JA, Smolders E, Journal of Food Composition and Analysis, 83,
2019, 103277

METHOD DEVELOPMENT FOR IDENTIFICATION OF OXYGENATED
COMPOUNDS IN BIO-OIL: EXTRACTION USING DLLME
Lavínia C. Rodrigues (UG)a, Ana Luiza G. Mendes (PG)a, Lisiane O. Diehl (PG)b,
Darliana M. Souza (R)a, Fernanda de Castilhos (R)b, Paola de A. Mello (R)a*
b Federal University of Santa Maria, Chemical Engineering Department, Santa Maria, RS, Brazil, 97105-900
The continuous increase in energy demand has driven search for alternative sources in the
production of fuels, chemicals, and materials.1-2 In this scenario, biomass is an alternative source of
energy with abundant and renewable resources. Biomass pyrolysis can produce bio-oil which means
a highly oxygenated liquid material that can be easily transported, stored and converted into
materials of higher added value, as well as can be used as fuel. The composition of the bio-oil can
change according to the biomass used as raw material and the type of the conversion process
applied. The knowledge regarding the main components of bio-oil is very important to assign its
applications. Mass spectrometry (MS) has been used as a tool to understand this composition2. To
enable a better performance for mass spectrometry coupled to gas chromatography, the bio-oils
need adequate sample preparation. Time-consuming and volume-intensive methods, such as liquid-
liquid extraction, are usually applied, and one strategy is to employ miniaturized methods, such as
liquid-liquid dispersive microextraction (DLLME).2 The objective of this work is to develop a method
for the speciation analysis of oxygenated compounds in bio-oils from lignocellulosic biomass. Gas
chromatography MS was used in the multidimensional gas chromatography (MDGC) configuration
with a flame ionization detector in the first dimension and a triple quadrupole mass spectrometer in
the second dimension (model MDGC/GCMS 2020, Shimadzu, Japan). Acetol, acetic acid, eugenol,
furfural, syringol, guaiacol, m-cresol, o-cresol, p-cresol, vanillin, hydroquinone, 2-cyclopenten-1-one,
2-(5H)-furanone, furfuryl alcohol, 5-methylfurfural, phenol and levoglucosan were evaluated. The
heating program used follows the NREL/TP-5100-658893 method and was modified to allows the
separation of the compounds in this study, starting at 45 °C for 10 min, a heating rate of 3 °C min-1
up to 100 °C, maintained for 2 min and then heating at 6 °C min to 260 °C, finally remaining for 5
min. Better performance was observed for the column Rtx-1701 (60 m x 0.25 mm x 0.25 μm) in the
first dimension and the Elite-5MS (30 m x 0.25 mm x 0.25 μm) in the second dimension to setup the
MDGC-MS method. To develop the DLLME method the first investigation was regarding the
combinations for the drop formation. Several solvents were investigated for this purpose (hexane,
dichloromethane, chloroform, acetone, methanol and acetonitrile). For DLLME procedure, 1000 µL
of sample were added to a 15 mL glass vessel and 500 µL of the mixture of extracting and dispersing
solvent was added. Finally, a centrifugation step for 3 min at 3400 rpm was carried out and the drop
was collected and diluted in acetonitrile up to 1.5 mL for MDGC-MS. An experimental design (DCCR
23) was carried out in order to determine the combination of extracting solvent volume, dispersing
solvent volume and sample volume. Based on the results, the DLLME method in conjunction with
MDGC-MS demonstrated to be useful for the speciation analysis of some oxygenated compounds
in bio-oil.
1 Michailof CM, Kalogiannis KG, Sfetsas T, Patiaka DT, Lappas A.A, Energy Environmental, 5, 2016, 614.
2 STAS M, Auersvald M, Kejla L, Vrtiska D, Kroufek J, Kubicka D, Trends in Analytical Chemistry, 126, 2020, 115857.
3 Christensen E, Ferrell J, Olarte MV, Padmaperuma AB, Laboratory Analytical Procedure (LAP), National Renewable
Energy Laboratory (NREL), 2016.
[PRH-ANP, CAPES, FAPERGS, CNPq, UFSM]

MICROWAVE-INDUCED COMBUSTION IN DISPOSABLE VESSELS AS A GREEN
ALTERNATIVE FOR DETERMINATION OF Ni, V AND Co IN PETROLEUM
Jaime S. Mandlate (R)a, Letícia C. Brudi (PG)b, Eduarda L. Vendruscolo (UG)b,
Paola C. Crestani (PG)b, Paola A. Mello (R)b and Fábio A. Duarte (R)b
a University Eduardo Mondlane, P.O. Box 252, Maputo, Mozambique
b Universidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul, Brazil, 97110-970
*e-mail: fabio.duarte@ufsm.br
In the last 25 years, as a result of economic development and population growth, global energy
consumption has grown by about 50% and is expected to increase more 28% up to 2040. Petroleum
is still one of the main energy sources in modern society.1 Its composition is a complex mixture of
hydrocarbons containing small amounts of heteroatoms including N, S, O and some elements such
as Co, Ni and V. The knowledge of the concentration of some elements in crude oil allows to identify
the source of oil extraction, as well as change the processing and quality of its derivatives, mainly
due to the negative influence of metals such as Ni, V and Co on fuel degradation, deformation and
cracking of storage reactors.2 Since the beginning of industrial petroleum exploration in the 19th
century, continuous effort to develop suitable and efficient sample preparation and detection
techniques for the characterization of petroleum have been proposed.2 On the other hand, the
digestion of the petroleum is hindered by the stability of the aromatic compounds present in this
matrix, requiring high cost instrumentation able to operate at extreme conditions of pressure (199
bar) and temperature (300 ºC). Microwave assisted digestion (MAD) in closed vessels is commonly
used as a routine for petroleum digestion, mainly it reaches high temperatures, small reagent
volumes, minimized risk of contamination and analyte loss by volatilization.3 In this perspective, the
aim of this work was to develop a fast, safe and low cost sample preparation method with subsequent
determination of Co, Ni and V by inductively coupled plasma mass spectrometry. The proposed
microwave-induced combustion in disposable vessels (MIC-DV), using a domestic microwave oven,
polypropylene (PP) digestion vessels and quartz holders made of high-purity quartz.4 The
experimental conditions evaluated for MIC-DV for determination of Ni, V and Co were: i) type and
concentration of the absorbing solution; ii) volume of the absorbing solution; iii) sample mass; iv)
volume of NH4NO3 as combustion igniter and v) purging time with O2. Ten crude oil samples of
different physical and chemical characteristics and origins were selected. The results after digestion
by MIC-DV were compared with those obtained by MAD. Comparing the results, the agreement (p>
0.05) for Ni, V and Co ranged from 92 to 107%, from 96 to 106% and from 93 to 107%, respectively.
In order to evaluate the agreement of the proposed method with the 12 principles of Green Analytical
Chemistry (GAC), the AGREE software was used, which allows to establish a score on a scale from
0 to 1, where 1 is highly green (the color is changed from 1 to 0 from dark green to red). MIC-DV
was considered greener than MAD, presenting a score of 0.68 comparing to 0.44 for MAD. Thus,
the developed method provided an alternative to high-cost and tedious methods, presenting a simple
and easy operation, with accurate and precise results for the determination of Ni, V and Co in
petroleum. The LOQ values achieved by this method were 0,28, 0,15 and 0,040 µg g-1 for Ni, V and
Co, respectively. The concentration values for Ni, V and Co ranged from 3,07 to 33,7 µg g-1, from
1,23 to 44,6 µg g-1, from 0,135 to 1,41 µg g-1. In addition, MIC-DV complies with the principles of
GAC, showing greater efficiency with less reagent consumption and decreasing the environmental
pollution by using a single disposable vessel for the overall analysis.
1 Imam A. et al, TrAC Trends in Analytical Chemistry, 118, 2019.

2 Acter T. et al., TrAC Trends in Analytical Chemistry, 151, 2022.
3 Flores EMM et al., AMSTERDAN: Elsevier B. V, 2014.
4 Duarte AF et al., Analytical Chemistry, 92, 2020.

IMPLEMENTATION OF DUAL-SORBENT DYNAMIC HEADSPACE
EXTRACTION IN WINE ANALYSIS
Olga Vyviurska*a, Adriano A. Gomesa,b, Ivan Špánika
aSlovak University of Technology in Bratislava, Faculty of Chemical and Food Technology, Institute of Analytical
Chemistry, 81237 Bratislava, Slovak Republic
b Institute of Chemistry, Federal University of Rio Grande do Sul,
Bento Gonçalves Avenue, 9500, 91501-970 Porto Alegre, RS, Brazil

*e-mail: olga.vyviurska@stuba.sk
A limited extraction capacity is a critical point of miniaturized sample preparation techniques, which
could be overcome with a combination of a few extraction principles or increased amount of acceptor
phase. A dynamic headspace method (DHS) represents a viable solution for this problem, due to
relatively large sorbent amount (180 mg) and a short extraction time. At the same time, multi-volatile
mode based on a few DHS steps allows to cover a wide range of target compounds. It is achieved
through a variation of sorbent types and extraction temperatures. In our study, such approach was
used to estimate trace wine volatiles in botrytized wines. In order to ensure the best performance of
the developed method, critical extraction parameters as incubation time, incubation temperature,
purge flow and purge volume were optimized with multivariate design of experiment, i.e. central
composite design. The responses of target volatiles were evaluated with Pareto charts to determine
statistically significant effects. In general, purge volume, quadratic incubation time and quadratic
incubation temperature improved extraction efficiency of wine volatiles. Optimum experimental
conditions were determined with surface response methodology based on a second order polynomial
function and a total desirability function. The selected parameters were set as follows: a 54 °C
incubation temperature, a 20.18 min incubation time, a 344.3 mL purge volume and a 16.0 mL/min
purge flow. Consequently, the quantification method with 0.2–2.0 µg/L LOD and 0.5–5.0 µg/L LOQ
values were developed for volatiles present at trace concentration level. The increased levels of
linalool oxide, ethyl phenylacetate, γ-hexalactone and α-terpineol were observed in the samples, that
correlated with botrytized wine technology.
[Government of Slovakia, Ministry of Education, contract VEGA 1/0521/19]

COMBINING MICROWAVE-INDUCED COMBUSTION AND ION SELECTIVE
ELECTRODE FOR FLUORIDE DETERMINATION IN ELASTOMERS
Renata V. Corrêa (UG)a, Cristian R. Andriolli (PG)a, Vanessa R. Nascimento (PG)a,
Beatriz R. Rigui (UG)a, Rochele S. Picoloto (R)a, Cezar A. Bizzi (R)a, Darliana M. Souza (R)a*
*e-mail: darliana.souza@ufsm.br
Elastomers are polymeric materials with special properties, such as high elasticity, strength,
stiffness, abrasion, and tear resistance.1 To improve mechanical, thermal, and flame resistance
properties, as well as modify the color and appearance characteristics, organic and inorganic
additives, are used, including halogenated compounds.1,2 In this sense, for the determination of
halogens, especially fluorine, due to its utilization to provide more chemical stability and mechanical
resistance in elastomers, the development of viable analytical methods is increasingly necessary. A
convenient sample preparation step is required because polymers are, in general, more difficult to
digest or dissolve when compared to other samples. In the present study, microwave-induced
combustion (MIC) was investigated for further fluorine determination in elastomers by potentiometry
using an ion-selective electrode (ISE). A microwave oven (Multiwave 5000 Anton Paar, Austria)
equipped with 8 quartz vessels (80 ml of capacity), highest temperature and pressure of 280 °C and
80 bar, respectively, was used for the MIC method. Fluoride determination was performed by a
potentiometer (Model 7.1 pH/Ion meter, Metrohm, Switzerland) using an ion-selective electrode
(Model 6.0502.150, Metrohm) and an Ag/AgCl reference electrode (Model 6.0726.100, Metrohm).
Also used ionic chromatography (IC) to compare the results obtained with ISE, the chromatograph
is equipped with an anion-exchange column (Metrosep A Supp 5, 250 x 4 mm d.i.), guard column
(Metrosep A Supp 4/5 Guard), conductivity detector, and a chemical suppressor module. Using MIC
it was possible to digest 100, 150, 200, 250, and 300 mg of sample. Fluoride was absorbed in 6 ml
of water, 25, 50, 100, 150, and 200 mmol L-1 NH4OH solution. Suitable results were obtained with
300 mg of sample and using 100 mmol L-1 NH4OH as the absorbing solution. The carbon content in
solutions obtained after MIC was lower than 25 mg L-1 avoiding interferences during the
determination. The accuracy was investigated by comparing the results obtained by ISE with those
obtained by IC after the proposed MIC method and no statistical difference (t-test, 95%were as
observed. For the determination by ISE after MIC, the limit of quantification (LOQ) was 20.8 µg g1.
The proposed MIC method combined with potentiometry proved to be fit for the purpose presenting
several advantages for routine analysis, such as the use of diluted extraction solution, low risk of
contamination, and low amount of reagents required.
1 Moraes DP, Pereira JSF, Diehl LO, Mesko MF, Dressler VL, Paniz JNG., Flores ÉMM, Química Analítica and Bioanalítica,
397(2010), 563-570.
2 Mello PA, Barin JS, Duarte FA, Bizzi CA, Diehl LO, Muller EI, Flores EMM, Analytical And Bioanalytical Chemistry, 24,
2013, 405.

NUTS AND FRUITS FROM THE BRAZILIAN CERRADO:
PROMISING PROTEIN SOURCES
Mikael K. de A. Mendes (PG)*a, Darlisson S. N. Silva (PG)a, Carlos A. Galinaro (PG)b,
Christian B. S. Oliveira (PG)a, Carla M. S. Medeiros (UG)a, Ana R. A. Nogueira (R)c,
Emanuel Carrilho (R)b, Cícero A. L. Júnior (R)a, Edivan C. Vieira (R)a.
aUniversidade Federal do Piauí, Departamento de Química, Teresina, Piauí, Brasil, CEP: 64049-550
bInstituto de Química de São Carlos, Departamento de Química, São Carlos, São Paulo, Brasil, CEP: 13560-970
cEmbrapa Pecuária Sudeste, São Carlos, São Paulo, Brasil, CEP: 13560-970
*e-mail: mikaelkelvin@hotmail.com
Proteins are macromolecules formed by polymers of amino acids and are essential for the well-being
and longevity of the population. The production and consumption of protein concentrates of plant
origin have emerged as meat substitutes, aiming to reduce the consumption of foods from animal
origin. However, most meat substitutes currently come from soy (Glycine max), which this may
become environmentally unsustainable1. So, other sources of plant protein need to be figure out.
Brazilian cerrado fruits such as Brazil nut (Bertholletia excelsa), Buriti (Mauritia flexuosa), Cashew
nut (Anacardium occidentale L.), Guabiraba (Campomanesia xanthocarpa), Murici (Byrsonima
crassifolia), Pequi (Caryocar brasiliense), Tucum (Astrocaryum vulgare) and Tuturubá (Pouteria
macrophylla) have been little explored. In this aspect, the objective of this work was to obtain protein
concentrates from different plant matrices from the Brazilian Cerrado. Regarding the sample
preparation, ca. 200 g of each sample was freeze-dried. Later a 5g portion was placed to lipid
extraction in a Soxhlet system following AOAC 920.39 according to Zanqui et al.2. So, the sample
was collected, and the solvent was evaporated to a measure the lipid content. A volume of 1.5 mL
of 10 mmol Tris buffer, pH 7.4, was added to 15 mg of the sample to extract the soluble proteins in
the defatted flours. The mixture was vortexed for 5 min. Then, the extract was centrifuged and the
supernatant was collected and stored at -80 ºC until the subsequent steps. Next, the Bradford
method quantified the proteins using a 96-well microplate reader. Cashew and Brazil nuts were the
concentrates that showed the highest levels of total protein soluble (Fig. 1), being higher than those
found in commercial oat protein concentrates1. Tuturubá and pequi also showed considerable
protein levels. Except for murici, the evaluated Brazilian cerrado fruits are good protein sources.
Therefore, this study suggests that the nuts and fruits of the Brazilian cerrado can be a suitable
substitute for foods of animal origin. For future studies, these protein concentrates will be used to
generate peptides with different biological activities. fruits from the brazilian cerrado.
Figure 1. Content of lipids and total soluble proteins in nuts and fruits from the Brazilian cerrado. Brazil nut
(Bn), Buriti (B), Cashew nut (Cn), Guabiraba (C), Murici (M), Pequi (P), Tucum (Tm) and Tuturubá (Ta).
1 Ramos-Diaz JM, Kantanen K, Edelmann. JM, Jouppila K, Sontag-Strohm T, Piironen V. Current Research in Food
Science, 5, 2022, 858.
2 Zanqui AB, da Silva CM, Ressutte JB, de Morais DR, Santos JM, Eberlin MN, Cardozo-Filho L, da Silva EA, Gomes
STM, Matsushita M. Journal of Supercritical Fluids, 157, 2020, 104686.
[CNPq, CAPES]

DEVELOPMENT OF A GREEN DIGESTION METHOD FOR THE DETERMINATION OF
ELEMENTAL IMPURITIES IN CONTRACEPTIVE PHARMACEUTICALS
Thiago C. Pereira (PG)a, Daiana K. de Oliveira (PG)a, Vitoria H. Cauduro (PG)a,
Alessandra S. Henn (R)b, and Érico M.M. Flores (R)a*.
b Federal University of Technology – Parana, Department of Chemistry, Medianeira, Paraná, Brazil, 85884-000
*e-mail: ericommf@gmail.com
In recent years, there has been increased interest in elemental impurity determination in
pharmaceuticals. The quality control parameters are based on globally recognized documents such
as the ICH Q3D guidelines1. In this guideline, elemental impurities are divided into classes 1 (As,
Cd, Pb, and Hg), 2A (Co, Ni, and V), 2B (Ag, Au, Ir, Os, Pd, Pt, Rh, Ru, Se, and Tl), and 3 (Ba, Cr,
Cu, Li, Mo, Sb, and Sn) according to their toxicity, route of administration, and probability of
contamination in the pharmaceutical product. Considering pharmaceuticals of prolonged use, such
as contraceptive drugs, quality control is especially important to prevent diseases caused by
exposure of the human body to elemental impurities. In this sense, microwave-assisted wet digestion
(MAWD) is usually the sample preparation method of choice, as modern microwave equipment
enables precise control of pressure and temperature inside digestion vessels, as well as providing
the direct heating of the samples2. Moreover, greener MAWD methods can be achieved with the
utilization of diluted acids in an oxygen-rich atmosphere, in which nitric acid can be recovered due
to reactions between nitrogen oxides and oxygen3. In this sense, this study aimed to propose a
sample preparation method based on MAWD for further determination of elemental impurities of
classes 1, 2A, 2B, and 3 in commercial contraceptive tablets by inductively coupled plasma optic
emission spectrometry (ICP-OES). Three contraceptive samples were used in this study, containing
the active ingredients ethinylestradiol (ETD) and levonorgestrel (LGV) with dosages of 0.05 mg of
ETD and 0.25 mg of LGV (S1) or 0.03 mg of ETD and 0.15 mg of LGV (S2 and S3). For the
optimization of the method, sample S2 was used. For the MAWD procedures, 300 mg of samples
were digested in quartz vessels with concentrated (14.4 mol L-1) or diluted nitric acid (0.5 to 3 mol
L-1). For the digestions with diluted HNO3, 1 or 2 mL of 50% H2O2 was used as oxygen source (final
concentrations of 8.3 and 16.7% in the digestion vessels, respectively). A 45 min irradiation program
was used. The accuracy of the proposed method was evaluated using certified reference materials
(CRMs) and standard addition experiments. The CRMs used consisted of dogfish liver (DOLT-4)
and lobster hepatopancreas (TORT-2). Standard addition experiments were performed according to
the permitted daily exposure of each analyte from ICH guidelines1. The optimized condition consisted
of 0.5 mol L-1 HNO3 with 2 mL of H2O2, which resulted in clear digest solutions, with a low residual
carbon concentration of 345 ± 30 mg L-1 and residual acidity of (25 ± 1%). Hence, no interferences
were observed during the determination step. Finally, the proposed MAWD method presented
advantages such as the use of diluted acids, being considered suitable for the decomposition of
commercial contraceptive drugs, allowing the determination of elemental impurities at trace levels
by ICP-OES. It’s important to mention that, considering green chemistry principles, the proposed
method could also help to minimize the impact of analytical procedures used in the quality control of
pharmaceutical products, as well as reducing reagent use and the risks associated with concentrated
acids.
1 ICH Guideline for Elemental Impurities Q3D(R1). Available online:
https://www.ich.org/products/guidelines/quality/qualitysingle/article/guideline-for-elemental-impurities-copy-1.html
(accessed on 26 July 2022)
2 Cauduro VH, Henn AS, Picoloto RS, Muller EI, Mesko MF, Flores EMM, Sustainability, 14, 2022, 422
3 Bizzi CA, Flores EMM, Barin JS, Garcia EE, Nóbrega JÁ, Microchemical Journal, 99, 2011, 193.
[CNPq, FAPERGS, CAPES, UFSM]

DETERMINATION OF CHLORIDE AND SULFUR IN PETROLEUM AFTER
MICROWAVE-INDUCED COMBUSTION IN DISPOSABLE VESSELS
Jaime S. Mandlate (R)a, Paola C. Crestani (PG)b, Eduarda L. Vendruscolo (UG)b,
Letícia C. Brudi (PG)b, Érico M. M. Flores (R)b and Fábio A. Duarte (R)b*
aUniversity Eduardo Mondlane, P.O. Box 252, Maputo, Mozambique
bUniversidade Federal de Santa Maria, Santa Maria, Rio Grande do Sul, Brazil, 97105-900
*e-mail: fabio.duarte@ufsm.br
This work aims the development of a simple low-cost sample preparation method for subsequent
determination of Cl and S in petroleum by inductively coupled plasma optical emission spectrometry
(ICP-OES). Petroleum is a complex mixture of hydrocarbons originated from the decomposition of
organic materials high pressure and temperature by the action of bacteria. Although it is mostly
composed by hydrocarbons, crude oil can contain some impurities, depending on the place.1 Taking
into account the elements contamination in crude oil, Cl and S compounds can be found in crude oil
itself as well as in some fractions. These elements can cause some problems during the refining of
the crude oil, such as poison of catalysts, incrustation and corrosion of pipes. In this case, the
determination of these elements can estimate the level of the contamination and provide better
conditions for petroleum refining.2 There are a few sample preparation methods that allow a
“complete” sample digestion. The microwave-induced combustion (MIC) is a method that digest the
organic matrix by sample combustion in a rich O2 atmosphere and the subsequent analytes
absorption in a suitable solution for elemental determination techniques. The MIC combines the
advantages of the microwave assisted digestion and combustions methods (as relatively high
sample amount, low residual carbon content and negligible blanks).3 Even though this method is
widespread in the literature, a few parameters can be improved in this system. The microwave-
induced combustion in disposable vessels (MIC-DV) is a greener and low cost version of MIC that
uses a domestic microwave oven, which reduces the errors during sample preparation4. MIC-DV
allows the sample digestion in polypropylene (PP) vessels using a quartz holder inside which
containing a filter paper with a combustion initiator. Petroleum was directly weighted in the filter
paper, followed by addition of NH4NO3 as igniter. After the combustion, the analytes are retained in
a suitable absorbing solution. Some parameters as the type and concentration of the absorbing
solution (H2O; H2O2 1%; 10, 25, 50 and 100 mmol L-1 of NH4OH; 25, 50 and 100 mmol L-1 of
(NH4)2CO3), volume of the absorbing solution (2,5; 5,0; 7,5 and 10 mL), sample mass (5, 10, 15 and
20 mg), concentration (6 and 10 mmol L-1) and volume (10, 20, 30, 40 and 60 μL) of NH4NO3,
purging time with O2 (20, 40, 60 and 80 s), were evaluated. The use of consecutives combustions
for analytes preconcentration were also evaluated. The proposed MIC-DV method was applied to
10 petroleum samples with different physical and chemical characteristics. The limit of quantification
(LOQ) for the proposed MIC-DV/ICP-OES method was 105 and 4 µg g-1 for Cl and S, respectively,
whereas the analytes concentration ranged from 194 to 14547 µg g-1 for Cl and from 1961 to 6529
µg g-1 for S. It is important to mention that the results from the proposed method were compared with
those achieved by MIC and detection by ICP-OES and ionic chromatography (IC). In this case, the
agreement ranged from 95 to 103% for Cl and from 91 to 104% for S. To conclude, the MIC-DV and
subsequent determination of Cl and S by ICP-OES showed to be a suitable method for petroleum
digestion, providing accurate results. Even though, comparing MIC-DV to MIC, MIC-DV is greener
considering the Green Analytical Chemistry (GAC) principles.
1 FARAH MA. Petróleo e seus Derivados: Definição; Constituição; Aplicação; Especificações; Características de
Qualidade. Rio de Janeiro: LTC, 2012.
2 PEREIRA, JSF. et al. Spectrochimica Acta Part B, 64, 2009, 554-558.
3 FLORES EMM. et al. Spectrochimica Acta Part B, 62, 2007, 1051-1064.
4 DUARTE FA. Analytical Chemistry, 92, 2020, 8058-8063.
[UFSM, CNPq, FAPERGS, CAPES]

MINERAL COMPOSITION OF RICE, CARROTS, AND CHAYOTE AFTER
MICROWAVE ASSISTED DECOMPOSITION USING DILUTED NITRIC ACID
Wellen K. M. Serafim (PG)1*, Briyitte S. S. Torres (PG)1, Taciana H. Kunst (PG)1,
Iago J. S. da Silva (PG)2, Ana P. S. Paim (R)1
1 Universidade Federal de Pernambuco, Departamento de Química Fundamental, Recife, PE, Brazil,50740-560
2 Universidade Federal de Pernambuco, Departamento de Engenharia Civil e Ambiental,
Laboratório de Saneamento Ambiental (LSA), Recife, PE, Brazil, 50740-560
*email: wellen.karen@ufpe.br
Food is the fuel for human existence, depending on the diet it helps to live a long and healthy life (1).
In 2014, the Ministry of Health published a food guide for the Brazilian population, in which it
recommends the consumption of some fresh foods (2). However, even consuming foods of plant
origin, there is a risk of damage to health, as they can be contaminated by different sources, from
the cultivation process to the way of preparation (3). Therefore, it becomes important to develop
analytical methods to evaluate the composition of foods. This work aimed to develop a sample
decomposition method to prepare rice, carrot, and chayote samples in a raw and cooked state, using
an aqueous solution of nitric acid and hydrogen peroxide with microwave radiation (Start D,
Milestone). Two decomposition methods were studied varying only the amounts of reagents used
(HNO3 and 30% H2O2). A factorial design (22) was also carried out to know the effect of variables on
digestion efficiency. The elements Na and K were quantified by Flame Atomic Emission
Spectrometry (DM-61, Digimed) and Al, Cd, Cr, Cu, Fe, and Pb by Inductively Coupled Plasma
Optical Emission Spectrometry (Optima 7000 DV 9, Perkin Elmer). Based on the analysis of residual
acidity and dissolved organic carbon, the condition that presented the best decomposition results
was using 5 mL of 0.5 mol L-1 HNO3 and 2.5 mL of H2O2 for 100 mg of sample. The limits of
quantification ranging from 0.14 (Cr) to 3.6 µg g -1 (Na), and the correlation coefficient were close to
one in all cases. The Cu concentration in the food samples decreased after the cooking process,
while Al, Cd, and Cr concentration increased in all food after being cooked. This upward and
downward trend can be explained by the method of preparation, where the pan and water used may
have increased the content of elements in foods. Normative Instruction No. 88/2021 of ANVISA
(2021)(4) establishes the maximum limits (LMT) of contaminants in foods, including the levels of Cd
and Pb for rice, carrots, and chayote. The Pb concentration was higher than the tolerable limit by
ANVISA (0.10 – 0.20 µg g-1) in the raw chayote sample (1.25 ± 0.67 µg g-1) and after cooking (1.12
± 0.28 µg g-1) and the Cd concentration was also higher than that allowed by legislation (0.05 µg g -
1
) in the cooked chayote sample (0.56 ± 0.09 µg g-1), while the carrot and rice samples were below
the tolerated limit. The validated method based on residual acidity and dissolved organic carbon, is
viable for determining the mineral composition of rice, carrot, and chayote. The procedure has the
advantages of using a diluted reagent and low sample consumption. Furthermore, the proposed
method could assess contamination by toxic elements or loss of essential elements during cooking.
Future studies will investigate this effect, in which a more representative sampling of cooked foods
will be carried out.
1 Brewer SH; Kellow J.. Eat Better, Live Longer: Understand What Your Body Needs to Stay Healthy. DK Publishing. 1
ed. 2018. Estados Unidos.
2 Basic Attention Notebooks. Food guide for the Brazilian population/Ministry of Health, Department of Health Care,
Department of Primary Care. 2014. 156 p.
3 Affonso CV; Sonati JG. Food Security. In: Roberto Vilarta. (Org.). Healthy Eating, Physical Activity, Quality of Life. 1 ed.
Campinas: Ipes Editorial, 2007, v. 1, p. 19-26.
4 ANVISA. National Health Surveillance Agency. Normative Instruction - No. 88, of March 26 2021. Official Union Gazette.
Body: Ministry of Health/National Surveillance Agency Health/College Board. Ed. 61. Section: 1. P. 226. 2021
[FACEPE/NUQAAPE, CNPq/INCTAA]

VALIDATION OF AN ANALYTICAL METHOD TO CONFIRM THE PRESENCE OF
COBALT IN HUMAN URINE BY LC-MS FOR ANTI-DOPING PURPOSES
Vanessa F. dos Santos (PG)a*, Gabriel R. A. Carneiro (PG)a, Sérgio de P. Machado (R)a,
Henrique M. G. Pereira (R)a
a Universidade Federal do Rio de Janeiro, Instituto de Química, Rio de Janeiro, RJ, Brasil, 21941-598
*e-mail: vanessa.farelo@pos.iq.ufrj.br
All World Anti-Doping Agency (WADA) accredited laboratories must follow a List of Prohibited
Substances and Methods1, updated every year. Cobalt was incorporated, in 2015, to the list, although
the use of cyanocobalamin (vitamin B12) is allowed. The supplementation with cobalt salts has effect on
stimulus of erythropoiesis via stabilization of the hypoxia-inducible factors. An exposure to 120 or 150 mg
day-1 of cobalt chloride, for example, results in the development of polycythemia, with a substantial
increase in hematocrit and hemoglobin up to 20% above pretreatment levels.2 In 2015, Thevis et al. first
reported products containing significant amounts of undeclared cobalt and nickel species in products
advertised as erythropoiesis-stimulating agents by inductively coupled plasma mass spectrometry (ICP-
MS).3 However, the analysis of cobalt in human urine is yet a challenge because of its incompatibility with
the classical strategies available in anti-doping laboratories [e.g. gas chromatography (GC) or liquid
chromatography (LC) coupled with mass spectrometry (MS)]. Here, the validation of an analytical method
to confirm the presence of cobalt in human urine by LC-MS through a complexation strategy with
diethyldithiocarbamate [DDC, (C2H5)2NCSS-] ligand is described. Briefly, the developed method
constituted in the addition of palladium, proposed as the first internal standard (I.S.), in 200 µL of urine
sample, which was hydrolyzed with HCl in a heating block (100°C, 1 hour). After neutralization, the
complexation with DDC and oxidation with citric acid was performed before the liquid-liquid extraction
(LLE) with tert-butyl methyl ether (TBME). To the organic phase, rhodium, proposed as the second I.S.,
was added before evaporation and sample reconstitution. The analysis was performed in an ultra-high
performance liquid chromatography coupled with high resolution mass spectrometry (UHPLC-HRMS).
Rhodium (second I.S.)
o Palladium (first I.S.)
o Acid hydrolysis
o Complexation with
200 µL DDC Evaporation UHPLC-HRMS
Then, the confirmation method was validated based on the Laboratory Technical Note on Analytical
Method Validation for Doping Control4 and WADA Technical Document–TD2021IDCR5. The following
parameters were evaluated: selectivity, limit of identification (LOI), carryover, recuperation, stability in
autosampler (20°C, 48 h), precision, and intermediate precision. Fourteen representative urine samples
from different volunteers were used, divided equally in two days of experiment. The method was selective
for the cobalt complex formed. LOI was estimated in 6.3 ng mL-1, and no carryover was observed when
cobalt is spiked until the concentration of 100 ng mL-1. For recuperation, 71% was obtained for spiked Co
at a concentration of 10 ng mL-1, and the samples were stable for 48 hours in autosampler in 20°C.
However, an elevated matrix effect was observed during validation, which resulted in a coefficient of
variation (CV%) of 35% for day 1 and 15% for day 2. Intermediate precision was evaluated considering
all 14 samples and resulted in 29%. An extra experiment with water, prepared in seven replicates
according to the proposed method, presented a CV% of 7.4%, reinforcing the impact of matrix in the
method outcome. As a conclusion, a method for detection of cobalt in human urine by LC-MS based on
its complexation with DDC ligand was proposed and validated according to the WADA guidelines. For a
quantitative determination, however, a digestion with a microwave device is proposed to minimize the
matrix effect.
1 World Anti-Doping Agency, https://www.wada-ama.org/sites/default/files/2022-01/2022list_final_en_0.pdf, 2022.
2 Lippi G, Franchini M, Guidi GC, Journal of Occupational Medicine and Toxicology, 1, 2006, 18.
3 Thevis M, Krug O, Piper T, Geyer H, Schänzer W, Int J Sports Med., 36, 2015, 82.
4 World Anti-Doping Agency, Version 01, 2020.
5 World Anti-Doping Agency, https://www.wada-ama.org/sites/default/files/resources/files/td2021idcr_final_eng_0.pdf,
2021.
[CNPq, ABCD, FAPERJ]

REVERSED PHASE DISPERSIVE LIQUID-LIQUID MICROEXTRACTION:
GREEN, SIMPLE AND FAST SAMPLE PREPARATION METHOD FOR
DETERMINATION OF Cu Fe BY IN INSULATING MINERAL OIL
Jéssica F. B. Wendt (PG)a, Alessandra S. Henn (R)b, Paulo R. S. Bittencourt (R)a,b,
Oldair D. Leite (R)a,b, Burkhard Horstkotte (R)c, Petr Solich (R)c, Eder L. M. Flores (R)a,b*.
a UniversidadeTecnológica Federal do Paraná, Campus Medianeira, Programa de
Mestrado em Tecnologias Ambientais, Medianeira, Paraná, Brazil, 85884-000
b Universidade Tecnológica Federal do Paraná, Campus Medianeira,
Departamento de Química, Medianeira Paraná, Brazil, 85884-000

c Charles University, Department of Analytical Chemistry,
Faculty of Pharmacy in Hradec Králové, Hradec Králové, Czech Republic

*e-mail: eder.utfpr@gmail.com
Mineral oil is widely used as dielectric insulator in power transformers. Contamination of Cu and Fe
in insulating mineral oil result in decrease of the dielectric strength and corrosion can occur in the
equipment, favoring its early aging and possible environmental impacts1,2. The determination of Cu
and Fe in insulating mineral oil is performed by ICP-OES after microwave assisted acid digestion
(MAWD) or after by dilution in xylene or kerosene as sample preparation step. These sample
preparation procedures have some drawbacks such as the use of expensive equipment and large
volumes of concentrated acids or toxic organic solvents. Furthermore, the direct introduction of
organic solvents in ICP-OES results in a decrease in the lifetime of the quartz torch and possible
matrix interference. In this work, reversed-phase dispersive liquid-liquid microextraction (RP-
DLLME) was proposed as a simple, fast and green sample preparation method for microextraction
and pre-concentration of Cu and Fe in samples of paraffinic-based mineral insulating oil and further
determination by atomic absorption spectrometry (F AAS). Sample preparation by RP-DLLME was
performed with a mixture of extracting (HNO3, 1 mol L-1; 300 μL) and dispersing (n-propanol; 700
μL) solvents, injected onto the sample, then the aqueous phase containing the analytes was
separated by centrifugation. The following RP-DLLME conditions were adjusted: concentration and
volume of extracting and dispersing solvent, heating temperature, sample mass, agitation and
centrifugation time. The proposed method showed analytes recovery ranging from 98 to 104% (RSD
< 7%) for Fe and from 98 to 106% for Cu (RSD < 3%). The method showed LOQs of 0.0037 µg g-1
for Cu and 0.0103 µg g-1 for Fe and LODs of 0.0061 µg g-1 for Cu and 0.0020 µg g-1 for Fe. Different
types of insulating mineral oil samples were analyzed (new oil, with little use, used and burned) and
compared with reference values obtained with the decomposition of analytes by MAWD
(determination by ICP-MS for Cu and MIP-OES for Fe). The evaluation of the greenness of the
proposed method was carried out based on the principles of green analytical chemistry, using the
AGREE software. The developed method was considered suitable for the determination of Cu and
Fe in insulating mineral oil samples, following the Green Analytical Chemistry Principles. The RP-
DLLME proposed method is fast and simple, allows the use of a high sample mass, requires low
consumption of reagents and generates less waste. Moreover, the proposed method shows high
sample throughput in comparison with the MAWD. This study is the first application of RP-DLLME
in the sample preparation of insulating mineral oil matrix.
1 Bruzzoniti MC, Carlo RM, Sarzanini C, Maina R, Tumiatti V. Talanta, 99, 2012, 703.
2 Ametican Society for Testing and Materials. ASTM D7151: Determination of Elements in Insulating Oils by Inductively
Coupled Pasma Atomic Emission Spectrometry (ICP-AES). West Conshohocken – PA, 2015.
3 Lourenço, EC; Eyng, E; Bittencourt, PRS; Duarte, FA; Picoloto, RS; Flores, ELM. Talanta, 199, 2019, 1.
[CEANMED, UTFPR/Campus Medianeira, Comtrafo Transformadores S.A., LAQIA/UFSM,

CAPES]

SAMPLE PREPARATION FOR METABOLOMIC STUDIES OF BACTERIAL
STRAINS OF THE GENUS BRUCELLA
Joane M. M. Corrêa (PG)a*, Mauro L. G. Oliveira (R)b, Patrícia G. Souza(R)c,
Paulo M. S. Filho (R)c, Adriana N. Macedo (R)a, Adriana F. Faria (R)a
aUniversidade Federal de Minas Gerais, Department of Chemistry, Belo Horizonte, MG, Brazil, 31270-901
bInstituto Mineiro de Agropecuária, Belo Horizonte, MG, Brazil, 31920-590
cLaboratório Federal de Defesa Agropecuária em Minas Gerais, Pedro Leopoldo, MG, Brazil, 33600-000
*e-mail: miaricorrea@yahoo.com.br
Brucellosis is a serious zoonosis caused by bacteria of the genus Brucella, which are listed by the
Centers for Disease Control and Prevention (CDC) as a bioterrorism agent, category B. According
to the CDC, pathogens in this category are moderately easy to spread, cause moderate morbidity
and low mortality rates.1 In addition, bacteria of this genus present environmental resistance and
difficult therapeutic treatment.2-3 Sample preparation is a critical step in the metabolomics workflow.
All procedures applied must be evaluated so as not to affect the metabolic profile obtained and not
to impair the quality of the data. Errors can result in an incomplete or even mistaken biological
interpretation. 3-4 In this study, bacterial sample preparation processes and a method of extracting
intracellular metabolites from the bacterium Brucella abortus were optimized. The steps of bacterial
strain sample preparation evaluated were: a) growth time of the Colony Forming Units (CFU) from
24 to 120 h; b) collection (1 to 3 levels); c) bacterial biomass sedimentation time (1, 2 and 3 min) by
centrifugation at 7000 xg; d) solution for bacterial inactivation (70% v/v methanolic solution and 70%
v/v ethanolic solution). The optimization of the extraction method was performed by testing five
protocols adapted from the literature. The extraction solvents tested were
acetonitrile/isopropanol/water, methanol/water, methanol/water/chloroform, methyl tert-butyl ether
(MTBE)/methanol/water (with phase separation), and methanol/water/methyl tert-butyl ether (single
phase). The extracts were subsequently derivatized and analyzed using gas chromatography
coupled to high resolution mass spectrometry (GC-QTOF-MS). The experiments were prepared in
biological triplicates. All biological and chemical assays were randomized. Data processing was
performed on the XCMS Online platform and statistical analysis was performed on MetaboAnalyst.
The extraction procedures were evaluated based on the number of extracted molecular features with
coefficient of variation (CV) under 30%. The five extraction protocols were also assessed using
multivariate analysis, as well as the calculated CV for the peak intensities of the internal standard
present in the bacterium extracts and blank samples. The optimized preparation protocol was 48 h
for CFU growth, 3 levels for collection, 1 min of centrifugation at 7000 xg and 70% v/v methanolic
solution for bacterial inactivation. The extraction solvent methanol/water/chloroform presented the
best results and demonstrated the smallest retention time deviation among the molecular features.
This optimized sample preparation will be used in subsequent metabolomic studies involving the
extraction of intracellular metabolites from the bacterium Brucella abortus.
1 CDC, Centers for Disease Control and Prevention, 2018, https://emergency.cdc.gov/agent/agentlist-category.asp,
Accessed 10 Jul 2022.
2 El-Sayed A, Awad W, International Journal of Veterinary Science and Medicine, 6, 2018, 31.
3 Belinato JR, Bazioli JM, Sussulini A, Augusto F, Fill TP, Química Nova, 42, 2019, 546.
4 Dudzik D, Barbas-Bernardos C, García A, Barbas C, Journal of Pharmaceutical and Biomedical Analysis,147, 2018, 149.
[UFMG, LFDA/MG, CNPq]

ELEMENTAL DETERMINATION AND BIOACESSIBILE FRACTION
IN EDIBLE MUSHROOMS
Nicolle L. Bandeira (UG)ª*, Sabrina H. Scherdien (PG)ᵇ, Daisa H. Bonemann (PG)ᵇ,
Charlie G. Gomes (PG)ᵇ, Solange Cadore (R)c, Adriane M. Nunes (R)ª
ªFederal University of Pelotas, Chemical, Pharmaceutical and Food Sciences Center,
Campus Capão do Leão, 96160-000, Capão do Leão, RS
ᵇFederal University of Pelotas, Master’s Program in Chemistry, Campus Capão do Leão, 96160-00, Capao do Leão, RS
cInstitute of Chemistry, University of Campinas, CEP 6154, 13083-970, Campinas, SP, Brazil.
*e-mail: niicolle.lima97@gmail.com
Due to the high demand for healthier eating habits nowadays, the consumption of mushrooms in the
country has increased because it has a lot of nutrients, fiber and proteins when compared to other
types of vegetables, besides had very few calories and zero cholesterol. Two decades ago, the
mushroom consumption in Brazil was around 50 grams per habitant, today it exceeds 288 grams
per habitant.¹ Mushrooms belong to the Fungi kingdom and they are considered bioaccumulating
organisms. Thereby, it is necessary to control its composition, including the elementary in order to
quantify essential elements, as well as elements harmful to the human organism. Not only controlling
its elemental composition is important, but also evaluating and studying its bioaccessibility to
understand what is released and absorbed by the gastrointestinal tract.²,³ Therefore, the objective of
this work is to determine the total concentration and the bioaccessible fraction in samples of Paris,
Portobello, Shimeji and Shitake mushrooms. For total concentration determination, 500 mg of dry
mass and 5.0 mL of HNO3 was used. The decomposition was performed using a digester block with
reflux system at 125 °C for 120 minutes. After cooling, 2 mL of H2O2 was added and it was taken to
the digester block for another 60 minutes in the same temperature. Subsequently, the sample was
cooled, transferred to polypropylene vials and the final volume of 20 mL was filled with deionized
water. For bioaccessible fraction studies, the procedure described by Minekus et al³ was employed.
The determinations were performed by microwave induced plasma optical emission spectrometry
(MIP OES). The accuracy of the method was evaluated using certified reference materials of
sugarcane (CRM-Agro C1005a) and tomato pulp (CRM-Agro C1004a) and the recoveries ranged
from 80 to 120%. For the bioaccessible fraction results, according to the obtained values and to the
mass balance, the recoveries ranged between 90 and 119%. The total concentrations obtained for
the mushroom samples ranged from 0.19 to 10.0 mg kgˉ¹ for Al; 2.8 to 61 mg kgˉ¹ for Ca; 0.43 to 7.1
mg kgˉ¹ for Cu; 2.1 to 26 mg kgˉ¹ for Fe; 1,195 to 4,005 mg kgˉ¹ for K; 69 to 174 mg kgˉ¹ for Mg; 0.42
to 2.4 mg kgˉ¹ for Mn; 9.6 to 165 mg kgˉ¹ for Na; 0.21 to 0.69 mg kgˉ¹ for Pb and 2 to 12 mg kgˉ¹ for
Zn. The values obtained from the bioaccessible fractions ranged from 1 to 53% for Al; 0 to 87% for
Cu; 0 to 79% for Fe; 59 to 95% for K; 42 to 82% for Mg; 7 to 84% for Mn; 41 to 73% for Pb and 20
to 55% for Zn. Overall, it can be verified that the method developed showed satisfactory results
through validation studies. The concentrations in the different samples showed a great variation in
the total concentrations and bioaccessible percentages, demonstrating the importance of nutritional
information of edible mushrooms. The studied samples exhibited essential elements, in addition to
concentrations for Pb, which is a potentially toxic element, which was found above the recommended
by the legislation in practically all the studied samples.
1 URBEN, A.F. et al. Mushroom production using modified Chinese technology: biotechnology and applications in
agriculture and health. Technical publishing company. 3.ed rev. e ampl., Embrapa: Brasília-DF, 2017, 274 p.
2 Pereira CC, Silva NE, Souza OA, Vieira AM, Ribeiro SA, Cadore S, Journal of Food Composition and Analysis, 68, 2018,
73-78.
3 Minekus M, Alminger M, Alvito P, Ballance S, Bohn T, Bourlie C, Carrrière F, et al., Food & Function, 5, 2014, 1113.

DETERMINATION OF PESTICIDE RESIDUES IN BANANA BY QuEChERS
METHOD AND UHPLC-MS/MS
Ana Paula F. Prochnow (PG), Rafael Bel P. da Silva (PG), José Dílson F. da Silva (PG),
Francielle F. Machado (PG), Osmar D. Prestes (R), Martha B. Adaime (R), Renato Zanella (R)*
*e-mail: renato.zanella@ufsm.br
In order to maintain a high production yield, the use of pesticides is a conventional agricultural
practice. Pesticides are used to eliminate pests in agriculture and to control the quality and quantity
of crop production.1 Despite their many merits, pesticides are some of the most toxic,
environmentally stable, and mobile substances in the environment. Pesticide residues, which remain
after an agricultural treatment, may penetrate plant tissues and be determined in the pulp and the
juice of the fruit.1 Pesticide residues represent one of the most important factors of chemical
contamination in fruits and vegetables. Bananas (Musa spp.) are one of the most popular fruits on
the world market and probably one of the fruits most consumed in Brazil. The aim of this work was
to investigate the occurrence of pesticides in banana samples. For that purpose, the QuEChERS
method was applied prior to ultra-high performance liquid chromatography tandem mass
spectrometry (UHPLC-MS/MS) determination. Three versions of the QuEChERS were evaluated. 2
Different sorbents (activated carbon, alumina, chitosan, C18, diatomaceous earth, fluorinated, GCB,
propylamine, PSA, silica, Oasis® HLB, florisil®, Celite® 521, StrataTM-X, and Supel™ QuE Z-Sep+)
were tested in the dispersive solid phase extraction (d-SPE) clean-up step. The QuEChERS citrate
method combined with d-SPE cleanup step performed with Oasis® HLB (25 mg) was chosen as the
sample preparation method. Pesticide analysis was performed using a XevoTM TQ UHPLC-MS/MS
triple quadrupole system (Waters, USA), equipped with an Acquity UPLC® liquid chromatograph and
BEH C18 column (50 x 2.1 mm, 1.7 ìm). The mobile phase consisted of (A) aqueous solution with
2%(v/v) methanol (v/v) and (B) methanol, both containing 0.1%(v/v) formic acid (v/v) and 5 mmol L-
1
ammonium formate. Elution was performed by gradient, using a flow rate of 0.225 mL min-1, with a
total run time of 10 min. The proposed method was validated for the determination of 82 pesticides,
which showed recoveries between 70 and 118%, RSD ≤ 20% and method LOQ levels between 1
and 5 µg kg-1. The method was applied to 16 banana samples, and four pesticides: carbendazim,
difenoconazole, imidacloprid and mecarbam were found in the concentration from 1.8 to 161.9 µg
kg-1. The QuEChERS citrate method with Oasis® HLB in the cleanup step was fast and easy for the
determination of more than 80 compounds in banana and can be used in routine analyzes.
1 Christia C, Bizani E, Christophoridis C, Fytianos K. Environ Sci Pollut Res Int, 22, 2015
2 Perestelo R, Silva P, Porto-Figueira P, Pereira JAM, Silva C, Medina S, Câmara JS, Analytica Chimica, 1070, 2019.
[UFSM, CAPES, CNPq, FAPEAM]

METHOD DEVELOPMENT FOR CHEMICAL SPECIATION OF AS IN SEAFOOD
AND CR IN AGRICULTURAL SAMPLES BY LC-ICP-MS
Ívero Pita de Sá (PG)a, Mykaelli Andrade Santos (PG)ab,Ana Rita Araujo Nogueira (R)a*
aEmbrapa Pecuária Sudeste, São Carlos, São Paulo, Brazil, 13560-970
bFederal University of São Carlos, Department of Chemistry, São Carlos, São Paulo, Brazil, 13565-905
*e-mail: mykaelliandrade97@gmail.com
Regarding species separation and detection, a combination of analytical techniques is usually

necessary to achieve adequate selectivity and sensitivity for chemical speciation analysis.
Hyphenated liquid chromatography with inductively coupled plasma mass spectrometry (LC-ICP-
MS) is the most suitable combination for speciation of As in fish and Cr in foods 1,2. In order to
establish a single extraction and separation condition for the different types of samples in their
respective methods, the LC-ICP-MS potential, and the need for chemical speciation of As and Cr in
agricultural samples, two methods were developed for the As species determination in fish and Cr
in agronomic samples. The mobile phase 10 mmol L-1 (NH4)2PO4 diluted in 1% ethanol (pH 8.0), and
the mobile phase 15 mmol L-1 Na2SO4, 5 mmol L-1 EDTA, 5 mmol L-1 NaH2PO4 (pH 7.0) provided
adequate separation and sensitivity data by LC-ICP-MS for As and Cr, respectively. The extraction
of the species was defined using experimental designs. The optimized conditions employed were 30
mmol L-1 HNO3 at 90°C for 45 min in digester block for As, and 45 mmol L-1, [EDTA], 90°C, 40 min,
100 mg of sample in digester block for Cr. Recoveries of 88 to 106% of arsenobetaine (AsB), arsenite
[As(III)], dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), arsenate [As(V)], Cr(III) and
Cr(VI) were obtained for all evaluated samples. The limits of quantification provided by this method
were 5.3; 52.1; 16.4; 2.8; 83.3; 113.8; 53.9 ng g-1 for AsB, As(III), DMA, MMA, As(V), Cr(III) and
Cr(VI) respectively. The veracity of the method was evaluated througn analyzes of the fish tissue
reference material (IPEN) and the CRMs of dogfish liver (DOLT-5), lobster hepatopancreas (TORT-
3), fish protein (DORM-4), oyster tissue (NIST-1566b), soil contaminated with hexavalent chromium
(NIST 2701). The sum of the species was in agreement with the total concentration of As or Cr(VI)
content.
1 Schmidt L, Landero JA, Novo DLR, Duarte FA, Mesko MF, Caruso JA, Flores EMM, A feasible method for As speciation
in several types of seafood by LC-ICP-MS/MS, Food Chemistry. 255, 2018, 340–347.
2 Séby F, Vacchina V, Critical assessment of hexavalent chromium species from different solid environmental, industrial
and food matrices, Trends in Analytical Chemistry, 2018.
[FAPESP, CNPq, CAPES, INCTAA]

DETERMINATION OF PHARMACEUTICALS IN RESIDUAL WATERS TREATED
FROM CONSTRUCTED WETLAND SYSTEMS BY SPE AND UHPLC/MS-MS
Rômulo de O. Schwaickhardt (PG)a, Júlia A. de Oliveira (PG)a, Marisa Demarco (PG)a,
Ênio L. Machado (R)b, Osmar D. Prestes (R)a, Martha B. Adaime (R)a, Renato Zanella (R)a,*
a Laboratoryof Pesticides Residue Analysis (LARP), Chemistry Department, Federal University of Santa Maria,
b Postgraduate Program in Environmental Technology, University of Santa Cruz do Sul,
Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96815-900

In recent years, the production and use of pharmaceuticals and personal care products (PPCPs) has
increased exponentially. The use of PPCPs associated with inefficient effluent treatment systems
makes it possible to find these compounds in concentrations that can vary from parts per trillion (ng
L-1) to parts per million (mg L-1) in different water bodies.1-3 Thus, new treatment alternatives such as
constructed wetlands emerge as a potential technology for removing these contaminants from
effluents. Therefore, to evaluate the efficiency of this and other types of effluent treatment systems,
it is necessary to develop multiclass methods to control the treatment. In this work, a multiclass
method was developed for the analysis of 37 compounds in raw and treated residual waters from
wetlands constructed in different configurations. The determination was performed by ultra-high
performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS).
Chromatographic separation was performed using an Agilent InfinityLab Poroshell HPH-C18 column
(2.1 x 50 mm, 1.9 µm). The composition of the mobile phase used consisted of: (A) water and (B)
methanol, both containing 1 mmol L-1 ammonium formate and 0.1% (v/v) formic acid. Total
chromatographic analysis time was 5 min maintaining a constant flow rate of 0.4 mL min-1 and a
column temperature of 35 ºC. The solid phase extraction method (SPE) was developed using Oasis
HLB cartridges, the sample was percolated at pH 5 with 0.01 mol L-1 of EDTA and was eluted with
methanol. A subsequent cleanup step by d-SPE was applied using PSA as sorbent. The method
showed recoveries between 70 and 120%, precision and intermediate precision (RSD ≤ 20%). The
limit of quantification (LOQ) was between 0.025 and 0.100 μg L-1 and the detection limit (LOD)
between 0.008 and 0.030 μg L-1. The validated method was applied to evaluate the efficiency
removal of pharmaceuticals present in the residual water after passing through a wetlands system
built with 4 different treatment configurations. The 14 compounds found in the raw effluent, only 8
were still detected and in lower concentrations after the constructed wetlands system in best
configuration.
1 Archer E, Petrie B, Kasprzyk-Hordern B, Wolfaardt GM, Chemosphere, 174, 2017, 437-446.
2 Liu JL, Wong MH, Environment International, 59, 2013, 208-224.
3 Yang Y, Ok YS, Kim KH, Kwon EE, Tsang YF, Science of the Total Environment, 596–597, 2017, p. 303-320.
[UFSM, UNISC, CAPES, CNPq]

CARBON NANOTUBES STABILIZED IN CHITOSAN SPONGE AS DISPERSIVE
SORBENT FOR PESTICIDE ANALYSIS IN EGGPLANT BY UHPLC-MS/MS
Cleusa F. Zanchin (PG)a, Bruna S. de Farias (PG)b, Luiz A.A. Pinto (R)b,
Tito R.S. Cadaval Jr. (R)b, Martha B. Adaime (R)a, Osmar D. Prestes (R)a Renato Zanella (R)a*
aLaboratory
of Pesticides Residue Analysis (LARP), Chemistry Department, Federal University of Santa Maria,
bSchool of Chemistry and Food, Federal University of Rio Grande, Rio Grande, Rio Grande do Sul, Brazil, 96230-000
The use of different classes (herbicides, insecticides, fungicides, acaricides, among others) in
agriculture has economic benefits, since they ensure the control and combat of pests and diseases,
protecting the quality of production. On the other hand, consumers may be exposed to these
chemical compounds through food.1 For that, the pesticide residues must comply with the maximum
residue levels established by regulations national and international. Eggplant has attracted the
attention of Brazilian farmers due to its low production cost, plant hardiness and increased market
demand. The consumption of this vegetable has been increasing in recent years, after scientific
evidence has shown its medicinal and herbal effects, especially related to the prevention and
treatment of diabetes and cardiovascular problems. It is a good source of minerals and vitamins. In
addition to being rich in fiber and having a low lipid content, it contains a variety of phytochemicals
such as polyphenols that provide important health benefits.2 This work describes the comparison of
different versions of an easy, rapid and low-cost sample preparation approach for pesticide
multiresidue determination in eggplant using the UHPLC-MS/MS. The final extraction step was
performed with 10 g of sample and 10 mL of acetonitrile, with the addition of MgSO 4 and NaCl. The
performance of different sorbents for the cleanup step was evaluated, and the carbon nanotubes
stabilized in chitosan sponge3 was the most efficient. The pesticide residue analysis was performed
with Acquity UPLCTM liquid chromatography coupled to a Xevo TQ® MS/MS triple quadrupole mass
spectrometer (Waters, USA) with positive and negative electrospray ionization interface in selected
reaction monitoring (SRM) mode. The chromatographic separation was performed using an Acquity
UPLC™ BEH C18 column (50 mm x 2.1 mm x 1.7 µm particle size). Mobile phase consisted of (A)
water:methanol (98:2, v/v) and (B) methanol:water (98:2, v/v), both containing 5 mmol L -1 of
ammonium formate and 0.1% (v/v) of formic acid. The injection volume was 10 μL and total run time
was 10 min. Therefore, 111 compounds were analyzed and validated in terms of linearity, accuracy,
precision and limits of quantification and detection. The results were satisfactory for most
compounds, with recoveries from 70 to 120% with RSD ≤ 20% at different levels (5, 10, 20 and 50
µg kg-1, n= 6). The method LOQ value was 5 μg kg-1 for the analyzed compounds. The validated
method was applied to different commercial eggplant samples. Of the pesticides evaluated in this
study, 27 were quantified and/or detected. Those pesticides that appeared most frequently were the
insecticides acephate (<LOQ to 0.504 mg kg-1), imidacloprid (until 0.074 mg kg-1), and thiamethoxam
(until 0.121 mg kg-1). Therefore, the QuEChERS method, using a cleanup step with a carbon
nanotubes stabilized in chitosan sponge, combined with UHPLC-MS/MS detection was considered
very efficient, as it allowed the determination of the selected analytes with high selectivity and
sensitivity.
1 Akanitapichat P, Phraibung K, Nuchklang K, Prompitakkul S. Food Chem Toxicol, 2010.
2 Lehotay, SJ; de Kok , A.; Hiemstra , M. et al. J. AOAC Int., 88, 2005.
3 Affonso, L.N., et al. Journal of hazardous materials 388 (2020): 122042.
[UFSM, FURG, CAPES, CNPq]

DETERMINATION OF VETERINARY DRUG RESIDUES IN POULTRY FEED
USING SIMPLE AND FAST EXTRACTION AND UHPLC-MS/MS
Lucila C. Ribeiro (PG), José Dilson F. da Silva (PG), Tífane S. Moureira (UG),
Franciele F. Machado (PG), Leonardo R. Amaral (PG), Martha B. Adaime (R),
Renato Zanella (R), Osmar D. Prestes (R)*
Animal feed is considered as any material that is suitable to be consumed as food, which meets the
standards of quality and identity, as well as sanitary and nutritional standards.1 It can be of vegetable,
animal or mineral origin, in its natural state, fresh or preserved. Also included are products from
industries (derivatives) with or without additives. The use of additives in animal feed products
purpose to improve and complement the nutritional needs and also the performance of animals.
However, these substances must be used correctly, only to achieve the expected effect, thus
ensuring the health of humans, animals, and the environment.2 Therefore, the aim of this study was
the development a multiclass method for the determination of veterinary drug residues in poultry
feed using simple and rapid sample preparation method and ultra-high performance liquid
chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The extraction step used
2 g feed with 2 mL of ultrapure water and 5 mL of acetonitrile acidified with formic acid. The samples
were shaken and centrifuged (10 min). An aliquot of 1 mL of extract was taken for d-SPE cleanup
step using 100 mg of C18 and 100 mg of PSA. The extract was diluted in mobile phase for injection
into the chromatographic system. The chromatographic separation was performed using Xevo TQ-
XS equipment (Waters, USA) with an Acquity UPLC BEH C18 column (100 x 2.1 mm, 1.7 μm),
mobile phase (A) ultrapure water and (B) acetonitrile, both with 0.1% (v/v) formic acid, and flow rate
of 0.4 mL min-1. The injection volume was 5 µL. The electrospray ionization source (ESI) was
operated in positive and negative mode, with the mass spectrometer operated in selected reaction
monitoring (SRM) mode. The method was evaluated for selectivity, limits of detection and
quantification, matrix effect, linearity, accuracy and precision. Solvent and matrix matched analytical
curves were prepared at concentrations from 0.7 to 20 µg L-1 and the determination coefficients were
>0.990 for all compounds. The matrix effect (-52 to 100%) was compensated using matrix-matched
analytical curves. The results of accuracy and precision tests, evaluated at the spike levels 5, 10, 40
and 70 µg kg-1 (n= 6), proved to be adequate since they showed recoveries between 70 and 120%,
with RSD ≤ 20% for most of the compounds. The method LOQ values were between 5, 10 and 40
µg kg-1. The developed method is suitable for regulatory purpose analyses of veterinary drugs in
animal feed.
1 BRAZIL. Normative Instruction No. 4 of February 23, 2007. Approves the technical regulation on the hygienic-sanitary
conditions and good manufacturing practices for manufacturing establishments of products intended for animal feed and
the inspection script. Available at: <http://www.agricultura.gov.br/assuntos/insumos-agropecuarios/insumos-
pecuarios/alimentacao-animal/arquivos-alimentacao-animal/InstruoNormativa04.2007.pdf>. Access on: 10 June 2022
2 BRAZIL. Additives. Published: December 30, 2016. Available at: <http://www.agricultura.gov.br/assuntos/insumos-
agropecuarios/insumos-pecuarios/alimentacao-animal/aditivos>. Access on: 10 June 2022.
[UFSM, CAPES, CNPq]

DETERMINATION OF HALOGENS IN EDIBLE SEAWEED USING VORTEX-
ASSISTED MATRIX SOLID-PHASE DISPERSION AND ION CHROMATOGRAPHY
Maiara H. M. Malinowski (PG)a, Paloma K. Maciel (UG)a, Enilin M. de Azevedo (PG)a,
Sergiane C. Barbosa (R)a, Ednei G. Primel (R)a, Fábio A. Duarte (R)b, Bruno M. Soares (R)a*
aUniversidade Federal do Rio Grande, Escola de Química e Alimentos, Rio Grande, RS, Brazil, 96203-900
b Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil, 97105-900
*e-mail: bm_soares@hotmail.com.br
Algae are a natural source of essential elements due to their ability to incorporate chemical species
from seawater.1 Few studies about the determination of halogens in algae have been reported in the
literature.2,3 Dry decomposition, pyrohydrolysis, microwave-induced combustion (MIC), and
microwave-assisted extraction (MAE) have been reported for the sample preparation aiming at the
determination of halogens in algae. However, the possibility of loss of halogens, risk of
contamination, low analytical frequency, and the need to use relatively high-cost equipment (in the
case of microwave ovens) are some disadvantages of the methods mentioned. Matrix solid-phase
dispersion (MSPD) is a simple method, based on sample maceration in the presence of abrasive
solid support, promoting the rupture of the physical structure of the sample. Therefore, this work
aims to develop an analytical method employing vortex-assisted matrix solid-phase dispersion (VA-
MSPD), which is a modification of the original MSPD, to extract F, Cl, Br, and I from samples of
edible seaweed with further determination by ion chromatography (IC). Important parameters that
influence VA-MSPD were evaluated, such as the type of solid support (diatomaceous earth, sand,
mussel shell, florisil, alumina, and without solid support), the type of extracting solution (water, a
mixture of 3.2 mmol L-1 Na2CO3 and 1.0 mmol L-1 NaHCO3, and 50 mmol L-1 (NH4)2CO3), sample
mass (0.1 to 0.25 g), solid support mass (0.25 to 1.0 g), and maceration time (1 to 5 min). VA-MSPD
conditions were optimized for the edible seaweed of the Nori (Porphyra spp.) type and applied to
samples of the Wakame (Undaria pinnatifida), Kombu (Laminaria ochroleuca), and Hijiki (Hizikia
fusiformis) types. Accuracy was evaluated in three ways: i) recovery assays; ii) CRMs of apple leaves
(NIST 1515) and peach leaves (NIST 1547); and iii) comparing methods using the determination of
Cl, Br, and I by inductively coupled plasma-mass spectrometry (ICP-MS) and the determination of F
by ion selective electrode (ISE), after decomposition of the samples by microwave-induced
combustion (MIC). The optimized conditions were: 0.1 g of sample; 1.0 g of sea sand (clean) as a
solid support, 5 min of maceration, and 10 mL of 50 mmol L-1 (NH4)2CO3, as extracting solution. The
method was linear (R2 > 0.99) within the evaluated range from 0.1 to 5.0 mg L-1 for F, from 1.0 to
100.0 mg L-1 for Cl, and from 0.1 to 10.0 mg L-1 for Br and I. No matrix effect was observed and the
calibration was performed with external calibration curves in solutions of 50 mmol L-1 (NH4)2CO3.
The detection limits (LODs) were 27, 26, 19, and 28 µg g-1 for F, Cl, Br, and I, respectively. The
analyte concentrations in the samples ranged from 161 to 427 µg g-1 for F, from 6528 to 101596 µg
g-1 for Cl, from 60 to 395 µg g-1 for Br, and from 61 to 3542 µg g-1 for l. The method showed good
agreement through recovery assays, with values between 92 and 108%. After the application of the
VA-MSPD/IC method in the CRMs, a good agreement (101% for NIST 1547 and 97% for NIST 1515)
with the certified values for Cl was obtained, with RSDs lower than 6%. In the comparison of the
results with those obtained by ICP-MS and ISE after MIC, agreements between 103 and 124% were
obtained, with RSDs lower than 6.5%. An eco-friendly alternative using VA-MSPD is suitable for the
extraction of halogens in different species of algae, presenting advantages such as simplicity and
low cost.
1 Pereira L, Marine Drugs, 18, 2020,65
2 Mesko MF, Toralles IG, Crizel MG, Costa VC, Pires NRX, Pereira CMPD, Picoloto RS, Mello PA, Química Nova, 37,
2014, 964.
3 Flores EMM, Mello PA, Krzyzaniak SR, Cauduro VH, Picoloto RS, Rapid Communications in Mass Spectrometry, 34,
2020, 1.
[CNPq, FAPERGS, FINEP]

EVALUATION OF IRON CONTENT IN EGGPLANT FLOUR AFTER ACID
DIGESTION ON A HOT PLATE
Anderson da S. Pereira (UG)a*, Aparecida M. S. Mimura (R)a
aUniversidade Federal do Vale do São Francisco, Colegiado de Licenciatura em Química,
São Raimundo Nonato, PI, Brazil, CEP 64770-000
*e-mail: anderson.silvapereira@discente.univasf.edu.br
Among the various types of anemia in the world, iron deficiency anemia is the most common and is
often related to an inadequate diet, with a low content of nutrients, such as iron. In order to minimize
anemia in Brazil, public policies were implemented, making iron fortification in wheat and corn flour
mandatory1. Although there are several methods of sample preparation of wheat flour2, only a few
studies have been reported regarding other types of flour. Recently, eggplant flour has been gaining
prominence in the market, being widely used by people who are intolerant or allergic to gluten. Thus,
the aim of this was to carry out the acid digestion of eggplant flour for consequent determination of
the iron concentration by atomic absorption spectrometry. The eggplant flour sample studied was
obtained from the local market in the city of São Raimundo Nonato-PI. Several sample preparation
methods were initially investigated using different mineral acids to verify the best digestion
conditions. The following method presented the best results. An acid digestion procedure was
performed on a hot plate at 100 °C for 40 min and 0.500 g of flour samples were weighed in a beaker.
The samples were digested by adding 5.0 mL of H2O, 5.0 mL of concentrated HNO3, and 2.0 mL of
H2O2 (35% v/v). After digestion, the volume was reduced, the contents were transferred into
volumetric flasks and the volume was completed to 15.0 mL with deionized water. All procedures
were performed in triplicate. Blanks were also prepared. Then, all the samples and blank solutions
were analyzed by flame atomic absorption spectrometry for the determination of Fe, at 248.3 nm. In
order to verify the accuracy, spike tests were performed at one concentration level (1.0 mg L-1 Fe).
The Fe concentration found in the eggplant flour samples was 6.6 ± 0.6 mg/100g. The method
presented good results regarding precision (RSD < 3.0%) and accuracy (recovery of 89.7%). Thus,
this sample preparation method was adequate for eggplant flour analysis. The iron content found
was accordingly to the data in the literature for wheat and corn flour and also accordingly to the
values recommended by ANVISA and Ministério da Saúde (between 4.0 and 9.0 mg/100g)3.
Therefore, eggplant flour consumption can contribute to the prevention of anemia as well as wheat
flour. The next steps of this work include analyzing other brands and evaluating the application of
ultrasound-assisted extraction for this type of sample.
1 Buzzo ML, Carvalho MFH, Tiglea P, Arauz LJ, Arakaki EEK, Matsuzaki R, Rev Inst Adolfo Lutz, 71, 2012, 645.
2 Martins CA, Cerveira C, Scheffler GL, Pozebon D, Food Anal Methods, 8, 2014, 1652.
3 ANVISA. Agência Nacional de Vigilância Sanitária. Resolução – RDC n° 344, de 2002; Resolução – RDC n° 269, de
2005; Resolução – RDC n° 150, de 2017. Disponível em: <http://www.portal.anvisa.gov.br >. Acesso em: 25 de julho de
2022.
[PRPPGI, UNIVASF, CNPq]

DETERMINATION OF ARSENIC SPECIES IN EDIBLE SEAWEED EMPLOYING
THE VORTEX-ASSISTED MATRIX SOLID-PHASE DISPERSION AND LC-ICP-MS
Ana C. B. Luckow (PG)a, Letícia C. Brudi (PG)b, Lisliane Kickofel (PG)a, Jean L. O. Arias (R)a,
Eliézer Q. Oreste (PG)a, Fábio A. Duarte (R)b, Bruno M. Soares (R)a*
aUniversidade Federal do Rio Grande, Escola de Química de Alimentos, Rio Grande, RS, Brazil, 96203-900
bUniversidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil, 97105-900
*e-mail: bm_soares@hotmail.com.br
Seaweeds are widely consumed and in several countries, due to their high nutritional value, since
they are considered a good source of fiber proteins and with antioxidant properties. However, the
bioaccumulation of potentially toxic elements, such as arsenic (As), can occur in edible seaweeds. 1
Liquid chromatography-inductively coupled plasma-mass spectrometry (LC-ICP-MS) has been a
powerful tool for arsenic speciation. Meanwhile, an efficient extraction method is necessary for
guaranteeing the total extraction of arsenic species. Ultrasound-assisted extraction (UAE) and
microwave-assisted extraction (MAE) have been widely used for the extraction of arsenic species.
However, these are methods that require relatively high-cost instrumentation. Matrix solid-phase
dispersion (MSPD) is a simple and low-cost method, based on sample maceration in the presence
of abrasive solid support, promoting the rupture of the physical structure of the sample. It is important
to point out that few studies have been reported in the literature using MSPD for extraction of
elements or metallic chemical species. Therefore, this work aims to evaluate the vortex-assisted
matrix solid-phase dispersion (VA-MSPD), to extract As(III), As(V), MMA, and DMA, from samples
of edible seaweed with further separation and determination by LC-ICP-MS. Parameters from VA-
MSPD were evaluated, such as the type of solid support (florisil, sea sand, alumina, diatomaceous
earth, silica, and maceration without solid support), solid support mass (0.25; 0.5; 0.75 and 1.0 g),
the type and concentration of extracting solution (0.01; 0.05; 0.1; 0.5; 1 mol L-1 HNO3, and ultrapure
water), and the type of heating (water bath at 50 °C for 10 min; ultrasonic bath for 10 min, and
ultrasonic probe for 5 min). Total As was determined in Nori, Kombu, and Hijiki algae using the ICP-
MS technique. To calculate the extraction efficiency, the samples were submitted to microwave-
assisted digestion (MAD) and the total As was determined by ICP-MS. The optimized conditions
were: 0.25 g of sample; 0.25 g of sea sand, 5 min of maceration, 10 mL of ultrapure water (as
extracting solution); mixture (sample and extracting solution) was placed in a water bath at 50 °C for
10 min; was vortexed for 1 min, centrifuged at 8000 rpm for 5 min, and the supernatant was filtered
with a syringe for total As determination by ICP-MS. The concentrations of total As obtained by ICP-
MS (after decomposition by MAD) were 18 ± 0.61, 69 ± 0.48 and 66 ± 4.55 µg g -1 for Nori, Kombu
and Hijiki algae, respectively. The extraction efficiencies were 103, 104 and 83% for Nori, Kombu
and Hijiki algae, showing the efficiency of the VA-MSPD/LC-ICP-MS method. The chromatographic
conditions were adapted to the work of Seege (2019). An anion exchange column (PRP-X100),
ammonium diphosphate 8.0 mmol L-1 was used as the mobile phase, and a flow rate of 1.5 mL min-
1
. Chromatograms with good resolution were obtained for both the reference solutions and the
samples. The method was linear (R2 > 0.99) in the range of 0.25 to 10 µg L-1. In the Nori seaweed,
the species As(III), DMA, MMA, and As(V) were detected in the range from 0.33 to 32.2 µg g -1. In
the Hijiki seaweed, As(III), DMA, MMA, and As(V) were identified. The concentrations ranged from
0.56 to 32.2 µg g-1. In the Kombu seaweed As(III), DMA, MMA, and an unknown species, which was
called UNK, were identified and the concentrations were from 0.47 to 32.6 µg g -1. The RDSs were
lower than 20%.
1 Reis, V. A. T.; Duarte, A. C. Trends in Analytical Chemistry; (102), 2018, 41.

2 Seeger, T. S. Dissertação. Santa Maria. 119, 2019.
[CNPq, FAPERGS, CAPES, FINEP]

MULTIRESIDUE DETERMINATION OF PESTICIDES IN BEES BY
MODIFIED QuEChERS METHOD AND UHPLC-MS/MS
Nathália Saibt(PG), José Dilson F. da Silva(PG), Martha B. Adaime(R),
Osmar D. Prestes(R), Renato Zanella(R)*
* e-mail: renato.zanella@ufsm.br
Brazil has favourable weather for extensive agricultural production, pushed by the huge demand for
food both on national and global scale. This demand, however, leads to the massive use of
pesticides. In 2020, 685,745,68 tons of pesticides were used, with the class of herbicides
representing 60.4% of all pesticides sold, fungicides account for 15.8% of the national market,
insecticides for 11.8% and other categories of pesticides for 12.1%.1 The pesticides are extensively
applied in agriculture to protect crops from different diseases, weeds, or insects, simultaneously
ensuring increased agricultural production. However, as is widely known, pesticides constitute a
group of contaminants of special concern. The detrimental effect of pesticides on bee health is a
topic of global concern.2 Since 2007, colony collapse disorder (CCD) has impacted honey bee
colonies in many parts of the world.2 Proposed explanations for the observed cases of CCD include
multiple stressors, such as reduced food availability, decreased flower diversity, parasites, modern
beekeeping practices, and continuous agrochemical exposure has often been on honey bee
exposure to different classes of pesticides.2 Honey bees can be contaminated with toxic substances
in many different ways, both during foraging flights and within the hive.3 They can come into contact
directly or indirectly with crop spraying treatments or dust dispersed during coated seed sowing, and
can ingest irrigation water, nectar and pollen contaminated.3 Therefore, the objective of this work
was to develop and validate a method for the determination of multi-residue of pesticides in bee
samples using ultra-performance liquid chromatography coupled to sequential mass spectrometry
(UHPLC-MS/MS). For sample preparation, the extraction solvents, partition salts and sorbents to
cleanup the extracts were evaluated. The method validated involves homogenization of the bee
samples with ultra-pure water, followed by extraction with acetonitrile containing 1%(v/v) formic acid,
partitioning with magnesium sulfate and sodium chloride and cleaning of the extract with magnesium
sulfate, PSA and C18. The obtained extract was diluted 1:2 (v/v) in ultrapure water, followed by the
addition of the internal standard and analysis in the chromatographic system. Pesticides were
analyzed using a XevoTM TQ-XS UHPLC-MS/MS triple quadrupole system (Waters, USA).
Separation was achieved in an Acquity UPLC® HSS T3 column (2.1 × 100 mm, 1.8 μm particle size).
The mobile phase consisted of water (phase A), and acetonitrile:methanol (1:1, v/v), both with 0.1%
(v/v) formic acid and 5 mM ammonium formate (phase B). The modified QuEChERS method proved
effective for the determination of 175 pesticides in bee samples (1 g). The quality and reliability of
the obtained results were warranted through the validation that ensured adequate precision and
accuracy. The linear determination coefficients of the calibration curves were ≥ 0.994, and the limits
of quantification were between 5 and 100 µg kg-1. The method showed excellent recovery ranging
from 70 to 120%, with RSD ≤ 18% for the spike levels 5, 10, 20, 50 and 100 µg kg -1 The proposed
method was applied to 16 samples of bees from different regions of the state of Rio Grande do Sul.
1 IBAMA, http://www.ibama.gov.br/relatorios/quimicos-e-biologicos/relatorios-de-comercializacao-de-agrotoxicos
2 Ardalani H, Vidkjær NH, Laursen BB, Kryger P, Fomsgaard IS, Chemosphere, 262, 2021, 127848
3 IBAMA, Manual de avaliação de risco ambiental de agrotóxicos para abelha, 2017
[CAPES, CNPq, FINEP]

ANALYTICAL METHOD DEVELOPMENT FOR PESTICIDE RESIDUES
DETERMINATION IN MANGO USING UHPLC-MS/MS
Franciele F. Machado (PG), Luana Floriano (PG), Martha B. Adaime (R),
Osmar D. Prestes (R), Renato Zanella (R)*
Mango is a fruit with high nutritional value besides the attractive flavor, aroma and exotic
appearance, has gained wide acceptance and led to its high demand in both domestic and export
markets, making it a product of increasing economic importance. To increase production yields and
obtain good quality fruits, pesticides are applied for pest and fungal control during the cultivation and
post-harvest stages.1 Inadequate agricultural practices for the use of these substances, however,
incur the presence of residues that, at levels above the maximum residue limits (MRL), can pose a
risk to consumer’s health. To monitor pesticides occurrence in the cultures, the optimal extraction
procedure is the one that allows the determination of a higher number of analytes, called a
multiresidue method.2 Thus, the aim of this study was to develop and validate an effective sample
preparation method for the determination of 60 pesticide residues in mango by ultra-high
performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS).
Chromatographic separation was performed using an Acquity BEH C18 column (50 mm x 2.1 mm,
1.7 µm) maintained at 40 °C and mobile phase consisted of an aqueous solution containing 2% (v/v)
of methanol (A) and methanol (B), both containing 0.1% (v/v) of formic acid and ammonium formate
5 mmol L-1. The flow rate was 0.225 mL min-1 with gradient elution. Nitrogen 6.0 (desolvation gas) at
600 L h−1 and argon 6.0 (collision gas) at 0.15 mL min−1 were used. The total run time was 10 min
and the injection volume was 10 µL. The preliminary tests were performed evaluating the three
QuEChERS versions (original, citrate and acetate). In addition, different sorbents such as C18, PSA,
GCB, activated carbon, Oasis® HLB and Strata™-X were evaluated in the dispersive solid-phase
extraction (d-SPE) step. The sample preparation procedure consisted of 10 g of sample and 10 mL
of acetonitrile, followed by vortexing for 1 min. Subsequently, unbuffered QuEChERS salts (4 g
MgSO4 and 1 g NaCl) were added to the tube, which was manually shaken during 1 min, and
centrifuged at 3500 rpm for 8 min. For the clean-up step, 2 mL of supernatant were transferred to a
tube containing 150 mg of magnesium sulfate and 50 mg of Strata™-X, a modified styrene
divinylbenzene sorbent. Then, the mixture was vortexed for 1 min and centrifuged at 3500 rpm for 8
min. The supernatant was filtered through a 0.22 μm nylon filter and diluted in ultrapure water (1:4,
v/v). Linearity of matrix-matched analytical curves was obtained from 0.5 to 20 µg L-1, with
determination coefficient (r2) > 0.99. Accuracy and precision were evaluated at the spike levels 5,
10, 25 and 50 µg kg-1, with recoveries from 70 to 120% and RSD ≤ 20%, for most of the compounds.
The method limits of quantification were between 5 and 10 µg kg-1 for most of the evaluated
compounds. The method was applied in 16 commercial samples, and the results showed the
presence of pesticide residues, such as azoxystrobin (6 mg kg -1, > Brazil MRL), and carbendazim
(forbidden for mango cultivation in Brazil).
1 Filho AM, Santos FN, Pereira PAP. Talanta, 81, 2010, 346-354.
2 Kemmerich M, Bernardi G, Prestes OD, Adaime MA, Zanella, R. Food Analytical Methods, 11, 2018, 556-577.
[UFSM, CAPES, CNPq]

DETERMINATION OF PESTICIDES RESIDUES IN ZUCCHINI USING
ACETONITRILE EXTRACTION AND UHPLC-MS/MS
Giovane S. dos Reis (PG), Martha B. Adaime (R), Osmar D. Prestes (R), Renato Zanella (R)*
In recent decades, the use of pesticides in Brazil and in the world has increased significantly, mainly
due to food demand. Brazil stands out in the international scenario, especially in the production and
cultivation of fruits, vegetables and grains, including zucchini, a food with high nutritional value, a
source of carbohydrates, vitamins and antioxidants1. Zucchini is among the ten most economically
valuable vegetables in the world with high annual productivity. Although it is necessary to employ
strategies aimed at increasing the quality and quantity of food produced, such as the use of
pesticides, the incorrect and indiscriminate use of these chemicals can have harmful effects on
human health and the environment. Thus, sanitary agencies and food inspection agencies establish
Maximum Residue Limits – MRLs for pesticides authorized for certain crops. Therefore, it is
necessary to develop multiresidue methods of pesticides to monitor these compounds in various
foods, including zucchini2. Thus, the objective of this work was to develop a method for the
multiresidue determination of pesticides in zucchini using the QuEChERS method and ultra-high
performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS).
Extraction consisted of 10 g of sample and 10 mL of acetonitrile. Partitioning was performed with 1
g of NaCl and 4 g of MgSO4, followed by manual shaking and centrifugation. The extract was diluted
(5x), filtered and analyzed by UHPLC-MS/MS. For the analyses, a Waters Acquity UPLC system
with a Xevo TQ mass spectrometer equipped with an electrospray ionization (ESI) source was used.
The chromatographic analysis was performed with Acquity BEH C18 (50 x 2.1 mm, 1.7 μm) column
with a mobile phase consisting of (A) aqueous solution containing 2% (v/v) methanol and (B)
methanol, both containing 0.1% (v/v) formic acid and 5 mmol L−1 ammonium formate at . The flow
rate was 0.225 mL min−1 and the total chromatographic run time was 10 min. The injection volume
was 10 μL and the column temperature was set at 40 °C. Nitrogen (desolvation gas) at 600 L h −1
and argon 6.0 (collision gas for MS/MS analyses) at 0.15 mL min−1 were used. Selected reaction
monitoring (SRM) was used as the acquisition mode, where the most abundant ion (1st transition)
was selected for quantification and the 2nd most intense transition for identification. The unbuffered
QuEChERS method without clean-up step was validated for 99 compounds. All the compounds
showed good linearity (r2 ≥ 0.990). Four spike levels (5, 10, 20 and 50 μg kg-1) in seven replicates
were evaluated and the recovery values ranged from 70 to 120% with RSD ≤ 20%, ensuring good
precision and accuracy. The method was applied in 20 zucchini samples from different cities of Rio
Grande do Sul, in which pesticide residues acetamiprid, azoxystrobin, carbofuran and imidacloprid
were found in concentrations from 6 to 84.2 µg kg-1. Based on these results, the proposed method
proved to be adequate for pesticide multiresidue determination and can be applied in routine
analysis.
1 Rodrigues RP, Farias WF, Goulart SM, Goulart AC, Santos JPV, Queiroz MELR, Química Nova, 41, 2018, 213.
2 Prestes OD, Adaime MB, Zanella R, Sci Chromatogr, 3, 2011, 51.
[UFSM, CNPq, CAPES]

METHOD DEVELOPMENT AND VALIDATION EMPLOYING SILLME AND GC-
MS/MS FOR ADSORPTION STUDIES OF PAHS IN BIOCHAR
Enedina S. A. Lima (UG)a, Anna C. N. Meireles (UG)a, Natiele Kleemann (PG)a,
Sergiane C. Barbosa (R)a, Ednei G. Primel (R)a, Jean L. O. Arias (R)a
aUniversidade Federal do Rio Grande - FURG, Laboratório de Análise de Compostos Orgânicos e Metais - LACOM, Rio
Grande, Rio Grande do Sul, Brazil, 96203-900
*e-mail: enedinasena.lima@gmail.com
Biochar refers to a material abundant in carbon, typically collected via the thermal treatment of
biomass, including sewage sludge. The physio-chemical characteristics of biochar, such as three-
dimensional reticulated and porous structure could serve as a lasting storage solution for carbon
while adsorbing and degrading pollutants1, such as Polycyclic Aromatic Hydrocarbons (PAHs). The
PAHs are a class of compounds with two or more condensed aromatic rings, resulting from the
incomplete combustion of organic, recognized as carcinogenic and mutagenic compounds2. To
evaluate the capacity of adsorption of PAHs in biochar, an analytical technique is required. In this
way, the Salting-out Induced Liquid-Liquid Microextraction (SILLME) is a technique that emerges as
an alternative method that follows the principles of Green Analytical Chemistry. Thus, this work
aimed at evaluating a method employing SILLME and Gas Chromatography Tandem Mass
Spectrometry (GC-MS/MS) for the determination of 16 priority PAHs in water for adsorption studies
in biochar. The extraction conditions were adapted from a previous method developed by our group3,
with modifications to adapt the extract for GC analysis. After optimization, the extraction was carried
out as follows: 5 mL water sample was transferred to a 15 mL polypropylene tube, then, 1 mL hexane
was added to the mixture, which was vortexed for 1 min. After that, 2 g MgSO 4 was added to the
mixture, which was agitated again, followed by centrifugation for 5 min at 7793 g. Finally, the upper
hexane layer was collected and transferred to a GC vial for further analysis, where 2 μL were injected
into the chromatographic system, whose conditions were adapted from a previous work2.
Experiments were performed in triplicate. The method was further validated, where the limits of
quantification (LOQ) were 1 μg L-1 to the sixteen PAHs under study. Linear ranges were from the
LOQ to 1000 μg L-1, with R2 higher than 0.99 for all the compounds. Recoveries were between 52 to
93%, with RSD ≤ 22%. Finally, the validated method was employed to evaluate the capacity of the
biochar from sewage sludge in the adsorption of PAHs. For this purpose, water samples were
fortified with the PAHs standard at the concentration of 100 μg L-1, being submitted to different
conditions of temperature, amount of biochar, and time. After that, aliquots of the water sample were
submitted to the validated SILLME method. Firstly, experiments were carried out under different
concentrations of PAHs (from 20 to 200 µg L-1) and temperatures (from 25 to 55 °C), with a large
amount of biochar fixed. A quick reduction of PAHs was observed, up to 100%. Thus, smaller
amounts of biochar (0.1 and 1.0 g) were evaluated at the initial concentration of 100 µg L-1, collecting
aliquots at different moments (from 30 seconds to 30 minutes). It was observed the reduction of the
concentration of PAHs over time, especially when 1.0 g of biochar was used. Finally, the temperature
(from 25 to 55 °C) and amount of biochar (from 0 to 4.0 g) were evaluated, and, similarly, reductions
close to 100% were observed. The analytical method employing SILLME and GC-MS/MS was
successfully applied to the determination of PAHs in water samples to perform adsorption studies in
biochar of sewage sludge. It was proven that the biochar has a high capacity for adsorption of PAHs
on its structure by using a simple method that follows the principles of Green Analytical Chemistry.
1 Qiu M, Liu L, Ling Q, Cai Y, Yu S, Wang S, Fu D, Hu B, Wang X, Biochar, 4, 2022, 19
2 Escarrone ALV, Caldas SS, Furlong EB, Meneghetti VL, Fagundes CAA, Arias JLO, Primel EG, Food Chemistry, 146,
2014, 597
3 Arias JLO, Rocha CB, Kupski L, Barbosa SC, Primel EG, Food Analytical Methods, 14, 2021, 1625
[FAPERGS, CNPq, FINEP, CAPES, CIA-FURG]

MULTI-CLASS METHOD FOR DETERMINATION OF ANTIMICROBIAL
RESIDUES IN CHICKEN MUSCLE USING SPE AND UHPLC-MS/MS
Luana Floriano (PG), Daniela K. Muenchen (PG), Pimpernelli J. dos Santos (PG)
Martha B. Adaime (R), Renato Zanella (R), Osmar D. Prestes (R)*
Chicken meat is one of the most consumed products globally due to its excellent nutritional value
and low production costs.1 Brazil is currently the leading broiler meat exporter, exporting 4.6 million
tons to over 150 countries in 2021; it also plays a significant role as the third-largest producer, only
behind the United States and China.2 Antimicrobials have been widely used in the prevention and/or
treatment of diseases within the broiler supply chain in order to maintain productivity. The
indiscriminate use of these veterinary drugs can, however, lead to the presence of residues in the
edible tissues of the treated animal, and thus pose risks to the consumer’s health. Therefore, the
development of analytical methods that allow the monitoring of these residues is very importance to
ensure food safety.3 In this work, a method was developed and validated for the multi-class
determination of 31 antimicrobials from different classes (including quinolones, sulfonamides,
tetracyclines, lincosamides, macrolides, pleuromutilins, and diaminopyrimidine derivatives) in
chicken muscle by ultra-high performance liquid chromatography coupled to tandem mass
spectrometry (UHPLC-MS/MS). All sorbents tested during sample preparation were evaluated in
terms of phospholipids removal efficiency. Furthermore, gas chromatography analysis was
performed in full scan mode to estimate the amount of the residual compounds in the extracts after
the three better cleanup procedures. The extraction step was carried out using 2 g of sample,
ultrapure water and acetonitrile acidified with oxalic acid, followed by stirring and centrifugation.
Afterwards, a clean-up step was performed using solid-phase extraction (SPE) in the pass-through
mode, in which an aliquot of supernatant was eluted directly through a Bond Elut C18 cartridge. The
proposed method was validated according to the Commission Implementing Regulation (EU)
2021/808. Two different protocols were found to be necessary to determine the unauthorised
substances in the lowest calibration level (LCL) and the substances with established maximum
residue limits (MRL). For substances with a MRL ≥ 100 μg kg -1, the extract was diluted 5 times in
ultrapure water. For unauthorised substances and for trimethoprim (MRL= 50 μg kg -1), 1 mL of the
extract was evaporated to dryness and reconstituted in water:acetonitrile (80:20, v/v). The validation
parameters evaluated were selectivity, matrix effect, linearity, trueness (obtained from recovery),
precision and decision limit. The procedures were performed using spiked blank matrix calibration
to compensate for the matrix effects (between -20% and +152%). All compounds showed linearity
in the working ranges evaluated, either from 0 to 2.0 times the MRL for permitted substances or from
0 to 4.0 times the LCL for unauthorised substances, with coefficients of determination greater than
0.97. The trueness, assessed at levels 0.5; 1.0 and 1.5 times the LMR; and 1.0; 2.0 and 3.0 times
the LCL, proved to be adequate, with recoveries between 90 and 117%. The method was applied to
22 chicken muscle samples collected in local supermarkets and 2 samples obtained directly from
farmers. Oxytetracycline was detected in one chicken sample, at a concentration below the lowest
calibration level for this compound (<25 µg kg−1). Despite these results, it is important to maintain
antimicrobial monitoring in a higher number of samples to guarantee the consumers’ food safety and
health protection.
1 Landoni MF, Albarellos G. The Veterinary Journal, 205, 2015, 21-27.
2 ABPA. Annual Report from the Brazilian Association of Animal Protein, 2022.
3 Khaled A, Gionfriddo E, Acquaro V, Singh V, Pawliszyn J. Analytica Chimica Acta, 1056, 2019, 34-46.
[CAPES, CNPq, UFSM]

MULTIRESIDUE METHOD FOR PESTICIDES DETERMINATION IN URINE
USING ACETONITRILE EXTRACTION AND LC-MS/MS
Marisa Demarco (PG), Leonardo R. Amaral (PG), Rômulo de O. Schwaickhardt (PG),
Luana Floriano (PG), Martha B. Adaime (R), Renato Zanella (R), Osmar D. Prestes (R) *
Pesticides have been widely used in food crops to control pests, weeds, fungi, etc. The indiscriminate
use of these organic compounds has contributed to the increase in cases of intoxication in humans,
which can lead to more serious health problems, becoming a case of public health. 1 Biological
samples, such as urine, are more common and easy to collect, in addition to providing recent
information on exposure to pesticides.2 In this sense, the use of methods that are capable of
determining different classes and the largest number of compounds in a single procedure are of
paramount importance to guarantee a more comprehensive result. Within this perspective, this work
developed and validated a method for multiresidue determination of pesticides in urine using the
unbuffered QuEChERS method and analysis by liquid chromatography coupled to tandem mass
spectrometry (LC-MS/MS). The parameters that influence the extraction efficiency were studied and
after finding the best extraction conditions, the method was validated. The proposed procedure
consisted of extracting 10 mL of urine (pH 5) with 2 mL of acetonitrile and the salting-out effect was
achieved by adding 4 g of MgSO4 and 1 g of NaCl. Afterwards, a cleanup step was carried out using
dispersive solid phase extraction, in which an aliquot of the supernatant was transferred to a tube
containing 150 mg of magnesium sulphate, 25 mg of primary-secondary amine and 25 mg of C18.
Subsequently, the extract was diluted in water (1:4, v/v) and filtered in PTFE 0.22 µm. Analyses were
performed by 6470 LC/TQ system (Agilent Technologies, EUA) employing electrospray ionisation in
positive and negative mode (ESI±) and selected reaction monitoring (SMR) acquisition. Separation
was carried out with a InfinityLab Poroshell HPH-C18 analytical column (150 x 3 mm; 2.7 µm)
maintained at 35 ºC and the mobile phase consisted of water (A) and methanol (B), both containing
5 mmol L-1 of formate ammonium and 0.1% (v/v) formic acid at a constant flow rate of 0.4 mL min -1.
The injection volume was 4 µL and the total analysis time was 20 min. The unbuffered QuEChERS
method was validated for 202 pesticides from different classes, proving to be selective and robust.
Linearity was evaluated using matrix-fortified calibration from 0.5 to 20 µg L-1 and showed r2 > 0.99
for all evaluated compounds. LOQs ranged from 0.5 to 5 µg L-1 (0.5 µg L-1 for 185 compounds) and
the LOD from 0.15 to 1.5 µg L-1 with adequate accuracy (70 to 120%) and precision (RSD ≤20%). In
the analysis of 50 urine samples collected from agricultural workers, residues of three neonicotinoids
pesticides were detected with concentrations ranging from < LOQ to 4.43 µg L-1. Considering the
results obtained, the use of the unbuffered QuEChERS method combined with LC-MS/MS allowed
the determination of a wide range of analytes with high selectivity and sensitivity, proving to be
efficient for multiresidue determination of pesticides in urine.
1 Béranger R, Hardy EM, Dexet C, Guidner L, Zaros C, Nougadère A, Metten MA, Chevrier C, Appenzeller BMR.
Environment International,120, 2018, 43-53.
2 Li P. et al. Chinese Journal of Analytical Chemistry, 45, 2017, 16481654.
[UFSM, CAPES, CNPq]

COMPARISON OF DIFFERENT SORBENTS FOR THE DETERMINATION OF
PESTICIDE RESIDUES IN MEAT BY GC–MS/MS
Pimpernelli J. dos Santos (PG)a, Sónia M. V. S. Cardoso (PG)a, Giséle P. Rosa (UG)a,
Mariana Baptistão (R)b, Martha B. Adaime (R)a, Renato Zanella (R)a, Osmar D. Prestes (R)a*
aLaboratory of Pesticides Residue Analysis (LARP), Chemistry Department, Federal University of Santa Maria,
bAgilent Technologies Inc., Barueri, São Paulo, Brazil, 06455-000
Meat is an important source of nutrition for many people worldwide. 1 Some pesticides can be used
to control pests and diseases in livestock and the animals can be contaminated by drinking water
and eating straw, grass, or grains.2,3 However, the application of pesticides in livestock raised the
concern about the presence of these compounds in food chain. Authorities around the world, 4
establish maximum residue limits (MRL) and good agricultural practices as strategies to protect the
public health. Furthermore, the development of analytical methods for determination of pesticide
residues in food, including bovine meat, is crucial for food safety. Considering the presence of lipid
and protein contents in bovine meat, a reliable and consistent sample analysis requires an efficient
matrix clean-up procedure. In this study, we evaluated three different sorbents to use as pass
through clean-up cartridges (Bond Elut C18, Bond Elut NH2 and Captiva EMR-Lipid) for 72
pesticides determination in bovine meat by GC-MS/MS. The sample preparation was based on a
simple solid-liquid extraction. The bovine meat was grounded and then, a portion was weighed into
50 mL polypropylene tubes. Acetonitrile was added for simultaneous protein precipitation and
pesticides extraction. An ultra-turrax homogenizer (10000 rpm) was used for better interaction of
solvent-meat samples. After, the centrifugation step was performed and the supernatants were
collected for the cleanup experiments. The GC-MS/MS analysis was performed using the Agilent
Intuvo 9000 coupled to 7010B triple quadrupole mass spectrometer. A planar capillary column HP-
5 MS UI (30 m x 0.25 mm x 0.25 µm), 1 µL injection volume (splitless mode) and helium as carrier
gas were employed. High efficiency source (HES) was used with electron source and dynamic MRM
(dMRM) as acquisition mode. Three spiking levels (10, 20, and 50 μg kg -1) of bovine meat samples
were evaluated in terms of recovery and relative standard deviation (RSD). Overall, using Captiva
EMR-Lipid for cleanup, recoveries for 56 pesticide residues from 62 to 119% with RSD ≤ 16% were
obtained. Peak shape and signal-to-noise (S/N) of pesticides were evaluated. Captiva EMR–Lipid
provided the highest S/N and the best symmetrical peak shape for most compounds when compared
to the other sorbents. A comparison of limit of quantification (LOQ) demonstrated that the sorbent
Captiva EMR-Lipid showed LOQs of 10 μg kg-1, value below the MRLs for most pesticides
determined by EU and Brazil regulations. Therefore, it was concluded that Captiva EMR-Lipid
showed better performance than the other clean-up sorbents.
1 Han L, Sapozhnikova Y, Lehotay SJ, Food Control, 66, 2016, 270.
2 Kang HS, Kim M, Kim EJ, Choe W-J, Food Science and Biotechnology, 29, 2020, 1573.
3 Souza R, Pareja L, Cesio MV, Heinzen H, Chromatographia, 79, 2016, 1101.
4Zhang H, Wang J, Li L, Wang Y, Food Analytical Methods, 10, 2017, 1826.
[UFSM, CNPq, CAPES, Agilent Technologies]

HALOGEN DETERMINATION IN EDIBLE INSECTS AFTER MICROWAVE-
INDUCED COMBUSTION: TOWARDS NEW NUTRITION SOURCES
Emanuele Frozi (UG)a, Vitoria H. Cauduro (PG)a, Alessandra S. Henn (PG)a,
Cristian R. Andriolli (PG)a, Thiago Castanho (PG)a, Érico M. M. Flores (R)a*
aUniversidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil
Research interest has increasingly been turned to the potential of edible insects to integrate human
diet, due to their high protein content.1 However, research is still lacking in terms of nutrient and
impurity determination in these alternative protein sources. In particular, there is no report of halogen
levels in this matrix.2 Due to the difficulty of halogen determination, it is necessary to develop
adequate sample preparation methods and detection techniques. In this sense, microwave-induced
combustion (MIC) is a promising method, as it allows the use of relatively high sample mass and
high temperatures are achieved (up to 1500 °C). Additionally, a reflux step can be applied, improving
analyte recovery into the absorbing solution.2 Hence, this study aimed to develop a MIC method for
edible insect decomposition for further halogen determination by inductively coupled plasma mass
spectrometry (ICP-MS). For the development of this study, insect samples of different species and
geographical origins were used, namely mealworm larvae (Tenebrio molitor) from 2 different origins
(Mealworm A and B), cricket (Acheta domesticus) from 3 different origins (cricket A, B and C), scarab
(Phyllophaga sp.), and grasshopper (Oxya yezoensis sp.). Samples were pressed as pellets using
a hydraulic press (3 tons for 1 min). Pellets were weighted and placed in the quartz holder containing
a filter paper wetted with 50 μL of igniter (6 mol L-1 NH4NO3). The holders were then introduced into
the reaction flasks containing 6 mL of absorbing solution. Samples were combusted in closed quartz
vessels pressurized with oxygen (20 bar), using a 5 min reflux step. Microwave assisted extraction
(MAE), was used as the reference method, using 200 mmol L-1 tetramethylammonium hydroxide
(TMAH) as extractant and a 50 min irradiation program.3 Sample cricket A was arbitrarily chosen for
method optimization. Parameters such as sample mass (100 to 600 mg) and absorbing solution
(NH4OH 25 to 200 mmol L-1 or ultrapure water) were evaluated, and results were compared to the
values obtained by the reference method. Method accuracy was evaluated through standard addition
experiments performed at two levels (50% and 100% of the sample concentration), as well as
decomposition of a standard reference material (DORM-5 fish protein). Additionally, stability of the
digests were evaluated for three weeks. The optimized condition was set using 500 mg of sample
and NH4OH 100 mmol L-1 as the absorbing solution. Quantitative recovery of Cl, Br and I (ranging
from 94% to 102%) obtained for standard addition experiments at both levels. Results obtained for
DORM-5 using the proposed MIC method showed no significant difference to certified values
(Student’s t test, 95% of confidence) and halogen concentration on the digests remained stable
during the whole stability test. By using the proposed MIC method for insect decomposition, it was
possible to determine Cl, Br, and I at trace levels, with agreement ranging from 95 to 105% with the
reference method. In comparison with MAE, the proposed method allowed lower relative standard
deviations (RSDs) and blank values. Additionally, MIC was significantly faster than the reference
method, and could be performed using a much safer reagent, also resulting in digests with lower
carbon content. Hence, the proposed MIC method can become a powerful tool for further halogen
determination in edible insects by ICP-MS.
1 Elhassan M, Wendin K, Olsson V, Langton M, Foods, 8, 2019, 95.

2 Mello PA, Barin JS, Duarte FA, Bizzi CA, Diehl LO, Muller EI, Flores EMM, Analytical And Bioanalytical Chemistry, 24,
2013, 405.
3 Novo DLA, Pereira RM, Henn AS, Caldeira V, Flores EMM, Mesko MF, Analytica Chimica Acta, 1060, 2019, 45-52.
[CNPq, FAPERGS, CAPES, UFSM]

MULTI-TECHNIQUE APPROACH FOR FEASIBLE HALOGENS SPECIATION
IN ANTARTIC SEAWEED
Filipe S. Rondan (PG)*a, Fernanda P. Balbinot (PG)a, Larissa C. A. Costa (PG)a,
Catarina F. S. Moraes (UG)a, Gabriela de Oliveira (UG)a, Paulina Pisarek (R)b,
Joanna Szpunar (R)b, Marcia F. Mesko (R)a.
aUniversidade
Federal de Pelotas, Centro de Ciências Químicas, Farmacêuticas
e de Alimentos, Pelotas, RS, Brazil, 96160-000
bCNRS/UPPA, Laboratoire de Chimie Analytique Bio-inorganique et Environnement, Hélioparc, 2, Pau, France
*e-mail: fsrondan@gmail.com
Given the importance of the Antarctic continent regarding to environmental variations, the knowledge
about chemical composition of the typical fauna and flora from that region, such as seaweeds,
becomes of great relevance, mainly with regard to climate changes. Since the chemical information
about Antarctic seaweeds reported in the literature is limited to a few organic species 1 and metals2,
the need for investigate the presence of halogens and their species on these organisms is
highlighted.3 In this work, several analytical methods were developed using a multi-technique
approach, using different sample preparation methods based on extraction and combustion, and
high sensitivite determination techniques. In this sense, comprehensive information about total
halogens concentration, distribution of bromine and iodine in different seaweed fractions (lipids,
water-soluble, proteins, carbohydrates, and residue) and several halogenated chemical species in
brown and red seaweed (ten species) is provided. Regarding the total halogens content in the
samples, the method was proposed using the microwave-induced combustion (MIC) as sample
preparation and the ion chromatography coupled to mass spectrometry (IC-MS) as determination
technique. On the other hand, for the halogen fractionation, a sequential extraction was employed
for sample preparation, using petroleum ether, water, driselase, protease, and sodium dodecyl
sulfate to separate the lipid, water-soluble, polysaccharide, and protein fractions, respectively. The
total Br, Cl, and I determination in the different fractions was also performed by MIC and IC-MS. For
the speciation of bromine and iodine in water-soluble fraction, the water extracts were analyzed by
size-exclusion chromatography coupled to inductively coupled plasma mass spectrometry (SEC-
ICP-MS) technique. Bromate was not present in any of the seaweed water extracts, while bromide
was detected in all samples. In all the brown seaweed and in one red seaweed were also detected
signals of unknown bromine species. Similarly, iodate was not detected in any seaweed sample, at
the same time that iodide was detected in all evaluated samples. Halogenated polyphenols were
determined by high performance liquid chromatography with electrospray ionization tandem mass
spectrometry (HPLC-ESI-MS/MS), after sample preparation using MeOH and NH4OH extractions.
Twenty-five halogenated polyphenols were identified in almost all samples, with four of them
reported in the literature. Finally, for the speciation of bromine and iodine reverse-phase HPLC
coupled to ICP-MS (RP-HPLC-ICP-MS) and anion-exchange HPLC coupled to ICP-MS (AE-HPLC-
ICP-MS) were used as determination techniques after sample preparation using pancreatin
extraction. Bromide and iodide were quantified in all seaweed species, while neither bromate nor
iodate were present in the extracts. Furthermore, iodinated amino acids, such as monoiodotyrosine
(MIT) and diiodotyrosine (DIT), were identified and quantified in the samples, in addition to unknown
bromine and iodine species.
1 Laturnus F Environmental Science and Pollution Research, 8, n. 2, p. 103-108, 2001.
2 Picoloto RS et al. Journal of Applied Phycology, 29, n. 2, p. 741-749, 2017.
3 Al-Adilah H et al. Phycology, 2, n. 1, p. 132-171, 2022.
[CNPq, CAPES, FAPERGS, UFPel, UPPA]

MICROWAVE-INDUCED COMBUSTION OF EDIBLE INSECTS SAMPLES FOR FURTHER As, Cd AND Pb
DETERMINATION BY ICP-MS
Gustavo Gohlke (UG)a, Thiago C. Pereira (PG)ª, Cristian R. Andriolli (PG)ª,

Vitoria H. Cauduro (PG)a,b, Alessandra S. Henn (R)b, Érico M. M. Flores (R)a*
a Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil, 97105-900
b Universidade Tecnológica Federal do Paraná, Campus Medianeira, Medianeira, PR, Brazil, 85884-000
Edible insects have been an alternative to the increasing demand for food. The breeding of these
arthropods demonstrates productive and environmental advantages such as lower water
consumption and reduction in the emission of pollutant gases when compared, for example, to
cattle.1 However, potentially toxic metals (such as As, Cd, and Pb), may be present in these insects
due to the substrate used in the production.2 Therefore, analytical methods for the determination of
As, Cd, and Pb in edible insects have to be developed to ensure consumers health and safety.
Inductively coupled plasma mass spectrometry (ICP-MS) is an analytical technique that presents
multielemental capabilities and low limits of quantification (LOQ). However, when operating with the
conventional sample introduction system, the ICP-MS equipment requires samples in a liquid form.
Therefore, a previous sample preparation step is required. Microwave-induced combustion (MIC) is
a sample preparation method that stands out for using dilute solutions and results in digests with low
residual carbon content when compared to conventional acid digestion methods.3 Therefore, this
work evaluates the use of MIC for the sample preparation of edible insects for further determination
of As, Cd and Pb by ICP-MS. For this, samples of edible insects (cricket, beetle, grasshopper, and
tenebrio molitor larva) were dried, ground and pressed into pallets for combustion. The maximum
sample mass (100 to 600 mg), the concentration of HNO3 used as absorbing solution (3 to 14.4 mol
L-1) and the reflux step time (5 to 15 minutes) were evaluated. Microwave-assisted acid digestion in
a single reaction chamber (MAWD-SRC) was used as a reference method. For MAWD-SRC a
sample mass of 300 mg and 6 mL of 14.4 mol L-1 HNO3 were used. In order to avoid interferences,
the residual carbon content of solutions obtained by both methods was determined by inductively
coupled plasma optical emission spectrometry (ICP-OES). Masses of 500 mg of cricket were
combusted by MIC without exceeding the maximum pressure of the system. Recoveries higher than
90% were obtained for As, Cd and Pb using 7 mol L-1 HNO3 as absorbing solution and a reflux time
of 5 minutes. The residual carbon content of solutions obtained after MIC was lower than
13,6 mg L-1, while in the MAWD-SCR (reference method) the concentration was about 150 mg L-1.
No interference was observed during ICP-MS measurements for both methods. There was no
significant difference (t-test, 95% confidence level) between results obtained by MIC and the
reference method for As, Cd, and Pb in all evaluated samples. Therefore, the MIC method proved
to be a promising sample preparation method for edible insects and further As, Cd and Pb
determination, presenting lower carbon content values, in addition to allowing the use of diluted
solutions and higher sample mass.
1 Silva LAJ. et al., Food Chemistry 311, 2020,126022.
2 EFSA Scientific Committee, EFSA Journal 13, 2015, 4257.
3 Flores EMM et al., Analytical Chemistry 76, 2004, 3525.
[CNPq, FAPERGS, CAPES]

GREEN SAMPLE PREPARATION METHOD USING REVERSED-PHASE
DISPERSIVE LIQUID-LIQUID MICROEXTRACTION FOR MULTIELEMENTAL
DETERMINATION IN EDIBLE COCONUT OIL
Leandra M. Prado (UG)b, Vitoria H. Cauduro (PG)b, Daneysa L. Kalschne (R)a,
Paulo R. S. Bittencourt (R)b, Oldair D. Leite (R)b, Eder L. M. Flores (R)b, Cristiane Canan (R)a*.
a Universidade Tecnológica Federal do Paraná – Campus Medianeira, Programa de Mestrado em
Tecnologia de Alimentos, Medianeira, Paraná, Brazil, 85884-000
b Universidade Tecnológica Federal do Paraná – Campus Medianeira, Departamento de Química,
Medianeira Paraná, Brazil, 85884-000

*e-mail: canan@utfpr.edu.br
Edible coconut oil is a versatile food, widely used in cooking as well as a nutraceutical, with numerous
health benefits. However, contamination with potentially toxic elements can occur in coconut oil from
the use of fertilizers, exposure of soil to environmental contamination and from the extracting and
refining processes of the oil. In this sense, the Brazilian Health Regulatory Agency (ANVISA) and
international agencies such as FDA and CE establish maximum limits for various metals in oily or
fatty foods. Coconut oil can be in liquid or solid states depending on its composition and ambient
temperature, which makes its handling difficult during sample preparation procedures, in addition to
the matrix being considered quite complex when using methods based on wet acid digestion in
closed flasks. In this study, a reverse-phase dispersive liquid-liquid microextraction method (RP-
DLLME) was proposed for the microextraction and pre-concentration of metals in coconut oil
samples. The analytes were determined by inductively coupled plasma optical emission
spectrometry (ICP-OES). In an innovative way, the green solvent d-limonene was first added in the
preparation of the samples, aiming at maintaining the liquid phase. Then, a mixture of two solvents,
a dispersant and an extractant (diluted HNO3), was added to the tube containing coconut oil sample
diluted with d-limonene (1:1, m/m), already heated at 70 °C. The samples were then shaken and
centrifuged (6000 rpm for 10 min) to extract the aqueous phase. The extracted/pre-concentrated
analytes (Al, Ba, Cd, Cu, Fe, Mg, Mn, Mo, Ni, Pb, Ti, V and Zn) were determined by ICP-OES using
external calibration with aqueous reference solutions. For the optimization of the method, the
following parameters were evaluated: type of dispersing solvent (ethanol, n-propanol or
isopropanol); proportion of dispersant:extractant solvents (50:50, 60:40, 70:30, 80:20, v/v), and
shaking time. The sample mass, and concentration and volume of extracting and dispersing
solvents, were also evaluated. The optimized RP-DLLME conditions were 5 g of sample diluted in 5
g of d-limonene, 1.0 mL of the dispersant:extractant solution consisting of n-propanol and HNO3 (1.0
mol L-1) in the proportion of 50:50 (v/v), and 1 min of shaking prior to centrifugation. The proposed
method resulted in analyte recoveries ranging from 81 to 106% (RSD < 6%). The obtained LOQs
and LODs for all analytes were lower than similar methods from the literature, proving the proposed
RP-DLLME method to be suitable for trace metal determination in different types of commercial
edible coconut oil samples (both liquid and solid products). The evaluation of the greenness of the
proposed RP-DLLME method was performed using the AGREE® software to establish Green
Chemistry metrics. The developed method was considered suitable for the determination of trace
metals in edible coconut oil samples, following the Green Analytical Chemistry Principles.
Additionally, the proposed RP-DLLME method is characterized by quick and simple operation, low
cost, possibility to use high sample masses, low consumption of reagents, less waste generation
and high analytical frequency. Finally, the proposed analytical method was viable, and this is the first
application of RP-DLLME for coconut oil combined with ICP-OES determination.
1 Kalschne DL; Canan C; Barin, JS; Picoloto RS; Leite OD; Flores ELM. Food Analytical Methods, 13, 2019, 230.
2 Kalschne DL; Canan C; Beato MO; Leite OD; Flores, ELM. et al. Talanta, 208, 2019, 120409.
3 Pena-Pereira F; Wojnowski W; Tobiszewski M; Analytical Chemistry, 92, 2020, 10076.
[CEANMED/UTFPR - Campus Medianeira, CAPES]

ULTRASOUND-ASSISTED EXTRACTION FOR FURTHER TOXIC ELEMENTS
DETERMINATION IN MARINE SEAWEED
Luana F. Rocha (UG)a, Gustavo Gohlke (UG)a, Emanuele Frozi (UG)a,
Giancarlo R. Machado (UG)a, Thiago C. Pereira (PG)a, Vitoria H. Cauduro (PG)a,b,
Alessandra S. Henn (R)b, Érico M.M. Flores (R)a*
a Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil, CEP 90105900
b Universidade Tecnológica Federal do Paraná, Campus Medianeira, Medianeira, PR, Brazil, 85884-000
*e-mail: ericomm@gmail.com
Marine algae have been extensively studied due to their bioremediation capacity. However, it is
essential to know the characteristics of the aquatic ecosystem and the interaction between
contaminants and algae.1 To correctly monitor the distribution and accumulation of contaminants in
the environment, it is necessary to apply analytical methods for elemental determination. These
methods can provide reliable results which can be used in routine procedures without generating
large volumes of residues. Among analytical techniques for metals determination are inductively
coupled plasma optical emission spectrometry (ICP-OES) and inductively coupled plasma mass
spectrometry (ICP-MS).2 However, these techniques require the sample in the form of a solution
containing the analytes. Therefore, a sample preparation step is required in order to convert the
sample matrix into a solution containing the analytes. Among the main sample preparation methods
used for the subsequent determination of metals in organic samples, such as seaweed, are those
based on the wet digestion using conventional energy sources (heating plates and digestion blocks)
or microwave heating. However, these methods present disadvantages such as the use of
concentrated acids, long time for digestion and low sample throughput.3 The use of extraction
methods based on ultrasound energy can be considered as promising alternatives for further
elemental determination.4 Thus, the objective of this work was to develop an ultrasound-assisted
extraction (UAE) method for further As, Cd and Pb determination in seaweed by ICP-MS. Initially,
the most suitable ultrasonic system was evaluated: bath systems (25 to 45 kHz, 37 to 80 kHz, and
37 to 180 kHz), probe systems (20 kHz and 750 W power and 20 kHz and 130 W power), and cup
horn systems (20 kHz and 750 W power and 20 kHz and 130 W power). Moreover, method
parameters such as the amplitude (30 to 90%), extraction solution (H2O and diluted HNO3 solutions),
sample mass (50 to 500 mg), extraction time (10 to 60 min), temperature (25 to 90°C) and extraction
solution volume (5 to 20 mL) were evaluated. Results obtained by UAE were compared to results
obtained after wet digestion (reference method). Agreement with the reference method was better
than 90% by using the following conditions: 25 kHz ultrasonic bath system, 70% of amplitude, 200
mg of sample, 70 °C of temperature, 10 mL of 2 mol L-1 HNO3 as extracting solution, and 30 minutes
of extraction. The UAE method was considered suitable for further As, Cd and Pb determination in
seaweed, being a promising alternative for monitoring the distribution and accumulation of
contaminants in the environment.
1 Vidotti, EC, Rollemberg MC, Quimica Nova, 27, 2004, 139.

2 Flores, EMM., Microwave-assisted sample preparation for trace element determination, Elsevier, 1st ed., Amsterdam,
2014.
3 Krug FJ, Rocha FRP., Métodos de Preparo de Amostras Para Análise Elementar, 3ª ed., São Paulo, 2016.
4Mesko MF, Costa, V.C, Picoloto, R.S, Bizzi, C.A, Mello, P.A, Journal of Analytical Atomic Spectrometry, 31, 2016, 1243.
[CNPq, FAPERGS, CAPES]

DEVELOPMENT OF GREEN METHODS FOR THE DETERMINATION OF
ELEMENTAL IMPURITIES IN COMMERCIAL PHARMACEUTICAL TABLETS
Ana P. F. Padilha (UG)a, Vitoria H. Cauduro (PG)a, Alessandra S. Henn (R)a ,
Rochele S. Picoloto (R)a, Edson I. Muller (R)a, Marcia F. Mesko (R)b, Erico M. M. Flores (R)a
aUniversidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil
b Universidade Federal de Pelotas, Centro de Ciências Químicas, Farmacêuticas e de Alimentos, Pelotas, RS, Brazil
For prolonged use of pharmaceuticals, such as those used to treat type 2 diabetes,1 quality control
concerning determination of elemental impurities is extremely necessary. In this sense, the ICH 3QD
guidelines2 separates elemental impurities different classes according to toxicity, route of
administration and abundance. For oral intake, Class 1 (As, Cd, Hg e Pb) and Class 2A (Co, Ni e V)
elements are most critical and should always be monitored.2 Plasma–based techniques, such as
inductively coupled plasma optic emission spectrometry (ICP-OES) allow sensitive detection of
these impurities. However, the sample needs to first go through a pre-treatment, usually performed
using concentrate reagents, which can result in high acidity in the digests, matrix effects, and
consequently high LODs.3 To overcome these limitations while maintaining digestion efficiency, the
aim of the present study was to develop sample preparation methods based on the use of diluted
acids, namely microwave-assisted wet digestion (MAWD) and microwave-assisted ultraviolet
digestion (MAWD-UV), for further elemental impurity determination in oral drugs by ICP-OES. Only
a small volume of diluted HNO3 and H2O2 were used as digesting solution. Six samples of
commercial tablets for type 2 diabetes were used, with no prior comminution, containing the active
ingredients metformin hydrochloride (MET), glibenclamide (GLIB), pioglitazone hydrochloride (PIO),
sitagliptin phosphate (SITA), canagliflozin (CANA) and repaglinide (REPA). Samples MET and
CANA were used for method optimization. For the MAWD method, the concentration of the digestion
solution (1-14.4 mol L-1 HNO3), the irradiation program (45 or 55 min), the simultaneous cooling
during irradiation and use of H2O2 as auxiliary reagent (8.3 and 16.7%) were evaluated. The
optimized conditions were 2 mol L-1 (or 3 mol L-1, CANA) HNO3, 8.3% H2O2, and a 45 min (or 55 min,
CANA) program. For the MAWD-UV method, digestion conditions were evaluated taking into
consideration the conditions in which a complete digestion was not observed for MAWD. Using the
MAWD-UV method, all samples could be decomposed using 1 mol L-1 HNO3, 8.3% H2O2 and the 55
min program. Accuracy of the methods was evaluated by standard addition experiments at the
specified permitted daily exposure (PDE) for each analyte considering the tablet with the highest
daily intake, and by decomposition of certified reference materials (CRMs) of dogfish liver (DOLT-
4), lobster hepatopancreas (TORT-2) and aquatic plant (BCR-60) mixed with sample matrix (GLIB
sample was randomly chosen for this experiment). Analyte recovery ranged from 90% to 109%
(MAWD), and from 92 to 110% (MAWD-UV) for all samples, with relatively low carbon and residual
acidity. Also, analyte determination in CRMs were in agreement with certified values. Hence, both
the proposed MAWD and MAWD-UV methods are suitable for decomposition of oral use drugs for
further elemental impurity determination by ICP-OES. Furthermore, considering the ICH Q3D limits,
the obtained LOQs for both methods were at least two times lower than the PDE for all analytes.
Thus, the proposed method allowed efficient digestion of commercial oral drugs for further
determination of elemental impurities using diluted reagents and no prior comminution, reducing the
risks associated with the use of concentrated acids and causing less waste generation.
1 Kahn, CR. Insulin action, diabetogenes, and the cause of type II diabetes. Diabetes 1994, 43, 1066–1085.
2 ICH Guideline for Elemental Impurities Q3D(R1).
3 Bizzi, CA, Flores, ELM, Nobrega, JÁ, Oliveira, JSS, Schmidt, L, Mortari, SR, Journal of Analytical Atomic Spectrometry,
29, 2014, 332–338.
[CNPq, CAPES, FAPERGS]

HOW COOKING METHODS AND UTENSILS AFFECT THE RICE MINERAL CONTENT
Iago J. S. da Silva (R)a,*, Micherlane M. da Silva (UG)b, Maria C. da Rocha (UG)a,
Mario T. Kato (R)a, Elvis J. França (R)c, Ana P. S. Paim (R)d, Maria J. F. Gomes(R)b
aUniversidade Federal de Pernambuco, Departamento de Engenharia Civil e Ambiental,
Laboratório de Saneamento Ambiental, Recife, PE, Brazil, 50740-530
bUniversidade Federal Rural de Pernambuco, Departamento de Química, Recife, PE, Brazil, 52171-900
cCentro Regional de Ciências Nucleares do Nordeste, Recife, PE, Brazil, 50740-545
dUniversidade Federal de Pernambuco, Departamento de Química Fundamental, Recife, PE, Brazil, 50670-901
*e-mail: iago.jose@ufpe.br
Rice (Oryza sativa) is considered an important component of the human diet, and the cooking
method of the grain is an essential factor for its mineral composition. However, despite the great
relevance of the subject, it has not been much addressed in the literature1,2; in most of the reported
works, the rice grain has been used for mineral determination in its raw form, which consumption is
not usual. Therefore, this work aims to evaluate the effects of cooking on the mineral composition
(Al, Ca, Cu, Co, Cd, Cr, K, Mg, Mn, Mo, Na, Ni, Pb, V, and Zn) of rice commonly commercialized in
Pernambuco state, using for analytical determination the inductively coupled plasma optical
emission spectrometry (ICP OES) and flame photometry. Raw rice was also analyzed for
comparative purposes. The rice samples were cooked on a domestic stove, applying a full 23 factorial
design taking into account (i) the pre-washing of the rice, (ii) the type of pan used (aluminum or
stainless steel), and (iii) the destination of the cooking water (evaporated or drained). Twenty-four
cooked rice samples were obtained and analyzed after applying the factorial design (eight of each
of the three types of rice, white, parboiled, and brown). Then, the samples were dried, milled, and
decomposed in a microwave oven. Subsequently, the samples' residual carbon content was
determined by means of a molecular absorption spectrophotometer in the visible region. A full 23
factorial design with a central point was also carried out for optimizing the decomposition of rice
samples that consisted of using 1 mol L-1 for HNO3 concentration, the volume of 2.5 mL for 30%
H2O2, and 5 min for digestion time. In terms of the quality of the analytical procedure, the method
was validated for linearity (r > 0.999), precision (DPR < 20%), and LOQ ranging from 0.032 mg kg-1
(Mn) to 60 mg kg-1 (Na and K). Also, the results of Al, Ca, Cd, Co, Cr, Cu, K, Mg, Mn, Mo, Na, Ni,
Pb, V, and Zn determined in SRM NIST 1568a Rice Flour, CRM NIES 10-b Rice Flour-Unpolished,
and RM IAEA-V-8 Rye Flour were in accordance with the certified and reference values at the 95%
confidence level. The statistical analysis was performed using the Statistica 10 (StatSoft) software.
Cooked rice samples showed lower concentrations of Ca, Cu, K, Mg, Mn, Mo, and Zn and higher
content of Na when compared to those of their raw form. Considering the culinary practices, the pre-
washing of rice had a significant effect (p < 0.05) on reducing the concentration of K and Mn. Also,
the use of aluminum pan significantly increased the Al concentration in white and parboiled rice
(Figure 1). Likewise, the use of stainless-steel pan has increased the Mn content in the cooked rice
of the three types (Figure 1). Finally, the evaporation of cooking water, contrary to draining,
significantly increased the concentrations of K, Mo, and Na. For the other evaluated minerals, such
as Cd, Co, Ni, Pb, and V, the concentration Figure 1. Cubic diagrams of the factorial design results for Al
-1
values were lower than the LOQ of the cooked (left) and Mn (right) content in parboiled rice, in mg kg .
and raw rice method. Therefore, cooking
significantly affected the mineral profile of rice
through elemental leaching caused by pre-
washing the grain and draining the cooking
water for the three types of rice (white,
parboiled, and brown). Also, probable partial
solubilization of metals from the pans and
subsequent absorption by the grain was
verified, especially for Al and Mn.
1Horner NS, Beauchemin D, Anal. Chim. Acta, 758, 2013, 28.
2Jaafar M, Shrivastava A, Bose SR, Felipe-Sotelo M, Ward NI, J. Food Compos. Anal., 96, 2021, 103748.
[FACEPE, LSA/UFPE, CENAPESQ/UFRPE, CRCN-NE]

EVALUATION OF DIFFERENT SAMPLE PREPARATION TECHNIQUES FOR
THE ANALYSIS OF NAPHTHENIC ACIDS IN PRODUCED WATER SAMPLES
Juliana Crucello (PG)a*, Natalia de Aguiar Porto (PG)a,
Naiara Mariana Fiori Monteiro Sampaio (R)a, Leandro Wang Hantao (R)a
aUniversidade estadual de Campinas - UNICAMP, Departamento de Química Analítica/Instituto de Química, Campinas,
São Paulo, Brazil, 13083-872
*e-mail: crucellojuliana@gmail.com
Naphthenic acids (NAs) are a class of petroleum-derived compounds comprised of a complex

mixture of hundreds of organic acids possessing varied structures1. Such species are linked with the
acute and chronic toxicity character of produced water (PW), the most significant by-product of oil &
gas industry2. Monitoring NAs is an urgent need regarding environmental concerns1–3. High-
resolution analytical instruments such as comprehensive two-dimensional gas chromatography
(GC×GC) combined with high resolution mass spectrometry (HRMS) are promising techniques to
achieve isomers separation and compounds identification. Sample preparation is one of the principal
bottlenecks of GC×GC-HRAM-MS analysis. Traditional methods relies on the use of o hazardous
solvents and multi-step, laborious and time-consuming procedures1. Here, a comparative study of
three new different extraction techniques, namely direct-immersion solid phase microextraction (DI-
SPME), vacuum assisted sorptive extraction (VASE)2 and liquid-phase hollow fiber SPME (LP-HF-
SPME)4, is shown. Different approaches for analytes derivatization are demonstrated, including
microwave-assisted and on-fiber derivatization. All evaluated methods exhibit better metrics
according to Green Analytical Chemistry (GAC) principles than conventional techniques such as
Liquid-Liquid Extraction (LLE), while also presenting similar or enhanced extraction performance to
it2,4. In addition, automation is possible for some of them. It is expected that this work showcases
different opportunities for the analysis of polar compounds in aqueous matrices.
1 Facanali R, Porto NA, Crucello J, Carvalho RM, Vaz BG, Hantao LW, Journal of Analytical Methods in Chemistry, 2021,
2021, 6078084.
2 Crucello J, Medeiros I, Carvalho RM, Hantao LW, Microchemical Journal, 180, 2022, 107581.
3 Crucello J, Carvalho RM, Hantao LW, LCGC North America, 39, 2021, 478-485.
4 Porto NA, Crucello J, Facanali R, Medeiros I, Carvalho RM, Hantao LW, Journal of Chromatography A, 1655, 2021,
42485.
[CNPq, CAPES, FAPESP, Nova Analítica, ThermoFisher Scientific, Petrobras]

DEVELOPMENT OF A MULTIRESIDUE METHOD FOR DETERMINATION OF
PESTICIDES IN GRAPE JUICES USING QuEChERS AND UHPLC-MS/MS
Karolaine da Silva Albarnaz (PG), Gabriel A. Barboza Prates (PG), Renato Zanella (R),
Martha B. Adaime (R), Osmar D. Prestes (R)*
Grape juice is an unfermented beverage obtained from the proper processing of the fruit, this product
is a food of high nutritional value, and can be compared with the grape itself. This happens because
all the main constituents of the matrix can be found in the product, such as sugars, minerals, organic
acids, phenolic compounds, vitamins and pectin.1 Grape production in Brazil covers approximately
78 thousand hectares, producing about 1.5 million tons of grapes per year.2 Currently, the use of
pesticides in agriculture has become widespread in order to prevent diseases and pests in
plantations, increase productivity and improve the quality of the obtained products. 3 Despite these
advantages, the indiscriminate use of pesticides in food can cause harmful effects to human health
when these substances are above the Maximum Residue Limits (MRLs). Based on that, a sample
preparation method was developed and validated for the multi-residue determination of 154
pesticides in grape juice. In this study, the initial tests were carried out based on previously published
works using the three versions of QuEChERS method (original, citrate and acetate). In the
gravimetric analysis, the experiments that resulted in a lesser amount of non-volatile coextratives
were the citrate and acetate versions with cleaning step. These tests were done using a “blank”
sample of grape juice. Several parameters of the sample preparation method were evaluated aiming
at the best conditions for the selected pesticides. The proposed extraction procedure is based on
the QuEChERS modified acetate method using the sorbent Celite in a step prior to the addition of
the extraction solvent. The determination of analytes was made with a ultra-high performance liquid
chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) system, model 6470
(Agilent Technologies, USA), composed of a triple quadrupole MS analyzer. Data acquisition was
done using the MassHunter system. The separation was performed through an Agilent Poroshell
120 EC C18 column (100 x 2.1 mm, 1.9 µm) with a mobile phase consisting of (A) water and (B)
methanol, both with 0.1%(v/v) formic acid and 5 mmol L-1 ammonium formate. The limits of
quantification of the method were from 5 to 10 µg L-1. The recoveries from the spike levels 1, 2, 4
and 10 µg L-1 showed recoveries between 70 and 120% with precision, in terms of relative standard
deviation (RSD), ≤ 20%. The method proved to be efficient for the determination of pesticides in
grape juice and pesticide residues were found in concentrations from 12.05 to 41.98 µg L-1. Four
pesticides were detected in 18 samples analyzed, namely the insecticide acetamiprid, and the
fungicides carbendazim, dimethomorph and methyl thiophanate.
1 Rizzon LA, Meneguzzo J. Suco de uva, Embrapa: Informação Tecnológica, 2007.
2 Embrapa. Inteligência e Mercado de Uva e Vinho. https://www.embrapa.br/cim-uva-e-vinho/a-viticultura-no-brasil.
3 Moraes RF. Agrotóxicos no Brasil: Padrões de uso, política da regulação e prevenção da captura regulatória, IPEA,
2019.
[UFSM, CAPES, CNPq]

COMPARISON BETWEEN QuEChERS AND MSPD METHODS FOR PESTICIDE
MULTIRESIDUE DETERMINATION IN STRAWBERRY BY UHPLC-MS/MS
Sónia M. V. S. Cardoso (PG), Pimpernelli J. dos Santos (PG), Theo Samborski (UG),
Strawberry is popular fruit rich in antioxidants as vitamin C and is widely consumed over the world. 1
The temperate and tropical areas are characteristic of Brazilian strawberry-producing regions, being
the state of Minas Gerais the largest producer.2 However, pesticides are applied in order to improve
the productivity and minimize the presence of pests and diseases in this type of crop. Consequently,
the establishment of maximum residue limits (MRL) of pesticides by Brazilian authorities, is a safety
precaution for strawberry consumers' health. In order to monitor the pesticides residues in food, it is
desirable to develop sample preparation methods that allow the extraction and detection of these
compounds with a wide range of polarities by using a simple procedure. The QuEChERS method3
is widely employed for pesticides multiresidue determination in food samples. This method involves
an acetonitrile extraction, the use of salts for partitioning of organic and aqueous phases and cleanup
extract by dispersive solid phase extraction (d-SPE) using sorbents such as C18 and primary
secondary amine (PSA). Moreover, the matrix solid phase dispersion (MSPD) enables extraction
and cleanup in a single step by using sorbents, followed by an elution step to extract the analytes.
In this study, a comparison between QuEChERS and a modified MSPD4 extractions were employed
for determination of 172 pesticides in strawberry samples by ultra-high performance liquid
chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The extraction with
QuEChERS method4 consisted with 10 g of sample, 10 mL of acetonitrile 1% acetic acid (v/v) as
extraction solvent and MgSO4 (4 g) and CH3COONa (1.7 g) as partitioning salts. The extracts were
not submitted for clean-up but were diluted (10x) in ultrapure water and filtered (PTFE, 0.22 µm)
before analysis. For modified MSPD,4 0.5 g of sample was vortexed with 1 g of C18 and ceramic
homogenizers for 1 min. Then, 2 mL of pure acetonitrile was added to promote the extraction of
analytes. After sonication and centrifugation, the supernatants were diluted (1:1, v/v) with ultrapure
water. The extracts were filtered (PTFE, 0.22 µm) and submitted for analysis. The UHPLC-MS/MS
analysis was performed with an Agilent 1290 Infinity II LC System and 6470 LC/TQ mass
spectrometer. The column Agilent Poroshell 120 EC C18 (100 x 2.1 mm, 1.9 µm) and mobile phase
A) ultrapure water and B) methanol both containing 0.1% formic acid (v/v) 5 mmol L-1 ammonium
formate at flow rate 0.3 mL min-1 were employed. The injection volume was 3 µL. Electrospray
ionization was operated in positive and negative mode (ESI+/-) and selected reactions monitoring
(SRM) as acquisition mode. The total run analysis was 17 min. Recovery studies were evaluated at
the spike levels 5, 10 and 20 µg kg-1. Both extraction methods were suitable for more than a hundred
of pesticides residues in strawberry, but the QuEChERS method provided better performance than
MSPD. In the lowest spike level (5 µg kg-1), 146 compounds showed recovery rates between 70 to
120% with RSD ≤ 20% and 155 compounds in the intermediate and highest spike levels (10 and 20
µg kg-1) for the QuEChERS method. Only 102 compounds were determined by MSPD. The
QuEChERS method based on acetonitrile extraction combined with UHPLC-MS/MS is simple, easy
and rapid to execute, being useful for routine analysis.
1 Universidade Estadual de Campinas, Tabela Brasileira de Composição de Alimentos - TACO, 4 ed, 2011.
2 Jordão WAF, Lima e Silva FL, Ciência Rural, 52, 2022.
3 Anastassiades M, Lehotay SJ, Stajnbaher D, Schenck FJ, Journal of AOAC International, 86, 2003, 412-431.
4 Kemmerich M, Demarco M, Bernardi G, Prestes, OD, Adaime, MB, Zanella, R, Journal of Chromatography A, 1612,
2020.
[UFSM, CNPq, CAPES, FINEP]

DETERMINATION OF PESTICIDE RESIDUES IN PERSIMMONS USING
QuEChERS METHOD AND UHPLC-MS/MS
Graziela I. Rocha (PG), Osmar D. Prestes (R), Martha B. Adaime (R), Renato Zanella (R)*
Pesticides are widely used in food crops and residues of these compounds may be present in these
crops. Despite their large-scale use in agriculture, pesticides can be dangerous to human health,
animals and the environment. These risks arising from the frequent use of pesticides demand the
establishment of adequate analytical methods, as well as monitoring programs to assess compliance
with the maximum amount of pesticide residues that may be present in food, called maximum residue
limits (MRLs). Persimmon is a fruit widely consumed in nature and sometimes used in the production
of by-products.1 Persimmon production stands out in Brazil, appearing in recent years among the
twenty most produced fruits. The state of São Paulo is the leader in production, but the state of Rio
Grande do Sul has been increasing persimmon production, being the second state that produces
the most.2 The export volume of the fruit is also high. There are numerous varieties of persimmon,
which is a very sensitive fruit, with a short harvest period and has conservation problems, resulting
in widespread use of pesticides.1,3 The simultaneous determination of pesticides of different classes
in different matrices is quite complex, so the use of a multiresidue method is essential. The
QuEChERS method has been shown to be very efficient for these cases, and allows several
modifications according to the characteristics of the matrices.4 Thus, this work aimed to develop and
validate a fast and efficient sample preparation method using unbuffered QuEChERS method for
multiresidue determination of pesticides in persimmon samples by ultra-high performance liquid
chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Some modifications were
made to the original QuEChERS method, such as replacing the d-SPE clean-up by a dilution step.
This procedure, generates a simpler, faster and cheaper analysis.The modified QuEChERS method
was validated for 86 pesticides according to the parameters of selectivity, linearity and working
range, sensitivity, matrix effect, limit of quantification (LOQ), limit of detection (LOD), accuracy and
precision. In this method, the extraction was performed with a 10 g persimmon sample and 10 mL
acetonitrile followed by vortexing for 1 min. Then 4 g MgSO4 and 1 g NaCl were added, followed by
manual stirring for 1 min to carry out the partition step. The tube was centrifuged for 8 min at 3400
rpm and, finally, the extract obtained was filtered and diluted 20 times in ultrapure water. The method
LOQ ranged from 10 to 40 µg kg-1. Recovery levels were between 70 and 120% with RSD ≤ 20%.
The matrix effect was between ±20% for all evaluated compounds. The analyzes by UHPLC-MS/MS
with ionization by electrospray showed high selectivity and sensitivity. The validated method was
applied to commercial persimmon samples, residues of five pesticides were found. Some of them
are forbidden to use in this fruit, such as the fungicide carbendazim and the insecticide dimethoate.
Thus, the results obtained showed that the proposed method is efficient and can be used routinely
for pesticide multiresidue determination.
1 Kluge, RA, Tessmer, MA, Caqui - Diospyroskaki. Exotic Fruits, Elsevier, 113-119, 2018.
2 IBGE, Agricultural production: production of persimmon. Production of persimmon, 2021.
3 Vieites, RL. Persimmon. Brazilian Journal of Fruiticulture, 34, 653-955, 2012.
4 Anastassiades, M, et al. Journal of AOAC International, 86, 2, 412-431, 2003.
[UFSM, CNPq, CAPES]

DETERMINATION OF PESTICIDE RESIDUES IN BRAZILIAN OLIVE OIL BY
ACETONITRILE EXTRACTION AND GC-MS/MS
Juliana D. B Kuntz (PG),Martha B. Adaime (R), Renato Zanella (R), Osmar D. Prestes (R)
Widely used in the Mediterranean diet, olive oil has become more and more frequent on the Brazilian
consumer's table. Although, per capita consumption is still small, the country already figures on the
international scene as the second largest importer of olive oil. In recent years, olive cultivation has
been growing in Brazil, mainly in the state of Rio Grande do Sul.1 However, there is still no well-
established legislation for olive cultivation and only maximum residue limits have been established
for nine pesticides in olives, and unauthorized pesticides end up being used to combat pests.2
Therefore, developing an analytical method that allows the determination of pesticides of different
classes in olive oil is essential to ensure the safety of these products before they reach the
consumer's table. Analytical problems associated with the analysis of pesticides in fatty foods, as
olive oil, are well known, especially when gas chromatography analysis is applied. Therefore, sample
preparation is a crucial step in the analytical procedure since even small amount of lipids can harm
columns, detectors or cause signal suppression.3 This work is focused on the development and
evaluation of a simple sample preparation strategy based on acetonitrile extraction with a clean-up
performed by dispersive solid phase extraction, followed by gas chromatography coupled to tandem
mass spectrometric analysis (GC-MS/MS). The extraction method was based on 1 g of olive oil, 5
mL of water, 10 mL of acetonitrile and 4 g of NaCl, followed by manual agitation for 1 min and
centrifugation for 5 minutes at 5000 RPM, to the partition step. The upper layer phase was frozen
for 1 hour at -18 ºC. After another centrifugation step, an aliquot of 6.5 mL was transferred to 15 mL
polypropylene tube with 800 mg of MgSO4, 150 mg of C18 and 150 mg of PSA, centrifuged again
and filtered before GC-MS/MS analysis. The method was validated for 79 compounds. The limit of
quantification (LOQ) ranged from 25 to 100 µg kg -1 and the limit of detection (LOD) from 7.5 to 30
µg kg-1Most of the analyzed compounds showed recovery range between 70 to 120% with RSD
<20%. The method was applied to 18 samples of olive oil produced in Brazil. From this samples, 7
showed pesticide residues of different classes at concentrations ranging from 8.9 to 689.4 µg kg-1.
1 BRASIL, Ministério do Desenvolvimento, Indústria e Comércio Exterior. Relatório de Análise de Azeite de Oliva
Extravirgem. Rio de Janeiro: Instituto Nacional de Metrologia, Qualidade e Tecnologia, 2015.
2 ANVISA (AGÊNCIA NACIONAL DE VIGILÂNCIA SANITÁRIA). Monografias Autorizadas. [s.d.]. Disponível em:
<http://portal.anvisa.gov.br/registros-e-autorizacoes/agrotoxicos/produtos/monografia-de-agrotoxicos/autorizadas>.
3 Rosalía LR, Jesús MS, Osmar DP, Roberto RG, Antonia GF, Critical Reviews in Analytical Chemistry, 2022, DOI:
10.1080/10408347.2022.2040352
[UFSM, CAPES, CNPq]

SIMULTANEOUS DETERMINATION OF ANTIMICROBIALS AND β-AGONISTS
RESIDUES IN PORK MEAT BY UHPLC-MS/MS
Daniela K. Muenchen (PG), Luana Floriano (PG), Vinícius P. de Souza (UG),
Santa Maria-RS, Brazil, 97105-900
In swine farming, veterinary drugs are mainly applied to provide animal’s health. However, its
application may result in residues in the edible tissues, and in view of this, the development of
analytical methods for the determination of these substances is extremely important. In this sense,
pork is a matrix particularly important for residues monitoring, as it is a widely consumed tissue and
has great relevance in the economic market.1 Thus, the aim of this work was to develop a multiclass
method for the determination of veterinary drugs residues in pork meat by UHPLC-MS/MS. More
than 30 different veterinary drugs included antimicrobials (lincosamides, macrolides, quinolones,
sulfonamides, tetracyclines and other minority families) and β-agonists. UHPLC-MS/MS analyses
were carried out on an Acquity UPLCTM system coupled to a Xevo TQTM MS/MS triple quadrupole
mass spectrometer (Waters Corporation, USA), with positive electrospray ionization interface in
multiple reaction monitoring (MRM) mode acquisition. The separation was achieved using a HSS T3
column (100 × 2.1 mm i.d., 1.8 μm particle size), at 60 °C. The mobile phase consisted of (A) water
and (B) acetonitrile:methanol (80:20, v/v), both containing 0.1% (v/v) formic acid, at a flow-rate of
0.4 mL min-1 in gradient elution. The total running time was 8 min and injection volume was 10 µL.
Sample preparation was based on the solvent extraction with a sorbent clean-up. The extraction was
performed with 2 g pork sample and 2 mL of aqueous solution containing 0.1% EDTA (w/v) and
1%(v/v) formic acid, followed by 8 mL acetonitrile. A partitioning step performed by the addition of
salts such as NaCl or (NH4)2SO4 was evaluated. In the cleanup step, different sorbents were
evaluated, such as C18, secondary primary amine (PSA) and EMR-Lipid®, in both dispersive solid-
phase extraction (d-SPE) or pass-through SPE mode. The presence of lipids co-extractives in the
extract was verified in full scan analysis by gas chromatography coupled to tandem mass
spectrometry (GC-MS/MS). The final sample preparation method consisted of extraction with a 2 mL
aqueous solution containing 0.1% (m/v) EDTA and 1% (v/v) formic acid followed by the addition of
8 mL acetonitrile, and clean-up by d-SPE using 50 mg C18 sorbent per mL of extract. Further, two
different protocols were required before the chromatographic analysis to determine the authorized
compounds at of maximum residue level (MRL from 50 to 400 µg kg -1)2 and prohibited or
unauthorized compounds at the lowest calibrated level (LCL = 5 µg kg-1), corresponding to dilution
of the extract in ultra-pure water (1:4, v/v) and evaporation of the extract, respectively. Method
validation was performed based on the European3 and Brazilian4 guidelines. Linearity was evaluated
using matrix-fortified calibration and showed r² > 0.99 for most compounds. The recoveries ranged
from 83 to 113% and RSD between 2 and 18%. Decision limit (CCα) concentrations ranged from 5.3
to 447.8 µg kg-1. The proposed method was applied to 25 pork samples from different sources and
residues of enrofloxacin, oxytetracycline and ractopamine were detected.
1 Associação Brasileira de Proteína Animal. Relatório Anual de Atividades 2021. 2022. Disponível em: http://abpa-
br.org/relatorios/.
2 European Commission. Commission Regulation 37/2010. Official Journal of the European Union, 15, 1-72, 2010.
3 European Commission. Commission Implementing Regulation 2021/808, Official Journal of the European Union, 180,
84-109, 2021.
4 Brasil. Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Manual de garantia da qualidade analítica –
resíduos e contaminantes em alimentos, 2011.
[UFSM, CNPq, CAPES, FAPERGS]

ANALYSIS OF PESTICIDE RESIDUES IN CHICKEN EGGS BY LIQUID
CHROMATOGRAPHY COUPLED TO TANDEM MASS SPECTROMETRY
Roberta Fabricio Loose (PG), Diogo Marchesan (UG), Luana Floriano (PG),
Renato Zanella (R), Martha B. Adaime (R), Osmar Damian Prestes (R)*
Eggs constitute an important and basic component in the human diet, since they are highly nutritious
and easily obtained, in addition to their affordable price.1 Over the past two decades, eggs have
represented the largest increase in protein demand among the world’s population. In 2021, the
consumption of chicken eggs in Brazil was estimated at 257 units per person/year. To meet this
demand, 54.973 billion units were produced, with 99.54% of this total destined to supply the internal
market.2 With the increasing production and consumption of eggs, their safety and quality has been
regarded. At present, the detection of synthetic substances in eggs is mainly focused on antibiotics
and other veterinary drugs. Although pesticides are less used in livestock, they can reach eggs
through animal feed consisting mainly of soybean and corn, as well as household products for animal
use and rural environments.3,4 Evidence suggests that pesticides can be transferred to both the yolk
and albumen of eggs.5 Thus, the determination of these residues in chicken eggs is therefore
necessary to ensure product safety. In this work, we developed and validated a sample preparation
method for the determination of pesticide residues in eggs using a modified QuEChERS method and
ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-
MS/MS). All sorbents tested during the cleanup step were evaluated in terms of removal efficiency
of phospholipids present in the final extract. UHPLC-MS/MS analyses of pesticides were performed
by Waters Xevo TQ (USA) with electrospray ionization in positive and negative mode (ESI±) and
were carried out by selected reaction monitoring (SRM) acquisition. The separation was performed
using a Acquity UPLC™ BEH C18 (50 x 2.1 mm, 1.7 µm) chromatographic column and mobile phase
composed by (A) water:methanol (98:2, v/v) and (B) methanol, both containing 5 mmol L -1
ammonium formate and 0.1% (v/v) of formic acid at flow rate 0.225 mL min-1. The injection volume
was 10 µL and total run time was 10 min. The optimised method is basically composed of four steps,
using acetonitrile 5% formic acid (v/v) as extraction solvent, MgSO4 and NaCl as partition salts, and
the Captiva EMR-Lipid® (Agilent Technologies, USA) for removing lipids in the cleanup step.
Linearity was evaluated using matrix matched analytical curves in the range of 0.5 to 20 µg L-1, and
showed r2 > 0,99 for most compounds. The results of the accuracy and precision tests, evaluated at
5, 50 and 100 µg kg-1 (n= 6), proved to be adequate, since they showed recoveries between 70 and
120%, with RSD ≤ 20%. The limits of quantification and detection of the method were between 5 and
100 µg kg-1 and 1.5 and 30 µg kg-1, respectively. The method was applied in the analysis of 41
commercial egg samples from different cities and regions of Rio Grande do Sul and Paraná states,
however no pesticides were detected.
1 Li Y, Chen Z, Zhang R, Luo P, Zhou Y, Wen S, Ma M. Chromatographia, 79, 2016, 1165-1175.
2 ABPA. Annual Report from the Brazilian Association of Animal Protein, 2022.
3 Zhang X et al. Journal of Chromatography A, 1593, 2019, 81-90.
4 Pereira MU et al. Journal of Food Science and Technology, 57, 2020, 1719-1730.
5 Souza MMR, Moreira CO, Lima TG, Aquino A, Dórea HS. Microchemical Journal, 110, 2013, 395-401.
[UFSM, CAPES, CNPq]

NEW HF-LPME FIBER FOR PASSIVE SAMPLING OF PESTICIDES
IN SURFACE WATERS
Nayara S. Fraga (PG)a*, Nathan S. Coelhoa (PG), Zenilda L. Cardeala (R)
and Helvécio C. Menezesa (R)
aFederal University of Minas Gerais, Department of Chemistry /ICEx, Belo Horizonte, M.G., Brazil, 31270-901
*e-mail: nayarasf@ufmg.br
Brazil sold domestically, approximately686 thousand tons of pesticides only in the year of 2020
(IBAMA).1 However, recent studies and important enforcement agencies show that some classes of
pesticides are extremely toxic, persistent and, bioaccumulative.2 In the context of the regulation of
national water sources, it is important to search for the development of procedures that are effective
in sampling, identification, and quantification of this type of pollutant.The work developed focused
on improving the HF-LPME system developed byValenzuela1 using polydimethylsiloxane (PDMS)
as an extraction agent inside the polypropylene hollow fiber for pesticide sampling and analyzing
them by gas chromatograph coupled to mass spectrometry(GC/MS). Polydimethylsiloxane was
synthesized from mixing Sylgard 184 silicone and the curing agent in a 10:1 m m-1 ratio. Hexane
was added to this mixture in a 1.5:10 m m-1 ratio to decrease the viscosity of the assembly. Hollow
polypropylene fibers Membrane Underlying Performance Co were cut to 8.0 cm, and they were filled
with the mixture prepared previously. The fibers produced (HF-LPME-PDMS) were placed in an
oven at approximately 80°C for 48 hours. After cooling to room temperature, the fibers underwent a
cleaning procedure in three sonication cycles of 30 min each with a 5% v v-1 methanol: ultrapure
water mixture.
HF-LPME-PDMS HF-LPME- Injection of 1,5 Temperature

fiber in a U-shape PDMS + 2,0 mL µL of the Programming
Sorption of the
+ 20,0 mL vial vial with Agitation in a extract into a 80 ºC (3 min) up to
analytes for 30
containing a acetonitrile for vortéx for 1 GC/MS 160 ºC at 20 ºC min-1,
min and 400
standard pesticide dessorption in min. Shimadzu 255 ºC at 5 ºC min-1
rpm.
solution sonicator for 5 GP2010 single and 280 ºC (1 min) at
(10 mg L-1) min at 65,0 °C. quadrupole. 20 ºC min-1.
Figure 1 – Extraction procedure, desorption, and analysis of the analytes by HF-LPME-PDMS and GC/MS.
Abundance
Relative
Retention Time (min)

Figure 2 – Total chromatogram of the ions after extraction with the HF-LPME-PDMS: 1-Phorate, 2-Lindane, 3-Diazinon,
4-Disulfoton, 5-Pirimiphos-methyl, 6-Fenthion, 7-Chlorpyrifos, 8-Procymidone, 9-Endosulfan, 10-Napropamid, 11-
Buprofezin, 12-Oxyfluorfen, 13-Kresoxim-methyl, 14-Endosulfan II, 15-4,4-DDD, 16-Methoxychlor, 17-Mirex.
This result above is promising, especially in the context of passive sampling, since thePDMS, after
being adhered to the hollow fiber, remains stable, allowing the sampling to be done without the
limitation of possible loss of the extractant throughout the monitoring. The work developed allowed
an efficient microextraction method usingpolydimethylsiloxane polymerized inside hollow fiber for
the extraction of pesticides in water. Thus, the system developed can be used for passive sampling
of these pollutants in environmental water, contributing to their monitoring and regulation.
1 Valenzuela EF, Doctoral Thesis, UFMG, 2021.

2 Campanale C et al., Trends in Analytical Chemistry, 144, 2021, 116423.
[CEMIG, CAPES]

MODIFIED CLOUD POINT EXTRACTION METHODOLOGY FOR Cd(II)
EXTRACTION USING MAGNETIC MORINGA OLEIFERA
Vanessa N. Alves (R)a*, Deborah C. Bagano Gonçalves (PG)a
aUniversidade Federal de Catalão (UFCAT), Instituto de Química, Catalão-GO, Brasil.
*e-mail: vanessanalves@gmail.com
The combination of sample preparation techniques stands out for contributing to the emergence of
new methodologies, using low volumes of reagents, contributing to the environment, in addition to
overcoming limitations of techniques already established within analytical chemistry. Although there
are several instrumentation techniques for metal analysis, such as Flame Atomic Absorption
Spectrometry (FAAS), some of this equipment has some limitations in sensitivity, detection, and
sample preparation. Cloud point extraction (CPE) and solid phase extraction (SPE) are known and
common techniques as sample preparation techniques for the extraction of various analytes1, as in
the case of this work, cadmium ion extraction. Therefore, the objective is the development of a
modified point cloud extraction (CPE) methodology, by combining this technique with solid phase
extraction (SPE) for Cd(II) extraction using Moringa oleifera magnetic as an adsorbent. The
methodology adopted was based on Falahnejad et al.,2 study and it was tested using magnetically
modified moringa seed husk3 as an adsorbent and Triton x-100 as a surfactant to cadmium
extraction. For adsorbent characterization, evaluation of zero charge point (PCZ), Infrared
Spectroscopy and Scanning Electron Microscopy with EDS (SEM) was performed. The ideal pH for
extraction and the mass of the adsorbent were studied in a univariate mode, with Cd(II) solutions at
pH ranging from 4 to 11 and mass from 2 to 20 mg. Subsequently, a 24 factorial design, with a central
point, was carried out, evaluating four variables, Triton x-100 and HNO3 surfactant concentrations,
sonication time and water bath time, 1 and 10 min. After the optimized procedure, an evaluation of
the effectiveness of the proposed methodology was carried out, from the comparison of the analytical
signals obtained with the techniques (CPE and SPE) separately. According to the procedure already
proposed, it was possible to notice that the metallic ion is extracted and preconcentrated, because
the analytical signal found was greater than the signal of the standard, which suggests that the
procedure is viable for extraction and pre-concentration of Cd2+. With the IR analysis, it is possible
to see that the functional groups present in the moringa structure, mainly hydroxyls and carboxyls,
remain unchanged after the magnetization procedure of the adsorbent material. The SEM analysis
shows that it is an amorphous and porous material, which contributes to the extraction process, in
addition to being possible to observe an increase in the percentages of iron in the magnetic material.
According to the analysis of pH and PCZ (6.4) and pH study, the pH chosen as optimal for extraction
was pH 6, and that the use of a buffer does not contribute to the extraction. Additionality, in this pH
the solubility product isn´t exceeded and cadmium ions don´t precipitate. From the factorial design,
it´s possible check that the sonication and heating times are significant in the extraction. In this way,
these variables are analysed in a univariate mode and the results shows de better values of
sonication and heating time are 5 and 6 minutes, respectively. From the adsorbent mass analysis,
we observed that the adsorption process is favoured with the use of greater adsorbent masses, and
this variable was fixed in 20 mg of adsorbent. When carrying out the study of the extraction procedure
with the separate and united methods, SPE and CPE, SPE + CPE, CPE + SPE, it can be seen that
the method proposed for the work has a higher analytical signal than the others studied, in addition
to an analytical signal greater than that of the standard used in the experiment, this therefore proves
that the combination of the techniques in this case is efficient for the extraction of Cd(II) to occur.
Therefore, the modified CPE procedure, using the junction with solid phase extraction, is a promising
and viable method with regard to the extraction and pre-concentration of Cd(II) using Moringa
oleifera magnetic as adsorbent.
1 Luiz, A. L.; Maciel, E. V. S.; Lanças, F. M. Scientia Chromatographica, v.3, n.7, p. 157-182, 2015.
2 Falahnejad, M.; Zavvar Mousavi, H.; Shirkhanloo, H.; Rashidi, A.M. Microchemical Journal, 125, 236–241, 2016.
3 Massart R. IEE Transactions on Magnetios, 1247-1248, 1981.
[UFCAT, IQ-UFU, FAPEG, CAPES, CNPq]

METHOD DEVELOPMENT FOR NONMETAL DETERMINATION IN SEAFOOD:
INFLUENCE OF pH AND DRYING METHOD IN ANALYTE CONCENTRATIONS
Fernanda P. Balbinot (PG)a, Filipe S. Rondan (PG)a, Jenifer H. Cerqueira (PG)a,
Tamires C. Ramires (UG)a, Marina S. Xavier (UG)a, Marcia F. Mesko (R)*a.
aUniversidade Federal de Pelotas, Centro de Ciências Químicas, Farmacêuticas e de Alimentos,
Pelotas, RS, Brazil, 96160-000
*e-mail: marciamesko@yahoo.com.br
The interest in assessing the presence of chemical elements such as halogens and sulfur in seafood,
which is a very important nutritional source1, has been increasing. In this work, an analytical method
for the determination of Br, Cl, F, I, and S, in seafood, by ion chromatography coupled to mass
spectrometry (IC-MS) in a single analysis, after decomposition by microwave-induced combustion
(MIC), was developed. For this, some important parameters of different steps in the analytical
sequence were evaluated. In the pre-treatment step, experiments were performed in order to
establish the most feasible drying method (oven-drying at 60 °C and 100 °C, and freeze-drying). In
the sample preparation step, the absorbing solution was optimized and the influence of pH values of
the digests in the stabilization of the analytes was evaluated. Shellfishes (Brown mussel, M1, and
Pacific oyster, O1), octopus (PO1), and fish (Argentine hake, P1) samples were evaluated. The MIC
method and chromatographic conditions were optimized using the sample “M1”. Standard recovery
tests, using three concentration levels, were performed in order to evaluate the most suitable
absorbing solution for the analytes. For this, NH4OH solutions (100, 150, and 250 mmol L-1) were
evaluated for all four types of samples. Additionally, certified reference materials were analyzed
using the optimized conditions. The relation between recovery values and the pH values of the
digests was assessed. The only solution that provided suitable recoveries for all analytes in the three
standard recovery tests was 250 mmol L-1 NH4OH, of which the pH value of the digests was 5. At
lower concentrations of the absorbing solutions, the recovery of the analytes in the three levels of
concentration was impaired, and the pH of the final digests was between 3 and 4. The pH of the
digests of the four samples using the three absorbing solutions, without analyte addition, was
measured. The pH values for each digest (from M1, O1, and PO1 samples) using the different
absorbing solutions (100, 150, and 250 mmol L-1 NH4OH) were, 3, 4, and 5, respectively. On the
other hand, the pH values of the digest from the P1 sample, using the different concentrations of
absorbing solutions, were 5. It was possible to observe a tendency for pH decreases as the
concentration of the analytes increases. It was observed in standard recovery tests and digests from
the M1, O1, and PO1 samples, which presented analyte concentration about ten-fold higher than
the digests from the P1 sample, especially regarding Cl and S, as well as a relation between low pH
values of the final digests and the unsuitability of the halogens and sulfur stabilization on the solution.
Further evaluations of the influence of the pH in the analyte retention using different concentrations
of (NH4)2CO3 as absorbing solution must be carried out. Suitable recoveries and agreements (92%
to 109%, considering the three levels of the standard recovery tests and CRMs analyses) were
obtained using 250 mmol L-1 NH4OH as the absorbing solution for all analytes. In addition, analyte
losses were evaluated according to the drying pre-treatments. The results for F concentration in all
samples (except O1, in which F concentration was below the LOQ – 0.3 mg kg-1) using oven-dry at
100 °C were statistically different from those obtained by drying at 60 °C or freeze-drying (Tukey
test, One-way ANOVA, confidence level of 95%). Thus, it was observed that about 30% of the F
concentration was lost when the samples were dried at 100 °C. Regarding the content of the analytes
in the samples, for M1, O1, and PO1, the concentrations of the analytes varied from 49 to 181 mg
kg-1 (Br), 19,970 to 24,180 mg kg-1 (Cl), < 0.3 to 8 mg kg-1 (F), < 0.6 to 7 mg kg-1 (I), and 14,250 to
18,000 mg kg-1 (S). On the other hand, in the P1 sample, the concentrations of the analytes were
about 7 mg kg-1 (Br), 1,285 mg kg-1 (Cl), 34 mg kg-1 (F), < 0.5 mg kg-1 (I), and 7,900 mg kg-1 (S).
1EFSA, EFSA Journal, 12(7), 2014.
[CNPq, CAPES, FAPERGS, UFPel]

STUDY OF MAGNETIC ADSORBENTS 2-AMINOBENZOTHIAZOLE FUNCTIONALIZED
IN THE SELECTIVE PRECONCENTRATION OF Cd(II) IONS AND TS-FF-AAS
Liriana M. Rovedaa, Pâmela C. de Oliveiraa, Lucimara R. P. Ruisa, César R. T. Tarleyb,c,
Daiane Romana, Marcela Z. Corazzab*
aFederal University of Grande Dourados, FACET, Dourados, MS, Brazil, 79.804-970.
b StateUniversity of Londrina, Department of Chemistry, Londrina, PR, Brazil, CEP 86057-970
cState University of Campinas, National Institute of Science and Technology (INCT) of Bioanalytical,
Campinas, SP, Brazil.

*e-mail: corazza@uel.br
Exploring the chemical and physical properties of nanomaterials, especially when associated with
magnetism, has yielded a series of improvements in analytical processes aimed at separation and
detection heavy metals at trace levels1. Magnetite and carbon nanotubes are among the
nanoparticles that have been used as a basis for the preparation of SPE adsorbents. In addition to
the attractive properties intrinsic to nanometric particles, such as high specific surface area and fast
kinetics of mass transfer, these materials can be easily modified by the insertion of functional groups
that direct their selectivity towards analytes of interest2,3. Aware of these properties, two novel
magnetic sorbents, based on silica coated magnetite (Fe3O4@SiO2) and other based on multiwalled
carbon nanotubes (MWCNT), both functionalized with 2-aminobenzothiazole (ABT), a ligand with
potential functional groups to adsorption Cd(II) ions, were synthetized and applied in the
preconcentration and separation of Cd(II) ions in water and food samples. The sorbents were
characterized by FTIR, TGA, BET, DRX, SEM and TEM. The FTIR spectra showed peaks in the
range of 3400 – 3100 cm-1, corresponding to the stretching vibrations of the –NH groups, which
confirm that the sorbents were successfully functionalized with the ABT ligand. Under the previously
optimized pH conditions, other parameters pertinent to the magnetic SPE system, such as mass
adsorbent, eluent type, preconcentration and elution times, were investigated using full factorial
design 24. To make the proposed methods even more sensitive, thermospray flame furnace atomic
absorption spectrometry (TS-FF-AAS) was employed as a detection technique. Preconcentration
factors of 19.36 and 10.10 times and limits of detection of 0.02 and 0.11 µg L-1 were achieved with
Fe3O4@SiO2-ABT and MWCNT-Fe3O4@ABT, respectively. The retention of Cd(II) ions in the
adsorbents was not affected in the presence of foreign ions, studied in different proportions. As proof
of concept, the analytical methods developed with these sorbents were applied to samples of water,
rice, mate tea and tobacco. The percentages of Cd(II) recovery at the 95% confidence level for the
two methods ranged from 91 to 109%. Accuracy was assessed using GFAAS as a reference
technique and the results obtained showed no statistical differences in relation to the proposed
methods. These results prove that the materials exhibit performances with satisfactory results when
applied to real samples. Thus, both should be considered promising in studies of contaminant
removal in analytical processes, aiming at the preconcentration of ions and their consequent
determination in aqueous and food matrices.
1 Bendicho C, Bendicho-Lavilla C, Lavila L, Trends in Analytical Chemistry, 77, 2016, 109

2 Giakisikli G, Anthemidis A N, Analytica Chimica Acta, 789, 2013, 1
3 Kazemi E, Dadfarnia S, Haji Shabani A M, International Journal of Environmental Analytical Chemistry, 97,2017,1
[UFGD, UEL, LAMAAE, CNPq, INCT]

ULTRASOUND-ASSISTED EXTRACTION AS A SIMPLE STRATEGY FOR THE
DETERMINATION OF MACRO AND MICROELEMENTS IN CAJUÍNA
EMPLOYING ICP OES AND MULTIVARIATE ANALYSIS
Tiago L. S. Coelho (PG)a*, Tomas M. C. Silva (PG)a, Darlisson S. N. Silva (PG)a, Luiz B. Souza
Filho (PG)a, Jardel M. Rocha (PG)a, Raimundo R. Gamela (PG)b, Ivero P. Sá (PG)c, Julymar M.
Higuera (PG)c, Cícero A. Lopes Júnior (R)a, Ana R. A. Nogueira (R)d, Edivan C. Vieira (R)a
aUniversidadeFederal do Piauí, Departamento de Química, Teresina, Piauí, Brasil, CEP: 64049-550
bInstituto
Superior Politécnico de Gaza, Chokwe-Gaza, Moçambique, 1204
cUniversidade Federal de São Carlos, Departamento de Química, São Carlos, São Paulo, Brasil, 13565-905
dEmbrapa Pecuária Sudeste, São Carlos, São Paulo, Brasil, 13560-970
*e-mail: tiagolinusquimico@gmail.com
The cashew apple has been relevant in the food industry to produce several products such as
sweets, jellies, and juices. Among these, Cajuína, a typical Northeastern Brazilian beverage with a
Geographical Indication (GI) certificate from the State of Piauí and is registered as a Brazilian
Cultural Heritage by the Institute of National Historical and Artistic Heritage (IPHAN) 1. Cajuína is
prepared from cashew juice, without alcohol, clarified, and sterilized. It has an amber-yellow color
resulting from the caramelization of the juice's natural sugars2. Despite being nationally widespread,
vitamin C reduction in industrialized cashew juice and Cajuína was reported3. Furthermore, Cajuína
still has an unknown inorganic composition. In this sense, this study focuses on determining
inorganic constituents in the Cajuína, an essential task for nutritional reasons and product quality. In
this context, a fast, safe, simple, and low-cost analytical method was developed for the determination
of macro and microelements in Cajuína samples by inductively coupled plasma optical emission
spectrometry (ICP OES) using ultrasound-assisted extraction (UAE). The UAE was applied to extract
Ca, Cu, Fe, K, Mg, Na, P, S, and Zn in Cajuína juices produced in Piauí and Ceará, Brazil. A factorial
and face-centered central composite design were applied for the UAE optimization. It was possible
to observe significant variables and reach the optimal extraction conditions, which were 30 min of
sonication time and 4 mL of HNO3 8 mol L-1 plus 1.5 mL H2O2. This procedure allowed the
determination of Ca, Cu, Fe, K, Mg, Na, P, S and Zn with limits of detection (LOD) 0.103, 0.0012,
0.0038, 0.557, 0.002, 0.053, 0.087, 0.016 and 0.009 mg L-1, respectively. The precision expressed
in relative standard deviation (% RSD), < 5.5% (n = 3), and the recovery of the method, evaluated
by spike experiments in the UAE, presented results between 87 to 108%. The accuracy was
confirmed by comparison with the microwave-assisted digestion method. The analysis of 38 samples
of Cajuína juices presented an average of Ca, Cu, Fe, K, Mg, Na, P, S and Zn among 8.70. 0.10,
0.97, 1223.33, 111.79, 60.05, 83.97, 65.10, and 0.99 mg L−1. Therefore, the proposed analytical
method is a good alternative for simultaneously determining macro and microelements in Cajuína
and juice similar matrices.
1 IPHAN, Institute of National Historic and Artistic Heritage. http://portal.iphan.gov.br/noticias/detalhes/286.

2 Galvão LMV, Sousa MDM, Nascimento AMDCB, Souza BVC, Nunes LCC, Food Sci. Technol. 42, 2022, 1–6.
3 Lima ES, Silva EG, Neto JMM, Moita GC, Quimica Nova, 30, 2007, 1143–1146.
[UFPI, CNPq, Embrapa Pecuária Sudeste]

DEVELOPMENT OF A METHODOLOGY FOR TRAPPING OF PAHS IN VEHICLE
COMBUSTION USING THE IT-FEX DEVICE FOR ANALYSIS BY GC-BID
Henrique G. P. Menezes (PG)a, Ísis G. S. Silva (PG)a, Jhonatan B. Oliveira (PG)b,
Venilton M. V. Ferreira (PG)c, Patrícia S. Rezende (R)a, Patterson P. Souza (R)c
aFederal Center for Technological Education of Minas Gerais State, Technology of Products and Process Post-
GraduateProgram, Belo Horizonte, MG, Brazil, 30510-000
b Federal University of Minas Gerais State, Department of Chemistry, Belo Horizonte, MG, Brazil, 31270-010
cFederal Center for Technological Education of Minas Gerais State, Multicenter Chemistry Post-Graduate
Program,Belo Horizonte, MG, Brazil, 30510-000

*e-mail: henriquegiacomo96@gmail.com
Polycyclic aromatic hydrocarbons (PAHs) are a class of highly dangerous compounds, mainly due
to their genotoxicity, in addition to having a long half-life, causing biomagnification in living beings.
Their usual formation and subsequent atmospheric emission occur by the burning of fossil fuels,
mainly by diesel engines [1]. Miniaturized sampling techniques have presented great advances,
suchas in tube solid phase microextraction (IT-SPME), which allows the capture and concentration
of analytes in a fast way (minutes), even in less availability of the compounds in complex matrices.
It aims to adsorb the compounds on sheets of adsorbent polymeric fibers inside the tube, trapping
them. Subsequently, these are thermally desorbed and, finally, available for chromatographic
analysis [2]. In order to innovate in this technique, the In-Tube Extraction device (IT-FEx) was
developed – patent # BR1020180731670 – being a borosilicate glass tube with the inner walls
coatedwith a polymeric film of PDMS, adsorbing the compounds even in a liquid phase such as in
headspace. Thus, the chemical species are retained by the film to its maximum, reaching phase
equilibrium. However, changes in sampling with IT-FEx are desirable, allowing the compounds to
betrapped in a dynamic process (without reaching system equilibrium), in order to mimic vehicular
combustion, with complete passage of the matrix (carrying the analytes) through the fiber.
Therefore,the work proposed a potential method of dynamic sampling in the gas phase, using the
IT-FEx to capture PAHs, which is coupled to a portable sampler at the exit of the vehicle's exhaust
for 30 s, then, inserting the device in the inlet of gas chromatograph, providing a thermal desorption
of compounds from the fiber and subsequent analysis. The devices were made according to the
studiesof Caldeira (2020) [3] and characterized by thermogravimetry and infrared spectroscopy. A
Sigma-Aldrich standard mixture of 15 PAHs (1000 µg mL-1) was used. The gas chromatograph
was a Shimadzu Nexis-2030 (with a BID detector), containing a HP-5MS fused silica capillary
column (30 m x 0.25 mm x 0.25 μm). The chromatographic method started at 50 °C (1.0 min), then
20 °C min-1until 160 °C (7.0 min), and finally 20 °C min-1 until 315 °C (4.25 min). The method
proved to be satisfactory in the separation of 9 compounds, with peak resolution greater than 1.5.
Only 3 pairs ofPAHs could not be separated due to their very high structural similarity, being then
analyzed in a summed way by the non-separated pairs (quantitative results) without significant
losses to the paper’s purpose. The adsorption selectivity of the compounds in aqueous phase
(reaching the equilibrium in 25 min) was analyzed, obtaining marked selectivity for medium-sized
PAHs (3 and 4 rings) in comparison to the others analyzed (1, 2, 5 and 6 rings). Thus, the sampling
test in the dynamic flow for a laboratorial bench level was performed using a heating apparatus set
at 75 °C,in which 1.0 mL of a standard solution of PAHs was inserted in a chamber and sealed,
passing, in the gaseous phase, only through the IT-FEx (attached to the chamber lid), and
conducted to the chromatograph. Finally, tests were carried out on real samples of exhaust
emissions from diesel engine vehicles, which proved to be promising. Peaks referring to PAHs
were observed with satisfactory separation for quantitative analysis, to be accomplished in
following studies subsequent to this paper.
1 Genaro LAT, VA M, Borra E. Atmospheric Environment, 43, 2009, 5944
2 Fan W, Almirall J. Analytical and Bioanalytical Chemistry, 406, 2014, 2189
3 Caldeira GS. Federal Center for Technological Education of Minas Gerais, 2020, 100
[FAPEMIG, CEFET-MG]

A GREENER METHOD FOR DETERMINATION OF As, Cd, AND Pb IN SOIL AND
PLANT TISSUE BY ICP-MS EMPLOYING NADES IN SAMPLE PREPARATION
Sabrina S. Ferreira (PG)a*, Thomas A. Brito (PG)a, Floriatan S. Costa (PG)b,
Taciana G.S. Guimarães (PG)c, Daniel F. Andrade (R)d, Rafaela S. Lamarca (R)a,
Andrea Oliveira (R)b, Clarice D.B. Amaral (R)b, Mario H. Gonzalez (R)a
a São Paulo State University (UNESP), Department of Chemistry and Environmental Science, National Institute for
Alternative Technologies of Detection, Toxicological Evaluation and Removal of Micropollutants and Radioactives (INCT-
DATREM), São José do Rio Preto, SP, Brazil, 15054-000
b Federal University of Paraná, Department of Chemistry, Curitiba, PR, Brazil, 81531-980
c Federal University of São Paulo, Laboratory of Integrated Sciences, Diadema, SP, Brazil, 09972-270
d Federal University of São Carlos, Department of Chemistry, São Carlos, SP, Brazil, 13565-905
*e-mail: sabrina.s.ferreira@unesp.br
The principles of green analytical chemistry have been widely applied in the development of
analytical procedures1, with three main objectives: reduced use of reagents and toxic compounds;
increase in energy efficiency; and the development of safe compounds using raw materials from
renewable sources2. Natural deep eutectic solvents (NADES) are considered green solvents due to
attractive characteristics, including biodegradability, low toxicity, low cost, simple preparation and
handling, low volatility, and, importantly, adjustable physicochemical properties (density, viscosity,
and polarity)5. These solvents are composed of natural precursors whose interaction occurs between
a hydrogen bond donor compound (HBD) and a hydrogen bond acceptor compound (HBA)4. These
solvents have been widely used in green sample preparation methods, with the aim of obtaining
reliable and accurate results, while prioritizing the sustainability of the analytical process. In this
work, NADES based on carbohydrate and organic acids were synthesized, characterized, and
evaluated in terms of their suitability for the determination of inorganic contaminants in soil and plant
tissue samples. NADES based on citric acid/malic acid/water (CA-MA) and citric acid/xylitol/water
(CA-Xyl) were synthesized according to a ratio of 42:13:45 (% w w−1) under stirring and heating6,
followed by the determination of the viscosity and density values. The NADES were employed in a
microwave-assisted extraction (MAE) method optimized for the extraction of As, Cd, and Pb,
according to a Doehlert Design. The analytes concentration present in the soil and plant tissue
extracts were determined by inductively coupled plasma mass spectrometry (ICP-MS) standard
mode and a kinetic energy discrimination cell (KED) mode to avoid possible interferences. The
density values for the CA-Xyl and CA-MA NADES were 1.26 ± 0.0126 and 1.26 ± 0.0034 g mL-1,
respectively, while the corresponding viscosity values were 7.54 ± 0.030 and 5.20 ± 0.010 mPa.s.
The optimization of the extraction conditions considered the effects of the differents temperature,
extraction time, and sample mass/NADES volume ratio. Recoveries in the range from 80 to 110%
were obtained for 111Cd+ and 208Pb+, when the CA-MA and CA-Xyl NADES were used for the
extraction of a soil-certified reference material (EMBRAPA, Agro E2002a), as well as for 75As+ and
208
Pb+, when the CA-Xyl NADES was used to extract a forage grass CRM (Brachiaria brizantha cv.
Marandu; EMBRAPA, FO-01/2012). The solvents proved to be highly promising as extractors for the
samples evaluated. The use of experimental design assisted in the optimization of the MAE method,
obtaining an efficient analytical method that was following the GAC.
1 Sajid, M., Płotka-Wasylka, J. Talanta, v. 238, 2022.
2 La Guardia, M.; Garrigues, S. Challenges in Green Analytical Chemistry v. 2, 2020.
3 Nowak, P. M., et al. TrAC Trends in Analytical Chemistry, v. 138, p. 116223, 2021.
4 Dai, Y. et al. Analytica Chimica Acta, v. 766, pp. 61–68, 2013.
5 Fernández, M. de los Á. et al. Analytica Chimica Acta,v.1038, pp. 1-10, 2018.
6 Santana, A.P.R. Talanta, v. 216, 2020.
[CAPES, FAPESP, CNPq, INCT-DATREM]

IONIC LIQUID INTERCALATED IN MONTIMORILLONITE (IL-MMT) FOR
EXTRACTION OF PAHs BY PIPETTE-TIP SOLID PHASE EXTRACTION
Débora de Azevedo Domingues (PG)a*, Rafael Garrett Dolatto (PG)a, Diana Castillo
Gómez (PG), Milton Hernando Rosero Moreano (R)b and Marco Tadeu Grassi (R)a
a Universidade Federal do Paraná (UFPR), Departamento de Química, Curitiba, Paraná, Brazil, 81531-980.
b Universidad de Caldas, Departamento de Química, Grupo de Investigación en Cromatografía y Técnicas Afines,
GICTA, Caldas, Manizales, Colombia, 170004.
*e-mail: deboraazevedod@ufpr.br
Determination of polycyclic aromatic hydrocarbons (PAHs) in environmental aqueous matrices is an

analytical challenge due to physical-chemical characteristics such as solubility in water,
heterogeneity, and hydrophobicity, among others. In addition, PAHs often occur at trace levels in
environmental samples. Therefore, sample preparation is a crucial step for the detection and
reliability of the analytical protocol¹. Among the methods of microextraction, pipette-tip solid phase
extraction (PT-SPE) stands out, because it requires fewer material (sample, solvents, sorbent, etc.)².
As an example of sorbent phase, IL-MMT has been reported as an attractive alternative, for the
extraction of low-polarity organic compounds in water³. In this work a method based on PT-SPE
using IL-MMT as the sorbent for extraction of priority PAHs in seawater, for first time, with the
determination step carried out by gas chromatography-mass spectrometry (GC-MS). A self-made
extraction device was assembled re-using two pipette tips (1250 and 200 µL), glass wool, and the
IL-MMT. Aqueous solutions of PAHs at 1 µg L-1 were used and the following variables were studied
through univariate design: amount of sorbent (5, 10, 15, 20 mg), solvent type (hexane, acetonitrile,
acetone), solvent volume (250, 500, 1000 µL), number of desorption cycles (3, 10, 15, 20), sample
volume (2.5; 5; 10 mL), and number of extraction cycles (5, 10, 15, 20). For light PAHs (Naphthalene
to Fluorene) 20 mg of the IL-MMT exhibit higher peak areas being adopted for the following steps.
The best efficiency for solvent was obtained when 500 µL of hexane was used as eluent, and its use
led to higher analytical frequency, eliminating the evaporation and resuspension steps. Also, after
10 cycles of extraction and 15 of desorption the efficiency reached an apparent equilibrium, which is
time-dependent and can be attained after certain extraction/desorption cycles². Finally, the best
extraction efficiency was reached when the smaller sample volume (2.5 mL) was used. Afterwards,
a two level (-1, +1) fractional factorial design 26-2, with 16 experiments, was defined. The variables
and working ranges were the amount of IL-MMT (15-20 mg); solvent type (acetone-hexane); solvent
volume (250-500 µL); number of desorption and extraction cycles (10-15) and sample volume (2.5-
5.0 mL). Based on these conditions it was possible to estimate enrichment factors of 20 times,
determined according to an instrumental analytical curve. Protocol repeatability was evaluated
based on five extractions of a PAH aqueous solution and the uncertainty levels proved to be
satisfactory (RSD 5.6-10%). Results of the application of the PT-SPE procedure in aqueous samples
will be shown.
1 Sajid M, Nazal MK, Ihsanullah I. Analytica Chimica Acta, 2021.
2 Zhang Y, Zhao YG, Chen WS, Cheng HL, Zeng XQ, Zhu Y. Journal of Chromatography A, 2018.
3 Fiscal-Ladino JA, Obando-Ceballos M, Rosero MF, Montaño DF, Cardona W, Giraldo LF, Richter P. Analytica Chimica
Acta, 953, 2017.
[UFPR, CAPES, CNPq, FINEP, PETROBRAS, INCTAA]

COMPARISON STUDY OF EXTRACTION PROCEDURES FOR METALS
DETERMINATION IN SEDIMENTS WITH DIFFERENT CHARACTERISTICS
Bruna Silva Correa (UG)a, Larissa D. Roriz (UG)a, Suellen G. Cordeiro (PG)a,
Maiara Krause (PG)a, Luiza, V. Vieira(R)a, Geisamanda P. Brandao(R)a,
Maria Tereza W. D. Carneiro(R)a*
aFederal University of Espirito Santo, Chemistry Department/CCE, Vitoria, ES, Brazil, 29075-910
*e-mail: jmariacarneiro@hotmail.com
Currently, atomic spectrometric techniques are frequently used to determine trace elements in
sediment samples. However, usually, the samples are introduced as an aerosol liquid gas and the
sample should be present as a liquid. So, sample preparation steps are often necessary. Microwave-
assisted acid decomposition is widely used as a sample preparation procedure in which different
acid mixtures may be used. The extraction may be total or partial, depending on the acids used. The
process can decompose the entire sample matrix, making available all the analytes previously
trapped in the crystalline lattice, or decompose part of the sample, making available a significant
amount of the analytes. It is not simple to choose acids and methods of extraction to determine the
concentrations of trace elements in complex matrix samples, such as sediments.1 The present study
aimed to compare the effectiveness of three extraction methods, the USEPA 3051A HNO3, US EPA
3051A HNO3+HCl, and the USEPA 3052 HNO3+HCl+HF to determine Al, Fe, Mn, and Zn in 17
marine sediment samples with different granulometric and compositional characteristics. The marine
sediment samples used for the study had compositional variations based on carbonate content and
heavy minerals. Samples with carbonate content greater than 55% m/m, are called carbonate, and
samples with less than 15% m/m, are called siliciclastic. Samples with >1% m/m of heavy minerals
(HM) were called high HM and samples with levels <1%m/m were named low HM. The metals were
determined using an inductively coupled plasma optical emission spectrometry (ICP OES). The
precision of the methods was evaluated through the RSD values, which ranged from 0.7 to 19.9%
for the most accurate data (RSD<20% was considered acceptable).2 The accuracy of the methods
was evaluated using the reference material NIST 2702 (marine sediment) and the observed
recoveries ranged from 15.9 to 87.1%. The content of Al, Fe, Mn, and Zn was compared using the
different methods in all the samples and it was expected the increasing order of extraction: EPA
3051A HNO3, EPA 3051A HNO3+HCl, and EPA 3052. The nitric acid only does not completely
decompose sediment, as does HF, but the presence of HCl stabilizes Al and Fe in the solution. This
work showed that in general, the EPA 3051A HNO3+HCl extracted more when compared to the EPA
3051A HNO3 method for all elements, in all sample classes. However, for the EPA 3052 method, the
expected only occurred for siliciclastic sands and for Mn and Zn in carbonate samples. To improve
the performance of EPA 3052 method, the volumes of the acids used, and the method of addition of
H3BO3 were changed. The recoveries of the CRM were satisfactory for all elements, except Zn. The
results obtained for the samples showed a better extraction of Al and Fe in the carbonate ones, but
not for Mn and Zn. So, the results of this study showed that the most appropriate extraction method
depends on the compositional characteristics of the sediment and the elements intended to
determine.
1 Ščančar J., Milačič R, Horvat M. Water, Air, and Soil Pollution, v. 118, n. 1, p. 87-99, 2000.
2 Chen M., & Ma LQ. American Society of Agronomy, Crop Science Society of America, and Soil Science Society of
America, 1998.
[CNPq, CAPES, FAPES, PPGQUI-UFES, LabPetro/UFES, FEST-Renova]

COMPARATIVE ANALYSIS OF SAMPLE PREPARATION METHODS FOR
DETERMINATION OF COPPER AND SILVER IN INTRAUTERINE DEVICE (IUD)
Clara C. S. Pereira (PG)a*, Santos A. V. Neto (R)a, Cristiane Barata-Silva (R)a,
Silvana C. Jacob (R)a, Josino C. Moreira (R)a, Lisia M. G. Santos (R)a
1Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (Fiocruz), Departamento de
Química/ Setor de Elementos Inorgânicos, Rio de Janeiro, Rio de Janeiro, Brasil, CEP: 21.040-900
*e-mail: clara.caruso@outlook.com
Intrauterine devices (IUDs) and contraceptive implants, also called long-acting reversible
contraceptives (LARC), are the most effective reversible contraceptive methods and rea great
alternative for families to carry out family planning 1. Currently, different types of IUDs are available
on the national market, but the one made available by the Unified Health System (SUS) is Copper-
containing IUDs2. Despite being scarcely used in Brazil (less than 5% of sexually active women use
IUDs), this method is highly effective, low cost, and easy to use3. Due to the need to control the
quality of this device, ANVISA published legislation RDC nº 552 (2021)4 which determines the
minimum content of 99.9% of Cu and does not define the minimum allowed content of Silver (Ag) in
IUDs marketed in Brazil. The aim of the present study was to compare different sample preparation
methods for the determination of Copper (Cu) and Silver (Ag) in an intrauterine device. The 15
samples were acquired from the same batch, marketed in Brazil and each sample were weighed
(0.05 g) to perform acid digestion in an open system, at 3 different times: 15 minutes, 24 hours and
48 hours, and acid microwave digestion using a SpeedWave microwave (Berghof, Germany).
Samples were analyzed by Inductively Coupled Plasma Optical Emission Spectrometry (ICP OES),
model Optima 8300, Perkin Elmer. Statistical analysis was performed using the Past® Software
version 3.26. The mean of Cu and Ag concentration for the closed system acid digestion method
was 81.2 ± 5.9% and 8.1 ± 0.5 %, respectively. In the open system, the concentration of Cu and Ag
was 81.0 ± 4.6 % and 8.1 ± 0.4 % for the time of 15 minutes, 83.4 ± 1.5% and 8.4 ± 0.2% for 24
hours, and 82.3 ± 0.7% and 8.2 ± 0.1% for 48 hours, respectively. With a confidence interval of 95%,
the Analysis of Variance (ANOVA) indicates that there is no difference between the averages
obtained. In this way, the study indicates that the sample preparation techniques evaluated produce
similar results, thus allowing the analyst to choose the way in which he will prepare the sample,
according to his reality in the laboratory for quality control in health.
1 Penna IAA, Brito MB. A importância da contracepção de longo prazo reversível. Universidade Federal Fluminense (UFF)
– Niterói (RJ), Brasil. FEMINA. FEBRASGO – Federação Brasileira das Associações de Ginecologia e Obstetrícia. Volume
43. Suppl. 1. 2015.
2 BRASIL. Portaria número 3.265, de 1 de dezembro de 2017. Altera o Anexo XXVIII da Portaria de Consolidação nº
2/GM/MS, de 28 de setembro de 2017, que dispõe sobre a ampliação do acesso ao Dispositivo Intrauterino TCu 380 (DIU
de cobre) no âmbito do Sistema Único de Saúde (SUS). Ministério da Saúde.
3 Machado RB, et al Long-Acting Reversible Contraception. Rev. Bras. Ginecol. Obstet. 39 (06) June 2017.
4 BRASIL. Resolução da Diretoria Colegiada número 552, de 30 de outubro de 2021. Dispõe sobre registro, fabricação,
controle de qualidade, comercialização e uso de Dispositivo Intra-Uterino (DIU) contendo cobre. Brasília - DF, 2021.
[Ministério da Saúde, CAPES]

DEVELOPMENT OF A DISPERSIVE LIQUID–LIQUID MICROEXTRACTION
METHOD FOR THE DETERMINATION OF FIVE ANTIMICROBIALS IN MILK
USING LC-QTOF-MS ANALYSIS
Maurício Perina *, Louise Janka, Fabiano Barretoa
aLaboratório Federal De Defesa Agropecuária – LFDA/RS. Ministério da Agricultura, Pecuária e Abastecimento, Porto
Alegre, RS, Brazil, 91780580
*e-mail: mauricio.perin@apoio .gov.br
Drug residue is the most serious problem in food safety. The polypeptides bacitracin A (BTC),
virginiamycin (VIR), and the macrolides, azithromycin (AZI), clindamycin (CLI) and tilmicosin (TIL)
are antimicrobials which are commonly used veterinary medicines for the promotion of growth,
prevention and control of diseases in farm animals. Several countries and Brazil have set values of
maximum residues limits (MRL)1 for these compounds. Milk was included in National Residue and
Contaminants Control Program (PNCRC) and the polypepetides and macrolides class are critical
group for risk assessment. The sample preparation techniques that have been used previously to
extract antibiotics compounds from milk (SPE and LLE) are time consuming and are not
environmental friendly2. In the present study, we developed a simple, rapid, and eco-friendly method
(DLLME) for extraction of BTC, VIR, AZI, CLI and TIL from milk samples, which can be used as a
routine extraction method in food quality control laboratories. The sample preparation procedure is
based on an miniaturized liquid-liquid extraction (DLLME) as shown in Figure 1.
Figure1: Schematic of the sample preparation procedure using DLLME for the determination of macrolide antimicrobials
in a milk.
The effect of several parameters on DLLME efficiency was evaluated, among them, a study to
remove proteins of milk with trichloroacetic acid. When different concentrations of the acid 1-10%
(w/v) were added to 2g of fortified milk, the best results were obtained in volume of 8 mL with
concentration of the 5% (w/v) acid. The recoveries for milk samples spiked at concentration levels
(LMR) ranging between 50 to 200 ng g−1, depending on the compound, were between 73 and 108%,
except for the CLI (40%), for all compounds relative standard deviations lower than 20%. Simplicity,
rapidity and reliability are important advantages of the proposed method, while the sample
preparation step can be regarded as environmentally friendly.
1 ANVISA, Instrução Normativa N° 51, de 19 de dezembro de 2019, 2019.

2 Campello N, et al., Dispersive liquid–liquid microextraction for the determination of macrocyclic lactones in milk by liquid
chromatography with diode array detection and atmospheric pressure chemical ionization ion-trap tandem mass
spectrometry, Journal of Chromatography A, 2013.
[CNPq, MAPA, LFDA-RS]

DEVELOPMENT OF A METHOD FOR ANALYSIS OF DISINFECTION BY-
PRODUCTS IN DRINKING WATER USING HS-SPME-GC-MS
Maísa D. Habenschusa, Eduardo T. Cunhaa*, Amanda G. Nitolia,
aCompanhia de Saneamento Básico do Estado de São Paulo (SABESP), Departamento de Controle da Qualidade dos
Produtos Água e Esgotos (TOQ), São Paulo, São Paulo, Brazil, CEP 07082-240
*e-mail: ecunha@sabesp.com.br
Disinfection By-products (DBPs) are substances formed by oxidizing agents, such as chlorine, during
the water disinfection process. Some of the most commonly DBPs formed from chlorination are the
Trihalomethanes (THMs, Chloroform, Bromoform, Bromodichloromethane and
Dibromochloromethane), Haloacetic Acids and Halophenols [2,4-Dichlorophenol (DCP) and 2,4,6-
Trichlorophenol (TCP)]1. This work proposed the development and validation of an analytical method
for the simultaneous monitoring of DCP, TCP and THMs in drinking water using the headspace solid-
phase microextraction sampling technique (HS-SPME), followed by gas chromatography (GC)
coupled with mass spectrometry (MS), HS-SPME-GC-MS. The extraction parameters are shown in
Table 1.
Table 1- Extraction Parameters used during SPME.

Fiber PDMS/DVB 65 µm
Incubation Temperature (ºC) 65
Incubation Time (min) 3
Extraction Time (min) 10
Desorption Temperature (ºC) 250
Desorption Time (min) 2
The developed method was validated according to the National Institute of Metrology, Quality and
Technology (INMETRO) guidelines2. The parameters verified during the validation were the
selectivity, carryover, matrix effect, precision and recovery, limit of quantification (LOQ), linearity and
repeatability. Selectivity showed no interference peaks and no carryover was observed.
The matrix effect was studied using a sample of drinking water, without any analyte, collected in a
40 mL amber bottle, preserved with HCl (1:1 v/v, pH ≤2) and dechlorinated with sodium thiosulfate
p.a (75 mg L-1). The results of the F test and t test (Student) showed that the matrix had no effect
on the precision and recovery of the method at the three concentration levels (low, medium and high)
studied. The obtained results are shown in Table 2.
Table 2- Obtained results.
LOQ Linear Range Repeatability - Intermediate Precision -
Analyte Recovery (%)
(µg L-1) (µg L-1) RSD (%) RSD (%)
THM 12.0 12 to 120 4.1 to 16 82.1 to 107.7 4.2 to 16.1
DCP and TCP 3.0 3 to 30 2.1 to 7.7 94.1 to 98.5 7.4 to 13.7
In conclusion, the method proved to be suitable for laboratory application. Furthermore, it also
presented economic and technical advantages when compared to traditional methods: not needing
to use large volumes of samples, organic solvents and a considerable reduction in sample
preparation time.
1 GILCA AF. et al. Emerging disinfection byproducts: A review on their occurrence and control in drinking water treatment
processes. Chemosphere, v. 259, p. 127476, 1 nov. 2020.
2 INMETRO. Orientação sobre Validação de Métodos Analíticos- DOC-CGCRE-008. Available in:
<http://www.inmetro.gov.br/Sidoq/pesquisa_link.asp?seq_tipo_documento=5&cod_uo_numeracao=00774&num_docum
ento=008>. Acesso em: 4 abr. 2022.

ENVIRONMENTAL
CHEMISTRY
312
ASSESSMENT OF CONCENTRATION LEVELS OF 16 GASEOUS ATMOSPHERIC
POLLUTANTS/CONTAMINANTS IN 5 BRAZILIAN CITIES
Franciele O C da Rochaa*, Vânia P Camposb, Gisele O da Rochab, Marcos A Bezerrac, Indiara
S Sampaiob e Anne Valesca S Britob
aUniversidade Federal de Roraima, Centro de Ciências e Tecnologia, Departamento de Química, Boa Vista, Roraima,
Brazil, 69310-000
bUniversidade Federal da Bahia, Instituto de Química, Departamento de Química Analítica, Salvador, Bahia, Brazil,
40170-115
cUniversidade Estadual do Sudoeste da Bahia, Departamento de Ciências e Tecnologias, Jequié, Bahia, Brazil, 445205-
490
*e-mail: francieleocr@gmail.com
The growing population in urban centers is often exposed to concentrations of gases and particles
that often exceed established air quality standards. In order to understand the main processes that
may be involved in the variation of pollutant concentrations in the air in Brazilian cities, the
concentrations of 16 gaseous pollutants/contaminants (SO2, O3, NO2, NOx, NO, NH3, BTEX, HCOH,
CH3COH, HCl, HNO3, HCOOH and CH3COOH), were determinate after passive sampling. Possible
sources of emission and the effect of the sampling period (dry and rainy) were discussed from
backward air mass trajectories (NOAA HYSPLIT model), wind roses (WRPLOT View model) and
multivariate statistics. Six sampling campaigns were carried out using the AnaliseAr kit, for
consecutive periods of 7 and 14 days, in Salvador, São Paulo, Belo Horizonte, Rio de Janeiro and
Londrina. BTEX compounds were quantified by gas chromatography with flame ionization detection;
HCOH and CH3COH by high performance liquid chromatography with UV-Vis detection; NOx, NO2,
NH3 and H2S by UV-VIS molecular absorption spectrophotometry and SO2, O3, HCl, HNO3, HCOOH
and CH3COOH by ion chromatography. Although without exceeding the limit legislated by CONAMA
Resolution 491/20181 (160 µg m-3), O3 had the highest mean concentration in São Paulo (58 µg m-
3
), ranging from 5.3 to 139 µg m-3 in all urban centers considered in this study, followed by Belo
Horizonte (BH).This corroborated with the intensity of the vehicular fleet in the two cities, combined
with meteorological parameters and NOx concentrations levels, which exceeded the European
Union recommendation2 as critical levels for the protection of vegetation (30 µg m-3). The
concentration of formaldehyde and acetaldehyde exceeded international recommendations in all
urban centers and benzene concentrations showed values > 1.7 μg m-3, a level associated with the
probability of leukemia, according to the World Health Organization3. NH3 concentrations indicated
a significant contribution of anthropogenic emissions from the use of biofuels, industrial sectors or
the use of fertilizers. The urban centers influenced by the marine aerosol had lower concentrations
of HNO3. This can be explained by the occurrence of the reaction of this acid with NaCl from the sea
spray, producing particulate matter and gaseous HCl. The discharge of sewage in the Pinheiros river
- SP and in the Pampulha lagoon - BH, close to the sampling points, contributed to higher levels of
H2S, toluene and NH3 in these urban centers. Additionally, the BH atmosphere was also influenced
by emissions from 2 landfills, located about 10 km from the sampling site. Organic and inorganic
acids presented a similar percentage of contribution to the atmospheric acidity. Ratios < 1 between
the concentrations of formic acid and acetic acid in 71% of the cases demonstrated the influence of
vehicle emissions on the atmospheric levels of these contaminants. Strong positive correlations
between nitrogen pollutants (NOx, HNO3, NH3), conventional pollutants (SO2, NO2), xylenes and
aldehydes indicated a significant influence of vehicular emissions for the observed concentrations
levels. In general, multivariate analysis showed that VOCs and organic acids are more influenced
by temperature and solar radiation parameters. The strongest direct relationship of NOx was with
wind direction, showing competition between local emission and photochemical aging of these
compounds brought from other locations.
1 Brazil, Resolution CONAMA 491/2018.
2 European Union - Directive 2008/50/EC.
3 WHO - World Health Organization, Air Quality Guidelines-2005
[FAPESB, INCT E&A, CIEnAm-UFBA, UEL-PR]
––––––––––––––––––––– Environmental chemistry –––––––––––––––––––––

313
DETERMINATION OF MERCURY (TOTAL AND INORGANIC) IN THE WATER
OF LAGOON MUNDAÚ (Maceió/AL, Brazil)
Sângela Ferreria da Silva (PG)*, Aryanna S.P.N. Costa (PG),Emerson Freire (PG), Ellen dos
Santos Silva Barros (PG), Daniela S. Anunciação (R), Josué Carinhanha C. Santos (R)a
Universidade Federal de Alagoas, Instituto de Química e Biotecnologia, Maceió - AL, Brazil, 57072-900
*e-mail: sangela.silva@iqb.ufal.br
The Mundaú lagoon (Maceió, AL) makes up the Mundaú Manguaba Lagoon Estuarine Complex
(CELMM), responsible for a large part of the population density of the place; besides, it is a source
of income and food for about 260 thousand people.1 The Mundaú lagoon has a direct connection to
the Atlantic Ocean, characterizing it as brackish water (class 1) used for recreation, aquaculture,
irrigation, and fishing activities by the local population.2The present work aimed to evaluate levels of
total [Hgtotal] and inorganic mercury in the cationic form [Hgino = Hg(II)] in water samples and to assess
physicochemical parameters in the study area. Thus, four samplings were carried out from July/2021
to July/2022, so they were in rainy and dry periods. Therefore, 11 selected points were chosen
considering the discharge of domestic and industrial effluents, high and low population occupation,
and regional fishing activity. For the determination of Hgino the samples were collected in amber glass
flasks (without acidification) because some organomercury compounds are photosensitive, while for
the determination of Hgtotal, the samples were collected in glass flasks previously acidified with HNO3
10%(v/v). For each sampling point, the physical and chemical parameters [pH, dissolved oxygen
(DO), and temperature] were determined, comparing the values obtained with those recommended
by the national legislation 3 for brackish water (class 1), the DO is below the minimum limit of 5 mg
L-1 due to the organic matter amount in the medium, ranging from 1.1 to 4.7 mg L-1. In addition, pH
and temperature ranged from 7.14 to 8.37 and 22 to 32 °C, respectively. The Hg measurements
were performed using the cold vapor atomic fluorescence spectrometry (CV AFS) technique, and
the samples were previously filtered with a cellulose acetate membrane (0.45 m), ensuring that the
measurement would correspond to Hg dissolved. This way, to determine Hgtotal was added in the
samples KBr / KBrO3 (0.1 / 0.017 M) in an acidic medium (HCl 0.6 M) to promote organic mercury
compounds oxidation. In the determination of Hgino only HCl (0.6 M) was used. Finally, the organic
mercury (Hgorg) concentration was estimated through the difference between the concentration of
Hgtotal and Hgino. In this period, the concentration intervals found in the collections between July/2021
to March/2022 for Hgtotal, Hgino, and Hgorg are respectively 0.34 - 6.01; 0.02 - 1.64, and 0.2 to 5.45 µg
L-1. Since the Hgtotal level is higher than the maximum concentration (0.2 µg L-1) allowed by national
legislation.3 The highest concentration of Hgorg found was about 90% higher than Hgino, which is
worrying because it has a more significant toxicological potential, which can affect the health of the
local population and biota. The results of the July/2022 collections resulted in the lowest
concentrations (less than 0.05 µg L-1) of Hgtotal and Hgino, possibly due to the period of floods with
rainfall above 400 mm. Thus, according to the results for Hgtotal and DO obtained and the criteria
established by the competent environmental agency, systematic monitoring is necessary for the
region to assess water quality as a function of time and the influence of different environmental
events.
1 Wanderley et al., Quim. Nova, Vol. 43, 2, 206-211, 2020
2 Silva, R.F et al., Ecotoxicol. Environ. Saf. 219, 2021, 112337
3 CONAMA n° 357, de 17 de março de 2005.
[UFAL, IQB, PPGQB, CAPES, CNPq, FAPEAL]

314
PESTICIDES INVESTIGATION IN SURFACE WATERS AND SEDIMENTS IN THE
HYDROGRAPHIC BASIN OF IJUÍ RIVER (RS)
Rafaela Roberta Morelatoa* (UG), Laila S. Cardosoa (UG), Júlia V. T. Ferreiraa (UG), Mateus
Martellia (UG), Jonas S. Dugattoa (R), Liziara C. Cabreraa (R)
a
Federal University of Fronteira Sul – Campus Cerro Largo, Cerro Largo, RS, Brazil, 97900-000
*e-mail: rafaelarobertamorellato@hotmail.com
Pesticides are being identified in surface waters and sediments in studies taken place in the state of
Rio Grande do Sul (RS)1, as well as all over the world. In this context, the purpose of this work was
to evaluate the possible presence of multi-residues of pesticides in surface waters and sediments in
twelve sampling points (Pn), distributed in spring of 2021 and summer of 2022 seasons, in the Ijuí
River Basin – RS. For the extraction of different pesticides, the solid phase extraction (SPE) with
C18ec 500 mg/3 mL was used for water and QuEChERS acetate method for sediment. The
determinations were conducted with a Shimadzu 20-20 liquid chromatography mass spectrometry
(LC/MS). The method showed accuracy and intermediate precision with recoveries between 70 and
120% and RSD ≤ 20%, for the 28 and 24 pesticides in water and sediment, respectively. The limit of
quantification (LQ) was between 0.02 µg∙kg-1 a 1.00 µg∙kg-1 for sediment and 0.01 µg∙L-1 a 1.00 µg∙L-
1
for water. In spring, no pesticide was detected above the LQ for surface waters or sediments.
However, in summer the herbicide 2,4-D was detected in P6 and P8 sampling points. These points
use the surface water for animal husbandry, fishing and irrigation. CONAMA 357/2005 sets that the
maximum allowable value (MAV) of this compound in class 2 surface water is 4.0 µg∙L -1, thus
complying with the legislation in the country2. The 2,4-D is possibly carcinogenic to humans and
presents environmental hazards3-4. As to sediments, the studies quantified five compounds above
the LQ. The herbicide bentazone was found in P 6.1 collection point with concentration of 0.13 µg∙kg -
1
, while fungicide epoxiconazole was determinated in P 5.1 point in 0.022 µg∙kg-1. The herbicides
imazapic, imazethapyr and metsulfuron-methyl were also quantified: imazapic (in P1 and P4, 0.02
and 0.057 µg∙kg-1 respectively), imazethapyr (P3, 0.027 µg∙kg-1), and metsulfuron-methyl (with
concentrations of 0.18 a 0.13 µg∙kg-1 in P3 and P4 respectively). There aren’t laws about the MAV
of pesticides in sediment. The predominant crops at collection times were corn and soybeans, which
the detected compounds are applied to. It should be noted that the rainfall during the summer
collection months ranged from 57.65 to 102.362 mm5 and the pesticides could have reached the
water body due to the occurrence of rains and respective drainage of areas where these pesticides
were applied. Finally, through this study, it the importance of constant monitoring of pesticides and
further studies in the region was evidenced due to the environmental health risks caused by these
compounds.
1 Chiarello, M., et al. Química Nova, 40, 2017, 158.

2 CONAMA. Conselho Nacional do Meio Ambiente. Resolução CONAMA n° 357, de março de 2005.
3 IARC – International Agency for Research on Cancer. Lyon, França, 2022. Available in:
https://monographs.iarc.who.int/agents-classified-by-the-iarc/. Access in: 5 jun. 2022.
4 IBAMA. Instituto Brasileiro de Meio Ambiente. Portaria nº 84, de 15 de outubro de 1996.
5 WEATHER UNDERGROUND. Dados meteorológicos para o município de Cerro Largo/RS. 2022. Available in:
https://www.wunderground.com/dashboard/pws/IRIOGRAN38/graph/2019-12-31/2019-12- 31/monthly. Access in: 13 jun.
2022.
[IC-UFFS, financial resource approved in notice 121/GR/UFFS/2021]

315
DETERMINATION OF TOTAL LEAD CONCENTRATION IN ARBOREAL RINGS
ON NEOTECTONIC FAILURE
Jaqueline D. Oliveira* (UG), Miquéias L. Portugal (UG), Kevin F. Ramos (PG),
Marcos F. S. Teixeira (R)
Department of Chemistry and Biochemistry, School of Science and Technology, São Paulo State University (UNESP),
Presidente Prudente, SP, Brazil.
e-mail: marcos.fs.teixeira@unesp.br
The use of arboreal rings as environmental indicator in the identification and quantification of
potentially toxic metals has been studied as a tool for spatiotemporal evaluation of different
geological environments1. The presence of toxic metals in the environment does not come
exclusively from anthropogenic activities, but also from regions with probable geological faults [2].
Such areas with neotectonic faults are prone to the emanation of radon gas ( Rn) from the 222
successive decay of uranium, whose most stable element in its decay line is the isotope of lead-
206. The present research studied the concentration of total lead ( Pb, Pb, Pb and Pb) in tree
210 208 207 206
rings of Oiti (Moquilea tomentosa) in the municipality of Presidente Prudente. For this, a collection
area was delimited between two geological faults. The arboreal rings were extracted using a metal
probe adapted to a motorized equipment. To perform dendrochronology, used an electron
microscope coupled with digital camera. The digestion of the samples was based on the method
adapted from USEPA 350. For the quantification of the metal in the rings, the differential pulse anodic
stripping voltammetry technique (DPASV) was used for an electrochemical cell containing three
electrodes: hanging mercury drop electrode as the working electrode, Ag/AgCl (3M KCl) electrode
as a reference and auxiliary Pt electrode2. Figure 1 illustrates histograms of total lead concentration
for a time scale of 16 years (Y1 to Y16) obtained from two Moquilea tomentosa trees with different
locations in the study area.
Figure 1. Histogram of total lead concentration obtained in the samples of the arboreal rings of Moquilea tomentosa. Y1
to Y16 = year 1 to year 16.
Analyzing the histograms, we can verify that for both trees present a similarity in the
dendrochronological cycle of concentration. However, the magnitude of lead concentration between
the two sets of results is different due to the georeferencing of the sampling site. Currently, other
dendrochronological cycles of concentration are being studied with trees located at a distance from
the geological fault to carry out a kernel density mapping.
1 Aquino, S. M. F. et al. Journal International of Science, 1, 2011, 77.
2 Moreira-Silva, M. R. et al. Environmental chemistry letters, 15, 2017, 481.
[CNPq]

316
REDOX SPECIATION AND DETERMINATION OF TOTAL Sb IN WATER AND
BIVALVE FROM LAGUNA MUNDAÚ (MACEIÓ-AL)
Ellen S.S. Barros (PG)a*, Aryanna S.P.N. Costa (PG)a, Emerson Freire (PG)a, Sângela
Ferreira(PG)a, Francisco Cunha (R)b, Daniela S. Anunciação (R)a, Josué Carinhanha C.
Santos (R)a
Universidade Federal de Alagoas, Instituto de Química e Biotecnologia, Maceió - AL, Brazil, 57072-900
a
Universidade Federal da Bahia, Instituto de Química, Salvador - BA, Brazil, 40170-115

b
*e-mail: ellen.barros@iqb.ufal.br
Laguna Mundaú (Maceió - AL) is included in the Mundaú-Manguaba estuarine complex (CELMM) and is
considered an environment with high environmental impact; however, productive due to the sururu (Mytella
charruana) fishing, a bivalve characterized as filter organism and constituted the primary source of income for
the neighboring population1. In this scenario, antimony compounds are associated with the process of
environmental degradation derived from human intervention, and exposure to them can cause several
damages to health. Thus, this work aims at the redox speciation of Sb in estuarine water and the determination
of SbTOTAL in sururu. Initially, the influence of previous acidification of water samples collected at Laguna
Mundaú (Maceió, AL) with HNO3 (final concentration of 2.5% (v/v)) was evaluated, aiming at the determination
of SbTOTAL and redox speciation through atomic hydride generation atomic fluorescence spectrometry (HG
AFS). Thus, to determine the SbTOTAL has used a pre-reducing solution [50% KI (m/v) + 10% ascorbic acid
(m/v)], and no statistical difference was observed between vials types. Meanwhile, 8-hydroxyquinoline 0.02%
(m/v) [masking agent to Sb(V)] was employed to determine Sb(III), observing the decrease in the concentration
level of the species in the acidified flasks. In both cases, a paired t-test was used at 95% confidence. Therefore,
SbTOTAL and Sb(III) results were expressed using acidified and non-acidified samples. In the 11 sample points
evaluated at Laguna Mundaú, the Sb(III) concentration ranged from 0.03 to 0.26 μg L -1, while for SbTOTAL from
0.19 to 0.74 μg L-1 for samples collected in July/2022 (Fig. 1).
Figure 1. Concentrations of SbTOTAL and Sb(III) in estuarine water

samples. Besides, the content of Sb(III), the most toxic
inorganic species, in the analyzed samples is
similar to or exceeds 50% of the SbTOTAL
concentration in 5 points, despite all SbTOTAL
concentrations being less than 5 μg L-1, the
maximum value allowed by the National
Council for the Environment.2 Additionally, it is
important to mention the significant reduction
of SbTOTAL in this ecosystem concerning the
values obtained by Santos et al. (2021)1 in
July/2017, which ranged from 0.06 to 3.00 μg
L-1.
This difference is possible due to the dilution increase of the analyte in the medium related to the high rainfall
from May to June 2022 (an average of 433 mm for these two months). The determination of Sb TOTAL in the
bivalve was carried out, where the samples underwent lyophilization procedures, acid digestion (HNO 3 14 M)
with H2O2 (30%, v/v) and quantification by atomic absorption spectrometry with electrothermal atomization (GF
AAS) using Pd(NO3)2 as a chemical modifier. The concentration range varied from 0.94 to 3.0 mg kg-1 in dry
matter (0.18 to 0.62 mg kg-1 in wet matter). Although it is not yet a legislated element for this type of food in
Brazil, these values can be considered high compared to other limit values for other elements. 3 Thus, the
variability of Sb concentration in water and food helps to understand the system's dynamics and compare with
the levels of concentrations established by the legislation.
1 Santos et al. Journal of the Brazilian Chemical Society, 32, 2021, 1762.
2 CONAMA, Resolução no 357 de 17 de março de 2005.
3 ANVISA, Regulamento técnico Mercosul sobre limites máximos de contaminantes inorgânicos em alimentos. Resolução
RDC n° 42, de 29 de agosto de 2013.
[UFAL, IQB, PPGQB, CAPES, CNPq, FAPEAL]

317
STUDY OF THE SPATIOTEMPORAL DISTRIBUTION OF LEAD IN CONCENTRIC
RINGS OF TREES AS BIOMONITORING ON NEOTECTONIC FAILURE AREAS
Kevin F. Ramos (PG)a*, Marcos F. S. Teixeira (R)b
a São Paulo State University “Júlio de Mesquita Filho” (UNESP), Department of Chemistry and Biochemistry, School of
Science and Tecnology, Presidente Prudente, SP, Brazil.
The study of the spatiotemporal distribution of potentially toxic metals, such as lead in tree growth
rings, is an important tool in the biomonitoring method, since the environmental variables and the
storage of contaminants are recorded in the tree core1. Lead, present in the radon-222 decay series,
may come from anthropogenic and natural sources, such as neotectonic faults2. The aim of the
present research was to evaluate the spatiotemporal distribution of total lead in arboreal tree growth
rings Moquilea tomentosa over areas with geological faults. The study area, delimited for the
collection of samples, is integrated between two geological fault lineaments. Using a metallic probe
coupled to a motorized equipment, cylindrical samples were extracted from the growth rings of the
tree species under study. The dendrochronological study was determined using an electron
microscope, coupled to a digital camera and application software. The sample digestion method was
adapted from the USEPA 350 system. To determine the total lead concentration in the samples, the
anodic stripping differential pulse voltammetry (DPASV) technique was used. From a statistical point
of view, the Principal Component Analysis (PCA) (Fig. 1) was obtained through the values of lead
concentration in each sample and the data of annual environmental variations.
2007
Temperatura
PC 2 (25.10%)
2014
2006
2009
2015
[Pb]
Precipitação
2019
2005
2010
2013
Umidade
2018
2016
2012
2012
2011
2008
2004
PC 1 (64.03%
Figure 1. Principal Component Analysis obtained in the samples of the arboreal rings of Moquilea tomentosa.
Through the results obtained, it was verified that the annual variations of the concentrations of total
lead in the tree species, correlates inversely with the environmental variations, mainly precipitation,
and directly with the location and direction of the probable geological fault in the region, resulting in
the emanation of effective radon gas. and intense, coexisting with a high concentration of lead.
1 Badamasi, H. Mayfeb J Environ Sci, 2, 2017, 27.
2 Amaral, P.G.Q. et al. Applied Radiation and Isotopes, 70, 2012, 808.
[CAPES]

318
CHARACTERIZATION OF INDOOR AIR QUALITY IN A UNIVERSITY LIBRARY:
IMPLICATIONS ASSOCIATED WITH POLLUTANT EMISSIONS FROM NEW AND
OLD BOOKS AND CHEMICALS USING GC × GC-TOFMS
Thairine Lima dos Santos (PG)a, Vinicius Barreto Pereira (PG)a, Jessica Lima Caetano
Ribeiro (UG)a, Manoel Mendes Alves Junior (UG) a, Marcello Campos Azevedo (UG)a
Adriana Gioda (R)b, Celeste Yara dos Santos Siqueira (R)a*
aUniversidade Federal do Rio de Janeiro, Química Analitica/Instituto de Química, Rio de Janeiro, RJ, Brazil, 21941-909
bPontifical Catholic University of Rio de Janeiro (PUC-Rio), Department of Chemistry
*e-mail: celesteyara@iq.ufrj.br
Comprehensive two-dimensional gas chromatography (GC × GC) with time-of-flight (TOF) mass
spectrometry (MS) (GC × GC-TOFMS) has been applied to the analysis of Semi-Volatile Organic
Compounds (SVOCs) and Volatile Organic Compounds (VOC) extracted from an older (1930 - early
20th century) and more recent (2000 - early 21st century) book from the shelves of a library and indoor
air library. In order to assess the potentiality of GC × GC-TOFMS for accessing the compounds
produced during the natural degradation, papers from different chronological times and indoor air
were examined. The GC × GC-TOFMS system used was the Pegasus 4D (Leco, St. Joseph, MI,
USA), which was used along with an Agilent Technologies 7890 GC (Palo Alto, CA, USA), which
was equipped with a secondary oven, a nonmoving quad-jet dual-stage modulator, and a Pegasus
H11 TOFMS (Leco). The GC columns consisted of a DB-5 (Agilent Technologies, Palo Alto, CA,
USA) with 5% phenyl–95% methylpolysiloxane (30 m × 0.25 mm i.d., 0.25 µm df) as the first
dimensional column (1 D) and a DB-17 (Agilent Technologies) with 50% phenyl–50%
methylpolysiloxane (1.2 m, 0.10 mm d.i., 0.10 µm df) as the second dimensional column (2 D). This
GC × GC-TOFMS method enabled the characterization of several organic compounds such as
furfural, 5-methyl furfural, vanillin and guaiacol that can be considered as relevant carbohydrates
and lignin degradation compounds. Their relative abundance has been followed during ageing
treatments in order to evaluate their potential role as lignocellulosic degradation markers. GC × GC-
TOFMS appears to be a suitable method for investigating volatile and semi- volatile organic
compounds extracted by a book, and for distinguishing some relevant degradation compounds.
Moreover, the method allowed us to monitor their relative abundance. It would now be worthwhile
trying to detect Volatile Organic Compounds (VOCs) and, likewise to suggest a new approach to
evaluate the state of deterioration of old books.

319
DEVELOPMENT OF 3D GRAPHENE-BASED O-DGT DEVICES FOR
PROPRANOLOL SAMPLING
Mayara B, Leão (PG)a,b*, Marco T. Grassi (R)c, Carolina F. M. Jauris (R)b,
Andreia N. Fernandes (R)a
aFederal University of Rio Grande do Sul, Institute of Chemistry, Porto Alegre, RS, Brazil, 91501-970
bFederal University of Pampa, Environmental Science and Technology Center, Caçapava do Sul, RS, Brazil, 96570-000
cFederal University of Paraná, Chemistry Department, Curitiba, PR, Brazil, 81531-980
*e-mail: mayarabitencourtleao@gmail.com
A relevant analytical tool in determining contaminants in situ is based on diffusive gradients in thin
film (DGT). These passive samplers have been widely used to determine inorganic and organic
contaminants when the technique receives the acronym o-DGT. One of the focuses applied to these
devices is to test and validate new binding layers to increase the range of target analytes. Due to
their properties, nanomaterials have been tested; however, no work has reported the use of three-
dimensional graphene-based (3D-rGO) in this application. Thus, this study aimed to conduct
preliminary tests to develop an o-DGT device with a modified binding layer, using a 3D-rGO, to
sample the drug propranolol (PPL) in water. Material synthesis was realized through an
environmentally friendly and low-cost route1. The adsorption study was carried out in two stages.
First, the kinetic study was performed using ~6 mg of hydrogel and 5 mL of PPL 50 mg L -1. The
determination of the concentration of the solution was performed at predetermined time intervals, up
to 450 min. The results were evaluated according to the general order and pseudo-second-order
kinetic models. For the construction of adsorption isotherms, ~12 mg of hydrogel was kept in contact
with 5 mL of PPL (5-100 mg L-1). At the end of 5 h, the final concentration in the solution was
determined, and the adsorptive capacity was calculated. To evaluate the PPL's desorption from the
material, 5 mL of PPL 50 mg L-1 was kept in test tubes containing 3D-rGO for 24 h. Desorption was
evaluated using the solvents ethanol (EtOH), acetonitrile (ACN), acetone (ACT), and methanol
(MeOH), individually or combined in different proportions. The first desorption step consisted of
adding 3 mL of solvent to the material, followed by sonication for 3 min. Afterward, the sample was
centrifuged for 5 min at 3000 rpm. An aliquot of 1 mL was taken from each sample and stored. This
process was repeated 3 times, resulting in a 3 mL sample. All tests were performed in triplicate, with
constant ionic strength (CaCl2 0.01 M), and the samples had their concentrations determined by
molecular fluorescence (λem = 340 nm). The results showed an adsorption equilibrium time of 360
min, with a better fit by the general order kinetic model. The Sips model provides the best fit for the
adsorption isotherms, which predicts that monolayer adsorption is found at high concentrations and
an adsorptive capacity of 23.6 mg g-1. The desorption tests showed that using ACT and MeOH
solvents was not viable due to analytical limitations. In this sense, only the desorption results were
considered for the solvents EtOH, ACN, EtOH/ACN (2:1, v/v), ACN/EtOH (2:1, v/v), and ACN/EtOH
(1:1, v/v). It was found that the desorption efficiencies for these solvents were 38.8% ± 4.8; 34.2% ±
6.4; 30.3% ± 0.3; 25.8% ± 5.5 and 24.3% ± 7.6 for ACN/EtOH (1:1, v/v), EtOH/ACN (2:1, v/v),
ACN/EtOH (2:1, v/v), ACN and EtOH, respectively. Therefore, it appears that using a combination
of acetonitrile and ethanol solvents in different proportions is more efficient than using these solvents
individually. Thus, we observed that 3D-rGO could adsorb and desorb PPL efficiently. Although
desorption does not happen as efficiently, correcting the values using an elution factor can ensure
the reliability of the results obtained later. Therefore, the material evaluated can be considered
promising for application as a binding agent in o-DGT devices.
1 Leão MB, da Rosa PCC, Dalla Corte CL, Matos CF, Materials Chemistry and Physics, 288, 2022, 126408.
[CNPq, FAPERGS, INCTAA, INCT Nanocarbon]

320
CHEMICAL FRACTIONATION OF RARE EARTH ELEMENTS IN SODA LAKES
OF NHECOLÂNDIA (PANTANAL, BRAZIL) USING DGT AND ICP-MS
Melina B. T. Zanatta (R)a*, Carlos Eduardo Eismann (PG)a, Luiz Felipe P. P. Moreira (PG)a,
Amauri A. Menegário (R)a, Anne H. Fostier (R)b
bUniversity of Campinas (UNICAMP), Institute of Chemistry, Campinas, SP, Brazil, 13083-970
*e-mail: melinabt@alumni.usp.br
The rare earth elements (REE) form a group of 17 elements, which 15 belong to the group of
lanthanides (La to Lu) together with scandium and yttrium, due to the similar properties and
components between them1. The Nhecolândia region is characterized by a system of several lakes
of different shapes. Many complex and diverse processes that occur in the lakes of Nhecolândia
generate a variety of conditions that influence the distribution of chemical species of the many
elements present in the region2. These conditions constitute a constant scientific challenge. The
impacts and biogeoavailability assessment of REE are poorly studied due to the lack of data and the
fact that these elements are in very low concentrations in the environment. Therefore, the present
study aims to evaluate the chemical fractionation of REE in water from the Nhecolândia sub-region
using the Diffusive Gradients in Thin Films (DGT) technique followed by determination by Inductively
Coupled Plasma Mass Spectrometry (ICP-MS). Water from two different saline-alkaline lakes (green
and black lakes) were collected in the Pantanal of Nhecolândia. To evaluate the total content of REE,
the samples were digested in a microwave oven following the EPA 3015A protocol. For the dissolved
fraction, a 0.45-µm PTFE filter was used. For DGT devices, the Chelex® was employed as binding
layer and the polyacrylamide hydrogels were used as diffusive layers. Some parameters such as
pH, conductivity, temperature and resistivity were measured in the water samples before and after
the DGT immersions. To overcome the spectral interferences commonly found in these elements
determination by ICP-MS, the APEX system for aerosol desolvation was used and the MO+/M+ ratio
was less than 0.003 using the optimized procedure. DGT diffusion coefficient and elution factor
values used for each REE were based in a previous work 3. The results are show in Table 1.The
black lake (SR06) was the one with the highest concentration of the REEs and the results agree with
light rare earth elements (LREEs) being more abundant in the earth’s crust than heavy rare earth
elements (HREEs). The analytes speciation was performed using the CHEAQS Next software, which
indicated that most of the REE would be in the precipitated form as phosphates and hydroxides.
However our results showed that more than 70% of the REE are in filtrated fraction (0.45 µm).
Therefore, it is suggest that the REE are in the form of solid colloids smaller than 0.45 µm. The
values obtained for the labile concentration (DGT) were considerably lower as compared with filtered
fraction, corroborating the fact that the species are in the form of inert colloids in the system under
study. The use of the DGT technique associated with the APEX system for interference reduction
proved to be efficient for chemical fractionation of REEs in lakes that present extreme conditions.
Table 1. REE concentration in µg L-1 in soda lakes of Nhecolândia.
SR04 (green) La Ce Pr Nd Sm Eu Gd Dy Ho Er Tm Yb Lu
Total 0.76 0.91 0.22 0.46 0.24 0.24 0.23 0.20 0.16 0.17 0.16 0.19 0.21
Dissolved 0.56 0.64 0.17 0.33 0.18 0.18 0.17 0.13 0.11 0.12 0.11 0.12 0.16
DGT 0.13 0.28 0.063 0.14 0.082 0.114 0.067 0.067 0.065 0.070 0.076 0.079 0.076
SR06 (black) La Ce Pr Nd Sm Eu Gd Dy Ho Er Tm Yb Lu
Total 7.63 14.80 2.23 8.77 2.00 0.62 1.96 1.66 0.38 1.01 0.18 0.93 0.18
Dissolved 6.19 11.50 1.74 7.03 1.57 0.46 1.64 1.39 0.27 0.79 0.11 0.75 0.13
DGT 0.16 0.38 0.065 0.22 0.070 0.056 0.066 0.053 0.029 0.041 0.025 0.034 0.023
1 Amiel N, et al., Environ. Pollut. 284, 2021

2 Barbiero L, et al. Sci. Total Environ. 619–620, 2018
3 Huang J, et al., Talanta. 230, 2021

321
VALIDATION OF LOW COST MEHODOLOGIES FOR ENVIRONMENTAL
MONITORING OF RIVERS
Giovanna J Ghellere (UG)a, Silvia S Silva (PG)b, Marcelo N Kapp (R)a, Marcela Boroski (R)b*
aFederal University of Latin American Integration (UNILA), ILATIT, Foz do Iguaçu, PR, Brazil, 85867-970
aFederal University of Latin American Integration (UNILA), PPGIES/ILATIT, Foz do Iguaçu, PR, Brazil, 85867-970
*e-mail: marcela.boroski@unila.edu.br
One of the impacts caused by human activity concerns water quality, its use and inadequate
occupation in watercourse protection strips. Monitoring water quality is extremely important in order
to provide (1) a comparison between the quality of water resources from different sources and to
assess (2) the presence of temporal contamination by using parameters determined and analyzed
through the concentration of inorganic and organic compounds present in the water. The goal of this
work is to validate the methodologies of colorimetric analysis for ammonia1, nitrite2, orthophosphate3,
organic matter4, and iron5 to apply the low-cost river monitoring kit, using a smartphone app. For
each analysis, a well-known methodology was employed in a miniaturized and modified way. For
validation purposes, analytical curves, intermediate precision, repeatability, the limit of detection
(LD), and of quantification (LQ) were monitored in a UV-VIS spectrophotometer and using the
smartphone application AQUA. The AQUA application [6] is in the testing phase and, in addition to
colorimetric analysis, it performs data georeferencing. To capture the images, a box with a battery-
powered LED apparatus was used to regulate the intensity of ambient light to which the sample is
exposed during image recording. The efficiency of the curve construction was measured by the
coefficient of determination, which remained above 0.990 in all analyzes carried out in the
spectrophotometer, with the exception of organic matter, which had its value equal to 0.954. In the
other hand, the results for the application remained between 0.838 and 0.985. The evaluation of
repeatability and intermediate precision was performed considering an acceptance criterion of 11%
for the coefficient of variation (CV). All analyzes performed in the spectrophotometer had values
within the expected range. In the AQUA application, for nitrite and ammonia, they were within the
expected value, while for orthophosphate, organic matter and iron, the average of the coefficients of
variation was above the established value. LD and LQ values were higher in AQUA. In conclusion,
the tests can be performed remotely through the AQUA application and using the equipped box with
significant results regarding the construction of the analytical curve of the adapted methodologies.
The LD and LQ presented values above for AQUA compared to UV-VIS.
1 Batista, F. G. A. et al. Revista da Universidade Estadual de Alagoas/UNEAL. 11, 2019.
2 APHA – American Public Health Association. Standard Methods for the Examination of Water and Wastewater, 26th
ed. AWWA, WPCF, Washington, D.C., 2017.
3 Associação Brasileira de Normas Técnicas. NBR 12772: Água - Determinação de fósforo - Método de ensaio. Rio de
Janeiro: ABNT/CEE Comissão de Estudo Especial, 1992, 9.
4 Associação Brasileira de Normas Técnicas. NBR 10739: Água - Determinação de oxigênio consumido - Método do
permanganato de potássio. Rio de Janeiro: ABNT/CEE Comissão de Estudo Especial, 1989, 3.
5 Lourenço, E. C. et al. Revista Virtual de Química, 13, 2020, 192.
6 Baldasso, B. et al. Desenvolvimento de Aplicativo para Monitoramento Participativo de Mananciais. Trabalho de
Conclusão de Curso de Engenharia Química. UNILA, 2021.
[UNILA/PRPPG, Fundação Araucária de Apoio ao Desenv. Cient. e Tecnol. do Estado do

Paraná, CAPES, SETI]

322
MICROPLASTICS AND ORGANIC CONTAMINANTS ON THE SURFACE WATER
OF RIVER GUAÍBA
Ellen de Souza A. Duarte (PG)a*, Mariana A. Dias (PG)b, Cassiana C. Montagner (R)b,
Crislaine Bertoldi (R)c, Andreia N. Fernandes (R)a
bUniversity of Campinas, Institute of Chemistry, Campinas, SP, Brazil, 13083-970
cFederal University of Paraná, Departament of Chemistry, Curitiba, PR, Brazil, 81531-980
*e-mail: ellenvrf@gmail.com
Microplastics can cause several problems, especially for living organisms.1 In addition, the capacity
of microplastics to interact with contaminants in aquatic water increase their toxicity. 2 Thus, the
purpose of this study was to investigate the presence of microplastics and organic contaminants on
the surface water of the River Guaíba (southern Brazil), which is the main source of water supply for
Porto Alegre city. The microplastics and organic contaminants samples were collected in June 2022
at six different points along the river. Microplastic samples were collected by trawling a zooplankton
net (60 µm), and 1 L of water was collected to quantify organic contaminants. In the laboratory,
QA/QC measures were performed to avoid and control the contamination of potential microplastic
during sample treatments. The microplastic samples were filtered through a sieve of 60 µm, and the
content was transferred with 100 mL of filtered deionized water to a beaker and dried oven at 60 °C.
The dried samples were submitted to density separation with a potassium iodide solution (1.6 g cm-
3
) and then to an oxidative treatment with Fenton for one hour at 60 ºC. The physical characterization
of the microplastics was carried out with a microscope (magnification of 4x and 10x), considering
their color and shape and the polymeric composition using µ-ATR-FTIR. All the identified particles
were considered as potential microplastics until the characterization of the polymeric composition.
The organic contaminants were extracted by SPE using an Oasis HLB cartridge (Waters) and
quantified by LC–MS/MS. The compounds investigated include pharmaceuticals, hormones,
pesticides and derivatives, and industrial wastes. So far, only one microplastic sample (P6) was
processed, and only its physical characterization was concluded. The preliminary results indicated
a concentration of microplastics of 14.3 items m-3 in site P6. Microplastics were predominantly
colorless, corresponding to 46.9% of the total found. The predominance of colorless microplastics
could be related to the high production of plastic materials in this color and the fading of microplastics
through the weathering in the environment.3 Concerning the shapes, 56.7% were fragments and
43.3% were fibers. The minor fiber content may be due to the significant sewage collection in Porto
Alegre city, reaching 70% and reducing the amount of fibers in the water body.4 The fragments found
have characteristics of secondary microplastics that result from larger plastics' fragmentation. The
results about organic contaminants shows the presence of eleven contaminants among the sampled
sites. Caffeine was the compound with higher concentrations in all samples, except for P2 (45,4 ng
L-1), with 99.7, 155.9, 160.7, 137.3, and 133.5 ng L-1, respectively, P1, P3, P4, P5, and P6. The high
concentration of caffeine could be related to the significant consumption in the region of yerba mate
and coffee5. Caffeine is an indicator of anthropogenic activity, then the results demonstrate that
domestic sewage may reach the River Guaíba. The presence of microplastics and organic
contaminants causes great concern since many studies have already proved their toxicity in the
environment, and they can impact the quality of the water used for human consumption. More studies
will be carried out with the intent to correlate the organic contaminants detected to the presence of
microplastic in the River Guaíba.
1 Queiroz AFS et al., Science of the Total Environment, 839, 2022, 156259
2 Wang Y et al., Chemosphere, 299, 2022, 134370
3 Zhao X et al., Environmental Science & Technology, 56, 2022, 9161-9163
4 DMAE; DEP; DMLU. Plano Municipal de Saneamento Básico,1, 2015
5 Machado KC et al., Science of the Total Environment, 572, 2016, 138-146
[UFRGS, CAPES, CNPq, INCTAA]

323
ARSENIC BIOACCUMULATION AND BIOTRANSFORMATION IN
CYANOBACTERIA (Anabaenopsis sp. and Arthospira sp.) FROM THE SALINE-
ALKALINE LAKES OF THE PANTANAL, BRAZIL.
Marianna A. Canciana (PG), José Lucas M. Vianaa (PG), Thierry A. Pellegrinettib (PG), Marli de
Fátima Fioreb (R), Anne H. Fostiera (R)
a University of Campinas (UNICAMP), Institute of Chemistry, Campinas, São Paulo, Brazil, 13083-862
b University of São Paulo, Center for Nuclear Energy in Agriculture, Piracicaba, São Paulo, Brazil, 13400-970
marialvescancian@gmail.com
Saline-alkaline lakes in Pantanal, Brazil present high dissolved arsenic concentrations (> 3 mg L-1),
mostly in the form of As (V)1. Despite extreme conditions, such as pH in the range of 8 to 10, these
lakes shelter an abundant microbial community, such as cyanobacteria that are able to tolerate and
metabolize As. The species of As frequently reported in aquatic environments are the inorganics
As(III) and As(V) and organic forms as monomethylarsonic acid (MMA) and dimethylarsinic acid
(DMA), for aquatic organisms, the other organic forms, such as arseno-sugars, are also found2. In
this work, we aimed to investigate the bioaccumulation and speciation of As in the cyanobacteria
Anabaenopsis sp. and Athospira sp., previously isolated from saline-alkaline lakes. The
cyanobacteria were cultivated in a medium (Z8) enriched with 1 mg/L As(V). Cultures were carried
out in triplicate and collections were performed at the beginning of the experiment, T1, and after 21
and 42 days. Control cultures without As enrichment were also performed. Determination of total As
concentration (AsTot) was performed by Inductively Coupled Plasma Mass Spectrometer (ICP-MS)
and speciation using High-Performance Liquid Chromatography (HPLC) coupled to ICP-MS. The
preliminary results show that AsTot in the culture medium without cyanobacteria (Fig. 1) didn’t vary
significantly over the 42 days cultivation. For the cultures of both cyanobacteria, the AsTot after 42
days was maintained when compared to the control cultures (enriched with As but without
cyanobacteria) (Fig.2). Speciation analysis of the culture medium (supernatant) without and with the
inoculation of cyanobacteria and with As enrichment and analysis of total As and speciation of the
cyanobacterial biomass will be carried out to evaluate the bioaccumulation and biotransformation of
As in the cyanobacteria.
Figure 1. Total As concentration in the culture medium Figure 2. Culture medium a) with Anabaenopsis sp. and
without and with As enrichment (1 mg/L) on different days. without As enrichment; b) with Anabaenopsis sp. and As
enrichment (1 mg/L); c) with Arthospira sp. and without
As enrichment (1 mg/L); d) with Arthospira sp. and As
enrichment (1 mg/L);
1 7Richter, L. et al. Science of the total environment, v. 687, p. 917-928, 2019.

2 Anawar, H.M. Talanta, v. 88, p. 30- 42, 2012.
[FAPESP]

324
THE STUDY OF DROPLET SIZE DISTRIBUTION TO RECOVER OIL FROM
SPENT DRILLING FLUIDS USING HYDROCYCLONE
Laine B. Pereiraa*, Cristina M. S. Sada, Márcio F. Martinsb , Eustáquio V. R. de Castroa,
Valdemar Lacerda Jr.a
a Federal University of Espirito Santo, Chemical Department, Vitória, Espirito Santo, Brazil, 29075-910
b Federal University of Espirito Santo, Mechanical engineering Department, Vitória, Espirito Santo, Brazil, 29075-910
*e-mail: lainebadaro@gmail.com.br
Spent drilling fluids are one of the most generated residues from oil Exploration & Production (E &
P) activities. Drilling fluids are defined as a complex mixture, chemically stable, of synthetic or natural
compounds responsible for performing many essential functions in well drilling. They are classified
according to their continuous phase, which can be water-based, oil-based, synthetic-based, or gas-
based.1 It is noteworthy that water-based drilling fluids are the most used because of their low
production cost and high biodegradability. The recirculation of drilling fluid between the well and the
platform occurs several times during the drilling of an oil well until it reaches the reservoir when they
lift to the surface with oil contaminants. The resulting waste is a mixture of oil, drilling fluid, and drill
cuttings.2 After drill cuttings separation, remains an oil/drilling fluid emulsion. Oil generally is
dispersed in drilling fluid in droplets form. Among all the treatment methodologies described in the
literature3, hydrocyclones can be an excellent option to promote phase separation and enable the
recovery and reuse of all separated phases. The oil droplet size is one factor influencing the
hydrocyclone separation efficiency. Generally, Larger diameter drops tend to separate more easily.4
In this sense, an oil/drilling fluid emulsion was prepared with 20 wt% of heavy oil. Then, the droplet
size distribution and photomicrographs were obtained by optical microscopy of the emulsion. In
Figure 1a, it is possible to observe the photomicrograph of the prepared emulsion. And Figure 1b
shows the emulsion's size distribution of oil droplets.
Figure 1. Photomicrograph and (b) Droplet size distribution of oil/drilling fluid emulsion with 20 %wt of oil.
It is possible to observe a significant variability of droplet size in Figure 1a, which can be confirmed
in the Gaussian curve behavior (Figure 1b). It is highlighted that the average droplet diameter was
54.36 μm with a standard deviation of 32.28 μm. Braga (2015), in a study, showed that it is possible
to obtain a granulometric efficiency (i.e., oil droplets that separate and exit through the overflow) of
95.6% for 40 µm, reaching 98.4% for 50 µm. In this sense, developing a hydrocyclone is a promising
alternative to promote oil separation from the spent drilling fluid.
1 Mitchell RF, Miska SZ, Fundamentals of Drilling Engineering, Society of Petroleum Engineers, 2011.
2 Elnenay AEMH, Nassef E, Malash GF, Magid MHA, Egyptian Journal of Petroleum, 26, 2017, 203-208.
3 Pereira LB, Sad CMS, Castro EVR, Filgueiras PR, Lacerda Jr. V, Fuel, 310, 2022.
4 Braga ER, Tese de doutorado, Escola de Química, UFRJ, 2015.
[UFES, LABPETRO, LFTC, CNPq, CAPES, FAPES]

325
EVALUATION OF SUGARCANE BAGASSE FOR EXTRACTION AND RECOVERY
OF GOLD FROM ACID AQUEOUS SOLUTION
Gabriel R. Bento (UG)*, Dairon P. Fuentes (PG), Lucas E. O. Porto (PG), Pedro V. Oliveira (R)
Instituto de Química, Universidade de São Paulo, São Paulo, SP, Brazil, 05508-900
*e-mail: gab_bento551@usp.br
Gold is an essential metal in diverse fields. Currently the demand of this valuable metal is increasing
due to its vast applications in electrical and electronic products, pharmaceutical drugs, and
manufacturing of printed circuit boards (PCIs). The most commercially used process for extraction
and recovery of gold is based on complexation with cyanide, due to its efficiency, simplicity and low
cost. However, it has been gradually replaced because of its poor selectivity and severe
environmental impacts.1 For this reason, the development of new techniques to recycling and
recovering gold have received significant attention, with increasing amount of electronic waste, which
cover all electrical or electronic devices at the end of their useful life. 2 The present work was
conducted to evaluate the potential of cellulose-lignin mixture, present in sugarcane bagasse, as
adsorbent material for separation of Au(III) from aqueous solutions. The material of cellulose-
lignin was prepared without modifications; the sugarcane was washed with distilled water and dried
in an oven at 60°C. Thereafter, the dried sugarcane was ground in a knife mill, washed with 1 mol L-
1
HCl solution and filtered under reduced pressure. The material was then dried and sieved at 400
μm pore size. Adsorption isotherms were obtained in batch, the concentration range from 1 to 200
mg/L of Au(III), constant mass of 30 mg sugarcane bagasse, volume of 5 mL, pH=2, at 24±1oC. Gold
was monitored by inductively coupled plasma optical emission spectrometry (ICP OES). The
linearized model of Langmuir was applied, resulting qmax = 18 mg Au/g of adsorbent. The adsorption
experiments were made using fixed-bed columns with 150 mg of the material. To investigate the
effect of pH on adsorption solutions, 10 mg/L of Au(III) were prepared at pHs from 0 to 8, at a flow-
rate of 1.0 ml/min. For desorption of gold, different solutions were tested (5 mL): 0.5 to 2,0 mol/L of
HCl and 0.1 mol/L of thiourea, at pH 2. The eluting solutions from column, at different pHs, were
collected and analyzed by ICP OES. The adsorption of Au(III) on the column material was more
effective at pH~2, reaching at 99% of adsorption efficiency. At higher pHs (6 and 8), the adsorption
decreases, with adsorption close to 0% in pH=8. In HCl solution (pH~2), Au(III) is mostly present in
the form of tetrachloroaurate(III) (AuCl4-), which can be reduced to metallic gold by the hydroxyl
groups of cellulose-lignin of sugarcane bagasse.3 Considering the presence of Au(0) adsorbed on
the material, desorption using 0.5 to 2,0 mol/L of HCl was not effective. On the other hand, desorption
using thiourea solutions proved to be very effective, reaching a recovery of gold near to 96 %. The
performance of thiourea can be explained by the high formation of complexes Au(I)-thiourea
[Au(CS(NH2)2]+, K = 9.1 x 1021.4 The presence of other ions such as Cu(II) and Fe(III) in solution can
act as oxidant of Au(0) to Au(I) promoting the leaching from the adsorbent in the column. This
procedure was used for separation and recovery of Au(III) from hydrochloric acid solution with
efficiency higher than 90%.
1Qin, H.S., Ren, Z, J. Qinghai Environ, 10,2000,78
2 STEP. Solving the E-Waste Problem, 13, 2014, 3576,
3 Ogata, T., Nakano, Y,Water Research 39, 2005, 4281
4 Nicol, M.J.; Fleming, C.C.; Paul, R.L, S. Afr. Inst. Min. Metall., Monogr.Ser., M7,1987, 831-906

326
ESTIMATED GREENHOUSE GASES EMISSION FROM BELO MONTE
HYDROELECTRIC PLANT-PA – PRE-CONSTRUCTION PHASE.
Henryck A. Fernandes (UG)a, Isaque W. S. Brandão (PG)a, Heronides A. Dantas Filho (PG)a*,
Augusto C. F. Saraiva (PG)b
a Universidade Federal do Pará, Departamento de Química pesquisa/Instituto de Ciências Exatas e Naturais, Grupo de
Espectrometria Analítica Aplicada, Belém, Pará, Brasil, 66075-110.
b ENVIRON, Gestão e Engenharia Ambiental, São Paulo, São Paulo, 03114001
*e-mail: heronides@ufpa.br
Greenhouse gases (GHG) emissions causes the elevation of the average temperature on earth
surface. According to the Intergovernmental Panel on Climate Change (IPCC) hydroelectric plants
are responsible for GHG emissions.1 In 2019, Brazil emitted more than 2.2 billion tons of carbon
dioxide equivalent (t CO2 e) to the atmosphere.2 In the amazon region the Belo Monte hydroelectric
plant, shows itself as one of the biggest contributors to this amount, once the Xingu watershed, that
feeds the plant, is a large tributary on the right bank of Amazon River. 3 This present study aims to
analyze the emission of methane and carbon dioxide gases from the hydroelectric plant of Belo
Monte during the pre-construction phase, verifying mathematical models that describe the behavior
to these gases emissions and applying geostatistical methods of interpolation to evaluate the spatial
and seasonal variation of data. The collect of the samples was made in a diffusion chamber in 17
different collection sites in the Xingu watershed, in four different campaigns which happened in
March, July and November months of 2012 year and January month of 2013 year. The analysis of
the collected samples was made using gas chromatography technique. The results reveals that the
gas mostly emitted from the Belo Monte plant was the dioxide carbon, and its biggest value to the
flux emission was 7500 mg m-2d-1, during the period of January month of 2013 year. To the methane
results, the biggest value to the flux emissions was 1200 mg m-2d-1, during the July month of 2012
year. It was verified that the linear model showed the best performance in the data description of the
data. The geostatistical analysis in this present study reveals little spatial and seasonal dependence
of the data. The study concludes that gases emissions from the Belo Monte plant contributes to the
greenhouse effect, and the literature needs more researches related to the temporal and seasonal
variability, using interpolation techniques in hydroelectric plants and amazon rivers, which allows a
better understating about these gases’ emissions.
1 IPCC – Intergovernamental Panel on Climate Change. Climate Change 2013: the Physical Science Basis. Cambridge
University Press, New York, 2013.
2 Costa, D. J., Gueges, F., Júnior, A. A. B., Faria, W. R. Enaber – Associação Brasileira de Estudos Regionais, 2021.
3 ANA-PHMDA. Plano Estratégicos de Recursos Hídricos dos Afluentes de Margem Direita do Rio Amazonas:
Introdução. Brasília-DF: Agência Nacional de Águas, 2012.
[UFPA, GEAAp, CAPES]

327
SUGARCANE BAGASSE AS ADSORBENT FOR METAL IONS REMOVAL FROM
WASTEWATER OF TEACHING LABORATORIES
João J. A. Fujiwara (UG)*, Dairon P. Fuentes (PG), Lucas E. O. Porto (PG), Pedro V. Oliveira (R)
Universidade de São Paulo, Instituto de Química, São Paulo, SP, Brazil, 05508-000
*e-mail: joao.jou@usp.br
Water is an essential resource for life, but also for chemistry laboratories. The awareness of its use
is a topic of growing relevance inside and outside the scientific environment, and it is of extremely
importance the treatment of the water we use on a daily basis in the laboratories of research and
industry. Millions of litters of wastewater are produced by the students in the many classes given in
the didactic laboratories. Chemical waste should be diminished to minimize environmental pollution,
and it is important that the amount of waste be minimized in chemistry teaching laboratories to reduce
the negative impact on environment, as well as the disposal costs of university.1 The wastewater
composition from teaching laboratories is diverse, containing organic and inorganic components and
normally are treated using evaporation methods or disposed in residual sludge, being designated to
proper landfills. Although evaporation is very cheap, it just reduces the volume of liquid waste and
do not eliminate the problem. This work proposes the usage of natural sugarcane bagasse (SCB)
as a bioadsorbent, analysing the removal of metallic ions from the wastewaters produced in
analytical chemistry classes. The SCB is cheap, widely available and renewable. Its composition is
formed mainly of cellulose and lignin (~90%) and has been used for many applications, such as
substrate for elements mitigation.3 The SCB was cleaned, washed, milled and sifted, resulting in a
fine (<400 μm) very hydrophobic powder. Fixed-bed column was prepared with 200 mg of the in
natura SCB and using SCB modified with TAN (1-(2-Thiazolylazo)-2-naphthol). Although the
complexity of laboratory waste is high, only 10 elements (Ag, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn)
was chosen for this research. The determination of elements was done by inductively coupled
plasma optical emission spectrometry (ICP OES). Reference solutions of 1 mg/L of the elements in
pH range of 2 to 8 were tested using SCB and SCB-TAN with a flow through the column of 1 mL/min.
For SCB, at pH = 2, the adsorption of ions was lower (around 25%), while at pH 8, the adsorption
increased for all elements, ranging from 66% (Mn) to 100% (Cu and Cr). Good results were obtained
for pH = 6 using SCB-TAN, with adsorption around 100% for almost all elements. Volumes of 10 ml
of wastewater from the laboratory of qualitative classes had their pH adjusted from 1 to 8. A
precipitate was formed in all solutions, increasing with the pH, and the supernatant was analysed to
determine the elements concentrations that remained in solution. Significant amounts of many
elements presented in the wastewater were precipitated at pH = 7 (Co, Cr, Cu, Fe, Ni and Zn),
however not sufficiently for discharging in the natural water. Then, this solution (pH = 7) was flowed
through the SCB-TAN column at 1 mL/min and between 80 to 100% for almost all elements were
separated. Another experiment was done using 10 mL of laboratory wastewater without any previous
treatment (pH~1) was flowed subsequently by 3 columns with in natura SCB, at 1 mL/min.
Determination of elements at the eluate from the third column showed acceptable removal of
elements (ranging from 28% for Ni to 75% for Pb). The results obtained with this research so far,
showed that the combination of precipitation with separation with SCB-TAN column were very
efficient for the removal of potentially toxic elements from the residues of the teaching laboratories.
1 Hui YG, Wei WCW, Yue YO, Journal Chemical Education 97, 2020, 87-96.
2 Novaes SD, Oliveira PV, Petri DFS, Environmental Science and Pollution Research, 2022
https://doi.org/10.1007/s11356-022-20345-4.
3 Iwuozor KO, Oyekunle IP, Oladunjoye IO, Ibitogbe EM, Olorunfemi TS, Sugar Tech, 24, 2021, 1167-1185.

328
ANATOMICAL AND CHEMICAL CHANGES IN LYCOPERSICON ESCULENTUM
(TOMATO) CAUSED BY SOIL CONTAMINATION WITH FLUORESCENT LAMPS
Gabriela do Nascimento Camargoa, Thiago Silva Diasa, Rodolfo Santos Barbozaa, Camila
Vicente de Fariasb, Lívia Gonçalves Leida Soaresc, Manuel Castro Carneiroc, Fernanda
Veronesi Marinho Pontesa, Jéssica Frontino Paulinoa
aUFRJ - Universidade Federal do Rio de Janeiro
bSENAI CETIQT - SENAI Centro de Tecnologia da Indústria Química e Têxtil
cCETEM - Centro de Tecnologia Mineral
The fluorescent lamps (FL) residues contain Hg in their compositions, leading it to the "class I"
classifying (Dangerous).1 Beyond Hg, there are others Potentially Toxic Elements (PTE), such as
Cu, Pb, Cd, Mn, Ni and Zn. In Brazil, in the last decade it was estimated that 100 million FLs were
consumed per year and only 6% of them were recycled being common the improper disposal of this
material, contaminating soil, water, and air.2 Human health risk assessment determined that
vegetables cultivated in Cd contaminated soil are a potential hazard to children and adults and that
residual Hg is a high risk to children.³The mobility of PTE from FL powder to the soil can promote
bioaccumulation in plants and, consequently, modify their development and metabolism.
Polyphenolic compounds are one of the major classes of secondary metabolites produced by the
species Lycopersicon esculentum Miller (tomato) (Solanacea).4 The present study investigates the
cultivation of tomato under FL powder contamination stress that were stated in 10 and 20 g of FL
powder per 300 g of soil (level I and II, respectively), during 60, 90 and 120 days, in triplicate and
using control samples (samples cultivated without FL powder - Blank). The main parameters adopted
to evaluate the influence of soil contamination on plant development were the anatomical
characteristics, accumulation of metals and main alterations in the production of phenolic substances
in the cultivated specimens. The plants were collected and cleaned, then separated into roots, stems
and leaves, followed by the drying process at 40 °C. In general, the masses per surviving species of
roots, stems and leaves of the contaminated plants were statistically similar to the control, except for
the roots with 120 days of age which was observed larger mass for the control. After solubilization
of the samples (in HNO3), the metals Mn, Ni, Cu, Zn, Cd and Pb were quantified by ICP-MS. At both
levels, Cd was found in all parts of the plants; Cu and Zn were present in roots and stems, but not
in leaves; Mn was found in roots and leaves; Pb were found in stems and leaves; and Ni only in the
roots. For the determination of phenolic substances, the leaves (5 mg) were extracted with
MeOH/H2O 3:1 (v/v) (1 mL) and partitioned with hexane (3 × 1 mL). Defatted dry extracts were
solubilized in 100 µL of MeOH and analyzed by thin-layer chromatography (TLC) under monitoring
conditions for polyphenolic substances (flavonoids, phenolic acids, among others)3. TLC
chromatographic profiles suggested the selective production of a flavonoid from specimens grown
under two-level contamination conditions at 90 and 120 days. In addition, the supposed flavonoid
and other substances showed prominent antioxidant activity potential through the DPPH reaction.
Thus, a solid phase extraction (SPE) method was developed and optimized for the isolation of
substances present in the extracts under study, being isolated a flavonoid (quercetin-3-O-α-L-
rhamnopyranosil(1’’’→6’’)-β-glucopyranosideo (routine)), and at least two alkaloids (still in the
process of structural characterization). It is important to note that due to the small amount of plant
material obtained (leaf mass), it has not yet been possible to structurally characterize the flavonoid
produced exclusively by specimens cultivated under contamination. However, a larger-scale
experiment is being carried out for this purpose.
1 ABNT. NBR 10.004, Resíduos sólidos – Classificação. Rio de Janeiro: ABNT, 2004
2 Laruccia, M. M. et al. International Journal of Business Administration, 2, 2011, 101–112
3 De Farias, C. V. et al. Chemosphere, 261, 2020
4 Kisa, D. Russian Journal of Plant Physiology, 64, 2017, 876–882

329
EVALUATION OF THE ENVIRONMENTAL MOBILITY OF METALS BY
SIMULATING A SOIL CONTAMINED BY BATTERY WASTE
Júlia C. Guimarães (UG)a*, Ricardo G. Cesar (R)b, Jéssica F. Paulino (R)a, Fernanda V. M.
Pontes (R)a
a UFRJ, Departamento de Química Analítica, Instituto de Química, Rio de Janeiro, RJ, Brazil, 21941-909
b UFRJ, Instituto de Geociências, Rio de Janeiro, RJ, Brazil, 21941-909
*e-mail: juliaconcordido@gradu.iq.ufrj.br
Soils can be contaminated with potentially toxic metals due to mining, fertilizers, metallurgical
industry, urban residues and other anthropogenic contributions.1 Battery waste is a type of urban
residue that, when discard incorrectly, could contribute to this environmental problem, because their
internal components gradually leak due to the external case degradation. 2 Knowing the total
concentration of this metals is not enough to evaluate their environmental availability, since it
depends on the way they are linked to the matrix and how available to the environment.3 Therefore,
sequential extraction schemes, such as Community Bureau of Reference (BCR) protocol, are
applied simulating different environmental conditions. BCR Protocol consists in 4 steps: acid-soluble
(CH3CO2H), reducible (NH2OH.HCl), oxidizable (H2O2 + NH4CH3CO2) and residual (HNO3 + HF),
each step more drastic than the former one.3 The present study aims to determine zinc and
manganese mobility by simulating a contaminated oxysol scenario with waste batteries (0,5 g of
residue per 100 g of contaminated soil) by applying BCR sequential extraction protocol. The
concentrations of metals were determined by flame atomic absorption spectrometry. The values
obtained in oxidizable, for both elements, and residual fraction, for zinc, were below the first analytic
curve point. Figure 1 shows extraction percentage in each step. Both elements were considered
available to the environment, because of their high extraction in the first steps. Their high percentage
in the acid-soluble step could be explained by their increased solubility in low Ph.4,5 Mn was more
extracted in step 2 because of the low redox potential that favored the reduction of insoluble
manganese oxides increasing Mn mobility 4. Some physical and chemical properties of the studied
soil are: 58% of clay, 6% of silt, 36% of sand, 0.22 % of organic matter and 2.3 cmol/dm,3,6 these
parameters could have contributed to the increased percentage extracted in the first two steps. A
low cation exchange capacity, when compared to other tropical soils, could be responsible for their
increased environmental availability since this characteristic increase Zn and Mn solubility.5,7 In
addition, a low organic matter also contributes to an increase in Mn availability.4
Figure 1. Extraction percentage for each step for each element
1 Zemiani, A.; Ahmann, F. S. Utilização de técnicas de FRX e AAS para análise de metais pesados em solo sob
aplicação de dejetos oriundos da suinocultura. 77 p. TCC (Graduação em Engenharia Ambiental) - Universidade Federal
do Paraná, Francisco Beltrão, 2014.
2 Agourakis, D. C. et al. Quim. Nova, 29, 2006, 960-964.
3 Marin, B. et al. Analytica Chimica Acta, 342, 1997, 91-112.
4 Nádaská, G.; Lesný, J.; Michalík, I. Hungarian Electronic Journal of Science, 100702-A, 2012, 1-16.
5 Obrador, A. et al. Geoderma, 137, 2007, 432–443.
6 Cesar, R. et al. Environmental Monitoring and Assessment, 186, 2014, 1487–1497.
7 Facco, D. B. et al. Pedosphere, 32, 2022.
[IFRJ –Nilópolis Campus for the facilities]

330
PRELIMINARY STUDY CONCERN THE PRESENCE OF MICROPLASTICS AND
ORGANIC CONTAMINANTS IN THE ESTUARINE ZONE OF RIVER TRAMANDAÍ
Vanessa F. Fonseca (PG)a*, Mariana A. Dias (PG)b, Cassiana C. Montagner (R)b,
Crislaine Bertoldi (R)c, Andreia N. Fernandes (R)a
bUniversity of Campinas, Institute of Chemistry, Campinas, SP, Brazil, 13083-970
cFederal University of Paraná, Department of Chemistry, Curitiba, PR, Brazil, 81531-980
*e-mail: vanessa.fontana@ufrgs.br
Microplastics are ubiquitous in the environment, and studies have demonstrated their potential
toxicity to different ecosystems, especially in aquatic environments.1 In addition, they are considered
carriers of contaminants due to their sorption and leaching capacity.2 Therefore, this study aims to
investigate the presence of microplastics and organic contaminants on the surface water in the
Tramandaí river estuary (southern Brazil). The sampling points were selected considering different
salinities and occupation land. Point 1 (P1) was in the River Tramandaí with a predominance of
freshwater; point 2 (P2) was in the Tramandaí lagoon; point 3 (P3) was in the estuarine zone with
higher salinity. The sampling was carried out on April 2022, where 46 L of water was collected with
the aid of a bucket at each site. The water was passed through a polyamide filter with 60 µm mesh.
Before the analysis, QA/QC measures were performed to avoid and quantify potential contamination
during the sample treatments. In the laboratory, the filters were washed using a maximum of 50 mL
of ultrapure filtered water. After drying at 60 ºC, the samples were first submitted to density
separation with a saturated solution of NaI (1.6 g cm-3), then to oxidative treatment using H2O2 (35%)
under 60 ºC for six hours. The potential microplastics were counted and characterized according to
their shape (fibre or fragment), colour (colourless; blue; green; red/orange; black or yellow), and size
using a microscope (magnification of 4x and 10x). The presence of organic contaminants was
investigated in samples of 1 L of water by LC-MS/MS, including pharmaceuticals, hormones,
pesticides, and industrial wastes. The preliminary results showed the presence of 278 potential
microplastics (6.5 items L-1) in sample P1, being 65.8% of fibres and 34.2% of fragments, with the
predominance of colourless (55.0%) and blue (32.7%) particles. This value shows the significant
contamination by microplastics in the River Tramandaí, being also noteworthy the considerable
number of colourless fibres representing 46.8% of the total potential microplastics. The main source
of fibres in the environment is domestic sewage, mainly due to the wear of fabrics, which maybe
explain the high value found in the sample for this shape of microplastics. Since the river’s banks
are surrounded by crops and it’s a traditional point of fishing, other possible sources of microplastics
are the degradation of plastic materials used to cover plants and fishing nets. The results on
microplastics to the other samples are still in analysis. Concerning organic contaminants, 10
compounds were detected in P1, 11 in P2, and 9 in P3. The higher concentrations were found to be
caffeine in P2 (146 ng L-1), bisphenol A in P3 (102.4 ng L-1), and desetilatrazine in P1 (94.2 ng L-1).
The occurrence of caffeine could indicate the presence of domestic sewage in the water body, while
Bisphenol A is used in the manufacture of some plastics, so it may be released by fishing baits and
nets. Thus, the presence of microplastics and organic contaminants in the water represents a great
concern since all these compounds have already demonstrated harmful behaviour to the
environment and probably reaches the Atlantic Ocean. Further studies will quantify and characterise
the microplastics in all samples, investigate the sorption of contaminants on their surface and relate
the presence of these compounds in water with the microplastics.
1 Trifuoggi M et al., Environmental Research, 179, 2019, 108815
2 Kinigopoulou V et al., Journal of Molecular Liquids, 350, 2022, 118580

331
HUMAN HEALTH RISK EVALUATION IN A SCENARIO OF CONTAMINED SOILS
BY INCORRECT BATTERY DISPOSAL
Júlia C. Guimarães (UG)a*, Letícia F. Pereira (UG)a, Vanessa S. Lisboa (UG)a, Bernardo J. M.
Miranda (PG)a, Daniel A. Barcelos (PG)b, Jéssica F. Paulino (R)a, Fernanda V. M. Pontes (R)a
a UFRJ, Departamento de Química Analítica, Instituto de Química, Rio de Janeiro, RJ, Brasil, 21941-909
bUniversidade de Lisboa, Centro de Química Estrutural, Instituto Superior Técnico, Lisboa, Portugal, 1049-001
*e-mail: juliaconcordido@gradu.iq.ufrj.br
In Brazil, 1.2 billion batteries are commercialized annually, almost 6 units per person per year. 1
Moreover, these products contain potentially toxic metals and their incorrect discard could lead to
contamination of the air, water tables, soils and in the end, reach the food chain resulting in risks to
human health.2 The environmental mobility of Zn, Mn, Cu and Ni in irregular and Zn-C batteries
wastes was studied using sequential extraction schemes (SES) based on Community Bureau of
Reference (BCR) Protocol. BCR is based on four fractions (acid-soluble, reducible, oxidizable and
residual). A way to measure the potential impacts of this metals to the environment and the society
is a Human Health Risk Evaluation, a mathematical model which simulates their transportation in
soil, air and water. This model calculates the hazard quotient (HQ) for different elements, basing in
a dose above which the exposure becomes harmful to human health.3 An HQ ≥ 1 indicates hazard
of non-carcinogenic adverse effects.4 Using the sum, for each element, of the first three fractions
obtained previously, the present study aims to characterize the human health risk in a scenario of
contaminated soil with used batteries using RISC4® program. There is no information of HQ for
copper in RISC4®. Table 1 shows 11 cases with HQ ≥ 1 (in red), in which zinc represented the major
danger between the elements. The principal occurrence of hazard was in soil intake, followed by
vegetables intake. Children would be in more hazard in comparison with adults. Between the
batteries analyzed, the Zn-C battery contributed more for the hazard scenario. This result reinforces
the importance of the correct disposal of batteries, usually destinated to domestical trash.
Table 1. Hazard Quotient (HQ) obtained for irregular and Zn-C batteries
Children Adults
Element Dermal Contact Soil Vegetables intake Dermal Contact Soil Vegetables
Intake Intake intake
Irregular battery
Ni 0.00510 0.130 0.100 0.00160 0.0140 0.0440
Zn 0.240 6.00 180 0.0740 0.640 77.0
Mn - 11.0 - - 1.20 -
Zn-C battery
Ni 0.00170 0.0430 0.100 0,000520 0.00460 0.0150
Zn 0.470 12.0 340 0.140 1.30 150
Mn - 14.0 - - 1.50 -
1 Espinosa, D. C. R.; Mansur, M. B. Waste Electrical and Electronic Equipment (WEEE) Handbook. Woodhead
Publishing, 2a edição, 2019.
2 Fernández, A. A.; Pérez, C. B.; Fernández, G. E.; Falqué, L. E. The Analyst, v.125, p.1353 – 1357, 2002.
3 Cathcart, D.; Análise comparativa de modelos de avaliação de risco à saúde humana para o gerenciamento de áreas
contaminadas. TCC (Graduação em Eng. Sanitária e Ambiental) – Centro Tecnológico, UFSC. Santa Catarina, 2014.
4 USEPA. EPA/540/1-89/002: Risk Assessment Guidance for Superfund - Vol. 1: Human Health Evaluation Manual (Part
A). Washington D.C., 1989.
[NQTR, IQ/UFRJ]

332
MICROCRYSTALLINE CELLULOSE BEADS FOR REMOVAL AND RECOVERY OF
PB (II) FROM DIFFERENT WATERS: BATCH AND COLUMN ADSORPTION
STUDIES
Dairon P. Fuentes (PG)a*, Lucas E. O. Porto (PG)a, Denise F. S. Petria, Pedro V. Oliveira (R)a
a University of São Paulo, Department of Fundamental Chemistry/Institute of Chemistry, Sao Paulo, SP, Brazil, 05513-970
*e-mail: dairon.pfuentes@usp.br
Pb (II), as one of the typical heavy metal ions, exists in sewage from different areas such as leather,
mining, painting,1 battery manufacturing, combustion of automobile petrol, electroplating industry,2
etc. The maximum contaminant level of 0.010 mg L-1 in drinking water established by the World
Health Organization (WHO),3 upon that could cause serious injuries to the nervous, digestive,
immune, reproductive, and urinary systems.4 Remediation of potentially toxic elements ions and their
species from wastewater is essential because of their extreme environmental, public health, and
economic impacts.5 This study aims to investigate the removal of Pb (II) using microcrystalline
cellulose beads prepared in HNO3. Different concentrations of HNO3 (0.5 - 8 mol L-1) were
investigated to obtain the optimal material for adsorption. These materials were characterized using
FT-IR, SEM images, and potentiometric titration. The adsorption process in flow was optimized for
the removal of Pb (II), parameters such as pH and flow rate were evaluated, and the best adsorption
performance at pH 6 and flow rate 0.5 mL min-1. The maximum adsorption capacity was evaluated;
the adsorption process adjusts to the de Langmuir model obtaining a maximum adsorption capacity
of 108.4 mg g-1. Real water samples (Drinking water, Dam water, and Seawater), were evaluated to
assess the potential of the material for real applications, reaching recoveries in the range of 93.55%
to 101.76% under the optimized adsorption process. These results indicate that these
microcrystalline cellulose beads are a potentially effective and eco-friendly adsorbent for application
in wastewater remediation.
Figure 1. Graphical abstract
1 Li Y, Xie D, Xiao J, Wu W, Zhang L, Xiao H, Chen J, Journal of Cleaner Production, 258, 2020, 120867.
2 Feng J, Zhang J, Song W, Liu J, Hu Z, Bao B, Ecotoxicology and Environmental Safety, 203, 2020, 111002.
3 WHO, Guidelines for Drinking-Water Quality: Fourth Edition Incorporating the First Addendum, 2017.
4 Xu X, Ouyang X, Yang L, Journal of Molecular Liquids, 322, 2021, 114523.
5 Fakhre NA, Ibrahim BM, Journal of Hazardous Materials, 343, 2018, 324.

333
PAPER-BASED DEVICE ANCHORED ON DRONE FOR FAST COLORIMETRIC
MONITORING OF TROPOSPHERIC OZONE
Pedro P. E. Campos (UG)a*, Habdias A. Silva-Neto (PG)a, Vanderli G. Leal (PG)b, Lucas C.
Duarte (R)a, João F. S. Petruci (R)b and Wendell K. T. Coltro (R)a
aUniversidade Federal de Goiás, Instituto de Química, Goiânia, Goiás, Brazil, 74690-900
bUniversidade Federal Uberlândia, Instituto de Química, Uberlândia, Minas Gerais, Brazil, 38400-902
*e-mail: ppedro@discente.ufg.br
In recent years, unmanned aerial vehicles (UAVs; Drone) have emerged as a powerful and versatile
analytical approach for remote monitoring.1 Another thankful trend is the fabrication of “do it yourself”
analytical tools, such as paper-based analytical devices (PAD).2 The combination of PADs and
drones is an excellent alternative for sampling and monitoring trace gas in outdoor environments. In
this study, we propose for the first time the simple assembling of PADs into a drone to monitor the
gaseous ozone in the troposphere. For manufacturing the PAD, chromatographic paper, vinyl stencil
(Ø = 5 mm) and glass varnish binder were successfully utilized as materials. A commercial generator
is utilized as a standard source of O3 (~ 229 mg h-1, characterized by iodometric titration method).
Initially, the colorimetric reaction for ozone quantification was evaluated spotting the reaction zones
with 1.5 µL of 1 mmol L-1 potassium indigotrisulfonate (ITS), 1.5 µL of poly(ethylene glycol) 400
(PEG) and the presence of ozone was verified by changing color from blue to colorless. After the
reaction, the PADs were digitalized on a desktop scanner and the analytical signals were extracted
using CORELDRAW software. Aiming to improve the analytical performance of the colorimetric
analysis, optimization in terms of reaction time, order of addition of reagents and concentration of
PEG 400 were performed in the presence of 1 mmol L-1 ITS. Under optimized conditions, the
analytical curve of O3, repeatability and reproducibility experiments were also acquired. The obtained
relation between the mass of ozone and the color channel cyan exhibited acceptable values with a
linear range between 0.95 to 7.63 mg of O3 (R2 = 0.99), limit of detection (LOD) of 146 µg and relative
standard deviation (RSD) ranging from 1.6 to 3.4% for the reproducibility and repeatability studies.
A plastic holder was manufactured on a 3D printer tool aiming for anchored the PADs upon the drone
(Figure 1). The drone anchored with four different PADs was positioned ~ 20 meters from the ground
and the sampling and analysis of ozone was realized during ~ 2 min. The obtained colorimetric
analysis indicated amount of O3 ranged from 6.80.7 to 7.30.2 mg that is similar to found amount of
O3 reported in literature involving trace gas in the atmospheric air. Thus, we believe that the analytical
tool introduced herein has the potential for fast, remote, and spatial monitoring of trace gases in the
troposphere.
Figure 1. Schematic representation involving the PAD preparation, drone, and colorimetric analysis of ozone.
1 Lally H.T. et al., Science of the Total Environment, 670, 2019, 569.
2 Silva-Neto H.A. et al., Analyst, 146, 2021,3463.
[UFG, UFU, PETROBRAS, CNPq, CAPES, INCTBio]

334
DETERMINATION OF CL, F, AND S IN DRILL CUTTINGS SAMPLES FROM
OFFSHORE OIL AND GAS EXPLORATION WELL BY ION CHROMATOGRAPHY
AFTER PYROHYDROLYSIS SAMPLE PREPARATION
Renan G. Marim (PG)a*, Gilberto da S. Coelho Junior (PG)a, Paola C. C. de Azevedo (PG)a,
Daniel L. G. Borges (R)a, Tatiane de A. Maranhão (R)a, Fernando J. S. de Oliveira (R)b, Vera
L. A. F. Bascunãn (R)a, Eduardo S. Chaves (R)a
a Universidade Federal de Santa Catarina, Departamento de Química, Florianópolis, SC, Brasil, 88040-900
b Petróleo Brasileiro S. A., Gerência de Meio Ambiente, Rio de Janeiro, RJ, Brasil, 20031-004,
*e-mail: marimrenan@gmail.com
The oil and gas industry is one of the most important sectors of the world economy. However,
environmental issues associated with oil and gas exploration have to be considered due to the high
polluting potential associated with this activity.1 Taking into account that information about the
concentration of non-metals in drill cuttings from oil and gas exploration wells in deep waters is
scarce, the determination of S and halogens in these samples, as well as other elements, may offer
an important environment and geological data.2,3 However, monitoring the concentration of these
elements is still a challenge, since these elements, especially the halogens, are volatile and can be
lost during the sample preparation. Hence, for chemical analysis, the sample preparation is a critical
step.4 The pyrohydrolysis stands out as a sample preparation for being a relatively simple, efficient,
and low-cost technique. It is based on the pyrolysis of samples in the presence of water vapor at
high temperatures, where the non-metals are hydrolyzed forming their respective hydrogen halides,
which are subsequently condensed and collected in an alkaline absorbent solution.5 Thus, the
determination of Cl, F, and S concentrations in depth profile of drill cutting samples, from an offshore
oil and gas exploration well, was performed by using ion chromatography analysis after the sample
preparation using pyrohydrolysis. The parameters of pyrohydrolysis were optimized by a Box-
Behnken design and the optimal conditions were found using 300 mg of drill cuttings, 300 mg of
V2O5, temperature of 1000 °C, 25 min of reaction time, and a mixture of 50 mmol L-1 NH3 + 150 mmol
L-1 H2O2 as absorbing solution. During the process, air flow rate of 0.2 L min-1, and water flow rate of
0.1 mL min-1 were applied. The accuracy of the proposed method was evaluated by recovery tests
and analysis of two certified reference materials. The obtained recoveries ranged from 89 to 108%,
and a good agreement (85-103%) between the determined concentrations with the reference values
was obtained. The precision of the proposed method, evaluated by RSD (< 10%), and the limits of
detection (0.5 – 2.5 mg kg-1) were suitable for drill cutting analysis. The proposed method was
applied for the determination of Cl, F and S in drill cutting samples collected at different depths (4,119
– 6,256 m). The results showed that the concentration of Cl (23.72 – 486.96 mg g-1) and F (0.073 –
0.421 mg g-1) decreases as the drilling depth increases. However, the XRD analysis showed that
the composition of the samples at the depths of 4,119 m and 4,644 m, which present the highest
concentrations of Cl, is predominantly halite (NaCl). For S, the obtained concentrations varied from
3.23 mg g-1 to 31.09 mg g-1, and the concentration increases with the drilling depth. This is due to
the characteristics of the rock, which at greater depths presents barite in its composition (BaSO 4).
Therefore, the variation in the concentration of analytes as a function of depth is probably related to
the mineral composition.
1 Martins SSDS, Silva MPOAM, Silva VP, Holos, 31, 2015, 54.
2 Kendrick MA, Burnard P, Advances in Isotope Geochemistry, 1, 2013, 319.
3 Kusebauch C, John T, Barnes JD, Klügel A, Austrheim HO, J. Petrol., 56, 2015, 299.
4 Tjabadi E, Mketo N, TrAC - Trends Anal. Chem., 118, 2019, 207.
5 Flores, E. L. M.; Flores, E. M. M.; Antes, F. G.; Duarte, F. A.; Barin, J. S.; Dressler, V. L. In Métodos de preparo de
amostra para análise elementar; Krug, F. J.; Rocha, F. R. P., Eds.; Sociedade Brasileira de Química: São Paulo, 2019.
[CNPq, CAPES, Petrobras, UFSC]

335
ENVIRONMENTAL MONITORING OF LEAD IN WATER SAMPLES AND
SURURU ASSOCIATED WITH THE MUNDAÚ MANGUABA ESTUARINE
COMPLEX, AL - BR
Emerson dos Santos Freire (UG)*, Josué Carinhanha Caldas Santos (PG), Daniela Santos
Anunciação (PG)
Universidade Federal de Alagoas, Instituto de Química e Biotecnologia, Maceió, Alagoas, Brazil, 57072-970.
*e-mail: freireemerson02@gmail.com
The Lagoon Mundaú-Manguaba Estuarine Complex (CELMM) is one of the most important
ecosystems in Brazil, playing a significant role in maintaining microclimates, in addition to having
high potential for the development of fisheries.1 Monitoring studies of the quality parameters of the
different environmental compartments are of paramount importance, such as water bodies. Among
these parameters is the determination of potentially toxic metals such as lead (Pb), which is an
indicator of contamination by domestic sewage.2 Thus, the objective of this work was to monitor the
Pb content in samples of water and sururu (Mytella strigata) from CELMM. Four samplings were
carried out at 11 collection points along the Mundaú lagoon, the CELMM site that is most influenced
by human activities3. The conditioning and treatment of water and sururu samples followed protocols
already established in the literature.3 The water samples were filtered with 0.45 µm cellulose acetate
membrane and the sururu samples were lyophilized and submitted to a microwave-assisted acid
digestion procedure. The Pbtotal concentration was determined by electrothermal atomization atomic
absorption spectrometry (GF AAS). The concentration of Pbtotal quantified in the water samples from
the first collection (07/2021) ranged from 1.0±0.1 to 10.1±0.5 µg L -1, in which for the samples from
points P5, P6, P8, P9, P10 and P11 Pb levels were below the quantification limit (0.98±0.2 µg L -1).
In the second collection (10/2021), the levels of this element ranged from 1.3±0.1 to 9.8±1.0 µg L-1,
in which for the sample points P1, P2, P10 and P11 the levels of Pb were below the limit of
quantification. For the samples from the third collection (03/2022) the lead levels ranged from 1.0±0.3
to 9.5±0.2 µg L-1. And for the fourth collection (07/2022), the Pb content ranged from 1.0±0.2 to
7.1±0.4 µg L-1. Observing the Pbtotal values quantified in the samples of the four collections, it was
verified that for all samples the Pb content was below the maximum limit allowed for this element
according to CONAMA resolution 357/2005, whose maximum level allowed for Pbtotal in brackish
waters class 1 is 10 µg L-1.4 In the sururu samples, the Pb level ranged from 0.35±0.05 to 0.65±0.09
mg kg-1 for the first collection and from 0, 21±0.04 to 0.26±0.06 mg kg -1 for the second collection.
The sururu samples from the third and fourth collections are being processed for further analysis.
Based on the Pbtotal values in the sururu of collections 1 and 2, it is possible to analyze that all
samples were below the maximum limit allowed for this type of food, according to ANVISA resolution
42/2013, whose maximum content of Pbtotal allowed for bivalve molluscs is 1.5 mg kg-1.5 According
to the results obtained from the sampling carried out over 2 years, it is possible to conclude that the
concentrations of Pbtotal in the water and sururu samples were in accordance with the limits
established by the current Brazilian legislation. However, for the water samples from the first three
collections, the values were very close to the maximum allowed limit, evidencing the need to maintain
monitoring in order to monitor concentration levels and possible accumulations in this ecosystem.
The fourth sampling revealed a decrease in the concentration range of the samples due to the
dilution effect resulting from the rains in the period before the collection.
1 Freire ES, et., J. Braz. Chem. Soc., 32, 3, 2021, 684-693.
2 Nag R, Cummins E, Ciência do Meio Ambiente Total, 810, 2022, 151168.
3 Dos Santos MC, et al., J. Braz. Chem. Soc, 32, 9, 2021, 1762-1772.
4 CONAMA. Conselho Nacional do Meio Ambiente. Resolução No. 357, 18 de março de 2005.
5 ANVISA. Agência Nacional de Vigilância Sanitária. Resolução nº 42, de 29 de agosto de 2013.
[CNPq, FAPEAL, UFAL, PPGQB, IQB]

336
A MICRO-IMPINGER SAMPLING DEVICE FOR DETERMINATION OF
FORMALDEHYDE IN AIR
Marcus Augusto dos Santos Catai*, João Flávio da Silveira Petruci, Arnaldo Alves Cardoso
Instituto de Química de Araraquara, Dep. de Química Analítica, UNESP. Araraquara/SP, Brazil, CEP 14800-105
*e-mail: marcus.catai@unesp.br
Formaldehyde is ubiquitous in the atmosphere. Formaldehyde is formed in the atmosphere by

photochemical reactions involving hydrocarbons with a substituent methyl group. Formaldehyde is
widely used in composite wood products containing resins containing formaldehyde and is in building
materials and insulation, glues, permanent press fabrics, paints, lacquers, and other coatings. All
these materials emitted formaldehyde in the indoor atmosphere. It is a gas that can irritate a person's
eyes, nose, throat, and lungs, or trigger an asthma attack, even at low concentrations. Prolonged
exposure to formaldehyde can cause cancer. In this work, we developed a low-cost and portable
micro sampler. A disposable semi-micro acrylic cell (volume: 1.4 mL and path length 10 mm) was
used as a μ-impinger bubbler containing 600 μL of solution of 3-methyl-2-benzothiazoline hydrazone
(MBTH). The formaldehyde reacts with MBTH and forms the azine in about 20 min. The oxidation of
MBTH by Fe (III) and dye formation requires 3 min. The MBTH forms a blue cation that can be
measured at λ=628 nm. The absorbance is proportional to the sampling time and the analyte
concentration. The proposed method was evaluated using outdoor and indoor samples, and it was
shown to provide an accurate measure of total aldehyde. The μ-impinger determined formaldehyde
concentrations lower than 0,05 ppm using a 50 mL min-1 sample flow rate during 10 min. This
formaldehyde concentration is forty times lower than the OSHA (Occupational Safety and Health
Administration) air quality standard to STEL (Short Term Exposure Limit). Standard atmospheres
with formaldehyde concentrations from 50 ppb to 2 ppm were used in the calibration procedure. The
limit of detection was 50 ppb considering 3.3 times the standard deviation of the blank signal. The
detection limit could be improved further by using longer sampling times. The method is selective
and highly sensitive.
Figure 1. Blank on the left and increasing concentration on the right from 50 ppm up to 2 ppm.
[CAPES, CNPq, MASC]

337
SPECIATION OF Gd (III) FROM Gd-CHELATE USING MICROCRYSTALLINE
CELLULOSE BEADS FOR SEPARATION AND PRECONCENTRATION FROM
WATERS
Lucas E. O. Porto (PG)*, Dairon P. Fuentes (PG), Pedro V. Oliveira (R)
University of São Paulo, Chemistry Institute, São Paulo, SP, Brazil, 05508-900
*e-mail: lucas.eduardo.porto@usp.br
The growing worldwide demand for rare earth elements is related to various applications, particularly
in the high technology industry.1 Due to the characteristics and physicochemical properties,
gadolinium (Gd) is of interest to medicine and the environment. Gd has found use in many different
industries. Old electronics and other waste that end up in landfills are potential sources of
contamination that may occur through leaching into the environment over time. Furthermore, Gd
content is substantially higher in soil and plants close to the mining and processing sites. However,
the main environmental concern with respect to this element is related to Gd containing chelates
used as diagnostics tools. The extensive use in magnetic resonance imaging has led to increased
Gd levels in industrialized parts of the world, adding to the natural occurrences and causing
environmental and health concerns.2 There is a strong correlation between Gd concentration in the
environment and the level of the health care system as Gd is found in excessive amounts closer to
larger cities and hospitals performing magnetic resonance imaging. 2-4 In general, the concentration
of Gd is very low in natural water requiring high sensitive techniques for detection or separation and
preconcentration methods for determination. In this way, the aim of this work is to explore a method
for speciation of Gd (III) in presence of Gd-chelate using microcrystalline cellulose beads for
separation and preconcentration in natural water using ICP-OES as detector. Preparation of the
cellulose beads was carried out using a solution of 5% (w/v) microcrystalline cellulose + 7% (w/v)
NaOH + 12% (w/v) urea, at 10 ºC, under stirring (200 rpm) for 20 minutes. This solution was dropped
in an acidic coagulant bath of 2 mol/L HCl, at 25°C, using a peristaltic pump. Cellulose beads were
characterized using FT-IR and SEM images. Columns were prepared in a polypropylene tube (id of
0.6 cm) and porous polyethylene tips (d=0.6 cm; pore size < 1 µm) with 200 mg of beads. Solutions
of 1 mg/L of Gd(III) in different pHs were percolated through the column and results showed
preconcentration close to 100% at a large pH range (4 to 8). A solution of 1 mol/L HCl was very
efficient for desorption (100%) of gadolinium from the column. The addition of 100 ng/mL Gd (III)
presented recoveries of 98% in tap water and 96% in river water. Tests to evaluate the adsorption
of Gd-chelate over the microcrystalline cellulose beads column was done and showed no affinity of
this specie for the material. The addition of a mixture containing 5 ng/ml of Gd (III) + 5 ng/mL of Gd-
chelate in river water showed recovery of 100% for Gd (III), demonstrating good selectivity of bead
for Gd (III).
1 Balaram V, Geoscience Frontier, 10, 2019, 1285–1303.
2 Unruh C, Bavel NV, Anikovskiy M, Prenner, EJ, Molecules, 25, 2020, 5762.
3 Brünjes R, Hofmann T, Water Research, 182, 2020, 115966.
4 Telgmann L, Sperling M, Karst U, Analytica Chimica Acta, 764, 2013, 1-16.

338
ATRAZINE AND METABOLITES MONITORING IN BRAZIL AND PARAGUAY
STREAMS BY GC-MS
Bianca do Amarala*, Natalie P. Toyamaa, Diego A. Tavaresa, Gilcélia A. Cordeirob, Lourdes I.
Garayc, Gabriela S. Benegasc, Jussara E. Souzad
aItaipu Technological Park, Territorial Intelligence and Management, IT.DT, Foz do Iguaçu, PR, Brazil, 85867-900
b Federal University for Latin American Integration, ILACVN, Foz do Iguaçu, PR, Brazil, 85867-970
c Itaipu Binacional, Ciudad del Este, Departamento Alto Paraná, Paraguay
d Itaipu Binacional, MARR.CD, Foz do Iguaçu, PR, Brazil, 85867-970
*e-mail: bianca.amaral@pti.org.br
Pesticide consumption is increasing every year. According to the agri-environmental indicator of the
Food and Agriculture Organization of the United Nations (2022), 5.94 kg of pesticides were used per
hectare of area cultivated in 2019 in Brazil, relatively close to Argentina (6.07 kg/ha), Uruguay (8.03)
and Paraguay (4.68).1 The presence of pesticides in surface water concerns due to the chronic
effects on human and animal health, which are still poorly understood. Monitoring pesticides is
essential to support efficient water resources management, requiring information of pollutants in this
environment.2 Although this topic has been widely discussed in the scientific community, monitoring
data in Brazil and Paraguay streams are still scarce. In this context, this study aimed to investigate
atrazine and its main metabolites (DIA and DEA) in surface water. The studied region comprised
twenty-four streams in Brazil (BR) and Paraguay (PY), an area characterized by intensive
agriculture. Samples sites were classified as groups a, b or c according to anthropic influence
considered elsewhere.2,3 The analytical approach involved solid-phase extraction (SPE) followed by
gas chromatography with mass spectrometry (SIM mode). SPE method was optimized by 22 factorial
design using Oasis HLB® (200mg). The central point condition showed good recovery values (99%
to 113%) and it was selected: methanol and dichloromethane as the solvent and 200 mL of sample
amount. Matrix-matched were used to minimize the matrix effect. Good linearity was achieved using
the SPE-GC-MS method (R2≥0.98). The limits of quantification (LOQ) were 25 ng L-1 for DEA only
in BR and 50 ng L-1 for other compounds in BR and for all compounds in PY. Recovery ranged from
86.9 to 122%, with a relative standard deviation between 3 and 21%. The results of three monitoring
campaigns in 2019 and 2020 revealed the occurrence of ATZ and DEA. DIA was not detected in any
samples collected in Brazil and Paraguay. DEA was quantified in 7 events in different campaigns,
ranging from 31.8 to 140.5 ng L-1. The highest concentration was detected at MBR16c in November-
2019. Also, it was detected below LOQ in July-2019 samples (MBR12c, MBR16c, MBR17b, MPY06,
and MPY10b) and the November-2019 samples (MBR17b). Relationships with sediment transport
influence the results in sampling points MPY10, with a recent landfill event and MPY06 located
downstream of an artificial reservoir. ATZ was only found below LOQ (50 ng L-1) in November-2019
samples (MPY10b) and in October-2020 (MBR14b). These results coincided with the period of maize
cultivation in the region. Although the screening analysis2 confirmed the presence of these
compounds, the concentration must be lower than the detection limits reached by the quantification
technique. These limits also accomplish the regulatory limits for freshwaters in Paraguay (3.000
ng/L) and Brazil (2.000 ng/L). According to the ATZ and its metabolites’ physicochemical properties,4
the degradation process of ATZ increases the solubility and polarity of the newly formed compounds.
ATZ has a maximum solubility in water of 33 mg L-1, while 3200 mg L-1 for DEA, and 670 mg L-1 for
DIA, showing greater mobility than ATZ in water. Furthermore, the half-life (t1/2) of DEA, 52 days, is
longer than that of DIA, 36 days. Hence, continuous monitoring will allow the estimation of the
environmental susceptibility and potential correlation with effects on local biodiversity.
1 FAO. https://www.fao.org/faostat/en/#data/RP
2 Becker RW, Araujo DS, Sirtori C, et al. Microchemical Journal, 168, 2021, 106502
3 Ouchi-Melo LS, Amaral B, Tavares DA, et al. Limnologica, 90, 2021, 125904
4 Pesticide Properties DataBase. University of Hertfordshire. Available on http://sitem.herts.ac.uk/aeru/ppdb/
[Itaipu Binacional, Itaipu PTI, UNILA]

339
DEVELOPMENT AND VALIDATION OF A METHOD FOR THE
DETERMINATION OF TOTAL NITROGEN IN SURFACE WATER SAMPLES BY
ION CHROMATOGRAPHY
Joslaine Jacumazoa*, Diego A. Tavaresa, Natalie P. Toyamaa, Gilcelia A. Cordeiro, Bianca do
Amarala
aFundação Parque Tecnológico Itaipu - BR, Foz do Iguaçu, Paraná, Brazil, 85867-900
bUniversidade Federal da Integração Latino Americana - UNILA, Foz do Iguaçu, Paraná, Brazil, 85866-000
*e-mail: joslaine.jacumazo@pti.org.br
Nitrogen is one of the most abundant elements in the universe and essential for all living
organisms.1,2 In water samples, it can exist in both inorganic and organic forms. Inorganic nitrogen
is present in the oxidized (NO2−, NO3−) and reduced (NH4+) species. On the other hand, organic
nitrogen may be available as amino acids, proteins and urea1. The sum of both forms gives rise to
total nitrogen which can be determined by several methods. Combining the persulfate digestion
method with ion chromatography (IC) separation has become an interesting option for its
quantification, reducing the analysis speed with less waste generation and resulting in more reliable
results.1-3 The total nitrogen determination is essential due to the proper balance of environmental
nutrients; when in excess in the water body, it promotes the growth of algae, impacting the
environment, human health, and the economy.2 In this sense, the objective of this study was to
develop and validate a method for determining total nitrogen by ion chromatography, for its
quantification in surface water samples. For this, a Metrohm® 930 Compact IC Flex chromatograph
with 919 IC Plus autosampler and 945 Professional Vario conductivity detector was used. In addition,
for all analytical separations, the following conditions were used: Metrosep A Supp 16 column (250
x 4.0mm), Metrosep A Supp 16 S-Guard pre-column (5.0 x 4.0mm) with injection loop of 250 μL,
sulfuric acid suppressive solution (200 mmol L-1) and flow rate of 0.8 mL min-1. For the determination
of total nitrogen, an experimental design 22 was used, with the independent variables: temperature
(25 and 32 ºC) and concentration of the mobile phase (solution of sodium hydroxide 20 and 30 mmol
L-1). The digester solution consisted of a mixture of potassium persulfate (0.2 mol L-1), boric acid (0.5
mol L-1) and sodium hydroxide (0.001 mol L-1). In addition, the nitrate standard and surface water
samples were autoclaved for 1h at 121 ºC and later diluted at 1:100 V V-1 for analysis in the
chromatograph. During the digestion process, there is a decomposition of persulfate, becoming a
potential interference in the analysis due to the presence of an intense sulphate peak. As it is a
multivalent ion, when there is an increase in temperature, greater retention in the stationary phase
was observed, leading to overlap and/or coelution with the nitrate. However, increasing the mobile
phase concentration leads to a decrease in its retention time. Regarding nitrate, the temperature did
not influence the retention times. On the other hand, by increasing the concentration of the mobile
phase, there is a decrease in the retention time and a more efficient separation of the peaks. Thus,
considering the selectivity, the best separation between sulphate and nitrate was observed for
conditions of intermediate mobile phase concentration and lower column oven temperature. From
the definition of the best condition, linearity (30 - 1500 µg L-1), limit of detection (7.5 µg L-1), limit of
quantification (22.7 µg L-1), precision (RSD <5.0%), accuracy (94% - 107%) and robustness (UV-Vis
and IC) were determined. Furthermore, when using surface water samples it was observed a more
reproducible signal when compared with the data obtained by the spectroscopic method. Therefore,
ion chromatography associated with the method of digestion with persulfate proved to be an efficient
and fast technique in total nitrogen determination in surface water samples.
1Michalski R, Kurzyca I. Polish Journal of Environmental Studies, 15, 2006, 5-18.
2De Borba BM. Jack RF, Rohrer JS, Wirt J, Wang D. Journal of Chromatography A, 1369, 2014, 131-137.
3Jackson PE. Trends in Analytical Chemistry, 20, 2001, 6-7.
[Itaipu Binacional, Itaipu PTI, UNILA]

340
LABILITY OF RARE EARTH ELEMENTS IN SOILS IN A URANIUM MINING
RESTORATION SITE USING THE DGT TECHNIQUE
Lucas P. Eliasa* (PG), Amauri A. Menegárioa (R), Hendryk Gemeinera,b (R), Amália E. M.
Rosac, Fabio W. Rodriguesc, Melina B. T. Zanattaa (R), Luiz F. P. P. Moreiraa (PG), Leonardo
H. Zoiaa (UG)
a São Paulo State University (Unesp), Environmental Studies Center, Rio Claro, SP, Brazil, 13506-900
b São Paulo State University (Unesp), Institute of Geosciences and Exact Sciences, Rio Claro, SP, Brazil, 13506-900
c Nuclear Industries of Brazil (INB), Caldas, MG, Brazil, 37780-000
*e-mail: lucas.elias@unesp.br
The rare earth elements (REE) are a group of seventeen chemical elements including fifteen
lanthanoids (La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, and Lu) as well as Sc and Y.
The REE are scarcely distributed in the Earth’s crust, being commonly found in the environment as
trace elements in rock-forming minerals. Due to their physicochemical properties, their use is
constantly intensified in industrial and agricultural activities, being vital to several modern
technologies.1 As a consequence of their recent intense use, the release of REE into the environment
has increased over the years. Many REE are considered to be emerging contaminants and
information about REE bioavailability is lacking, mainly, for soil. For this reason, it is important to
investigate the biogeochemical behaviour of REE aiming to evaluate the environmental risk and the
possible toxicity of these elements.2 The INB Caldas Mining-Industrial Complex (INB Caldas),
located in Minas Gerais state, is a former uranium mining area undergoing decommissioning and
restoration. The industrial extraction operated between 1982 and 1995 (in form of open pit mining).3
Uranium ores can also contain REE, and radioactive elements such as U and Th associated with
REE present several problems related to waste processing and disposal.4 In these areas, structures
such as tailings dams, mine pits and waste rock piles can represent several sources of
contamination, especially with regard to acid mine drainage. The Diffusive Gradients in Thin films
(DGT) technique has been recognized as an effective tool for studying various environmental
processes, including soil dynamics, chemical speciation, and elemental lability (as consequence
their bioavailability) analysis.5 The objective of this study is to evaluate the lability of REE in topsoil
samples and thus seek to understand the possible bioavailability of these elements. Samples were
collected at a reference point (external area of the mine complex) and in the contaminated area near
the tailings pile. Total metal content in soils was determined by ICP-MS. The flux of REE in the soil
samples was measured by DGT (24 h deployment at controlled temperature (21ºC) and constant
water soil holding capacity). Pore water was extracted from soils sampled by using centrifugation
and filtration through PTFE-filters with a pore size of 0.45 μm. The filtered solution was acidified and
analysed by ICP-MS. The total concentrations of La, Ce, Pr, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm, Yb
and Lu measured in soils of spoil pile area were 16.6 mg kg-1, 10.4 mg kg-1, 1.80 mg kg-1, 4.29 mg
kg-1, 0.39 mg kg-1, 0.05 mg kg-1, 0.72 mg kg-1, 0.23 mg kg-1, 0.04 mg kg-1, 0.12 mg kg-1, 0.02 mg kg-
1
, 0.10 mg kg-1 and <LOD, respectively. The range of concentrations measured by DGT in the
contaminated area varied from 0.21 µg kg-1 for Lu to 330.34 µg kg-1 for La. The pore water content
from soils sampled by DGT devices showed significantly higher values with concentrations varying
from 1.53 µg kg-1 for Lu and 2262.01 µg kg-1 for La. Comparing pore water concentrations and
measurements obtained by DGT, it was possible to analyse replacement and flow conditions,
providing important information for evaluating the lability of these elements in the environment.
1 Balaram V, Geoscience Frontiers, 10, 2019, 1285.
2 Squadrone S, Brizio P, Stella C et al., Science of The Total Environment, 660, 2019, 1383.
3 Gemeiner H, Menegário AA, Williams PN et al., Environ Sci Pollut Res, 28, 2021, 57149.
4 Khan AM, Bakar NKA, Bakar AFA et al., Environ Sci Pollut Res, 24, 2017, 22764.
5 Zhang H, Davison W, Environmental Chemistry, 12, 2015, 85.
[FAPESP, CAPES, CNPq, INB]

341
ADAPTATION OF PORTABLE AND LOW-COST ANALYTICAL METHODS FOR
DETERMINATION OF NITROGEN DIOXIDE AND OZONE IN THE
ATMOSPHERE OF RONDONOPOLIS, MT
Walter Aparecido Bezerra*, Joao Flávio da Silveira Petruci, Arnaldo Alves Cardoso*
*Instituto de Química de Araraquara, Dep. de Química Analítica, UNESP. Araraquara/SP, Brazil, CEP 14800-105
Instituto de Quimica, Universidade Federal de Uberlândia (UFU), Minas Gerais, Brazil
*e-mail: walteraparecidobezerra@gmail.com
Nitrogen dioxide (NO2) is the most common nitrogen oxide found in the atmosphere. The NO 2
promotes photochemical reactions resulting in nitric acid, ozone, and fine aerosol. Also, the presence
of NO2 can contribute with the formation of high levels of gaseous ozone. In the environment, both
pollutants (NO2 and O3) act directly on people's health. In spite of this, very few cities in Brazil have
data on the concentration of atmospheric pollutants. Rondonópolis, MT with about 240 000
inhabitants, is an industrial pole of agribusiness products. It receives air masses from the Cerrado
and Amazon burning areas during the dry season. The lack of data on local air pollutants inhibits
any effective source control action by the local public. Therefore, low-cost analytical devices with
reliable results are essential to establishing emission control policies for these gases. In this work,
we propose the combination of a lab-made,photometer and passive sampler to enable NO2 and O3
sampling and analysis with low-cost and portability. The photometer (Figure 2) was built using an
Arduino Uno board, a digital light sensor (TSL2561), a LED, a potentiometer and a LCD display. The
permeation passive samplers1 were constructed using a 50 mL Falcon conical tubes and Teflon
tape. NO2 was collected with filter paper impregnated with triethanolamine and the Griess-Saltzman
reagent for colorimetric determination of NO2. Ozone was sampled and determined by the ozonolysis
reaction of indigotrissulfonate (ITS), resulting in a colorless product. After the sampling time (24h),
the reagents were extracted with water and the absorbance was measured using the portable
photometer. The data obtained in the city, in 6 sampling points, show the first results (Figure 1). The
data obtained are compatible with the emission sources of NO2 and O3 precursors.
Figure 1. NO2 and O3 concentration in sampling sites, Rondonópolis
The study brought useful data regarding the NO2

and O3 concentrations in the investigated region.
The good capability of passive samplers combined
with a photometer showed fluctuations in
atmospheric gas concentrations. These variations
are expected due to the different intensities of the
emission sources in each sampled region. The
proposed procedure is applicable for air monitoring
on a local scale.
Figure 2. schematic drawing of photosensor
1 Souza PAF, Francisco KCA, Cardoso AA., Quimica Nova, 40,2017, 1233
[CAPES, CNPq, SEDUC-MT]

342
ALTERNATIVE AND LOW-COST BIOADSORBENT OBTAINED FROM AÇAÍ
SEED FOR IRON(II) REMOTION
Ricardo R. Bandeira (PG)a, Paulo T. Garcia (R)a*
aFederal University of the South and Southeast of Pará, Marabá, Pará, Brazil, CEP 68507-590
*e-mail: ptarsogarcia@unifesspa.edu.br
In the present study, an simple and alternative strategy to use açaí seed as a powerful bioadsorbent
for iron(II) remotion is proposed. Açaí seed were collected in a local market in Açailândia city, washed
with deionized water and then dried at 80 °C for 48 h. The dry biomass was mashed and the powder
was separated in 35 Mesh granulometric sieve as previous described.1 Tests were performed to
determine the potential of zero charge (pcz) and the ideal conditions in terms of adsorbent mass,
pH, contact time, adsorption kinetics and adsorption isotherms. Absorbance measurements were
obtained at 511 nm using a UV-VIS T90+ spectrophotometer using 1,10-phenantroline method. The
better iron(II) percentual remotion was obtained using a relation equal to 1 g of bioadsorbent for 1 L
of solution, using 10 mg/L of iron(II) standard solution and pH 5,0 (figure 1-A). The ideal range for
equilibrium time was obtained between 40 and 60 min once the adsorbed amount was constant
(figure 2-B). The kinetics models indicate a chemical behavior in the adsorbent process for iron(II),
once pseudo-second order was satisfy2 (figure 3-C). Related to the adsorption isotherms, Freundlich
model presented the better adjust and thus suggest that the adsorption process occurs in
multilayers3 (figure 4-D).
(A) (B)
(C) (D)
Figure 1. Optimization of (A) bioadsorbent mass, (B) contact time, (C) kinects study and (D) adsorption isotherm.
After optimize the experimental conditions was possible remove more than 70% of iron(II), therefore,
the use of this biomass for this purpose can be considered satisfactory once it’s an low cost,
abundant material and is not necessary previous adsorbent modification. Subsequently, miniaturized
electrochemical sensors will be used for iron(II) determination and compared with the results using
spectrophotometric procedure in order to obtain an portable an low cost complete method for iron(II)
remotion and determination at point-of-need. Finally, the proposed bioadsorbent will be also
evaluated to remove other ions like cadmium and copper.
1 Lima LR. et al. Revista Virtual de Química, 12, 2020.
2 Tran TH. et al. Science of The Total Environment, 725, 2020.
3 Akbar AAM. et al. Separation Science and Technology, 2018, 26-34.

343
STUDY OF THE PHOTOELECTROREDUCTION OF CO2 ONTO NANOTUBES
Ti/TiO2 MODIFIED BY UiO-Zr-NH2 METAL-ORGANIC FRAMEWORK
Kallyni Irikuraa*, Lucas M. Silvaa, Regina C. G. Frema and Maria V. B. Zanonia
aSão Paulo State University, UNESP, Institute of Chemistry
Francisco Degni, 55, Quitandinha, Araraquara, São Paulo, 14800-060, Brazil

*e-mail: kallyni@gmail.com
Carbon dioxide is the main gas responsible for the greenhouse effect, currently considered one of
the most serious global environmental problems.1 To minimize these problems, the
photoelectrocatalytic (PEC) processes are widely used, since they can reduce the CO2 molecules to
high value-added compounds such as CO, CH4, CH3OH among others.2 For this, the development
of new semiconductors able to generate charge carriers (e-/h+) is of fundamental importance. Studies
have found significant improvements in the CO2 reduction performance using TiO2 electrodes
modified with metal organic frameworks (MOFS).1,2 This can probably be explained by synergy
between the photocatalyst and the MOFs avoiding the recombination of e-/h+ pair photogenerated1.
Thus, the aim this work was to investigate the performance of TiO 2 nanotubes electrode (TiO2NT)
modified with UiO-Zr-NH2 to CO2 reduction by PEC process. Initially, TiO2NT electrode was prepared
by anodization1 and modified with UiO-Zr-NH2 MOF. The MOF was synthesized onto the TiO2NT
used solution of 60 mL of diphenylformanide, 0.240 g of ZrCl4 and 0.186 g of 2-aminoterephthalic
acid at 90 °C for 24 h, under magnetic stirring.3 Characterizations were performed by scanning
electron microscopy (FEG-SEM) and energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction
(XRD), X-ray photoelectron spectroscopy (XPS), diffuse reflectance spectroscopy (DRS) and linear
sweep voltammetry (LSV). CO2 reduction was performed in 150 mL of 0.1 mol L-1 Na2SO4 saturated
with CO2 using a reactor of two compartments, three electrodes (TiO2NT or TiO2NT/UiO-Zr-NH2
work, Ag/AgCl (4 mol L-1 KCl) reference and DSA® counter) with a bias potential of -0.5 V and
irradiation Xe arc lamp of 300 W for 3 h. The methanol generated was quantification by gas
chromatography with flame ionization detection (GC-FID) using the solid phase microextraction
technique performed with a DVB/PDMS fiber1. The FEG-SEM micrographs of the electrode showed
homogeneous TiO2 tubes, well distributed and UiO-Zr-NH2 particles with 37 nm of average size
dispersed on the entire surface of TiO2NT. EDS mapping analysis showed the presence of the Ti,
O, Zr, N and C elements. XRD diffractograms presented the characteristics peaks of anatase
crystalline phase and peaks characteristics of the UiO-Zr-NH2. XPS spectrum confirmed the
presence of C 1s, O 1s, N 1s and Zr 3d assigned to Zr 3d5/2 and elemental Zr 3d3/2 located around
182 and 185 eV respectively. Bad gap was estimated by Kubelka-Munk equation in a Tauc plot using
the DRS measurements, and the values found were of 3.2 and 2.9 eV to TiO 2NT and TiO2NT/UiO-
Zr-NH2 respectively. LSV data showed an increase in current under light incidence, presence of CO2
and a peak at around -0.5 V which can be attributed to CO2 reduction. The photoelectroreduction of
CO2 onto TiO2NT resulted in a methanol concentration below the quantification limit. On the other
hand, when TiO2NT was modified with UiO-Zr-NH2 generated 73, 153, 280 µmol L-1 of methanol per
hour of experiment, respectively. Methanol concentrations showed a linear formation ratio indicating
good electrode stability. Therefore, the better performance of the Ti/TiO 2NT/UiO-Zr-NH2 could be
explained by greater preconcentration of CO2 on the electrode surface and application of a bias
potential of -0.5 V, which minimized the rate of charge recombination, increasing the efficiency of
the process and the selectivity of the product formed.
1 Irikura, K; et al. Journal of CO2 Utilization 2021, 43, 101364.
2 Cardoso, JC; et al.. Applied Catalysis B: Environmental 2018, 225, 563.
3 Kandiah, M; et al. Journal. Material Chemistry. 2010, 20 (44), 9848.
[FAPESP]

344
DETERMINATION OF MICROPLASTIC AND HALOGENS IN FISH TISSUE BY
SINGLE MICROWAVE-ASSISTED ALKALINE DIGESTION METHOD
Débora P. Almeida (PG)a*, Gustavo Bittencourt (PG)a, Paola Mello (R)a, Cezar A. Bizzi (R)a
a Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, Brazil, 97105-900
*e-mail: debora.almeida@acad.ufsm.br
In 2020, the amount of waste plastics discharged into oceans was 460 Mt, and is estimated that this
will triplicate up to 2060.1 This problem has been increased with the Coronavirus pandemic (COVID-
19), which has led to a distinct in market demand trend for the types of polymers and increasing
demand for single-use products, which cannot be properly handled or recycled after disposal. As a
result, poorly mismanaged plastic waste (MMPW) is discharged into the environment, and can bring
risks after breaking into smaller fragments, called as microplastics (MPs, in general, size lower than
5 mm).1 Notably, with further degradation, the physical properties of MPs (such as colour,
morphology, size, crystallinity, and densities) are changed, which may influence their physical and
chemical actions in the environment. One of the risks that MPs can bring to the environment is the
leaching or adsorption of damaging chemicals, as well as their bio-accumulation in substances and
organisms consumed by humans.2 Thus, the purpose of this work is to develop a method for MPs
determination in fish tissue using a selective digestion of fish tissue without damaged MPs. A simple
approach was developed using alkaline reagents and microwave-assisted wet digestion, making
possible the determination of halogens contaminants in the same digests by inductively coupled
plasma optical emission spectrometry (ICP-OES) and ion chromatography (IC). For this, up to 2 g of
fish sample (shark specie) was spiked with 0.1 g of a mix of MPs (polyethylene terephthalate (PET),
polystyrene (PS), expanded polystyrene (EPS), polypropylene (PP), high-density polyethylene
(HDPE), low-density polyethylene (LDPE), polycarbonate (PC), and polyvinyl chloride (PVC)). The
digestion parameters evaluated were: alkaline digestion solutions (0.1 mol L-1 NH4OH, 10% w/v
NaOH, 0.1 mol L-1 KOH, and 0.1 mol L-1 TMAH), digestion temperature (30 °C to 200 °C), and sample
mass (0.5 to 2 g). After decomposition, the digests were filtered (pore size lower than 2 μm, Whatman
No. 589/3 Florham Park, MI) and the MPs content was gravimetrically determined. Suitable
conditions were obtained using 0.1 mol L-1 NH4OH at 180 °C and 20 min of heating time, making
possible to digest up to 1.5 g of in natura sample, without retained particles in the filtration process.
The recovery of MPs was from 93 to 98%. The proposed method allowed the gravimetric
determination of seven types of plastics (PET, PS, EPS, PP, HDPE, LDPE, PC and PVC) with
particle size higher than 0.5 mm. Because of the visually good digestion efficiency (without
suspended particles), the suitability of the proposed method for halogens contaminants
determination (F, Cl, Br, and I) was evaluated. The results obtained showed that Cl can be detected
by ICP-OES and IC. The concentration of Cl varying between 4013.6 and 4451.7 µg g-1, with no
statistical difference (t-test, confidence level of 95%) when comparing the sample with and without
MPs, showing that Cl was not leached by MPs. Further investigations are still needed to evaluate
the applicability and efficiency of the proposed method for the other halogens contaminants (F, Br,
and I). The proposed method can be considered suitable for the determination of seven types of
MPs in fish tissue. Besides, the same digest used for MPs determination can be used for the
determination of Cl, providing accurate and precise information about two different classes of marine
pollutants (MP and halogens contaminants).
1 Agrawala S., Lanzi E., Global Plastics Outlook Policy Scenarios to 2060, OECD – Evironment and Economy
Integration Division, Environment Directorate, 2022.
2 Haque F., Fan C., Journal of Cleaner Production, 2022.
3 Bitencourt G.R., et al., Science of the Total Environment, 749, 2020, 142301.

345
CHARACTERIZATION OF PHOTODEGRADED POLYAMIDE MICROPLASTICS
AND ITS INFLUENCE ON THE SORPTION OF E3 IN SIMULATED CONDITIONS
Larissa Z. Lara (PG)a*, Crislaine Bertoldi (R)b, Walter R. Waldman (R)c, Andreia N. Fernandes (R)a
aFederalUniversity of Rio Grande do Sul, Institute of Chemistry, Porto Alegre/RS, Brazil, 91509900
bFederal University of Paraná, Institute of Chemistry, Curitiba, Paraná, Brazil, 81531-980
cFederal University of São Carlos, Center for Science and Technologies for Sustainability, Sorocaba/SP, Brazil,
18052780
*e-mail: larissa_lara10@hotmail.com
Microplastics are a common type of small particles found in aquatic environments and due to their
characteristics, such as different degrees of crystallinity and hydrophobicity, they can act as vectors
for a wide range of contaminants.¹ In this sense, it is extremely important to understand the
mechanisms and the influence of environmental factors on the interaction between contaminants
and microplastics. Endocrine disrupting compounds (EDC), such as estriol (E3), are a class of
emerging contaminants widely detected in water bodies and have the potential to interact with
microplastics.² Therefore, this study aimed to characterize photodegraded polyamide microplastics
and evaluate the influence of photodegradation on the sorption behavior of E3 under a simulated
freshwater medium by applying organic matter. Firstly, polyamide pellets were milled on a cryogenic
analytical mill to ensure that plastic debris was smaller than 350 μm. Then, microplastics were
uniformly placed in Petri dishes to proceed with the photodegradation. The samples were added into
a custom photodegradation chamber equipped with a mercury lamp (350 W) and sampled after 32,
64, and 96 h of exposure. The photodegraded (96h) and pristine microplastics were characterized
by X-Ray diffraction, contact angle, BET surface area, and attenuated total reflection Fourier
Transform Infrared (32, 64, and 96h). The characterization was performed to evaluate the changes
in the polymer that occurred by the degradation. The kinetic study of E3 interaction was performed
by adding a known mass of microplastics in Erlenmeyer flasks with a solution of E3 at a fixed
concentration. They were shaken at room temperature (298K), varying the contact times (1, 5, 15,
30, 60, 120, 240, 480, 1440, 2880 min). The equilibrium study was performed in the same conditions
as the kinetic study with the optimized contact time and different E3 concentrations (1.0, 2.0, 3.5,
5.0, 7.5, and 10.0 mg L-1). The studies were performed in both ultrapure water (UP) and ultrapure
water with acid fulvic, as a proxy of the dissolved organic matter, and CaCl2 to simulate freshwater
conditions (FW). The E3 concentration in the solution was determined with a spectrofluorometer (ʎ
= 310 nm). Photodegraded microplastics presented a significant difference in their hydrophobicity
compared to pristine ones. The interaction results suggested that the E3 sorption process is affected
by the degradation of microplastics as evidenced by the changes between the pristine and degraded
sorption efficiencies. Despite the change in hydrophobicity of polyamide microplastics, other
characteristics such as surface area and crystallinity remained unchanged after the
photodegradation. Noteworthy to say that this work used a limited time of degradation so further
degradation levels or longer degradation times should be considered for a more thorough knowledge
of the degradation effects of these characteristics. In addition, the presence of organic matter
increased the sorption of the E3 in the degraded microplastics, probably due to the introduction of
oxygenated groups on the polymer surface during the photodegradation. This behavior influences
the interaction of E3 in the microplastics through bridges among the polymer particles, organic
matter, and E3. Therefore, the results indicate that polyamide microplastics can potentially sorb and
transport E3 in aquatic systems, and photodegradation influences the interaction with this
contaminant.
1 Crawford and Quinn. Microplastic Pollutants. Elsevier, 2016, Netherlands; United Kingdom and United States.
2 Peña-Guzmán, C; et al., Journal of Environmental Management, 237, 2019, 408.

346
PROBABILISTIC HUMAN HEALTH RISK ASSESSMENT ASSOCIATED WITH
BTEX AND FORMALDEHYDE EMISSIONS IN THE CITY OF SALVADOR, BAHIA
Mateus S. Moreira (UG)a, Lícia P. S. Cruz (R)*a, Franciele O. C. da Rocha (R)b, Vânia P.
Campos (R) a, Keliane S. P. P. Souza (UG)a, Rejane P. J. Santana (UG)a
a Federal University of Bahia, Analytical Chemistry Department/Chemistry Institute, Salvador, Bahia, Brazil, 40170-270
b Federal University of Roraima, Chemistry Department. Boa Vista, Roraima, Brazil, 69310-000
*e-mail: lila@ufba.br
The BTEX compounds (benzene, toluene, ethylbenzene, m, p-xylene and o-xylene) and
formaldehyde (FA) have a harmful impact on human health, because they are toxic and, in the case
of benzene and FA are classified as carcinogenic to humans (Group 1) by IARC.1 Benzene can
cause neurological, endocrine, reproductive disorders and some hematologic diseases such as
acute myeloid leukemia and myelodysplastic syndrome. Toluene can affect the reproductive and
central nervous systems. Ethylbenzene and xylenes can cause immunological, respiratory and
neurological effects.2 FA can cause nasopharyngeal cancer in humans and it is associated to
irritation of eyes and respiratory tract, nausea, headaches and fatigue.3 Health Risk Assessment
(HRA) is a tool used to determine the impacts caused by exposure to pollutants on human health.4
Thus, the objective of this study was to perform a HRA considering the lifetime carcinogenic (LCR)
and non-carcinogenic risks (expressed as hazard quotient - HQ) for groups of different age (0-5, 6-
19, 20-70 years corresponding to a G1, G2 and G3, respectively) associated with exposure to BTEX
and FA by inhalation in seven sites in the city of Salvador, Bahia, Brazil, in the dry and rainy periods,
using a probabilistic approach with Monte Carlo Simulation, and the these compounds
concentrations obtained in previous studies by the authors.3,5 The Crystal Ball software (version
11.1.2.4) was used to calculate the probability distributions of these risks, and a total of 10,000
repetitions were performed to simulate the Monte Carlo method and sensitivity analysis, which it was
carried out to determine the contributions of the input variables in the estimation of LCR values for
benzene and FA. The HQ values associated with BTEX and FA at the 95th percentile were below
the limit set by the United States Environmental Protection Agency – USEPA6 (HQ = 1) for all groups,
in the dry and rainy periods. However, the LCR values for benzene and FA at the 95th percentile
considering groups G1, G2 and G3 were 2.49x10-6, 3.56x10-6, 9.16x10-6 and 1.83x10-5, 2.53x10-5,
6.55x10-5 in the dry period, and 2.83x10-6, 3.94x10-6, 1.01x10-5 and 7.97x10-6, 1.02x10-5, 2.40x10-5
in the rainy period, respectively. The LCR values obtained for benzene were 2 to 10 times above the
limit allowed by the USEPA6 (1x10-6), and 7 to 65 times above for FA. For the three groups evaluated,
the LCR values for benzene were higher during the rainy period. However, the LCR values for FA
were higher during the dry period, when the concentrations were higher. The group G3 presented
the highest LCR values in all cases at the 95th percentile. Furthermore, even in the 5th percentile
(adopted as the lowest estimate of potential risk) all LCR values for G3 were above the acceptable
limit in both periods, showing that the group of the adult exposed population in the city of Salvador
is more susceptible to developing cancer, due to longer exposure time. The sensitivity analysis
showed that concentration was the variable that most contributed to LCR values in most groups. The
results obtained in this study showed the need to adopt measures to reduce the emissions of BTEX
and FA in the atmosphere of the city of Salvador, in order to reduce the health impacts of the exposed
population and on air quality.
1 IARC - International Agency for Research on Cancer. Agents Classified by the IARC Monographs, 2018.
2 Bolden AL, Kwiatkowsk CF, Colborn T, Environmental Science & Technology, 49, 2015, 5261-5276.
3 Santana FO, Campos VP, Cruz LPS, Luz, SR, Microchemical Journal, 134, 2017, 78.
4 Cruz LPS, Alves RS, Da Rocha FOC, Moreira MS, Santos Júnior A, Air Quality, Atmosphere & Health, 15, 2022, 159.
5 Cruz LPS, Santos DF, Santos IF, Gomes IVS, Santos AVS, Souza KSPP, Microchemical Journal, 152, 2020,104265.
6 USEPA - United States Environmental Protection Agency. Guidelines for Human Exposure Assessment. January,
2016.
[FAPESB, CAPES, UFBA/PERMANECER]

347
MEASUREMENTS OF THE CONCENTRATION AND ISOTOPIC COMPOSITION
OF Br AND Cl IN ANTARCTIC SEA ICE BY ICP-MS AND MULTICOLLECTOR-
ICP-MS
Jefferson Santos de Gois (R)a,*, Delphine Lannuzel (R)b, Frank Vanhaecke(R)c Paul
Vallelonga (R)d,e
aDepartment of Analytical Chemistry, Rio de Janeiro State University, Rio de Janeiro, RJ, Brazil, 20550013
bAntarcticClimate & Ecosystems CRC, Institute for Marine and Antarctic Studies, University of Tasmania, Australia
cGhent University, Atomic & Mass Spectrometry research unit, Campus Sterre, Ghent, Belgium
dPhysics of Ice, Climate and Earth, Niels Bohr Institute, University of Copenhagen, Copenhagen, Denmark
eUWA Oceans Institute, The University of Western Australia, Australia.
*e-mail: jefferson.gois@uerj.br
Sea ice plays an essential role in the exchange of energy, gases, and nutrients between the ocean
and the atmosphere in polar regions.1 The role of halogens in polar atmospheric chemistry has been
intensively investigated over the past few decades, but many key processes associated with sea ice
are still not fully understood due to the problems related to halogen and isotopic determination in
sea ice samples. We have recently demonstrated the feasibility of halogen isotope ratio
measurements in seawater by MC-ICP-MS using a conventional sample introduction system.2,3
Therefore, this work presents the first investigation of Br and Cl isotopic partitioning in Southern
Ocean sea ice. The sea ice samples characterized were collected during the Australian-led Sea Ice
Physics and Ecosystem eXperiment-2 (SIPEX-2) marine voyage in austral spring of 2012 (26 Sept
– 10 Nov), within the area bounded by 115–125°E and 62–66 S off the East Antarctic coast. Bromine
concentrations ranged from 710 ± 20 mg L-1 to 31440 ± 570 mg L-1 and chlorine concentrations
ranged from 218 ± 12 mg L-1 to 7945 ± 351 mg L-1. Bromine and chlorine concentrations were
positively correlated with a slope of 0.0038 (r=0.996, p<0.001), consistent with the seawater Br/Cl
ratio of 0.0035. Bromine isotopic compositions (denoted d81Br with respect to SMOB isotopic
reference) varied from -0.64 to 0.10‰ with a mean of -0.16‰ and standard deviation of 0.04‰.
Chlorine isotopic compositions (denoted d37Cl with respect to SMOC isotopic reference) varied from
-1.10‰ to 0.43‰ with a mean of -0.26‰ and standard deviation of 0.10‰. Despite their strong
concentration correlation, there was a much less consistent relation between Br and Cl isotopic
compositions (r=0.74, p<0.01) and great variability within individual sea ice cores. The samples
showing the largest degree of fractionation in their Br and Cl isotopic compositions were found
deepest in the sea ice samples. Multiple parameters may influence isotopic fractionation processes
in sea ice including ice texture, age, salt content, and algal biomass content (represented by
Chlorophyll-a concentrations); although no single factor was found to consistently correlate with the
bromine or chlorine isotopic signature. We consider the possibility of isotopic fractionation resulting
from brine rejection and desalinization processes.
1 Dieckmann GS, Hellmer HH, The Importance of Sea Ice: An Overview. In Sea Ice: Second Edition Blackwell Science
Ltd, 2020.
2 de Gois JS, et al., Journal of Analytical Atomic Spectrometry, 31, 537–542, 2016.
3 de Gois JS, et al., Analytical and Bioanalytical Chemistry, 408, 409–16, 2016.
[CNPq, CAPES, FAPERJ, UERJ]

348
A SIMPLE AND SELECTIVE METHOD FOR EXTRACTION OF BASIC NITROGEN
COMPOUNDS IN DIESEL OIL
Gabrielle D. Iop (PG)a, Alice P. Holkem (PG)a, Ana Luiza G. Mendes (PG)a, Oberdan C. Silva
(UG)a, Érico M. M. Flores (R)a, Paola A. Mello (R)a*
aUniversidade Federal de Santa Maria, Chemistry Departament, Santa Maria, RS, Brazil, 97105-900
Diesel oil can contain nitrogen compounds as contaminants, which can cause damage in the refining
and in the final product. These compounds can lead to instability in the commercial product, gum
formation, catalyst contamination, and others.1 Such nitrogen compounds contained in fuels can
present neutral or basic properties and, in general, neutral nitrogen compounds are more difficult to
remove from fuel using the conventional hydrotreatment process. In contrast, basic nitrogen
compounds are more reactive and may cause damage during the processing.2 To overcome some
of the problems that these compounds may bring, and to develop effective methods and conditions
to remove them, it is important to understand the structures and molecular types present in diesel
oil. On this purpose, the determination of basic nitrogen fraction is usually performed and can be
carried out following the UOP 269-10 method, using potentiometric titration with perchloric acid in
acetic acid medium. This work aims to develop a sample preparation method for fractionation of
basic nitrogen compounds from diesel oil, focusing on reverse-phase dispersive liquid-liquid
microextraction (RP-DLLME). The determination of total and basic nitrogen was performed following
the standard methods ASTM D 4629 and UOP 269-10, respectively. A diesel oil sample containing
154 ± 6 mg kg-1 of total nitrogen and 65.4 ± 3.3 mg kg-1 of basic nitrogen content was used for
optimization. The combination of solvents for drop formation, its amount and the sample mass were
investigated. Formic acid, acetic acid, ethanol, furfural, methanol, and furfurylic acid were evaluated
as extractant and chlorobenzene, chloroform, dichloromethane, carbon tetrachloride, 1,10-
dichlorodecane, ethyl acetate, acetone, hexane, isopropanol, toluene, undecanol and xylene were
evaluated as dispersant. Several conditions result in the drop formation, but the use of formic acid
as extractant solvent and isopropanol as dispersant solvent was chosen to continue the optimization.
With the use of 2.5 g of sample mass and the combination of 400 µL and 600 µL of extractant and
dispersant solvents, respectively, it was possible to extract 97% of the basic nitrogen and 55% of
total nitrogen, which represents a selective condition. After this selective extraction, nitrogen was
analysed in a total analyser following ASTM D 4629 and results agree with those by UOP 269-10.
The extracts were also analyzed by high resolution mass spectrometry (HRMS) and the main
compounds were pyridines, quinolines and anilines. Therefore, the sample preparation method
optimized allows an efficient approach for extraction of basic nitrogen compounds, as well as total
nitrogen. The RP-DLLME combined to the total analyzer can be quickly and easily performed in other
laboratories to determine basic nitrogen content in diesel oil samples without the need to carry out
the reagents and time consuming UOP method, minimizing also the waste generation.
1 Matar S, Chemistry of petrochemical processes, Gulf Publishing Company, 2000.
2 Batts BD, Fathoni AA, Energ. Fuel, 5, 1991, 1-21.
[CNPq, CAPES, FAPERGS, UFSM]

349
BIOREMEDIATION OF PETROLEUM COMPONENTS USING MICROALGAE:
EVALUATION EMPLOYING HS-SPME-GC-MS/MS
Anna C. N. Meireles (UG)a*, Enedina S. A. Lima (UG)a, Bruno G. Kubelka (R)b, Sergiane C.
Barbosa (R)a, Paulo C. Abreu (R)b, Fabiano Thompson c, Ednei G. Primel UGa, Jean L. O.
Arias (R)a
Universidade Federal do Rio Grande - FURG, Rio Grande, Rio Grande do Sul, Brazil, 96203-900
a
Universidade Federal do Rio Grande – FURG, Rio Grande, Rio Grande do Sul, Brazil, 96210-030
b
Universidade Federal do Rio de Janeiro – UFRJ, Rio de Janeiro, Rio de Janeiro, Brazil
c
*e-mail: annacnmeireles@gmail.com
Petroleum is an oil of fossil origin whose composition is a combination of aliphatic, alicyclic, and
aromatic hydrocarbons, being largely employed as a source of energy and in the obtention of
different derivatives. However, its exploration and transport may cause negative impacts on the
environment, especially when leakage happens.1 In this way, remediation strategies are constantly
required to reduce these substances’ impact on the environment. One of them is the cultivation of
microalgae in contaminated water. Thus, this work aimed to evaluate the cultivation of marine
microalgae as a tool for the purification of oil-stricken coastal waters. For each specie a comparative
analysis was performed by two treatments: i) without contaminated; ii) with contaminated water.
Microalgae growth was monitored daily, and 3 samples were collected for further analysis of PAHs
and n-alkanes. The treatments were simulated by mixing petroleum in natural saltwater, then the
undissolved phases were mechanically filtered. This simulated contaminated water was used for the
cultivation of two species of marine microalgae with high commercial value, Nannochloropsis
oceanica and Conticribra weissflogii. The determination of PAHs and n-alkanes was carried out by
using a Shimadzu Gas Chromatographic tandem Mass Spectrometry (GC-MS/MS) system. The
identification of each compound was based on its retention time by comparison with the standards
and the response used for the remediation experiments was the chromatographic area. For the
extraction of the compounds by automated headspace solid-phase microextraction (HS-SPME), 10
mL water samples were placed in 20 mL vials sealed with magnetic screw-capped lids lined with
PTFE-silicone septa. The samples were pre-heated at 80 °C with agitation at 250 rpm for 5 minutes,
and the 30 µm polydimethylsiloxane (PDMS) fiber was exposed in the headspace of the vial for 60
minutes in the same conditions, being thermally desorbed in the injection port of the GC system at
250 °C for 10 minutes. The chromatographic separation was carried out by a capillary column model
Rtx®-5MS (30 m x 0.25 mm x 0.25 µm). The carrier gas was Helium (99.999% purity) at a constant
flow rate of 1.35 mL min-1. The injector, interface, and ion source temperatures were 280, 290, and
250 °C, respectively. The separation started at 60 °C, remaining at this temperature for 1 minute,
followed by a heating rate of 5 °C min-1 up to 310 °C, being kept in the final temperature for 10
minutes, which totaled 61 minutes of chromatographic run. The EI ion source worked at 70 eV. The
Mass Spectrometer was operated in selective ion monitoring (SIM). For each compound, one ion
was used for quantification, while the other two ions were used for confirmation. For both species of
marine microalgae, it was possible to observe that they had grown when submitted to the water
contaminated with petroleum, being statistically different for Nannochloropsis oceanica when
compared to the control, in ten days of growing, being evaluated by optical density. After evaluating
the microalgae growth, they were submitted to remediation experiments, where was possible to
observe the reduction of the concentration of PAHs and n-alkanes up to 100 and 1000 times,
respectively, within ten days of the experiment. Finally, it was possible to observe that microalgae
cultivation is a very promising strategy for the remediation of environments impacted by petroleum
leakage.
1 Zacharias DC, Gama CM, Fornaro A, Marine Pollution Bulletin, 165, 2021, 112125
2 Escarrone ALV, et al., Food Chemistry, 146, 2014, 597

350
DETERMINATION OF PESTICIDE RESIDUES USING A GREENER APPROACH
IN VORTEX-ASSISTED MATRIX SOLID PHASE DISPERSION
Juliane M. dos Santos (PG)*, Sergiane C. Barbosa (R)a, Ednei G. Primel (R)a
* Universidade Federal do Rio Grande, Escola de Química e Alimentos/ Instituto de Oceanografia, Rio Grande/ São
Lourenço do Sul, Rio Grande do Sul, Brazil, CEP 96201-900
a Universidade Federal do Rio Grande, Escola de Química e Alimentos, Rio Grande, Rio Grande do Sul, Brazil, CEP
96201-900
*e-mail: julianemdsantos@gmail.com
The aquatic environment is one of the environmental matrices most affected by pesticide residues.
However, sediments represent the main compartment of contaminant accumulation. Most
contaminants released into aquatic environment will be incorporated in sediments, and these
sediments might later act as the major source of contaminants to water and biota.1 Given the
importance of evaluating pesticides in environmental matrices, it is essential to study sensitive and
selective analytical methods for the detection of these compounds. For this, sample preparation
techniques have been improved in order to develop miniaturized, faster, simpler and cheaper
methods that require less solvent consumption.2 Thus, this study aimed to assess a fast and low
cost method for the simultaneous extraction of 21 pesticides in sediment samples, using Vortex-
Assisted Matrix Solid Phase Dispersion (VA-MSPD) with determination by Liquid Chromatography
Tandem Mass Spectrometry (LC-MS/MS). Also, the presence of these compounds was investigated
in samples from São Lourenço do Sul, a coastal region of southern Brazil. Sample preparation
parameters were evaluated. Firstly, the solvent (methanol, ethanol, acetonitrile and ethyl acetate)
and the solid support (C18, mussel shell and without support) were standardized. After that, a
factorial design was carried out in which the sample mass (0.5, 1.25 and 2 g), homogenization time
(1, 3 and 5 minutes), volume of solvent (5, 7.5 and 10 mL) and vortexing time (1, 3 and 5 minutes)
were optimized. Best conditions were using 2g of sample, without support, homogenized for 5
minutes. Extraction was performed with 5 mL of ethanol, vortexed for 1 minute and centrifuged at
7000 rpm for 5 minutes. After, the method was validated. Values of LOD and LOQ ranged from 0.42
to 41.67 ng g-1 and from 1.25 to 125 ng g-1, respectively. Accuracy and precision were evaluated in
three levels of concentration, and recoveries ranged from 40 to 112% with RSD≤30%. Besides,
analytical curves presented values of correlation coefficient higher than 0.99 and matrix effect was
low for most of the compounds. Thus, the method meets the recommendation by the validation
guides.3 The optimized method was applied in sediment samples collected from the São Lourenço
Stream Watershed and clomazone (4.07 ng g-1), tebuconazole (2.6 ng g-1), difenconazole (< 1.25 ng
g-1) and trifloxystrobin (< 2.5 ng g-1) were detected. The proposed method employing VA-MSPD and
LC-MS/MS was suitable for the determination of the 21 pesticides. The use of VA-MSPD without
solid support and the use of reduced volume of solvent with less toxicity (ethanol) contribute to the
precepts of Green Analytical Chemistry, despite being efficient for extracting pesticides from
sediment.
1 Maqueda, C, et al., Science of the Total Environment 593, 2017, 787.
2 Caldas SS, et al., Química Nova 34, 2011, 1604.
3 INMETRO. DOQ-CGCRE-008. Orientação sobre validação de métodos analíticos-Revisão 09, Brasília, 2020.
[FURG, CNPq, FAPERGS]

351
ADSORPTION OF β-ESTRADIOL ON POLYPROPYLENE MICROPLASTIC: THE
INFLUENCE OF DIFFERENT CONDITIONS
Bianca Ferreira da Silva (PG)a*, Sonia Queiroz (R)b, Walter R. Waldman (R)c, Carla B. G.
Bottoli (R)a
a UNICAMP,Institute of Chemistry, Campinas, SP, Brazil, 13083-872
b EMBRAPA Meio Ambiente, Jaguariúna, SP, Brazil, 13918-110
c UFSCAR, Center for Sciences and Technology for Sustainability, Sorocaba, SP, Brazil, 18052-780
*e-mail: bianca.ferreirads@gmail.com
Because of the large amount of plastic waste generated and the incorrect disposal of this type of
material, an emerging contaminant of increasing environmental concern are the microplastics (MP),
which are plastic particles smaller than 5 mm. One of the negative results due to the presence of
these particles in the environment is due to their adsorption capacity, accumulating different types
of pollutants that are present in the environment.1,2 Once the substances are adsorbed on MP,
another concern that arises is the possibility of transferring, which means desorption of these
pollutants, for example, to aquatic biota. The presence of these products has long-term impacts that
are still unknown on human health and ecosystems.3 Several factors can influence the adsorption of
compounds on MP, like the type of plastic, the type of the environment, the type of the molecule,
and its concentration, among others.1,2 Hormones, another emerging contaminant, are a concern
because they are considered endocrine disruptors, which can cause adverse effects in humans, and
wildlife, like effects on the central nervous system and the reproductive system. 4 In this study, the
adsorption of β-estradiol (E2), a hormone molecule found in freshwater, on polypropylene (PP)
microplastics, was evaluated in different surface Brazilian freshwater conditions like pH value and
salinity. PP, often used as packing material, was chosen because of its relevance to the environment.
The MP was obtained by Cryogenic Freezer/Mill-6970 EFM, and their 250-500 µm were separated
to the next step. To obtain high adsorption in a PP MP, Design of Experiments tools to select
variables with major effects were used. A full factorial design with two levels for the three design
variables “concentration of E2” (0.25 to 1.25 µg/mL), “pH” (6 to 9), and “salinity” (0 to 0,05%) was
created. For the sorption, 25 mL of E2 solution with 50 mg of MP was mixed in a glass tube. Then,
the salinity was regulated by adding NaCl, and the pH values of the samples were adjusted with 0.1
mol L-1 sodium hydroxide and 0.1 mol L-1 hydrochloric acid. The samples were agitated (150 rpm)
on a horizontal shaker for 72 hours. Additionally, one sample without microplastics was treated
similarly, and their value was subtracted from the adsorption values for polymer materials. After 72
hours, an aliquot from the solution was collected and filtered in PVDF Millex® syringe filter (0.45 µm,
13 mm). The quantification was performed using high-performance liquid chromatography with
fluorescence and diode array detection (HPLC-FL-UV Shimadzu Class VP) on a C18 column
(Phenomenex Gemini, 5 μm, 110 Å, 250 x 4,6 mm). The data were processed using Design-Expert®
Software. Finally, the design was evaluated by analysis of variance (ANOVA), including second
interactions using “adsorbed the concentration” as the response variable. The results showed that
the adsorption onto PP microplastic is strongly governed by the concentration of E2. The effect of
salinity is comparably small, and pH does not have a significant effect on the adsorption capacity of
microplastics. The highest absorbed concentration of E2 was obtained when the factor concentration
of E2 was at high level and the factor salinity was at low level.
1 Lin L., et al., Process Saf. Environ. Prot., 2020, 140.
2 Goedecke C., et al., 2017, 4.
3 Rainieri S., et al., Environ. Res., 2018, 162.
4 Jarque S., et al., Sci. Rep., 2015, 5.
[CAPES, FAPESP, CNPq, INCTBIO, CNPEM]

352
SALT-ASSISTED SOLID-LIQUID EXTRACTION COUPLED WITH GC-MS/MS
AND UHPLC-MS/MS FOR PESTICIDES DETERMINATION IN WILD RODENTS
Pimpernelli J. dos Santos (PG)a, Luana Floriano (PG)a, Tomás M. Mac Loughlinb
Érica F. G. G. de Sá (PG)c, Pedro C. Estrela (R)c, Renato Zanella (R)a*
a Federal University of Santa Maria, Chemistry Department, Santa Maria, Rio Grande do Sul, Brazil, 97105-900
bEnvironmental Research Center - CIM (UNLP-CONICET), Faculty of Exact Sciences, National University of La Plata,
La Plata, Buenos Aires, Argentina, 1900
c Federal University of Paraiba, Systematics and Ecology Department, João Pessoa, Paraíba, Brazil, 58051-900
The indiscriminate use of pesticides leads their persistence in environmental compartments such as
aquatic, air, soil and living organisms, which can promote intentional or unintentional poisoning of
wild animals.1 Their direct and indirect ability to persist in wildlife through bioaccumulation and
biomagnification,2 contributes to ecosystems imbalance and compromise the health of living
organisms. As a result, monitoring the presence of pesticides residues in environmental samples,
including wild animal, is critical for determining the impact of these organic compounds on
ecosystems.3 The development of reliable, rapid and sensitive sample preparation methods using
chromatographic techniques is necessary to determine pesticides at low concentrations, which
usually are found in environmental samples. In Brazil, the Pantanal ecosystem is one of the largest
continuous freshwater floodplains in the world,2 rich in different plant and animal species. Some
herbivore wild rodents from Pantanal, consume food from agricultural crops nearby, which probably
are contaminated with pesticides. In this work, we developed and validated a method for pesticides
multiresidue determination by GC-MS/MS and UHPLC-MS/MS in liver and kidney wild rodents
Holochilus, Cavia and Euryoryzomys genus from Pantanal, Brazil. The method consisted in a simple
solid-liquid extraction combined with saturated saline solution and protein precipitation. The GC-
MS/MS analysis was performed with an Agilent Intuvo 9000 (USA), with a planar capillary column
HP-5 MS UI (30 m x 0.25 mm x 0.25 µm), 1 µL injection volume in splitless mode and helium as
carrier gas. High efficiency source (HES) was used with electron source and dynamic MRM (dMRM)
as acquisition mode and nitrogen collision gas. The UHPLC-MS/MS analysis was performed with
Waters Xevo TQ-XS (USA) using Acquity UPLC™ HSS T3 (100 x 2.1 mm, 1.8 µm) column and
mobile phase consisted of A) ultrapure water and B) methanol:acetonitrile (1:1, v/v) both with 0.1%
(v/v) formic acid 5 mmol L-1 ammonium formate at flow rate 0.500 mL min-1. The injection volume
was 5 µL. The ionization source by electrospray operated in positive and negative mode (ESI±) and
mass analyzer in selected reaction monitoring (SRM) mode. The salt-assisted solid-liquid extraction
was applied for 147 pesticides, where 37 compounds were analyzed by GC-MS/MS and 110
compounds by UHPLC-MS/MS. The linearity was expressed by seven concentrations using matrix-
matched calibration. Accuracy and precision were evaluated at the spike levels 10, 20 and 40 µg kg-
1
for GC-MS/MS and 5, 10 and 20 µg kg-1 for UHPLC-MS/MS. The recoveries ranged from 70 to
120% with RSD ≤ 20% for both techniques. The method showed limits of quantification values from
5 to 40 µg kg-1 and limits of detection from 1.5 to 12 µg kg-1 in both techniques. We applied the salt-
assisted solid-liquid extraction in 40 liver and 40 kidney samples from wild rodents. Seven pesticides
were detected with concentrations ranging from 1.6 (< LOQ) to 142 µg kg-1. Therefore, the developed
method was rapid, easy to perform, simple, sensitive and suitable for routine analysis in routine
laboratories.
1 Bertero A, et al., Science of the Total Environment, 707, 2020, 136129.
2 Vicente EC, Guedes NMR, Scientific Reports, 11, 2021, 5602.
3 Buah-Kwofie A, Humphries MS, Journal of Chromatography B, 1105, 2019, 85-93.
[UFSM, CNPq, CAPES, FINEP]

353
FRACTIONATION AND RISK ASSESSMENT OF METALS IN DRILL CUTTINGS
FROM OIL AND GAS EXPLORATION WELLS IN ULTRADEEP WATERS
Paola C. C. Azevedo (PG)a, Sindy R. Krzyzaniak (R)a, Fernando J. S. de Oliveira (R)b, Vera L.
A. F. Bascuñan (R)a and Eduardo S. Chaves (R)a*
a Universidade Federal de Santa Catarina, Departamento de Química, Florianópolis, SC, Brasil, 88040-900
b Petróleo Brasileiro S. A., Gerência de Meio Ambiente, Rio de Janeiro, RJ, Brasil, 20031-004,
*e-mail: eduardo.chaves@ufsc.br
The oil and gas industry has significant importance worldwide. Nevertheless, the activities of this
industry are responsible for the generation of considerable amounts of waste, including drill cuttings,
which are of high environmental concern.1 Although the composition of drill cuttings can vary
considerably, organic compounds and potentially toxic elements are generally part of its
composition.2 Thus, the effect of fractions of potentially toxic elements and their risk assessment is
not well understood. In this study, drill cuttings samples from an offshore oil and gas exploration well,
collected at depths of 5520 m, 5694 m and 5756 m were submitted to BCR sequential extraction
procedure3 to evaluate the fractionation and risk assessment of Ba, Co, Cu, Mn, Mo, Ni e Pb. The
physicochemical parameters, such as pH, conductivity, humidity, and organic matter content, were
measured. Additionally, the mineral composition of drill cutting samples was evaluated by DRX
analysis where it was possible to verify that the composition was mainly quartz, barite, calcite, and
hematite. The concentration of the analytes in acid-soluble (F1), reducible (F2), oxidizable (F3), and
residual (F4) fractions were determined by ICP-MS. Marine sediment PACS-2 was used as a
standard reference material for each extraction process and no statistical difference between the
results of sum of analytes were observed. Risk assessment codes (RAC) were used to estimate the
environmental risks based on analytes mobility. The percentages of analytes extraction in each
fraction are shown in Figure 1.
Figure 1. The percentage of metal fractionation in drill cuttings form oil and gas exploration well from different depth using
BCR sequential extraction procedure. A) 5520 m, B) 5694 m and C) 5756 m.
The total concentration of the analytes ranged from 4.41 to 56616 μg g-1. Considering the BCR
sequential extraction procedure, Ba, Co, Ni and Pb showed the greatest concentration in residual
fraction (F4), in all the analyzed drill cuttings. However, Mn and Mo were predominantly found,
respectively, in the reducible (F2) and oxidizable fraction (F3), except for sample collected at 5756
m (C), in which the greatest Mo concentration was found in F3. Considering the samples collected
at 5694 m (B) and 5756 m (C), the highest concentrations of Cu were found in the oxidizable fraction
(F3). The RAC values, for most of the elements determined in drill cuttings, suggested medium risk
(RAC 11-30%). In the samples collected at 5694 m (B) and 5756 m (C), the low RAC values (<1%)
suggested no risk from Ba.
1Jafarinejad S. Petroleum Waste Treatment and Pollution Control. 2016.
2Ismail, A. R. et al. Chemical Engineering Transactions, 56, 2017, 1351–1356.
3Kumkrong, P. et al. Talanta, 221, 2021.
[CNPq, CAPES, Petrobras, UFSC]

354
DEGRADATION OF ATRAZINE PRESENT IN WATER BY THE ADVANCED
OXIDATIVE PROCESS OF ELECTRO-PHOTO-OZONIZATION
Lucas Bohnenberger (UG)a, Renata Martins (UG)a, Jonas J. Inticher (PG)b, Manuela G.
Cardoso (R)a, Fernando H. Borba (R)a, Liziara C. Cabrera (R)a, Jonas S. Dugatto (R)a*
a Federal University of Fronteira Sul – Campus Cerro Largo, Cerro Largo, RS, Brazil, 97900-000
b West Paraná State University, Postgraduate Program of Chemical Engineering, Toledo, PR, Brazil, 85903-000
*e-mail: jonas.dugatto@uffs.edu.br
Annually in Brazil, tons of pesticides1 are sold for the control of the pests, prevent weed growth and
to obtain high productive crops.2 Most of these pesticides are associated with human health
problems, once through leaching and infiltration into the soil, they end up reaching natural water
sources intended for human consumption.3 Among these compounds, the atrazine has gained great
concern, since it is the sixth most sold active ingredient of pesticides in Brazilian territory, 2 has an
elevated persistence in the biosphere, such as water bodies and soils, and it is considered an
endocrine disruptor.4 As a result, several treatments have been investigated for the degradation and
mineralization of atrazine present in water.5 One of the most attractive technologies, is the advanced
oxidative process, which uses the hydroxyl radical, a strong oxidizing agent, to effectively degrade
organic pollutants.6 The aim of this work was investigate the degradation of atrazine present in water
by Electro-Photo-Ozonation, and study how the parameters of duration time (min), ozone
concentration (mg O3∙L-1), electric current intensity (A) and pH influence the process. Through
preliminary tests, the study range of the parameters and a central point was chosen. In the sequence,
an experimental design of DCCR type and a Response Surface Methodology was applied in order
to obtain the optimal operating conditions7 and to determine an equation model to the process. The
conditions of operation were: duration time of 15, 30 and 45 minutes, O3 concentrations ranging from
11.36 to 78.64 mg∙L-1, current intensity between 0.32 and 3.68 (A) and pH between 2.5 and 7.5. The
experiments were carried out with synthetic atrazine effluent at a concentration of 10 mg∙L -1, since
this value has been found in wastewater generated from washing packaging and pesticide factories.8
To verify atrazine degradation, a method was validated9 according to the parameters required by
INMETRO,10 monitoring m/z 216 (+) with a LC-ESI-MS (Shimadzu 2020). Separations were
performed with a 50 mm x 2,0 mm i.d., 2,8 µm particle size C18 analytical column, kept in a constant
temperature of 35ºC. The mobile phase were ultra pure water (A) and methanol grade HPLC (B), in
a proportion of 48% of B until 1 min, reaching 100% of B in 8 min, in a flow rate of 0.3 mL∙min-1. The
sample were diluted 1:1 with ultra pure water, filtered by 0,2 µm needle filters and injected in volume
of 10 µL. As results, the degradation rate obtained with 15 min varied from 3.88 to 92.07%, with 30
min was 27.61 to 98.43% and in 45 min the values were between 51.7 to 99.62%. The duration time
of 45 min was identified as the one with maximum degradation results, with O 3 concentration of 65
mg∙L-1, current intensity of 1 A and pH 5, with a residual concentration of 38 ppb. With the degradation
time of 15 min and through the adjustment of a full quadratic equation model, was possible to
identified that the parameters which influenced on the degradation of atrazine were: linear interaction
of O3 (O3) and linear interaction between O3 with electric current intensity (O3*A).
1 Siqueira DF, et al., Revista Brasileira em Promoção da Saúde, 26, 2013, 182
2 Relatórios de comercialização de agrotóxicos, Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais
Renováveis, IBAMA, Disponível em: <https://www.ibama.gov.br/agrotoxicos/>. Acesso em 09/08/2021
3 Loro VL, et al., 13, 2015, 569
4 USEPA – United States Environmental Protection Agency. National Primary Drinking Water Regulations - EPA 816-F-
09-004. Washington, D.C., 2009.
5 Marican A, Durán-Lara EF, Environmental Science and Pollution Research, 25, 2018, 2051
6 Metcalf L, Eddy HP, Tratamento de Efluentes e Recuperação de Recursos, AMGH, 5, 2016, Brasil
7 Montgomery DC, Design and analysis of experiments, John Wiley & Sons, 9, England
8 Jing L, Chen B, Wen D, Zheng J, Zhang B, Journal of Environmental Management, 203, 2017, 182
9 Inticher JJ, et al., Journal of Environmental Chemical Engineering, 9, 2021, 105883
10 Coordenação Geral de Acreditação. Orientação sobre a Validação de Métodos Analíticos, Instituto Nacional de
Metrologia, Qualidade e Tecnologia, INMETRO, 2020
[UFFS]

355
POTENTIAL RISK TO HUMAN HEALTH FROM THE CONSUMPTION OF
MELIPONINAE HONEY BY POLYCYCLIC AROMATIC HYDROCARBONS (PAHS)
Lucas C. Marcolin (PG)ª*, Jean L. O. Arias (R)a, Jônatas Külzer (UG)a, Enedina S. A. Lima
(UG)ª, Anna C. N. Meireles (UG)ª, Larine Kupski (R)ªb, Sergiane C. Barbosa (R)ª, Ednei G.
Primel (R)ª
a Universidade Federal do Rio Grande, Rio Grande, Rio Grande do Sul, Brazil, 96203-900
b Universidade Estadual de Maringá, Departamento de Tecnologia, Maringá, Paraná, Brazil, 87020900
*e-mail: lucas—marcolin@hotmail.com.br
Honey can be produced by beekeeping, (Apis mellifera), and stingless bee beekeeping
(Meliponinae).1 When bee visit flowers to collect nectar and pollen, they may be contaminated with
different types of pollutants such as Polycyclic Aromatic Hydrocarbons (PAHs), which are
compounds of concern in terms of safe food production, since they exhibit mutagenic and
carcinogenic activities.2 The United States Environmental Protection Agency (USEPA) has listed
sixteen PAHs as priority pollutants, while the European Union, has set maximum levels of some
PAHs in various types of food. However, for honey, maximum levels have not yet. 3 The objective of
this study was to evaluate the potential risk to human health by the consumption of honey from seven
Meliponinae species from two different locations, Ilópolis and Cruzeiro do Sul, in the state of Rio
Grande do Sul, Brazil. To extract PAHs from honey, original, citrate and acetate QuEChERS
methods were evaluated. Recoveries ranged from 78 to 96% with RSD < 11.9% in the original
QuEChERS, from to 112% with RSD < 16% in the citrate QuEChERS and from 71 to 96% with RSD
> 16.9% in the acetate QuEChERS. The obtained recoveries complied with values proposed by the
guidelines,4 however, the original QuEChERS method was chosen because it needs fewer reagents
than the others. Dietary Daily Intake (DDI), Carcinogenic potencies (B(A)Pteq) and Excess Cancer
Risk (ECR) via consumption of PAHs in Meliponinae honey were evaluated according to the study
of Moslen et al., 2019.5 The estimated DDI of PAHs ranged from 0.23 to 5.16 ng day-1. ECR resulting
from PAHs analyzed by this study was compared with the reference value considered acceptable by
the USEPA, which is 1 x 10-6. This acceptable value suggests cancer risk level of one in a million
(ECR = 10−6) in approximately 70 years of life; however, the risk of cancer of one in ten thousand or
more (ECR = 10-4) is classified into severe.6 ECR of PAHs in Meliponinae honey ranged from 6.21 x
10-7 to 1.17 x 10-4. Thus, it may be stated that concentrations of some PAHs are higher than the
acceptable risk level (anthracene, benz[a]anthracene, benzo[b]fluoranthene and indeno[1,2,3-
cd]pyrene) and severe (benzo[k]fluoranthene and dibenz[a,h]anthracene). These results indicate
risks to human health caused by the consumption of honey contaminated with PAHs throughout life.
Thus, it is essential to constantly monitor PAHs in honey, aiming at the production of safe food. The
ECR of Meliponinae honey was above the threshold classified as acceptable and severe by the
USEPA in some samples; thus, it poses health risks if consumed daily in large quantities.
Furthermore, results show that Meliponinae honey may also be used as an indicator of pollution by
PAHs and provide information on impacts of anthropogenic activities on the environment. Finally,
results of this study may help to establish maximum limits of PAHs in honey.
1 Sousa JMB, et al., LWT-Food Science and Technology, 65, 2016, 645-651.
2 Petrović J, et al., Food Additives & Contaminants: Part B, 12, 2019, 116.
3 EC, Regulation, Official Journal of the European Union L, 215, 2011, 4-8.
4 INMETRO, Instituto Nacional de Metrologia, Normalização e Qualidade Industria, DOQ-CGCRE-008: Orientações
sobre validação de métodos analíticos
5 Moslen M, Miebaka CA, Boisa N, Toxicology Reports, 6, 2019, 990-997.
6 Nie J, Shi J, Duan X, Wang B, Huang N, Zhao X, Journal of Environmental Sciences, 26, 2014, 432-439.

356
OCCURRENCE OF PESTICIDES AND PPCPs ON SURFACE AND TREATED
WATER SAMPLES FROM RIO GRANDE DO SUL
Paula R. P. Lima (UG)a*, Josilene N. Araújo (UG)b, Márcia H. S. Kurz (R)b, Manoel L. Martins
(R)b, Ednei G. Primel (R)a, Sergiane C. Barbosa (R)a
b Universidade Federal do Rio Grande, Santo Antônio da Patrulha, Rio Grande do Sul, Brazil, 95500-000
*e-mail: paulalima@furg.br
The occurrence of pesticides and pharmaceuticals and personal care products (PPCPs) have been
investigated in environmental matrices, as well as in treated water samples. Although there are
studies that have been carried out in Brazil, data of many regions are scarce. 1 Thus, the aim of this
study was the investigation of pesticides and PPCPs in surface and treated water samples from nine
cities of Rio Grande do Sul. Water samples were collected in Rio Grande, Santa Vitória do Palmar,
Camaquã, Santo Antônio da Patrulha, Tramandaí, Torres, Bento Gonçalves, Farroupilha and
Canela, from September 2019 to December 2020. Samples were collected in the water treatment
stations (WTS) from CORSAN company. Surface water and treated water samples were filtered and
extraction from PPCPs and pesticides was carried out using two different methods. For the extraction
of 28 PPCPs a solid phase extraction (SPE) extraction using polymeric cartridges with determination
by Liquid Chromatography tandem mass spectrometry was carried out.2 For the extraction of 57
pesticides, a SPE extraction using C18 cartridges and determination by Gas chromatography
tandem mass spectrometry was done. Twelve PPCPs (bisphenol-A, caffeine, carbamazepine,
diclofenac, ethylparaben, ibuprofen, lidocaine, enalapril maleate, methylparaben, propylparaben,
sulfamethoxazole and trimethoprim) were detected at least in one sample. Highest concentrations
were found to the plasticizer bisphenol A (163 – 585 ng/L). Caffeine was detected in all cities, in
concentration that range from <4.0 ng/L to 51 ng/L. Seventeen pesticides were detected (atrazine,
azoxystrobin, carbaril, carbendazim, cyproconazole, chlorantraniliprole, fenamidone, fipronil,
imazapique, imazetapir, imidacloprid, propiconazole, saflufenacil, simazine, tebuconazole,
thiamethoxam and tricyclazole), being 29% herbicides, 29% insecticides and 41% fungicides.
Samples collected in WTS from Rio Grande presented more pesticides (12 different compounds in
the six sampling campaigns). Atrazine was the most frequently detected pesticides, being found in
a least one sample from Rio Grande, Camaquã, Torres, Tramandaí, Santo Antônio da Patrulha,
Farroupilha, Bento Gonçalves and Canela cities. However, quantified concentrations were lower
than the maximum residue limit (MRL) (2000 ng/L) allowed in water for human supply established
by the legislation. Results indicate that surface water samples have been impacted by the presence
of anthropogenic discharges since caffeine was detected in sample from all cities, highlighting the
importance of monitoring studies to guarantee the quality of water that, after treatment, will be
consumed by the population. Pesticides were also detected, mainly in cities with more agricultural
activities near the surface water. However, for the detected compounds, concentrations were lower
than MRLs. This study contributes with data to a better understanding about the occurrence of
residues of pesticides and PPCPs in the environment and in the treated water, contributing to
subsidies for discussion and improvement of water quality services.
1 Montagner, CC et al, Journal of the Brazilian Chemical Society, 30, 2019, 614
2 Chaves, MJS et al, Science of The Total Environment, 734, 2020, 139374
[CORSAN, FURG, FINEP, FAPERGS, CNPq]

357
DETERMINATION OF CONTAMINANTS OF EMERGING CONCERN IN
SEAFOOD
Jônatas Külzer (UG)ª*, Marisa J. S. Chaves (PG)a, Paula R. P. de Lima (UG)ª, Sergiane C.
Barbosa (R)a, Lucas Cavagnoli Marcolin (PG)a, Ednei G. Primel (R)ª
*e-mail: jonataskulzer@outlook.com
Pharmaceuticals and personal care products (PPCPs) have become a contaminants class of major
concern because they are extensively used in human and veterinary medicine, resulting in their
continuous release to the environment. Besides, they have been detected in the environment, in
water, sediments and aquatic organisms.1 PPCPs include a diverse group of compounds, as
preservatives, UV filters, musk, veterinary drugs, etc. There are several input means for PPCPs to
the environment, as the inappropriate disposal of residential, industrial or hospital waste, effluents
from wastewater treatment plants.2 Contamination of seafood by PPCPs deserves attention mainly
because many times this kind of food is consumed in natura.3 Thus, aiming at verifying the
occurrence of PPCPs in seafood a method for the determination was studied in this work. After a
wide bibliographic review, 18 PPCPs (analgesics, anti-inflammatories, antibiotics, preservatives,
antiseptics, plasticizer, UV filters, antiepileptic and antidepressant) were chosen to be investigated
in seafood. For the determination, a liquid chromatograph tandem mass spectrometry (LC-MS/MS)
was used. Detection parameters as the ionization mode, cone voltage and collision energy were
evaluated as well as the chromatographic conditions such as the mobile phase and the stationary
phase of the analytical column. For PPCPs extraction, Original, Citrate and acetate QuEChERS
were evaluated. After, some parameters of the QuEChERS extraction that presented the best
accuracy were investigated (sample and salts mass, solvent volume, and the cleanup step). After all
extraction and determination conditions were chosen, the method was validated. Best results were
reached using an C18 analytical column (100 mm x 2.0 mm, 2.5 µm, Phenomenex) with a mobile
phase composed by acetonitrile and ultrapure water, both acidified with 0.1% de CH3COOH. For the
extraction, original QuEChERS was chosen using 2.5 g sample mass, 10 mL acetonitrile, 2.0 g
MgSO4 and 0.5 g NaCl. The cleanup step was carried out using chitin. The method was validated,
and Limits of quantification were between 2 ng/g and 400 ng/g, which are in agreement with
concentrations that have been found in seafood. For the accuracy and precision, samples were
fortified in three levels, and values for recovery between 70 and 120%, with RSD values lower than
20 were reached. Matrix effect was found for some compounds, thus matrix matched calibration
should be used to compensate this effect during quantification of samples. PPCPs occurrence in
seafood samples (salmon, mussels and oyster) acquired in the South of Brazil will be investigated.
A multi-residue method for the analysis of PPCPs in seafood based on QuEChERS extraction
followed by detection using LC-MS/MS was proposed. The method allowed simultaneous analysis
of 18 PPCPs belonging to different therapeutic families, and showed good performance in terms of
accuracy, precision and quantification limits.
1 Chaves, Marisa de Jesus Silva et al. Science of The Total Environment, 2020, 139374.
2 Xu, Xuming et al. Chemosphere, 301, 2022, 134721.
3 Serra-Compte, Albert et al. Food and Chemical Toxicology, 104, 2017, 3-13.
[FAPERGS, CNPq, FINEP, CAPES]

358
PHOTO-FENTON DEGRADATION OF SULFAMETHOXAZOLE USING
MAGNETITE AS CATALYST: THE ROLE OF THE SYNTHESIS PATHWAY
Raquel F. P. Nogueira*a, Jany H. F. Jesusa, David L. A. Silvaa
aSão Paulo State University, Institute of Chemistry, Araraquara, São Paulo, Brazil, 14800-900
*e-mail: raquel.pupo@unesp.br
Considering that biological treatment is not efficient for removing antibiotics, advanced oxidation
processes (AOP), mainly heterogeneous Fenton and photo-Fenton processes, are possibilities for
the treatment of effluents. Typically, the physical properties of synthesized catalysts are mainly
dependent on their specific area, particle size, and morphology, characteristics that vary according
to the synthesis route.1 Thus, the reactivity of magnetite in the photo-Fenton process may depend
on surface characteristics and properties, and Fe2+/Fe3+ ratio.2 In this scope, the present study
proposes to correlate the characteristics of magnetites prepared following different rout of synthesis
with their efficiency in the Fenton and photo-Fenton process for the degradation of sulfamethoxazole,
an antibiotic widely used in Brazil. The experiments were performed under UV irradiation (two 15 W
UV blacklights tubes, maximum emission wavelength at 365 nm) positioned around 3 cm above the
solution. The magnetites were synthesized through three different pathways: Fe3O4-S1,
coprecipitation method using both Fe2+ and Fe3+ salts as the precursors, Fe3O4-S2, using Fe3+ salt
as the precursor, Fe3O4-S3, using Fe2+ salt as the precursor, and were characterized by XRD, zeta
potential, and SEM-EDS. These three synthesized magnetites had their performance on Fenton and
photo-Fenton reaction compared with commercial magnetite (Fe3O4-C). The experiments were
performed at pH 5.5 due to the high stability of the catalysts at this pH, with no leaching of iron during
120 min of the degradation experiments (<LQ), and due to the advantages of the treatment under
circumneutral conditions. Besides, it keeps dissolved iron safely below the limits for wastewater
discharge according to CONAMA Framework Resolution 357/2005. Control experiments indicated
that the contributions of adsorption, photocatalysis, and photolysis of SMX did not exceed 6%, while
the contribution of H2O2 photolysis resulted in 69% of SMX degradation. However, the efficiency of
the Fenton and photo-Fenton process was strongly influenced by the type of catalyst. During the
dark Fenton process, Fe3O4-S1 and Fe3O4-S2 exhibited the highest catalytic performance, around
33 and 31%, for SMX removal, respectively. Fe3O4-S3 and Fe3O4-C were the less efficient catalysts,
with 6% and 10% of removal, respectively. Under irradiation of Fe3O4-S1 and Fe3O4-S2, degradation
of SMX was improved, reaching removal up to 88%. Once more, Fe3O4-S3 and Fe3O4-C decreased
SMX removal to 18% and 25%, respectively, compared to H2O2 photolysis process. With Fe3O4-S3
and Fe3O4-C, the turbidity of the suspension was likely to have caused the scattering of light,
reducing its availability to induce the photo-Fenton process.3 Besides, from the speciation diagrams
of SMX, the predominant species at pH 5.5 were neutral and anionic. From zeta potential, Fe3O4-S3
was the magnetite with the less positive charge, which can hinder the electrostatic interaction
between the target contaminant and the catalyst. To conclude, it was found that the synthesis
precursors of the Fe3O4 played an important role in the catalyst efficiency in Fenton and photo-
Fenton process. Extra characterizations are in progress to correlate the catalytic activity with the
surface properties of the materials.
1 N. Thomas, D.D. Dionysiou, S.C. Pillai, J. Hazard. Mater. 404, 2021, 124082.
2 Minella, M., et al. Appl. Catal. B Environ. 154–155, 2014, 102.
3 B. Iurascu, et al. Water Res. 1313–1322, 2009, 43.

359
PRELIMINARY STUDY OF THE STRUCTURAL MODIFICATION OF
DEGRADABLE BAGS SUBMITTED TO DIFFERENT ABIOTIC EXPERIMENTAL
CONDITIONS
Raimara de S. Gomes (PG)a*, Walter R. Waldman (R)b, Andreia N. Fernandes (R)a
aFederal University of Rio Grande do Sul, Institute of Chemistry, Porto Alegre, RS, Brazil
bFederal University of São Carlos, Department of Physics, Chemistry and Mathematics, São Carlos, SP, Brazil
*e-mail: raimara.gomes@ufrgs.br
Degradable polymers were developed to limit human-induced plastic pollution, but the level of
degradation and oxidation may vary depending on environmental conditions. Although comparative
studies claim that pro-degradant additives increase the rate of oxidation and fragmentation of
polymers, these parameters vary according to the type of additive and especially the abiotic
conditions to which they are subjected. In this study, the structural modification of degradable bags
compared to conventional bags, exposed to different abiotic conditions, was evaluated. One
conventional high-density polyethylene (HDPE) bag, three different HDPE oxo-biodegradable bags,
and one bag of HDPE biodegradable polymer were selected. For the accelerated degradation,
samples with a rectangular format (6.5 x 6.5 cm) were placed in an accelerated degradation box
equipped with three mercury lamps (36 W), with degradation periods of 432 and 632h, with blocks
of 8 h of exposition followed by 16h of a dark time. For the natural aging study, samples (5.0 x 2.0
cm) were exposed to the weather on two different slopes. Structural changes were evaluated through
visual changes and chemical characterization through attenuated total reflectance-Fourier transform
infrared spectroscopy (ATR-FTIR). After the accelerated degradation period, only 1/3 of the oxo-
biodegradable showed a slight colour change (it turned slightly yellow) and small cracks that followed
the process of fragmentation into small particles. For the natural-aged samples, only 1/3 of the oxo-
biodegradable bags showed colour change (it turned slightly opaque and yellowish). As the exposure
period increased, the conventional HDPE, the biodegradable, and 2/3 of the oxo-degradable
samples showed colour change and the presence of biofilms on the surfaces. Furthermore, both the
biodegradable and oxo-degradable samples had small cracks that followed until fragmentation.
These structural changes were more prominent in samples with a slope with greater exposure to
sunlight. The more exposure to the Sun added to the weathering actions corroborated for fade colour
and fragmentation of the particles. 2/3 of the oxo-biodegradable showed similar behaviour to the
conventional HDPE in all experimental conditions. The fragmented particles from the oxo-
biodegradable samples were classified within the microplastic scale (smaller than 5 mm). Structural
changes were observed using ATR-FTIR and showed that all samples (both natural and accelerated
aged) demonstrated structural changes. The ATR-FTIR spectra showed the characteristic bands of
HDPE represented by the methylene group between 2922 - 2910 cm-1 (CH2 elongation –
asymmetrical) between 2849 and 2840 cm-1 (CH2 elongation – symmetrical).1 It was also observed
in the region from 1800 cm-1 to 1650 cm-1 the increase in intensity, referring to the carbonyl region.2
Oxygenated groups may allow microplastic particles to interact with environmental contaminants.3
Oxo-biodegradable particles can be assimilated by microorganisms. However, studies show that
degraded plastic particles can interact with environmental contaminants.3 There is still no literature
that studies the interaction of microplastic particles from oxo-biodegradable bags with environmental
contaminants. The results showed that the different experimental abiotic conditions influence the
structural modifications of polymers and 2/3 of oxo-biodegradable bags do not behave as expected.
1 Charles J, Ramkumaar, G. R.; Asian Journal Chemistry. 21, 2009, 4477.
2 Benítez, A. et al., Polymers Degradation and Stability. 98, 2013, 490.
3 Hanun, J. N. et al., Journal of Environmental Chemical Engineering. 9, 2021, 106290.
[UFRGS, UFSCAR, CNPq, INCTAA]

360
THE RELATION BETWEEN TOXIC ELEMENTS IN BILE AND
BIOMORPHOMETRIC INDICES IN THREATENED RAYS FROM RJ, BRAZIL
Regina Fonsêca de Almeida (PG)a*, Amanda Pontes Lopes (PG)b, Natalia Guimarães (PG)c,
Marcelo Vianna (R)c,d, Tatiana D. Saint’Pierre (R)a, Rachel Ann Hauser-Davis (R)b
PUC-Rio, Departamento de Química, Rio de Janeiro, RJ, Brazil, 22451-900
a
FIOCRUZ, Instituto Oswaldo Cruz, Rio de Janeiro, RJ, Brazil, 21040-360

b
UFRJ, Instituto de Biologia, Rio de Janeiro, RJ, Brazil, 21941-902

c
AquaRio, IMAM, Rio de Janeiro, RJ, Brazil, 20220-360

d
*e-mail: refonseca25@gmail.com
Many anthropogenic activities result in severe aquatic chemical contamination, leading to several
deleterious biota effects.1 Elasmbranchs, comprising rays and sharks, are especially vulnerable to
chemical contamination, due to their slow maturation processes, low fecundity and long life
expectancy.2 Fish health can be assessed employing biomorphometric indices, such as the condition
factor (CF), the hepatosomatic and gonadosomatic indices (HSI and GSI, respectively), and
correlations with chemical contaminants, aiding in understanding deleterious health effects. In this
context, this study assessed, for the first time, statistical correlations between the CF, HSI, and GSI
and biliary As, Cd, Hg, Pb, and Se concentrations determined by inductively coupled plasma mass
spectrometry (ICP-MS) in four threatened ray species (Gymnura altavela, n=23, 4 females, 19
males; Dasyatis hypostigma, n=22, 8 females, 14 males; Rioraja agassizi, n=104, 9 females, 1 male,
Pseudobatos horkelii, n=36, 17 females, 19 males) from Rio de Janeiro, Brazil. Correlations were
assessed by Pearson and Spearman correlation tests. Regarding Gymnura altavela, moderate
positive correlations were noted between As and Se and As and Hg when grouping both sexes, as
well as a moderate negative correlation between As and the HSI and Se and the HSI, and a strong
positive correlation between Hg and Pb. Separating sexes, a moderate positive correlation between
As and Se, As and Hg, and Cd and Hg, and a moderate negative correlation between As and the
HSI and Se and the HSI were noted for males. No correlations were detected for females due to a
low sample number. For Dasyatis hypostigma, only one moderate positive correlation between Hg
and Pb was noted when grouping sexes. Regarding males, a moderate positive correlation was
noted between As and the HSI, Se and the HSI and As and Se, and a moderate negative correlation
was found between the CF and As. In females, a moderate positive correlation between Se and Cd
and between Cd and Hg and a strong negative correlation between Se and the HSI were detected.
Concerning Rioraja agassizi a moderate positive correlation was noted between Hg and the HSI, Se
and Cd, and Se and Hg, as well as a strong positive correlation between Cd and Hg. Females
exhibited a moderate positive correlation between As and Se and Se and Hg, a strong positive
correlation between Cd and Hg and a very strong positive correlation between Se and Cd. For
Pseudobatos horkelii, a moderate positive correlation was noted between the CF and Pb, Pb and
the GSI, As and Cd, and Cd and Hg, while a moderate negative correlation was noted between the
CF and Se, Hg and the GSI, Se and Pb, Cd and Pb, and Hg and Pb. In males, a moderate positive
correlation was observed between the HSI and As and Cd and Hg, in addition to a moderate negative
correlation between the CF and Se, and the CF and Cd. In females, a moderate positive correlation
between As and Cd was detected, as well as a moderate negative correlation between both As and
Cd and the HSI. These results indicate that all the toxic metals are associated to the calculated
biomorphometric indices for all ray species and metals in bile. Differences between sexes may
indicate different elemental metabolism dynamics and should be further investigated. The
correlations between toxic elements indicate similar exposure sources in the sampling areas, and
correlations between the toxic elements and Se suggest a protective effect of the latter against their
toxicity, demonstrating that biliary concentrations are valuable in assessing elasmobranch health.
1 Tiktak GP et al., Marine Pollution Bulletin, 160, 2020, 111701.
2 Zemah-Shamir Z et al., Fishes 2022, 7, 56.
[CAPES, FAPERJ, CNPq, MPF/RJ]

361
PRELIMINARY DETERMINATION OF POTENTIAL MICROPLASTICS IN
INTAKE POINTS FROM DRINKING WATER TREATMENT PLANTS
André J. da Rosa (UG)a*, Laura R. R. de Oliveira (UG)a, Raimara S. Gomes (PG)a,
Andreia N. Fernandes (R)a
*e-mail: andrerosa0901@hotmail.com.br
Nowadays, there is a rising concern about plastic debris in the 0.001 to 5 mm range, defined as
microplastics. These particles are ubiquitous throughout the ecosystem, with high polluting potential,
and can cause lethal damage to animals and humans.1 Despite Bertoldi and coauthors2 having
determined the presence of microplastics in River Guaíba, the primary supply source of Porto Alegre
city, the concentration of these particles in the intake points from drinking water treatment plants
(DWTP) was not carried out. Thus, in this study, the preliminary determination of potential
microplastics in intake points was performed from three different DWTPs (DWTP1, DWTP2, and
DWTP3). Samples were collected in January/2022 and March/2022. Before characterization, a
volume of 4 L of the sample was filtered through a 63 μm pore-sized microplastic sieve. The solid
was washed abundantly with filtered ultrapure water to eliminate the organic matter.3 Then, the solid
was transferred to a beaker using filtered ultrapure water and filtered by a cellulose ester membrane
(0.45 µm). To avoid cross-contamination, quality assurance and quality control (QA/QC) was carried
out, exposing a humid cellulose ester membrane to the air while the sample was exposed to air
during filtration. Potential microplastics were characterized and classified according to the shape,
color, and number of items per filtered volume using a microscope (4x and 10x magnification). The
concentration of potential microplastics varied between intake points and sampling month. The
highest values were observed in the DWTP3 (54.3 and 69.5 items L-1), followed by DWTP2 (29.5
and 44.3 items L-1) and DWTP1 (14.8 and 29.5 items L-1) for sampling in January/2022 and
March/2022, respectively. One of the main factors that can influence the distribution of microplastics
in aqueous matrices is the precipitation on the days before collection.2 According to the National
Institute of Meteorology,4 on March 30/22, the day before the sampling, the accumulated rain was
about 50 mm, which may have caused the suspension of sedimented particles in the River Guaíba,
increasing the level of contamination of potential microplastics in studied samples. In addition, the
highest concentrations of particles were related to areas of higher population density and human
activities close to intake points. Regarding the shape of potential microplastics found, the fragment
was the predominant shape in DWTP3 (60.1%) and fiber to DWTP1 (73.5%) and DWTP2 (56.5%).
The predominant color in all samples was white-transparent (62,7% for DWTP1; 49,5% for DWTP2;
67,5% for DWTP3), followed by blue and gray-black. The potential microplastics found in this work
are based on preliminary studies and require additional investigations to infer any level of
contamination since it is a public health issue. However, it is of great importance to determine the
concentration of microplastic particles present at the intake points from drinking water treatment
plants since these results can assist in the prediction of possible improvements in their systems.
1Leslie HA, et al., Environment International, 163, 2022, 107199.
2Bertoldi C, et al., Science of The Total Environment, 759, 2021, 143503.
3Jung, JW, Kim S, Kim YS, Jeong S, Lee J. Science of The Total Environment, 825, 2022, 154015.
4National Institute of Meteorology. https://portal.inmet.gov.br. Access day 15/05/2022.

362
Passive sampling of pesticides using hollow fiber liquid-phase
microextraction and comprehensive two-dimensional gas
chromatography
Zenilda L. Cardeal (R)*, Eduard F. Valenzuela (R), Helvécio C. Menezes (R)
Universidade Federal de Minas Gerais, Departamento de Química – ICEX, Belo Horizonte, Brazil, 31270-901
*e-mail: zenilda@ufmg.br
Brazil is considered one of the largest exporters of agricultural products in the world. In order for the
country to continue leading this economic panorama, pesticides have been used as a strategy to
increase the production of different crops. Although pesticides allow improving the agricultural
production, their adverse effects on the environment and human health have generated controversy
in their application. In this sense, research focused on the impact of pesticides has intensified in
recent years. The use of the hollow fiber liquid phase microextraction technique (HF-LPME) has
been reported as an alternative for extracting pesticides in water.1 On the other hand, passive
sampling has shown excellent results in the extraction of a high range of pesticides in water.2 This
tool unlike grab sampling seeks to minimize the number of samples needed to obtain a reliable and
representative analysis because it allows interaction with a larger volume of water since the process
is carried out in situ. The present work presents the determination of pesticides in water sources
through passive sampling and comprehensive two-dimensional gas chromatography (GC×GC). 38
pesticides commonly used in the main crops of Brazil were studied in order to develop the
determination method. A sampler based on the HF-LPME technique was developed for the passive
sampling of pesticides. The sampler was calibrated in the laboratory showing exposure times and
passive sampling rates in the ranges of 3 days to 18 days and from 0.17 mL d−1 and 23.15 mL d−1
respectively. The Sampler was applied to analyze environmental water from the San Francisco river
basin in the state of Minas Gerais in Brazil. The organophosphate compounds such as diazinon,
pirimiphos methyl and fenitrothion and organochlorine compounds such as 4,4-DDD, 4,4-DDE and
α-HCH were detected. Moreover, trifluralin and diazinon were the most detected pesticides.
Comparing the developed sampler with those found on the market, some advantages can be
highlighted; ease manufacture, low cost and simplicity of sample preparation. This work not only
allows to improve the determination of pesticides in Brazil but also in other Latin American
agricultural countries such as Colombia, Mexico, and Argentina where the use of pesticides is
common.
1 Menezes, H. C. et al. Journal of Analytical Methods in Chemistry, 2016. ISSN 2090-8865.
2 Grodtke, M. et al. Journal of Hazardous Materials, 410, 2021, 124538
[CNPq), CAPES, CEMIG]

363
METAL SPECIATION IN WATERS FROM A MINING SITE IN AMAZON BY
USING ATOMIC ABSORPTION SPECTROMETRY AND STRIPPING
VOLTAMMETRY
Cristina Trindade (PG)a, Arthur Jacques (UG)a, Gabriela Strieder (UG)a, Kasley Barreto (UG)a,
Luis Ferraz (R)a, Simone Noremberg (R)a, Leandro Carvalho (R)a, Lílian Amado (R)a
aFederal University of Santa Maria (UFSM), Graduate Program in Pharmaceutical Sciences, Santa Maria, RS, Brazil
a Federal University of Para (UFPA), Institute of Biological Sciences, Belém, PA, Brazil, 66075-110
*e-mail: cristina_trindade@live.com
Industrial processes have the potential to interfere on the quality of the surrounding aquatic
ecosystem, which can lead to detrimental effects on local biodiversity.1 So mining and ore processing
activities cause the mobilization and release of potentially toxic metals that can reach the aquatic
environment.2,3 In this work, water samples collected in rivers and tributaries located in a mining site
located near the city of Belém (PA) were analyzed in the dissolved (<0.2 µm membrane filtered) and
particulate (>0.2 µm membrane filtered) fractions for the metals Pb, Cd and Ni. The total content of
the metals was quantified by graphite furnace atomic absorption spectroscopy (GFAAS). For the
determination of Ni in the reactive (labile) dissolved fraction (<0.2 µm), the adsorptive cathodic
stripping voltammetry (AdCSV) with a mercury electrode (HMDE) was employed using
dimethylglyoxime (DMG) as a complexing agent in ammonium buffer (pH 9.0). In addition, the anodic
stripping voltammetry was employed for the determination of Pb and Cd using an acetate buffer (pH
4.5). The accuracy of the comparative methodologies by GFAAS, AdCSV and ASV were determined
by the measurement of standards reference materials (SRMs) from NIST. The use of the analytical
techniques allowed the speciation of the metals in the different water fractions (particulate, dissolved
and reactive dissolved). As example, figure 1 shows the distribution of dissolved and particulate
species of Cd and Pb within the sample sites from different rivers and their tributaries. The GFAAS
analysis allowed the determination of the dissolved fraction only for Cd and Pb, without enough
sensitivity to quantify Ni species. However, AdCSV allowed the quantification of dissolved and
reactive species of Ni, which was measured in the presence of dissolved organic matter (DOM)
without pretreatment of the sample. Thus, the complex Ni-DMG was able to remove nickel bound to
organic compounds from DOM by reacting with DMG at the HMDE in AdCSV.3 This work aims lastly
to establish a baseline study containing information on the species distribution of Cd, Pb and Ni in
the Pará river and tributaries, which will be part of the study of the mobilization of metals near the
mining industry in the Amazon region of Brazil.
Figure 1. Distribution of Cd and Pb species within the sampled sites in Pará river and tributaries
1Reza R., Singh G. International Journal of Environmental Science and Technology, 2010
2Luoma SN, Hydrobiologia, 1989
3Carvalho LM., Hollister AP., Trindade, CR., Gledhil,M., Koschinsky, A, Marine Chemistry, 2021.
[CNPq, FADESP]

364
SURVEILLANCE AND EARLY WARNING SYSTEM FOR COVID-19 BY
WASTEWATER-BASED EPIDEMIOLOGY (WBE)
Paulo Felipe N. Estrela (PG)*a, Geovana de M. Mendes (PG)a, Marcio N. de Souza Junior
(UG)a, Núbia N. de Brito (R)a, Andréa F. Arruda (R)a, Matheus R. Augusto (PG)b, Ieda C. M.
Claro (PG)b, Adriana F. A. Duran (PG)b, Aline D. Cabral (PG)b, Rodrigo de F. Bueno (R)b,
Gabriela R. M. Duarte (R)a
a Universidade Federal de Goiás, Instituto de Química, Goiânia-GO, Brazil, 74690-900
b Universidade Federal do ABC, Santo André-SP, Brazil, 09210-580
*e-mail: paulofne@hotmail.com
COVID-19 is currently the most spread disease worldwide. Thus, several tools have been used in
the last years to contain the spread of SARS-CoV-2. Among them, monitoring SARS-CoV-2 viral
load in wastewater composes a strategy defined by Wastewater-Based Epidemiology (WBE) that
can help to control the global health crisis.1 Fragments of viral RNA can be found in sewage, as
infected people, whether symptomatic, pre-symptomatic or asymptomatic, excrete the virus through
their biological fluids. In this way, the viral load found in wastewater can be correlated to the viral
infection level in the population. In addition, the WBE allows the anticipation of clinical cases due to
the time window between the viral infection and the clinical manifestation of symptoms (after clincal
testing).2 The study presents the data regarding SARS-CoV-2 RNA monitoring in Goiânia-GO
sweage from May 2021 to July 2022. On a weekly basis collection, sewage treatment plant samples
were pre-concentrated, the RNA was extracted and purified, followed by SARS-CoV-2 RNA
quantification by Quantitative Reverse Transcriptase-Polymerase Chain Reaction (RT-qPCR). After
evaluating 46 weeks, wastewater SARS-CoV-2 viral RNA load ranging from 105 to 109 genome
copies L-1 was detected in 86.96 % (40/46) of the weeks (Figure 1). Through statistical analysis of
prevalence using the Monte-Carlo model, it was possible to estimate the weekly viral load in
circulation in the population. Up untill now, it was issued two early warning notes, which subsequently
demonstrated an anticipated increase in clinical cases in the city, showing the method's ability to
promote an Early Warning System (EWS). The EWS can be useful to help health authorities to
manage the health crisis in advance. Thus, wastewater surveillance has demonstrated to be an
essential tool to help combat the pandemic's advance.
Figure 1. Correlation between SARS-CoV-2 viral load quantified in wastewater samples and the moving average of clinical
cases in the Goiânia-GO.
1 Hamouda, M et al. Science of The Total Environment, 759, 2021, 143493.

2 Panchal, D et al. Environmental Science and Pollution Research, 28, 2021, 22221-22240.
[CNPq, FAPEG, MPT, SANEAGO]

365
CHEMICALLY MODIFIED AÇAÍ SEEDS WASTES APPLIED AS BIOSORBENTS
FOR METHYLENE BLUE REMOVAL
Wyvirlany Valente Lobo (PG)a*, Flávio Augusto de Freitas (R)a,b
aFederal University of Amazonas - UFAM, Postgraduate Program in Chemistry, Manaus, Amazonas, 69077-000, Brazil.
bAmazon Biotechnology Center – CBA, Manaus, Amazonas, 69075-351, Brazil.
*e-mail: wyvir27@gmail.com
The presence of dyes in spring waters resulting from the inappropriate disposal of contaminated
effluents impairs the absorption of light by plants and animals, affecting the photosynthesis process,
in addition to causing several damages to human health.1 Given this scenario, the use of agro-
industrial residues appears as a sustainable alternative to reduce the high cost and increase the
effectiveness of procedures for the removal of these contaminants. 2 Açaí seed waste (ASW) was
evaluated as a low-cost, renewable biosorbent of methylene blue (MB) dye. The waste was treated
with 0.10 M NaOH solution to remove part of the polymeric component of the fibers, as well as to
change the surface groups and, consequently, to increase the efficiency in the removal of the dye
present in synthetic effluents. The biosorbent was characterized using Infrared (FT-IR) and PHZPC
techniques. To optimize the adsorption process, the pH and time parameters were varied. The
characterization of the biosorbent showed a pHzpc value of 6.3 and, through infrared spectroscopy,
the presence of the bands (–OH), (– C=O), (CH2), (–CO), (–C–O–C), and (C=C), was observed,
which are referents of the main functional groups. From the results of the batch adsorption tests, the
adsorption process was optimized at pH 8 and equilibrium time of 120 min, reaching a dye removal
of 99.41%±. The adsorption process of MB on ASW showed a better fit to the Langmuir model (R2=
0.9999; Qmax = 80,91 mg. g-1) and to the pseudo-second order kinetics (R2= 0.9999). The
thermodynamic parameters were also determined in different temperature ranges (25, 30, and 35
°C), where negative values of Gibbs free energy (∆𝐺°= -6961,02, -7029,47 and -7097,92,
respectively) were observed, which indicates the spontaneity of the adsorption process.3 Negative
enthalpy values (∆𝐻°= -4370,75) in the adsorption process indicate an exothermic nature of the
adsorption.4 Therefore, the alkaline treatment provided an interesting alternative to produce efficient
biosorbents from an abundant agro-industrial residue.
1 Xavier, CSF, et al. Res. Soc. Dev. 10, 2021.
2 Yaashikaa, PR, Senthil,KP, Varjani, S. Bioresour. Technol. 343, 2022.
3 Sun, Y. et al. Bioresour. Technol. 344, 2022.
4 Dada, AO, et al. Sci. Rep. 11, 2021.
[FAPEAM, INMETRO- PRONAMETRO]

366
EVALUATION OF PESTICIDES IN SAMPLES FROM IBIAPABA REGION AND
DISSEMINATION OF METHODS OF AGROECOLOGICAL PRODUCTION
Renata de Oliveira Silva (R)a*, Crisiana Andrade Nobre (R)b, Vitor Paulo Andrade da Silva
(R)c, Mairlane Silva de Alencar (UG), Denilson Gonçalves Queiroz (UG)
aCenter for Technology and Industrial Quality of the Ceará, Fortaleza, Ceara, Brazil,
b Federal University of Ceara, Chemical Engineering Departament/Technology Center, Fortaleza, Ceara, Brazil.
*e-mail: renata.oliveira@nutec.ce.gov.br
In the state of Ceará, in spite of large use of pesticides, few studies are verified regarding the problem
of pesticides. In Ibiapaba moutain range, one of the main fruit and vegetable production centers in
the state, agriculture is characterized by the presence of family farmers, who have been very
important to the economy of Ceará.1,2 The vegetable agribusiness has enabled the generation of a
large number of jobs due to the high demand for labor, from sowing to commercialization, showing
an unquestionable contribution to the development of the state. However, there is still a great lack
of data regarding the problem of pesticides, as well as their impacts in this region. The objective of
this work was to carry out research and compare the pesticide residues in food, water and soil in the
samples that represent the cultivation of the two agricultural models from the cities of the Ibiapaba
moutain range. Forty-four were analyzed pesticides of different classes were selected and that are
frequently investigated in pesticide residue monitoring programs, as well as the criterion of
bibliographic study of the region and current ordinances. The QuEChRES-Citrate method was used
in the preparation of soil and food samples.3 The water samples were used in polymeric phase
syringe-type SPE cartridges and later analyzed by liquid chromatography coupled to mass
spectrometry. In gas chromatography, extraction by SPME and fiber with polyacrylate phase was
used. Below is showing the pesticides list monitored in the period (2019-2021), and analyzed by gas
and liquid chromatography methods coupled with mass spectrometry (CG/MS and LC/MSMS). 1)
Molinate; 2) Trifluralin; 3) α-BHC; 4) Atrazine; 5) Chlorothalonil; 6) Alachlor; 7) Metalaxyl-M; 8)
Ametrine; 9) Heptachlor; 10) Malathion; 11) Metolachlor; 12) Chlorpyrifos; 13) Pendimentalin; 14)
Chlordane; 15) Endosulfan; 16) 4,4'-DDE; 17) Crezoxim-methyl; 18) Buprofenzine; 19) Dieldrin; 20)
Endrin; 21) 4,4'-DDD; 22) Triazophos; 23) Propiconazole; 24) 4,4'-DDT; 25) Tebuconazole; 26)
Bifenthrin; 27) Cyhalothrin; 28) Pyriproxyfen; 29) Myrex; 30) Permethrin; 31) β-Cyfluthrin; 32)
Cypermethrin; 33) Difenoconazole; 34) Deltamethrin; 35) Azoxythrombin; 36) Acephate; 37)
Dimethoate; 38) Aldicarb Sulfoxide; 39) Thiamethoxan; 40) Carbendazim; 41) Carbofuran; 42)
Diuron; 43) Imidacloprid; 44) Trifloxystrobin. The results showed that the analyzed foods were not
detected pesticides in organic foods, while 27% of samples from traditional crops were detected one
or more types of pesticides. In the water samples 21% of the samples were contaminated. And in
the soil samples 12% of the sample number was found one or more active ingredient.
1 Pignati et al, Ciência & Saúde Coletiva, 22, 2017.
2 Gomes et al, Ecotoxicology and Environmental Safety, 189, 2020.
3 Alcântara et al, Food Chemistry, 298, 2019.

367
A NEW APPROACH FOR EXTRACTING BIOACTIVE COMPOUNDS FROM
COFFEE SEEDS USING GREEN SOLVENTS
Aline T. Tocia,*, Letícia Z. Ronkob, Maria Alice Antoniassib, Karina M. Uedab, Fabiane O.
Fariasb, Luciana I. Mafrab, Marcos R. Mafrab
a Federal University of Latin American Integration (UNILA), Latin American Institute of Life and Nature Sciences
(ILACVN), Foz do Iguaçu, Paraná, Brazil, 85869-460
b Federal University of Paraná (UFPR), Department of Chemical Engineering, Curitiba, Paraná, Brazil, 81531-990
*e-mail: aline.toci@unila.edu.br
Coffee beans are rich in several bioactive compounds such as chlorogenic acids (5 to 10%), phenolic
compounds, caffeine (1 to 2.5%) and trigonelline (0.7 to 1%), and the variability depends on the
species, planting, climate, among other factors.1 Considering the 2030 Agenda for sustainable
development and the notorious demand for cleaner methodologies, deep eutectic solvents (DES)
have been increasingly used. The advantages of these solvents include non-toxicity (or low toxicity),
simple preparation, low costs and the flexibility to adjust their affinities with the analyte of interest.2
The present work presents an efficient method of simultaneous extraction of caffeine, trigonelline
and CGA, using green solvents. For the extraction optimization, a completely randomized design
(CRD) was used, with the following variables: type of solvent (four deep eutectic solvents based on
choline chloride, water and choline chloride aqueous solution (50% w/v); temperature (25, 45 and
65 °C) and extraction technique (solid-liquid extractions with agitated heating and assisted by ultra-
Turrax). The evaluation of the total amount of phenolic compounds was performed using the Folin-
Ciocalteu methodology and the FRAP and DPPH methods were used to evaluate the antioxidant
capacity. The levels of caffeine, trigonelline and chlorogenic acids were measured by high
performance liquid chromatography with diod-array detection.3 The analysis revealed that the extract
obtained with choline chloride solution (50% w/v) had a high content of total phenolic compounds
(TPC) and the highest antioxidant capacity, while the extraction with deep eutectic solvents (DES)
at a temperature of 65 °C was more effective in maintaining the antioxidant activity of the extracts.
Choline chloride aqueous solution (50% w/v) was the most suitable solvent for CGA extraction.
However, DES was more effective in preserving antioxidant activity. The results showed that the use
of green solvents, mainly DES, can be effectively used for the extraction of bioactive compounds,
maintaining the antioxidant activity of the biomolecules.
1 Toledo PRAB, et al. Comprehensive Reviews in Food Science and Food Safety, 4, 2017, 1.
2 Yoo ED, et al. Food Chemistry, 355, 2018, 357.
3 Toci AT, Farah A, Trugo LC. Química Nova, 29, 2006, 965.
[CNPq, Institutional Program of Aid for Researcher Integration/UNILA]

368
PRESENCE OF APHIDS (Aphidoidea) IN JAMBU (Acmella oleracea)
CONTAMINATED WITH LEAD
Pedro L. C. Sanches (UG)a, Danielle S. Nascimento (UG)a, Robert Soares (UG)a, Fabrício T.
Barroso (UG)a, Allan S. Cruz (PG)b, Kelly G. F. Dantas (R)b, Adriano B. Pacheco (R)a, Bianca
S. F. Alves (R)a,b*
a Universidade Federal Rural da Amazônia, Tomé-Açu, Pará, Brazil,68.680-000
b Universidade Federal do Pará, Programa de Pós Graduação em Química, Belém, Pará, Brazil, 66.075-110
*e-mail: bianca.alves@ufra.edu.br
The World Health Organization considers lead one of the 11 elements considered potentially toxic
and has this effect on the human body. In the soil, lead occurs naturally and is intensified by the
accumulation resulting from industrial processes. In plants, the lead content is conditioned to the
place of cultivation and the resistance of the culture to the absorption of the metal, a fact that
represents a risk to human health.1,2 The jambu (Acmella oleracea) is one of the main vegetables in
the State of Pará, known for the dampening of the mucous membranes, has its rudimentary
cultivation, carried out by small producers of family agriculture.3 It is not uncommon for jambu to be
affected by diseases that can damage production, such as the aphid (Aphidoidea), which are sucking
pest insects that may or may not have wings, but when they do, they are responsible for the spread
of the pest.4 The study aims to evaluate the presence of lead in aphids found in jambu cultivation.
The aphids obtained for the evaluation are the result of an experiment of planting jambu in
contaminated soil, in the test the sowing of jambu was carried out, using two seeds in each of the
256 cells (seedlings). Five seedlings with 2 to 4 definitive leaves were transplanted to pots with soil
contaminated with doses of lead, in this process 8 treatments were performed with 0, 30, 60, 120,
240, 480, 960 and 1920 mg kg-1 of Pb in triplicate (n = 3). The trial was followed for 52 days, ended
when each stem presented a capitulum (inflorescence), and at 33 days the beginning of aphid
infestation was observed. At the end of the test, the pests were removed from the jambu leaves
(Figure 1), placed in vials and frozen for later lyophilization, the dried samples were digested in a
microwave oven with cavity and the digests were analyzed using a microwave induced plasma
optical emission spectrometer (MIP OES). The result obtained shows that aphids absorbed the lead
present in the jambu and that the concentration presented is related to the Pb content added to the
treatment. Aphids from pots containing 0, 30, 60, 120 and 240 mg kg -1 of Pb are lower than the
detection limit and the pests removed from pots with 480, 960 and 1920 mg kg -1 are with respectively
27.9, 53.2 and 19.2 mg kg-1 of Pb. The information obtained is relevant because the aphids disperse
and can contaminate other crops. The addition and recovery test ranged from 98.8 to 110.4%.
Figure 1. a) Pb treatment curve in jambu cultivation b) Leaf used as host c) Microscopic image of aphids.
1 Kolesnikov S, Minnikova T, Kazeev K, Akimenko Y, Evstegneeva N, Water air soil pollut,18, 2022, 233
2 Lima FS, Nascimento CWA, Silva FBV, Carvalho VGB, Ribeiro Filho MR, Horticultura Brasileira, 27, 2009, 362
3 Boari AJ, Quadros AFF, Batista ICA, Kauffmann CM, Nechet KL, EMBRAPA, 2022, ISSN 1983-0491
4 Picanço MC, https://www.ica.ufmg.br/wp-content/uploads/2017/06/apostila_entomologia_2010.pdf
[UFRA, UFPA]

369
DETERMINATION OF METALLIC SPECIES IN SOIL USING TWO CALIBRATION
METHODS
Lúrian A. D. Cruz (UG)a*, Danielle C. Schnitzler (R)a
aFederal Technological University of Paraná, Academic Departament of Chemistry and Biology, UTFPR, Curitiba,
Paraná, Brazil, 81280-340.
*e-mail: lurian.2016@alunos.utfpr.edu.br
Soil is an extremely important natural resource, it is a source of food for plants and where agriculture,
livestock and forestry are developed, activities that are fundamental for the feeding of living beings.
One of the factors that impair soil quality is contamination by metals, often as a result of human
activities.1 Almost all metallic species have the potential for toxicity and can easily accumulate in the
soil, due to their non-biodegradability. Thus, it is necessary to efficiently determine the concentration
of metals in the soil, assessing whether the content is compatible with expectations or whether there
is contamination.2 However, soil is an extremely complex matrix, with several factors that make it so,
such as the organic matter content, which varies greatly between soils, ion exchange, pH,
granulometry, factors that can be combined in different ways.3 In this context, the need to find the
most suitable calibration method is very important. Therefore, the objective of this work was to
compare the external calibration method and the standard addition method, in order to choose the
most appropriate method for the determination of metallic species in soil. The samples were
collected in the Contestado Settlement in the city of Lapa/PR, in 2 different points, one in an
agroecological environment and another in a conventional environment. For the preparation, the
samples were taken to an oven for 48 hours at 50ºC, sieved in a 53 μm mesh and submitted to the
burning of organic matter according to EMBRAPA 1979.4 The extraction was carried out based on
the EPA 3050B5 protocol. The procedure was performed in triplicate and with the addition of different
concentrations of copper, zinc, manganese and iron to verify the recoveries. The calibration curve
for each metal was constructed in two ways, by external calibration and by adding standard, using 6
solutions with the metals at different concentrations. The analysis was performed by Inductively
Coupled Plasma Optical Emission Spectrometry (ICP-OES). The curves constructed by the two
calibration methods showed good linearity, with a correlation coefficient above 0.999. The recovery
study showed satisfactory results with recoveries above 90%. The parallelism study was applied to
verify if the matrix complexity influences the method of determination of metallic species and it was
possible to observe that the pattern addition method is the most suitable for these determinations.
1 Morillas, H. et. al. Chemosphere. 2019.
2 Qi, X. et. al. Chemosphere. 2022.
3 Skoog, Douglas A. et al. Fundamentos de química analítica. São Paulo: Cengage Learning, 2009.
4 EMBRAPA - Empresa Brasileira de Pesquisa Agropecuaria. Centro Nacional de Pesquisa de Solos. Manual de Métodos
de Análise de Solo. 2 ed. Rio de Janeiro, 1997.
5 USEPA. United States Environmental Protection Agency. Background Concentrations of Metal in Florida Surface Soils:
Comparison of Four EPA Digestion Methods and Baseline Concentration of 15 Metals. EPA, University of Florida,
Gainesville, FL 32611. May,1998.
[UTFPR, LEMASSA, Public Ministry of Paraná, Ministry of Labor, LAMAQ]

370
EVALUATION OF ANTHROPIC INFLUENCES ON INORGANIC CONSTITUENTS
LEVELS IN URBAN SOIL COLLECTED IN CITY OF ARACAJU, SERGIPE, BRAZIL
Rennan G. O. Araujo (R)a*, Samir H. dos Santos (PG)a, Sarah A. R. Soares (R)b, Antônio
Fernando de S. Queiroz (R) b, Silvânio S. L. Costa (R)c
aUniversidade Federal da Bahia, Inst. De Química, Dept o de Química Analítica, Salvador, BA, Brazil, 40.170-115.
bUniversidade Federal da Bahia, Inst. de Geociências, Dept o de Oceonografia, Salvador, BA, Brazil, 40.170-115.
cUniversidade Federal de Sergipe, LTMA, São Cristóvão, SE, Brazil, 49.100-000.
*e-mail: rennan@ufba.br
Soils in urban areas near residences avenues and roads can be contaminated by potentially toxic
elements over time, through human activities, such as vehicular traffic, sanitary landfills and human
waste and construction in cities.1 Thus, the determination of the concentration levels of inorganic
constituents in urban soil is important to evaluation the environmental quality of urban areas.2 In this
work, forty-three urban soil samples were collected in city of Aracaju, Sergipe, Brazil, from two depth
fractions [fraction (0-20 cm) and (20-40 cm)]. The simultaneous multielementary determination of Al,
As, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Mo, Na, Ni, P, Pb, S, Sb, Sr, U, V and Zn using inductively
coupled plasma optical emission spectrometry (ICP OES) was performed. The extraction procedure
of the chemical elements in soil samples was carried out through a closed digester block. The limit
of quantification (LoQ) obtained varied between 0.01 (Sr) and 1.5 µg g -1 (As) for the trace elements
and microelements, and varied between 0.2 (K) and 10 µg g-1 (S) for macroelements. The accuracy
of the analytical method was confirmed by analyzing six certified reference materials (CRM: BCR
142, BCR 032, NIST SRM 120c, NIST SRM 694, NIST SRM 695, NIST SRM 2710a), obtaining good
agreement ranging between 80.0 ± 0.75% (Mn) and 118.8 ± 3.6% (Pb). Precision was expressed as
relative standard deviation (RSD, n=3), ranging from 0.012 (U) to 10.0% (Cd), showing good
precision for quantitative analysis. According to the soil quality reference values (VRQ), established
by the Brazilian Council for the Environment (Conselho Nacional de Meio Ambiente - CONAMA)
through Resolution nº 420, of December 28, 2009 and the Sergipe Council for the Environment
(Conselho Estadual de Meio Ambiente - CEMA) through Resolution Nº 01/2019, of February 27,
2019, only samples 2S and 2F had concentrations of inorganic constituents lower than the quality
reference values (valores de referência de qualidade – VRQ) and in samples 13F, 15S and 18F
values were found greater than the prevention value (VP) (2.0 mg kg -1, Sb). Through granulometric
analysis and Pearson's correlation matrix, it was observed that the chemical elements are distributed
among very fine sand, silt and clay particles. A regional geochemical basis was established using
iron concentration as reference element. The enrichment factor (EF) indicated that only samples
11S, 12S and 12F showed severe enrichment by anthropic influences for Zn concentration. The
geoaccumulation index (Igeo) characterized sample 11S as moderately polluted and samples 12S
and 12F as moderately to heavily polluted. Principal component analysis (PCA) and hierarchical
cluster analysis (HCA) indicated that in the samples from urban areas where vehicular traffic was
more intense, the concentration of chemical elements was higher. In addition, a tendency was
observed in the formation of a group among the chemical elements considered from natural sources.
This work also shows the importance of studies related for inorganic constituents determination in
soil in urban areas, contributing with relevant information for new studies in the future, showing how
human activities, mainly vehicular traffic and urban waste, can cause great impacts to the
environment and human health.
1Adimalla N, Chen J, Qian H, Ecotoxicology and Environmental Safety, 194, 2020, 110406.
2Wang X, Birch GF, Liu E, Chemosphere, 297, 2022, 134155.
[FAPESB, CNPq, CAPES, LEPETRO/NEA, LTMA]

371
COMPARISON ON AIR QUALITY DATA BEFORE, DURING AND AFTER PARTIAL LOCKDOWN IN RIO DE JANEIRO:
A THREE YEARS STUDY
Soraya Nassar Sakalem (UG)a*, Igor Rabello Ferreira Henriques Pereira a (PG), Fabiano
Galdino Leal (PG)b, Julio Carlos Afonso (R)a, Celeste Yara dos Santos Siqueira (R)a
aFederal University of Rio de Janeiro, Analytical Chemistry Department, Chemistry Institute, RJ, Brazil, 21941-909
bFederal University of Rio de Janeiro, Civil Engineering, COPPE, RJ, Brazil, CEP 21941-611
*e-mail: snsakalem@coppe.ufrj.br
In March 2020, the World Health Organization (WHO) declared that COVID-19 had been
characterized as a pandemic.1 Since then, governments around the world have adopted lockdown
measures to minimize the spread of the virus. In Rio de Janeiro, schools, universities, stores, malls
and restaurants were closed. Recent studies indicate that the new Coronavirus pandemic had a
positive impact on air quality in several cities, such as Delhi in India,2 Barcelona in Spain3 and even
in São Paulo.4 In New York, in the United States, no major changes were observed in air quality
indicators.5 In the literature, it is also possible to find studies regarding the impact of the pandemic
on air quality in Rio de Janeiro,6 however, it is restricted to mostly residential regions (Bangu, Irajá
and Tijuca) and in a short period of time. Motivated by gap present in the literature about air quality
in Rio de Janeiro, it was decided to carry out a comparative study to evaluate the air quality before,
during and after the partial lockdown due to the pandemic, in the State of Rio de Janeiro. Three
collection sites were evaluated: (i) Campos Elíseos; (ii) Tijuca; (iii) Centro. Six different substances
were monitored: particulate matter (MP10), carbon monoxide (CO), nitrogen dioxide (NO 2), ozone
(O3), sufur dioxide (SO2) and nitrogen monoxide (NO). The samples were collected between
January, 1st, 2019 to December, 31st, 2021, totalizing 20,171,023 records were during these 3 years
of study. Due to the large volume of collected information, for this summary, it was decided to
approach only particulate matter (PM10) data. PM10 are inhalable particles, that can pass through
the nasal and oral cavities and penetrate to the larynx, being harmful to health. The graphs below
show the variation in the concentration of these particles over the years. In addition, the rates of
mobility at the Rio de Janeiro city as of March 2020, provided by Google, are also available. It is
worth mentioning that urban mobility data are only available from that date onwards, as they aim to
provide statistics on people's displacements, in order to assist in critical decisions in the fight against
COVID-19.
The data show that with the reduction of urban mobility, there is also a decrease in the PM10 content
for the Centro - RJ and for Tijuca, showing that in these regions the lookdown positively affected the
air quality. However, the same is not seen in Campos Elíseos, since industrial activities in this region
remained unchanged, even during the period of social isolation, as they are considered essential
activities.
1Cucinotta D., and Vanelli M., Acta Biomedica, 91, 2020, 157–160
2Mahato S., et al. Science of the Total Environment, 730, 2020, 139086
3Tobías A. et al., Science of the Total Environment, 726, 2020, 138540
4Freitas E. D. et al., 2020, doi: 10.20944/preprints202004.0515.v1.
5Zangari S., et al. Science of the Total Environment, 742, 2020, 140496
6Dantas G., et al. Science of the Total Environment, 729, 2020, 139085

372
DETERMINATION OF ANTIEMETIC RESIDUES IN SEDIMENTS USING
SALTING-OUT INDUCED LIQUID–LIQUID MICROEXTRACTION AND LC-MS
Jaqueline F. Reichert (R)*, Dinara S. Rodrigues (PG), Petronio S. Zinn (PG), Vanessa R. do
Nascimento (PG), Renata V. Corrêa (UG), Paola A. Mello (R), Ayrton F. Martins (R), Darliana
M. Souza (R), Érico M.M. Flores (R)
Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil, 97105-900
*e-mail: jaquereichert@yahoo.com.br
In recent years the occurrence of pharmaceuticals in the environment has been a matter of growing
concern. Methods for the analysis of pharmaceuticals in water, particularly in effluents from sewage
treatment plants, have been reported. In contrast, analytical methods for pharmaceuticals in solid
matrices, such as sludge and sediment, are scarce, impairing the evaluation of polluted areas. This
study aimed to develop a time and simple multi-residue method to extract and determine three
antiemetic drugs in sediment samples. The superficial sediment samples were collected at 10
strategic points, the first point being when the stream reaches the university and the last when the
stream is no longer part of the university area. Target analytes were ondansetron, metoclopramide,
and bromopride. The quantification was performed with liquid chromatography quadrupole mass
spectrometry – LC-MS (Shimadzu, Kyoto). The instrumental conditions for LC-MS were as follows:
Shim-pack CLC-OSD column (5 µm, 150 x 4.6 mm); column oven temperature at 35 °C; injection
volume of 10 μL; gradient elution mode for mobile phase (0.8 mL min-1 constant flow) composed by
(i): eluent A - 5 mmol L-1 of ammonium formate containing 0.1% formic acid and (ii) eluent B -
acetonitrile containing 0.1% formic acid; total running time of 16 min; electrospray ionization in
positive mode. The general procedure consisted of using 0.5 g of dry sediment, to which 5 mL of
water were added followed by stirring for 2 min, with posterior addition of 1.3 mL of solvent for
extraction, 2 g of salt, and a final step of 2 min of agitation, plus 5 min for centrifugation. The
supernatant was removed, evaporated under nitrogen flow at 40 ° C and resuspended in 1 mL of
acetonitrile for analysis by LC-MS. As extraction solvent, acetone, acetonitrile, ethyl acetate,
isopropyl alcohol, methanol, ethanol, and a mixture of these in a 1: 1 (v/v) ratio, were evaluated. The
best recoveries were obtained using a mixture of ethyl acetate and acetone. As the salt, anhydrous
magnesium sulfate, magnesium sulfate heptahydrate, ammonium chloride, potassium phosphate,
ammonium formate, sodium chloride, ammonium sulfate, anhydrous sodium sulfate, calcium
carbonate, potassium bicarbonate, dihydrate calcium chloride were studied to improve extraction.
Anhydrous sodium sulfate provided the best results for all substances. The quantitative factors that
significantly affect the extraction were identified using the Plackett-Burman design containing 16
experiments and 3 central points. In this way, the amount of sample, the proportion of the extraction
solvent mixture, the pH of the water, the amount of water, the amount of salt, the stirring time in both
stages, and the centrifugation time were optimized. Finally, the method was optimized through a
central rotational composite design. The developed method was suitable for the extraction of the
three pharmaceutical drugs. Metoclopramide was present in 4 samples, Bromopride in 3 samples,
while ondansetron was determined in 8 of 10 samples. All analytes were present at concentrations
below the limits of the quantification of the method. The developed method was suitable for the
extraction of the three pharmaceutical drugs and the method showed to be adequate for the
quantification of the three antiemetic pharmaceuticals in complex matrices, as sediment, which
showed 90% contamination with at least 1 of the investigated analytes.
1 Dongli Du, et al. Anal. Methods, 6, 2014 6973-6980
[CNPq, CAPES]

373
SYNTHESIS OF HYDROPHOBIC DEEP EUTECTIC SOLVENTS (hDES) FOR USE
IN ANALYTE EXTRACTION PROCESS IN ENVIRONMENTAL SAMPLES
Rafaela S. Lamarca (R)a*, Éder R. Paganini (PG)a, Sabrina S. Ferreira (PG)a, Karen C. Ferreira
(PG)b, Douglas F. Franco (R)c, Paulo C. F. de Lima Gomes (R)b, Mario H. Gonzalez (R)a
aSão Paulo State University(UNESP), Department of Chemistry and Environmental Science, São José do Rio Preto, São
bSão Paulo State University (UNESP), Institute of Chemistry, Araraquara, São Paulo, Brazil, 14800-060
c São Paulo State University (UNESP), Institute of Chemistry, Araraquara, São Paulo, Brazil, 14800-060
*e-mail: rafaela.lamarca@unesp.br
In the last decades, the concept green have emerged in different fields, as a way of preserving the
environment and reducing the negative impacts caused by human development. Therefore, in the
analytical chemistry area a current of study called Green Analytical Chemistry, started priorities to
reduce or eliminate the use of products, by-products, and toxic solvents to human and
environment.1,2 In this context, the development of green and sustainable solvents, such as
hydrophobic deep eutectic solvents (hDES) has been highlighted. The hDES are characterized as
solvents formed by hydrogen bond donor compounds (HBD) and hydrogen bond acceptor
compounds (HDA).3 These hDES have some particularities such as biodegradability, low cost,
simple synthesis and high efficiency in the extraction of water-insoluble volatile organic
compounds.4 In this study, two hDES with molar ratios 1:1 and 1:1:1 were prepared from a mixture
of DL-menthol and acetic acid; DL-menthol, acetic acid and pyruvic acid, under stirring at 220 rpm
and heating at 50 °C for 15 min. The formed solvents were characterized by density, viscosity and
infrared spectroscopy (ATR- FTIR mode). The density and viscosity of the solvent obtained in the
binary mixture were 1.023 g mL-1 and 4.195 mPa s, respectively. As for the ternary mixture, the
density were 1.653 g mL-1 and 3.902 mPa s, respectively. The vibrational characterization of the
precursor solvents via FTIR allowed the identification of vibrations frequencies of the main functional
groups, as well as shifts of the vibrational modes of the synthesized solvents. From the interpretation
of FTIR spectra of the synthesized solvents, it was possible to verify a shift of symmetric stretching
of carbonyl (C=O) group from 1704 to 1712 cm-1. The next step will focus on using the hDES
developed in the extraction of polybrominated diphenyl ethers (PBDEs) present in environmental
matrices and analysis by gas chromatography coupled to mass spectrometry.
1Armenta S, Garrigues S, de la Guardia M, TrAC Trends in Analytical Chemistry, 27, 2008, 497–511;
2Sajid M, Płotka-Wasylka J, Talanta, 238, 2022, 123046;
3Paiva A, et al, ACS Sustainable Chemistry & Engineering, 2 (5), 2014, 1063–1071;
4Florindo C, et al, Journal of Molecular Liquids, 297, 2020,111841.
[FAPESP, CAPES, CNPq, IBILCE-UNESP-São José do Rio Preto, INCT-DATREM]

374
ASSAYS OF INORGANIC ARSENIC BIOACCUMULATION AND
BIOTRANSFORMATION IN NILE TILAPIA (OREOCHROMIS NILOTICUS)
Nathalia S. Ferreira (PG)*a, Ívero P. Sá (PG)b, Ana R. A. Nogueira (R)b, Danilo G. H. Silva (R)c,
Mario H. Gonzalez (R)a
a São Paulo State University (UNESP), Department of Chemistry and Environmental Science, São José do Rio Preto,
SP, 15054-000, Brazil
b Embrapa Pecuária Sudeste, São Carlos, SP, 13560-970, Brazil
c Federal University of Mato Grosso do Sul, Department of Biology, Três Lagoas, MS, 79600-080, Brazil
*e-mail: nathalia.s.ferreira@unesp.br
Arsenic (As) is an inorganic element found naturally in soils and sediments, or as a contaminant
released from anthropogenic activities.1 Since fish lie at the top of the food chain in the aquatic
environment, they can bioaccumulate and biotransform As present in both inorganic forms, which
are more toxic, and methylated forms, which are less toxic.2 In this study, ecotoxicological assays
were performed with the As(III) and As(V) species, separately, using Nile tilapia under controlled
conditions in individual aquaria containing 10 liters of solution. Evaluation was made of the
bioaccumulation and biotransformation of As in different biological tissues (liver, stomach, gills, and
muscle). The fish were divided into 5 groups (control, G1, G2, G3, and G4) (n = 5). The control group
was not exposed to As, while groups G1 and G2 were exposed to concentrations of 5 and 10 mg L-
1
, respectively, during a 1-day exposure period. Groups G3 and G4 were exposed to the same
concentrations (5 and 10 mg L-1, respectively), but during a longer period of 7 days. Analyses of total
As in the tissues employed ICP-MS, and the accuracy of the method was assessed by analysis of
certified reference materials (DOLT-5 and DORM-4). Tables 1 and 2 show the total As
concentrations found in the fish tissues in the assays using As(III) and As(V), respectively.
Table 1. Concentrations of total As (µg g-1) in the different fish tissues for the bioaccumulation/biotransformation assay
with As(III) (n = 2, mean ± SD).
Concentration (µg g-1)
Groups Liver Stomach Gills Muscle
Control < LOQ <LOQ 0.08 ± 0.003 0.02 ± 0.02
G1 2.64 ± 0.17 0.69 ± 0.00 0.47 ± 0.02 0.46 ± 0.01
G2 15.38 ± 0.00 6.39 ± 0.00 4.39 ± 0.00 2.31 ± 0.002
G3 5.52 ± 0.00 5.57 ± 0.00 2.56 ± 0.03 2.10 ± 0.02
G4 21.24 ± 0.00 14.65 ± 0.00 4.90 ± 0.21 3.25 ± 0.04
Table 2. Concentrations of total As (µg g-1) in the different fish tissues for the bioaccumulation/biotransformation assay
with As(V) (n = 2, mean ± SD).
Concentration (µg g-1)
Groups Liver Stomach Gills Muscle
Control 0.10 ± 0.04 0.02 ± 0.05 0.10 ± 0.003 0.23 ± 0.003
G1 1.31 ± 0.061 1.09 ± 0.00 0.59 ± 0.004 0.48 ± 0.004
G2 9.07 ± 0.15 6.38 ± 0.00 3.55 ± .023 1.74 ± 0.02
G3 21.03 ± 1.53 5.55 ± 0.00 1.79 ± 0.85 1.91 ± 0.09
G4 11.72 ± 0.24 5.21 ± 0.00 3.52 ± 0.03 2.86 ± 0.05
In both assays, for the groups exposed to arsenic, the highest concentrations were found in the liver,
followed by the stomach, gills, and muscle. The groups exposed during 7 days generally presented
higher concentrations of As in the tissues, compared to the groups exposed to the same
concentration during 1 day. Chemical speciation analyses (in progress) will enable determination of
the distributions of arsenic species (As(III), As(V), MMA, DMA, and AsB) in the different tissues.
1 Greani S. et al., Ecotoxicology, 26, 2017.
2 Zhang W. et al., Environment International, 163, 2022.
[FAPESP, CNPq, INCT-DATREM]

375
WHAT GOVERNS THE PRESENCE OF MICROPLASTICS AND WHAT ARE
THEIR PHYSICAL CHARACTERISTICS IN RIVER GUAÍBA?
Crislaine Bertoldi (R)a*, Larissa Z. Lara (PG)a, Andreia N. Fernandes (R a
*e-mail: cris.bertoldi@hotmail.com
Microplastics (0.001 to 5 mm) have been found in different matrices, such as biota, water, sediment,
air, food, and even human organs. In this context, considering the potential impact of microplastics
on the ecosystems, this study investigated the presence of these particles in River Guaíba, which is
the main source of water supply to Porto Alegre city. The samples were collected with a manta net
(60 μm mesh size). The abundance and physical characteristics of the microplastics were evaluated
in 10 sites collected in 8 campaigns (2018 to 2020). Before characterization, samples were submitted
through extraction and clean-up following the main steps: sieving, wet peroxide hydrogen oxidation,
density separation, and physical characterization. The concentration of microplastics varied between
2.9 and 61.2 items m-3, and the distribution was, in general, positively influenced by the population
density, precipitation, and the hydrodynamic conditions of each site. The smallest amplitude in
microplastic abundance was determined in the spring, with concentrations between 5.0–15.8 items
m-3. On the other hand, the greater amplitude was determined for campaign 1, in the winter (11.9–
61.2 items m-3) and 4, in the autumn (4.8–53.8 items m-3). The increase of microplastics in the waters
after periods of precipitation may be a result of the release of microplastics downstream, street
runoff, atmospheric deposition, and resuspension of microplastics already deposited in the
riverbed.2,3 In addition, depth played a crucial role in the distribution of microplastics in water since
the points with the best correlation with precipitation are in regions of greater depth. The fragment
was the majority shape of microplastics, representing 88% of the particles counted, followed by fibers
with 7% and microbeads with 5%. This profile demonstrated that the microplastics present in River
Guaíba are predominantly secondary, produced unintentionally. This study observed the potential
formation of nanoplastics from some microplastics found in the river. Among these fragments, the
formation of cracks with a size below 10 μm on the surface of the particles was observed. Through
the analysis of non-metric multidimensional scaling (nMDS), it was possible to confirm the similarity
between the distribution of shapes for points 5, 8, and 9. This fact demonstrates that these points
may be subject to the influence of similar factors, possible contamination of domestic sewage. The
white-transparent color category was predominant (47%), followed by blue (15%), yellow (14%), red
(13%), green (8%), and black (3%). The high proportion of white-transparent microplastics may be
due to the large number of items produced in this color, such as plastic bags for transport, food
boxes, and another packaging. In addition, the possible discoloration of colored plastics in the
environment due to exposure to ultraviolet light and other stressors can increase the proportion of
this coloring class. The information collected in this study made it possible to observe that
microplastics are ubiquitous in the water source studied. The data demonstrate the direct impact of
human beings on the contamination of aquatic bodies. Therefore, while the impact of microplastics
on freshwater ecosystems is still largely unknown, this study shows evidence of the widespread of
this contamination, and further investigations are needed to improve the understanding of its impact
on the environment and human beings.
1Millican JM and Agarwal S, Macromolecules, 54, 2021, 4455–4469.
2Oni BA, Ayeni AO, Agboola O, Oguntade T, Obanla O, Ecotoxicology and Environmental Safety, 198, 2020, 110656.
3Xia W, Rao Q, Deng X, Chen J, Xie P, Science of the Total Environment, 732, 2020, 139065.

376
APPROACHES FOR DETERMINATION, FRACTIONATION AND SPECIATION
OF RARE EARTHS ELEMENTS BASED ON QICP-MS AND DGT TECHNIQUES
Amauri A. Menegário (R)a*, Melina B. T. Zanatta (R)a, Luiz F. Moreira (PG)a, Raquel de C.
Gradwohl (PG)a, Lucas Pellegrini Elias (PG)a, Johanna Irrgeher (R)b, Shaun Lancaster (R)b,
Ulrike Moser (R)b
a UNESP, Centro de Estudos Ambientais, Rio Claro, São Paulo, Brazil, 13506-900
b Montanuniversität Leoben, Department für Chemie, Leoben, Austria, 8700
*e-mail: amauri.antonio-menegario@unesp.br
Rare earth elements (REEs) comprise a group of 15 elements (called lanthanides) plus Sc and Y.
Recently, these elements have become extremely important for high technology applications and
processes due to their unique properties. REEs have low solubility and mobility in the terrestrial
crust, so their patterns of abundance can be used for the interpretation of geological and chemical
processes. The determination of REEs can be performed by various analytical techniques, but
inductively coupled plasma mass spectrometry (QICP-MS) has been considered the most effective
technique due to its sensitivity, selectivity and multi-element capability. Despite these advantages,
the accurate determination of REEs by conventional ICP-MS (QICP-MS) until now is complex due
to the low concentrations of the analytes found in the environment and spectral interferences.
Questions to be addressed during their analyses by QICP-MS is how to handle oxide and hydroxide
ions formed from the presence of Ba and light-REEs in the samples.1 In this talk, recent approaches
for the determination, fractionation and speciation of rare earths elements based on the use of QICP-
MS and diffusive gradients in thin films (DGT) techniques will be present and discussed. Special
emphasis will be given to environmental samples, such as river water, soil and soil porewater.
Initially, the discussion will focus on overcoming spectral interferences on the determination of La,
Ce, Pr, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm, Yb, Lu and Th by QICP-MS, ICP-MS coupled with
membrane and/or nebulisation desolvation system (DQICP-MS) and triple quadrupole ICP-MS
(TQICP-MS). In this part, data related to the magnitude of spectral interferences from analyses using
QICP-MS and DQ-ICP-MS in real river water samples will be compared. In sequence, determination
of La, Ce, Pr, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm, Yb, Lu and Th by QICP-MS and DQICP-MS after
DGT sampling will be discussed. Consistent data, particularly, for overcoming BaO+ interference on
determination of Eu+, Sm+ and Nd+ will be shown. Finally, emphasis of discussion will be headed on
fractionization and speciation of La, Ce, Pr, Nd, Sm, Eu, Gd, Dy, Ho, Er, Tm, Yb, Lu and Th. The
fractionating related to dissolved and labile fractions will be shown. Comparison between river water
of the filtrated phase (using 0.45 µm syringe filter) and labile fraction (obtained by using in situ DGT
measurements) with a speciation software will be presented and discussed. Trying to understand
the behaviour of REEs from a contaminated area, data about its contents in soil pore water, as well
as its flux from the soil solid phase (obtained by using in situ DGT measurements) will be shown. In
addition, unpublished data related to a new binding phase for DGT measurements will be presented.
Broadly speaking, this talk will discuss innovative data that will be attractive for researchers
searching for analytical approaches for the determination and understanding of the biogeochemical
behaviour of REEs.
1Trommetter G, Dumoulin D, Billon G, Spectrochimica Acta Part B, 171, 2020, 105992.

377
METAL REMOVAL FROM PRODUCED WATER SAMPLES OBTAINED FROM
CRUDE OIL EXTRACTION USING MICROORGANISMS
Larissa M. A. Oliveira (PG)a*, Marianne S. Schaeffer (PG)b, Juliana P. Souza (PG)a, Jhulia L.
Boff (UG)a, Marilene Vainstein (R)b, Roberta S. B. Rodrigues (R)a, Juliana S. F. Pereira (R)a
aUniversidade Federal do Rio Grande do Sul, Departament of Chemistry, Porto Alegre, RS, Brazil, 90650-001
b Universidade Federal do Rio Grande do Sul, Biotechnology Center, Porto Alegre, RS, Brazil, 91501-900
*e-mail: lary.mayar@gmail.com
Produced water (PW) obtained from crude oil extraction is a mixture of dispersed components such
as oil and solids and dissolved compounds as water-soluble organic compounds, dissolved salts,
chemical additives that makes this sample a complex mixture. Nowadays, PW represents the largest
volume of waste associated with the oil and gas industry, and it is estimated that about 85% of this
waste is currently discharged to the sea. In this sense, regulatory requirements have become more
stringent concerning the discharge of this effluent. On the other hand, PW could be recycled and
reused, but for this purpose it must be treated to remove some metals (e.g. Ba, Ca, Fe and Sr) in
order to avoid to form insoluble salts, clogging filters, among other problems.1 For this purpose, the
use of microorganisms was evaluated in this work to promote metals removal from produced water
obtained from crude oil extraction. The aim of this work was to remove some metals that in the
presence of high concentration of sulfate or hydroxides could form insoluble salts. Four samples of
PW with variable characteristics were selected and they were digested by a procedure combining
microwave and ultraviolet radiation for subsequent metals determination by inductively coupled
plasma optical emission spectrometry (ICP OES) and flame atomic absorption spectrometry (F
AAS).2 The samples contain variable organic carbon content and metals, ranging from 12 to 155 mg
L-1 (dissolved organic carbon), 0.12 to 0.43 g L-1 total solids content and metals content ranging from
1.72 to 9.00, 445 to 3551, 1.06 to 41.2, 127 to 1046 and 24.7 to 138 µg g -1 for Ba, Ca, Fe, Mg and
Sr, respectively. The results obtained for metals content were used to the firsts experiments to
produce synthetic samples to evaluate the performance of metals removal from the microorganisms
selected. The microorganism selected was the commercial Bacillus subtilis (ATCC 6633) and two
lines of this microorganisms that were isolated from one of the PW samples and were further
identified by DNA sequential. Parameters such as the time of interaction of the microorganisms with
the metal and the amount of cells were evaluated, together with the microorganisms lines (Bacillus
subtilis). After evaluation of experimental conditions, removals about 51, 90 and 15% were obtained
for Ba, Fe and Sr, respectively, for the synthetic solutions investigated. For Ca and Mg the efficiency
of removal was very low (lower than 5%). Based on the results obtained for Ba, Fe and Sr,
experiments were conducted in the real samples, in the same conditions. In this case, experiments
were conducted varying the cells content and sample mass and time where the pH and temperature
were kept at constant values. After the contact of the microorganisms with the samples and
consequent analytes removal, the microorganism was removed from the solution by centrifugation.
Metals were determined in the solution obtained after microorganisms removal and in the
microorganisms after sample digestion. Additionally, metals were also determined in the solution
obtained after washing the microorganisms cells with water. Analytes removal efficiency ranged from
63.1 to 91.1%, 23.3 to 49.0% and from 6.86 to 8.97% for Fe, Ba and Sr, respectively, depending on
the produced water sample used in the experiment and using the commercial line of Bacillus subtilis.
Similar results of metals removal were obtained using 2 other lines that were isolated from the
produced water samples. The results obtained for bioremediation were promising to promote the
removal of some metals from PW samples obtained from crude oil extraction in combination with
other processes.
1Neff J, Lee K., DeBlois E. M., Produced Water: Overview of Composition, Fates, and Effects in Produced Water, Springer,
2011
2Souza JP, et al., Journal of Analytical Atomic Spectrometry, 32, 2017, 2439
[UFRGS, CNPq, CAPES, FAPERGS]

378
GREEN METRICS FOR EVALUATION OF SAMPLE PREPARATION METHODS
USING NADES
Mario H. Gonzalez (R)a*, Sabrina S. Ferreira (PG)a, Thomas A. Brito (PG)a, Rafaela S.
Lamarca (R)a, Taciana G. S. Guimarães (PG)b, Karen C. Ferreira (PG)c, Ana P. R. Santana
(R)d, Andrea Oliveira (R)e, Clarice D.B. Amaral (R)e, Paulo C. F. Lima. Gomes (R)c
a São Paulo State University (UNESP), Department of Chemistry and Environmental Science, São José do Rio Preto,
SP, Brazil, 15054-000
b Federal University of São Paulo, Diadema, SP, Brazil, 09972-270
c São Paulo State University (UNESP), Institute of Chemistry, Araraquara, SP, 14800-060, Brazil
d Federal University of Minas Gerais, Department of Chemistry, Belo Horizonte, MG, 31270-901, Brazil
e Federal University of Paraná, Department of Chemistry, Curitiba, PR, Brazil, 81531-980
*e-mail: mario.gonzalez@unesp.br
The principles of green analytical chemistry (GAC) guide the development of analytical procedures
for sustainability. The crucial issue in GAC is the evaluation of the extent to which a process or
product is environmentally friendly.1 Different metrics have been created for the evaluation of
greenness, however the determination of the green nature of new analytical methods remains a
challenge, including for extraction methods that involve the use of natural eutectic solvents (NADES).
In this work, the environmental and sustainability characteristics of three different methods for the
preparation of plant samples were evaluated: microwave-assisted extraction (MAE), ultrasound-
assisted extraction (UAE) and microwave-assisted acid digestion (MW-AD). These methods2
employed different NADES based on xylitol, malic acid and citric acid as extraction solvents, as well
as dilute nitric acid as an oxidizing agent, for determination of As, Cd, Pb and V by ICP-MS. The
evaluation was carried out using five green metric tools: National Environmental Methods Index
(NEMI), Green Analytical Procedure Index (GAPI), Analytical Eco-Scale, Analytical AGREE
(AGREE) and White Analytical Chemistry (WAC). The particularities of each metric are presented,
highlighting the possibility to adapt the metric of choice based on the objective of the evaluation
which fits the desired application. The NEMI metric found no differences between the MAE-NADES
and UAE-NADES methods. The GAPI metric found differences among the MAE-NADES and UAE-
NADES methods and identify the disadvantageous aspects of each step of the methods. The
Analytical Eco-Scale and AGREE identified the MAE-NADES method as the greenest, while the
WAC indicated the UAE-NADES method as the greenest procedure. Divergences in the scores were
observed since these metrics are subdivided into qualitative, semi-quantitative and quantitative, and
some measure more aspects than others. It was also possible to identify differences related to the
types of solvents, showing that the use of xylitol/citric acid/water based NADES was the most
ecological, sensitive and accurate method. With the exception of NEMI, all the tools, with different
levels of complexity, were found to be useful for evaluating and comparing the greenness of sample
preparation methods, according to the objectives of the evaluator, and were able to include the main
GAC principles.
1 Płotka-Wasylka J., et al. Current Opinion in Green and Sustainable Chemistry, 31, 2021.
2 Santana, A.P.R. et al. Talanta, 216, 2020.
[CAPES, FAPESP, CNPq, INCT-DATREM]

379
EVALUATION OF ALKANES AND PAH LEVELS IN PM2.5 SAMPLES COLLECTED
DURING WET AND DRY SEASONS AT AN INDUSTRIAL REGION OF RIO DE
JANEIRO, RJ, BRAZIL
Nívia de M. Nascimento (PG)a*, Jéssica de M. Soares (UG)a, Bruno C. S. Rocha(R)a, Adriana
Gioda (R)b, Marcelo C. Bernardes (R)a
aFluminense Federal University, Geochemistry Departament, Niterói, Rio de Janeiro, Brazil, 24020-007
b Pontifical
Catholic University of Rio de Janeiro (PUC-Rio), RJ, Brazil 22451-900
*e-mail: nivianascimento@id.uff.br
This study analyzed alkanes and polycyclic aromatic hydrocarbons (PAH) in PM2.5 samples collected
in the district of Santa Cruz, metropolitan region of Rio de Janeiro during the wet (WS:January-April)
and dry seasons (DS:June-September) of the year 2019. Santa Cruz is an urban center located on
the coastal plain with an industrial complex. The methodology involved sampling through High
Volume Air Sampler (Hi-Vol), ultrasonic extraction with dichloromethane, clean-up by open
chromatography column filled with silica gel, analysis in gas chromatography (GC-FID and GC-MS)
and data treatment by statistical R Studio program.1 Regarding results for two seasons, the central
tendencies of total PAH (ng.m-3) was slightly higher than these alkanes results (ng.m-3). There was
a predominance of the odd long chain n-alkanes and the high molecular weight PAHs (5 and 6 rings).
Table 1. Medians diagnostic ratios of PAH in both seasons.4
Ratios Values Sources WS Median DS Median
Flt/(Flt+Pyr) 0.4-0.5 Fossil Fuel Combustion 0,42 0,41
>0.5 Diesel
Flu/(Flu + Pyr) 0,11 0,002
<0.5 Gasoline
0.2-0.5
Petroleum Combustion
Ind/(Ind+Bpe) 0,37 0,57
Grass, wood, coal combustion
>0.5
Abbreviations: Flt (Fluoranthene);Flu (Fluorene);Pyr (Pyrene); Ind(Indene[1,2,3-c,d]pyrene); Bpe(Benzo[g,h,i]perylene).
Furthermore, the values of Carbon Preferential Index (WS-1.22; DS-1.45), Cmax (WS-C31; DS-C31)
and diagnostic ratios of PAH (Table 1) pointed out as possible polluting sources traffic and industrial
emissions, biomass burning and road dust. Through the identification of pristane, phytane and
unresolved complex mixtures (UCM), it was noticed the presence of petroleum residues as well. 2,3
1Pereira et al. Air Qual. Atmos. Health 10(4), 2017, 457–68.
2Caumo et al. Atmosphere, 11(6), 2020, 1-19.
3Ceratti et al. Wat. Air and Soil Poll. 232(1), 2021.
4Tobiszewski and Namieśnik. Environ. Pollut. 162, 2012. 110–19.
[FAPERJ]

380
SIMULATION OF THE ADSORPTION CAPACITY OF PET MICROPLASTIC
EXPOSED TO HIGH METAL CONTAMINATION BY NI, CU AND CO UNDER
AQUEOUS SYSTEM
Cassiano A. R. Bernardino (R)a *, Claudio F. Mahler (R)a, Julia O. Fernandes (PG)b, Claudio S.
C. Lopes (PG)b, Bernardo F. Braz (R)c, Renata C. Borges (R)a, Ricardo E. Santelli (R)c,d,
Fernando H. Cincotto (R)c,d
a Universidade Federal do Rio de Janeiro, Programa de Engenharia Civil, Rio de Janeiro, RJ, Brazil, 21941-909.
b Universidade Federal do Rio de Janeiro, Programa de Pós-graduação em Química, Rio de Janeiro, RJ, Brazil, 21941-
909.
c Universidade Federal do Rio de Janeiro, Departamento de Química Analítica, Rio de Janeiro, RJ, Brazil, 21941-909.
d Instituto Nacional de Ciência e Tecnologia em Bioanalítica, Campinas, SP, Brazil, 13083-970
*e-mail: cassiano@coc.ufrj.br
Throughout the history of mankind plastic materials have been a part of modern society, and
according to previous reports a colossal scale production of several plastic based materials such as
polypropylene (PP), polyvinyl chloride (PVC), low-density polyethylene (LDPE), high density PE
(HDPE) and polyethylene terephthalate (PET) was already achieved during the 1950s with an annual
production around 5 million tons.1 Nowadays the continuous use of plastic materials has generated
a total of 6.300 million tons of plastic waste worldwide between 1950 and 2015. 2 Due to failures in
the recycling chain, the marine environment is one of the final destinies of plastic materials, leading
to a variable yet progressive scale of degradation of these plastic wastes, and finally generating
small plastic fragments called microplastics (particle size < 5 mm).3 The potential hazardous effect
of microplastic accumulation in the soft tissue of living being can be just one of the downsides of
microplastic deposition in the environment since other studies have reported the adsorption
properties of microplastic towards metal contaminants.4 Therefore, the objective of the present study
is to evaluate the levels of adsorption of PET microplastic when exposed to high amounts of Ni, Cu
and Co. The PET was collected in Guanabara Bay in the State of Rio de Janeiro and characterized
as to surface area, porosity, Fourier transform infrared spectroscopy with attenuated total reflectance
and scanning electron microscopy. The results showed a surface area of 0.24 m2 g-1, and the
presence of macro-and mesopores. Also, the presence of holes in the PET structure and functional
groups such as C-H stretching of the CH2 group, C=O and C-O stretching of the terephthalate group
and aromatic ring may have contributed to the adsorption of metals on the PET surface. The
construction and shape of the adsorption isotherms confirmed the presence of mesoporosity and
macroporosity on the PET microplastic surface. The Freundlich and Langmuir isothermal models
were used to study the adsorption process, and data processing and interpretation were
accomplished using pseudo 1st order and pseudo 2nd order kinetic models. The results indicated
that the adsorption process was well described by the Langmuir isotherm and pseudo-second order.
The removal rates by the PET microplastic varied from 24 to 93 % for Ni, 6.0 to 19 % for Cu and 6.0
to 38 % for Co after a period of 5 days. Also, the adsorption process was predominantly chemical
and extremely fast, indicating that the presence of microplastics in the environment can lead to a
rapid metal accumulation which elevates the hazards potential of microplastic in living beings.
1Napper, I. E.; Thompson, R. C. ;Glob. Challenges 4, 2020, 1900081.
2Geyer, R.; Jambeck, J. R.; Law, K. L. ;Sci. Adv. 2017, 3.
3Webb, H.; Arnott, J.; Crawford, R.; Ivanova, E. ;Polymers (Basel). 5, 2012, 1.
4Gao, X.; Hassan, I.; Peng, Y.; Huo, S.; Ling, L. ;J. Clean. Prod. 319, 2021, 128777.
[CNPq, FAPERJ]

381
WATER QUALITY FOR DRINKING WATER SUPPLY IN THE TAQUARI-ANTAS
WATERSHED, RS, BRAZIL
Juliana R. Rodrigues (UG)a*, Aline Teichmann (R)b, Demis P. Faqui (PG)c, Luana B. C.
Bender (PG)c, Fabiane Bartz (R)d, Mateus Guterres (R)d, Eduardo A. Lobo (R)b,c, Adilson B.
da Costa (R)b,c
aUniv.
de Santa Cruz do Sul, Depto de Eng., Arq. e Comp., Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96815-900
bUniv.de Santa Cruz do Sul, PPG em Sist. e Proc. Industriais, Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96815-900
cUniv. de Santa Cruz do Sul, PPG em Tec Ambiental, Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96815-900
dPhilip Morris Brasil, Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96835-749
*e-mail: juh.reinke@outlook.com
This study aimed to develop a diagnosis of the quality of water supply of 101 rural properties in the
in the Baixo Taquari Hydrographic Basin, central region of the state of Rio Grande do Sul, Brazil.
Water samples were collected directly at the point of use, after the residential reservoir, when
available. All analytical and sample collection methodologies used followed the standards described
in APHA (2005). In each of the samples, the following variables were determined: pH, temperature,
turbidity, hardness, electrical conductivity, total dissolved solids, carbonate alkalinity, bicarbonate
alkalinity, chloride, sulfate, nitrate, fluorine, iron, magnesium, manganese, sodium, potassium and
zinc. The analytical results indicate that 59 properties (58.4%) had problems with drinking water
supply. Critical potability variables are manganese (2 properties), sodium (10 properties), pH (16
properties), fluoride (40 properties), nitrate (2 properties) and sulfate (2 properties). The excessive
fluoride concentration is the main water potability problem, affecting 39.6% of the investigated
properties (40 properties), considering the limit established by Ordinance nº 10/1999, RIO GRANDE
DO SUL (1999), Figure 1. Despite being an important agent for the inhibition of dental caries, when
in adequate concentration (between 0.6 and 0.9 mg L-1), the consumption of water with excessive
concentration of fluoride is pointed out as the cause of a pathology called dental fluorosis, which
mainly affects children. The fluoride value in this set of samples reached values as high as 5.7 mg
L-1, 6.3 times higher than the maximum value allowed for human consumption. In addition to the
sanitary aspects, these water characteristics have resulted in a series of problems in the distribution
system, causing clogging, encrustation and burning of water heating equipment.
1 Standard Methods for the examination of water and wastewater, American Public Health Association, 21, 2005, USA.
2 Rio Grande do Sul. Portaria nº 10 de 16 de agosto de 1999.
[FAPERGS, CNPq, CAPES, Philip Morris Brasil]

382
MULTIVARIATE OPTIMIZATION OF HS-SPME/GC-MS PARAMETERS FOR
DETERMINATION OF 2-METHYLISOBORNEOL AND GEOSMIN IN WATER
Nicole H. Lazzari (PG)a*, Clara V. Miranda (UG)a, Tânia M. Pizzolato (R)a
aUniversidade Federal do Rio Grande do Sul, Instituto de Química, Porto Alegre, RS, Brazil, 91501 - 970
*e-mail: nicolehlazzari@gmail.com
2-methylisoborneol (2-MIB) and geosmin are cyanobacterial and actinobacterial secondary

metabolites which affect aesthetic quality of water. These compounds present taste and odour
described as earthy and musty, leading consumers to mistrust water treatment efficiency. Despite
the fact that there are no reported toxicological effects on humans or other animals caused by these
odorous compounds, they pose a social risk when it comes to economic burden, due to the
population expenditures resulting from switching to bottled water and the costs for alternative
operations in drinking water treatment plants¹. 2-MIB and geosmin events have been reported
worldwide in superficial waters, often in the order of nanogram per liter concentrations above their
odor threshold concentrations – 15 ng/L and 4 ng/L, respectively². Analytical methods with limits of
quantification in this order of magnitude are desired to detect the occurrence of these compounds.
Headspace solid phase microextraction followed by gas chromatography coupled to mass
spectroscopy (HS-SPME/GC-MS) analysis is an adequate extraction procedure for this kind of
analyte. This work presents the optimization of the parameters of HS-SPME/GC-MS for the
determination of 2-MIB and geosmin through a multivariate approach. A two-level fractional factorial
design with center point was used to determine the optimal conditions. The parameters were:
temperature (40°C, 60°C), fiber exposure time (20 minutes, 60 minutes), sodium chloride
concentration (10%, 30% w/v), sample volume (5 mL, 10 mL) and agitation level. Also, previously
three different fiber coatings were evaluated in eight extraction conditions in order to select the most
efficient. Divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) fiber coating was
selected and used in the subsequent essays. The limits of quantification (LOQs) of the optimized
method were 1 ng/L (geosmin) and 5 ng/L (2-MIB). External calibration was executed in the range
of the LOQs concentrations to 250 ng/L and the coefficients of determination (R²) obtained were
greater than 0,997 for both analytes. The methodology was used to analyse water samples from a
drinking water treatment plant in Porto Alegre, Rio Grande do Sul. The samples were collected from
Guaíba Lake at a location with previous reports of taste and odour events.
1Wang C, et al., Water Research, 172, 2020, 115507.
2Mustapha S, et al., Environmental Monitoring and Assessment, 193, 2021, 204.
[CAPES, PPGQ-UFRSG, IQ-UFRGS]

383
IMPACT OF COVID-19 PANDEMIC ON PATICULATE MATTER (MP10)
CONCENTRATIONS IN CURITIBA-PR
Luiz H. M. Rêgo (UG), Luana M. Milarski (UG), Victor B. Oliveira (UG), Mara F. Oliveira (PG),
Nilton L. Junior (UG), Roberta C. P. R. Domingues (R), Erika P. Felix (R)*
Federal University of Technology – Paraná, Academic Department of Chemistry and Biology, Curitiba, Paraná, Brazil,
81280-340
*e-mail: erikafelix@utfpr.edu.br
Particulate Matter (PM) is a complex mixture of solid and liquid particles of special concern due to
its adverse effects on human health and the environment.1 It should be highlighted that the
implementation of lockdown measures related COVID-19 pandemics has led to an improvement in
air quality worldwide. In this sense, although several studies have been reported to assess changes
in air pollution levels, studies in Brazil are scarce.2 Our research was performed to determine the
mass levels of particles (PM10) in different phases: pre, during and pos the restrictions period
(January 2020 to June 2022) in Curitiba – PR. The sampling site was chosen due to possible
pollutant sources such as traffic and industrial activities. PM10 samples were collected monthly by
using a high-volume air sampler (HiVol, Energética) at mean flow rate of 1.13 m3 min-1 for 24 h (total
volume collected of 1,63 x 103 m3) and glass fiber filters (Whatman, 20.3 cm × 25.4 cm). Prior to and
following the sampling procedure, the filters were kept in a desiccator for a 24-h period before they
were weighed three times using an analytical balance with the reading precision of 0.1 mg. After
sampling and weighing, the filters were stored at in an ultra freezer (– 80 ºC) until extraction and
chemical analysis for determination of cations and anions. Figure 1 shows the average PM10
concentrations at the sampling site.
Figure 1. PM10 concentrations from January 2020 to June 2022
It was not possible to quantify PM10 in March 2020 when restrictions started due to the outbreak of
the COVID-19 pandemic. This can be explained by the suspension of economic activities, schools,
universities, restricted social contact, restricted vehicles’ movement, among others. It can be noted
that after this period PM10 concentrations were not reduced, which agrees with research carried out
in the city of Rio de Janeiro in 2020 during the partial lockdown.3 Other studies also show a low
reduction for PM10. Since August (except for September) PM10 concentrations in 2021 were
generally higher than in 2020. The 24-hour standard established by CONAMA (50 μg m-3) was
exceeded in only five samples. In contrast the annual averages from 2020 (26.56 μg m -3) and 2021
(41.41 μg m-3) exceeded the annual standard (20 μg m-3). Meteorological data are being collected
and evaluated in order to establish their possible correlation with PM10 concentrations.
1Mukherjee A, Agrawal M, Environmental Chemistry Letters, 15, 2017, 283.
2Pérez-Martínez PJ, et al., Journal of Transport Geography, 101, 2022, 103349.
3Dantas, G, et al. Science of Total Enviroment, [s.l.], 28 abr. 2020.
[MPT-PR, APREA, SIMPEPAR]

384
LEVOFLOXACIN PHOTODEGRADATION ON Ag∕TiO FILMS 2
ARAUJO, K. C. M. (PG)a,b*, ARAÚJO, A. B. (R)c, TANAKA, A. A. (R)d,e

a UFMA, BIONORTE Graduate Program, São Luís, MA, Brasil, CEP 65080-420
b IFMA, Scholl Board/IFMA – ZD, Zé Doca, MA, Brasil, CEP 65365-000
c IFMA, Scholl Board/IFMA – CH, São Luís, MA, Brasil, CEP 65010-030
d UFMA, Department of Chemistry, São Luís, MA, Brasil, CEP 65080-805
e Instituto Nacional de Ciência e Tecnologia em Bioanalítica, C.P. 6154, Campinas, SP, Brazil, CEP 13083-970
*e-mail: karla.caroline.ma@gmail.com.br
Fluoroquinolones (FQs) are antibacterial drugs often found in different environmental matrices,
particularly aqueous1. Even at low concentrations, they can induce bacterial resistance, causing
damage to human and animal health2. In view of this, more efficient methods of water treatment,
such as heterogeneous photocatalysis, have been shown to be quite effective in removing FQs 3,
especially with the use of TiO2 4. However, in most studies, TiO2 is used in the form of a suspension,
with loss of catalyst in reuse during the lengthy filtering stage, making treatment more expensive
and, in many cases, making its use unfeasible on a large scale5. In view of this, the present work
presents a study of the efficiency in the reuse of Ag/TiO2 films synthesized by the sol-gel method
and immobilized on borosilicate glass6 support, assisted under UV-A irradiation (365 nm), for the
remediation of aqueous matrices containing levofloxacin (LEV), a FQ present at trace level 3.
Analyzes of degradation were also performed by HPLC with a fluorescence detector and the results
obtained showed that remediation by photolytic treatment is not an effective process for the
elimination of LEV in aqueous matrices 1, 3, 4, due to the low removal rate by a long exposure time to
UV irradiation (~12 hours). On the other hand, the photocatalytic process using Ag/TiO2 films
supported on borosilicate glass proved to be a great alternative for the treatment of water
contaminated with LEV, with degradation rates above 90% after ~90 min. In contrast, by photolysis,
the removal was only 26%. Furthermore, it was observed that the Ag/TiO2 films did not undergo
leaching during the experiments carried out, a fact confirmed by the reproducibility of the removal
rates throughout the work. Therefore, the use of these films proved to be much more viable when
compared to suspended catalysts, since the possibility of extensive reuse and ease of use would
result in considerable cost savings in the process of removing FQs in water treatment systems.
1 Rodrigues-Silva C, Maniero MG, Peres MS, Guimarães JR, Química Nova, 37, 2014, 868.
2 Khan NA, et al., Trends in Analytical Chemistry, 129, 2020, 1.
3 Pretali L, Fasani E; Sturini M, Photochemical and Photobiological Sciences, 2, 2022, 899.
4 Pretali, L, Maraschi F, Cantalupi A, Albini A, Sturini M, Catalysts, 10, 2020, 1.
5 Varshney G, et al., Coordination Chemistry Reviews, 306, 2016, 43.
6 Araújo AB, Jorge SMA, Padilha PM, Bozano DF, Castro GR, Florentino AO, Global Journal Inc., 15, 2015, 17.
[FAPEMA, CNPq]

385
EVALUATION OF PESTICIDES IN CAMBISOL SOIL
Pietra T Expósito (UG)a*, Danielle C Schnitzler (R)a
aFederal Technological University of Paraná, DAQBi, Curitiba, Paraná, Brazil, 81280-340
*e-mail: Pietratridaexp@gmail.com
Pesticides are synthetic chemical products used to kill insect larvae, fungi, among others, under the
justification of controlling the diseases caused by these vectors and regulating the growth of
vegetation1. Brazil is one of the biggest consumers of pesticides in the world, and the second biggest
buyer when it comes to pesticides already banned in Europe.2 In 2020 alone, 493 chemical
compounds were released by Anvisa.3 The International Labor Organization (ILO) states that
pesticides cause 70,000 acute and chronic poisonings per year and that progress to death in
developing countries. There are also more than seven million cases of non-fatal acute and chronic
diseases recorded1. Thus, it is necessary to efficiently determine the presence of these pesticides
even at low concentrations, since they can be toxic at any concentrations. In this scenario, the
objective of this study was to evaluate the contamination in ppb levels of Atrazine, Trifluralin, Alachlor
and Chlorpyrifos in soil samples. The samples were collected in the Contestado Settlement of
Lapa/PR, on the surface and at a depth of 20 cm in cambisol. For extraction, the samples were
previously sieved in an 18 mesh opening (0.250 mm). The extraction procedure was performed by
ultrasound (40 kHz) at room temperature, with 20 mL of ethyl acetate (HPLC grade), for 45 min.
Then filtered (0.45 μm membrane, Macherey Nagel, 47 mm diameter, GF-1) under vacuum. The
filtered samples were collected in 20 mL beaker and passed through the clean-up in florisil
cartridges. After being collected in a glass flask, the solvent was evaporated in a rotary evaporator
(IKA), at 923 mbar, 100 rpm, with a bath at 40ºC, and a water circulation system at 10ºC.
Reconstitution of analytes was performed in 1 mL of ethyl acetate and extracts were filtered through
0.22 μm PVDF/PTFE (13 mm diameter, Filtrilo), and stored in an amber vial. The procedure was
performed in triplicate. Samples were prepared and analyzed on the same day and kept in a freezer
until injection into GC-MS/MS. Table 1 shows the concentrations of pesticides in the samples after
analysis in GC-MS/MS. The values were obtained through an external calibration curve.
Table 1. Concentration values in ppb for Alachlor, Atrazine, Chlorpyrifos and Trifluralin, at the surface and at a depth of
20cm for the soil sample.
Sample Trifluralin Atrazine Alaclhor Chlorpyrifos
Surface 3.78±0.17 1.66±0.67 2.63±0.42 2.10±0.2
20 cm 4.9±1.07 1.57±0.17 3.31±0.28 3.14±0.6
Source: own authorship
Considering the values of Limit of quantification (LQ) 0.59; 0.5; 1.67 and 1.16 ppb for Trifluralin,
Atrazine, Alachlor and Chlorpyrifos respectively, it is observed that the method is sensitive, reaching
concentrations in the range of 1 ppb, which are far below those obtained in the literature.
1National Cancer Institute. Pesticides, according to the WHO, 20,000 deaths are recorded a year due to the consumption
of pesticides. Accessed on: July 28, 2022.
2Aranha, A. et al. Brazil is the 2nd largest buyer of banned pesticides in Europe, which imports food produced with these
chemicals. Public Agency.
3Soares, G. Release of pesticides and industrial components set a new record in 2020. Power 360.
[DAQBi, LEMASSA, LAMEAA, LAMAQ, UTFPR]

386
QuEChERS EXTRACTION FOR MYCOTOXIN DETERMINATION IN
WASTEWATER
Lima da Silva, Juliane (PG)a, Garda-Buffon, Jaqueline (R)a*
aUniversidade Federal do Rio Grande, Escola de Química e Alimentos, Rio Grande, RS, Brazil, 96203-900
*e-mail: jaquelinebuffon@furg.br
Mycotoxins are secondary metabolites produced by filamentous fungi and are known to infect food
and feed worldwide under favourable conditions. The most influential parameters favouring
mycotoxin production are moisture content and water availability in storage. Mycotoxins have
harmful effects on animals and humans, which may cause carcinogenic, teratogenic, hepatoxic,
nephrotoxic, immunotoxicity and other effects.¹ Although these compounds occurrence has been
extensively studied in food and feed, little is known about their environmental fate.² However, the
evaluation of their environmental exposure represents an important part of their hazard assessment.
As a start in closing this gap, the aim of this study was to develop an analytical method for the
determination of deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZEA) and aflatoxin B 1
(AFB1) in wastewater collected at Lagoa dos Patos – RS. In order to reduce the matrix effects by
reducing the interferences from the extraction step, the method QuEChERS was tested. Mycotoxins
are mostly soluble in polar and slightly polar solvents, thus, polar solvents and mixtures of solvents
such as acetone, acetonitrile, chloroform, dichloromethane, ethyl acetate or methanol with small
amounts of acids or water are commonly applied in the extraction. In this study, extracting solvent
was acetonitrile for DON and AFB1, and acetonitrile with 1% acetic acid (v v-1) for OTA and ZEA.
Wastewater spiked with mycotoxins was agitated in vortex (30 sec) and ultrasound (3 min), followed
by 30 min of rest. Afterward, 10 mL of extracting solvent was added and the mixture was subjected
to orbital agitation (200 rpm, 10 min). In the mixture was added 4g of MgSO4 and 1g of NaCl, vortexed
(30 sec) and subjected to an ultrasonic bath (3 min), followed by 5 min of centrifugation (3220 g).
Finally, an aliquot of 2.5 mL of the organic fraction was dried and resuspended in 1 mL of mobile
phase (water: acetonitrile 90:10 (v v-1) for DON and AFB1 and acidified water: acetonitrile 40:60 (v v-
1
) for OTA. Mycotoxins were quantified by liquid chromatography coupled to a fluorescence detector
(OTA, ZEA and AFB1) and array of diodes (DON). Parameters validated for the mycotoxin
determination were linearity, analytical curve, correlation coefficient, limit of detection (LOD), limit of
quantification (LOQ), precision and accuracy. Mycotoxins LOQ were 0,5 µg mL-1; 0,15 ng mL-1; 0,4
ng mL-1 and 0,02 µg mL-1 for DON, AFB1, OTA and ZEA, respectively. In general, it is recommended
that the LOQ of the trace contaminant analysis method be equal to or lower than the maximum limits
allowed by legislation. However, there is no legislation worldwide that limits the contamination of
wastewater for mycotoxins. It was possible to observe good linearity in the studied concentration
range since all mycotoxins showed correlation coefficients of at least 0.999. The reliability of the
mycotoxin extraction method and equipment quantification were evaluated according to the
responses in terms of recovery. The levels evaluated were 1, 5 and 10 times the LOQ. The recovery,
expressed in %, were 90 ± 3, 79 ± 2 and 96 ± 11 for DON, 76 ± 6, 74 ± 4 and 75 ± 7 for AFB1, 82 ±
1, 87 ± 3 and 80 ± 3 for OTA and 70 ± 13, 86 ± 0,5 and 91 ± 0,8 for ZEA at the levels 1, 2 and 3,
respectively. Recoveries are considered adequate and satisfactory for trace analysis in foods by
parameters described in validation guides. As a conclusion and future perspective, the method
validated is able to evaluate mycotoxins occurrence in wastewater of Rio Grande region.
1 Chandravarnan P, Agyei, D, Ali, A., Trends in Food Science & Technology, 2022.
2 Juraschek LM, Kappenberg A, Amelung W, Science of the Total Environment, 2022.
[CAPES, CNPq, FAPERGS]

387
EVIDENCE OF HIGH BIOACCESSIBILITY OF GADOLINIUM-CONTRAST
AGENTS IN NATURAL WATERS AFTER HUMAN ORAL UPTAKE
Laís A. Souza (R)a*, Rodrigo M. A. Pedreira (PG)a, Manuel Miró (R)b, Vanessa Hatje (R) a
aUniversidade Federal da Bahia, Instituto de Quimica, Salvador, Bahia, Brazil, 40170-115
b Universitat des les Illes Baleares, Departamento de Química Analítica, Palma de Mallorca, Illes Baleares, Spain, E-
07122
*e-mail: lais2.araujo@gmail.com.br
Considering the large occurrence of anthropogenic Gd concentrations in natural waters, its

continuous usage increase in technology developments and products and the lack of data on
potential Gd human exposure due to ingestion of contaminated waters, it is urgently needed to
understand how gadolinium contrast agents (Gd-CAs) reacts in the human digestive system.
Considering the occurrence of anthropogenic Gd concentrations in tap water and beverages,1,2,3 the
lack of data on potential Gd-CAs lowdosage exposure due to long-term ingestion of contaminated
waters, and the absence of legal maximum threshold values available, it is urgently needed to
understand how Gd-CAs reacts in the human digestive system. Here, we aimed to i) identify through
in vitro bioaccessibility tests whether Gd-CAs can be potentially assimilated by humans after orally
ingestion; ii) evaluate if there is a significant difference in the oral bioaccessibility values between
contrast agents; and iii) estimate the bioaccessibility of anthropogenic Gd already reported for tap
waters worldwide. Gd-CAs are highly bioaccessible (77 to 112%). The macrocyclic complexes pose
the highest potential risk, because there are more stable than linear complexes in the gastrointestinal
tract and, as such, tend to remain in solution and thus might bring Gd at the intestinal barrier making
it potentially bioavailable. The estimated range of potential intake of Gd varied from 13 to 4839 μg in
a lifespan of 70 years. The high bioaccessibility of anthropogenic Gd in tap waters calls for
appropriate actions to develop better practices to treat wastewater contaminated by Gd-CAs in order
to safeguard population and ecosystem health. With the increasing use of RMI worldwide, which has
escalated even more because helps diagnose heart damage during the COVID-19 pandemic, our
results (high Gd bioaccessibility) suggest that anthropogenic Gd should be evaluated more routinely
in tap and superficial waters. In addition, appropriate actions to develop better practices to treat
wastewater contaminated by Gd-CAs are called for in order to safeguard the population and the
ecosystem's health.
1 Kulaksız, S., Bau, M., Appl. Geochem. 26 (11), 2011, 1877–1885.
2 Tepe, N., Romero, M., & Bau, M. Appl. Geochem. 45, 2014, 191–197.
3 Schmidt, K., Bau, M., Merschel, G., Tepe, N. Sci. Total Environ. 687, 2019, 1401–1408
[GASBRAS/FINEP, CNPq, AEI/MICINN]

388
ANALYTICAL CHEMISTRY &
ENERGY
389
A NOVEL APPROACH FOR DETERMINATION OF PROTEIN IN FOODS
EXPLOITING MICRODESTILATION AND SMARTPHONE-BASED
COLORIMETRY
Isabela C. Gonçalves*a, Sâmara Soaresa, Fábio R. P. Rochaa

aCenter for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil, 13416-000
*e-mail: isabela.camargo.goncalves@usp.br
One of the most usual analyses in food quality control is the determination of proteins, due to their
nutritional importance and interest by the food industry. The Kjeldahl method1, the most usual for
protein determination and a reference to other indirect methods, is time-consuming and generates
significant amounts of waste. In this work, a simple, cost-effective, and environmentally friendly
procedure based on microdistillation of ammonium from Kjeldahl digests, absorption on an acid-base
indicator solution, and digital-image colorimetry is proposed for protein determination in food. The
lab-made microdistillation system2 consists of a 1.5 mL Eppendorf tube containing the acceptor
solution suspended inside a 50 mL Falcon capped tube containing 500 μL of the sample acid digest
and 1.0 mL of sodium hydroxide 10% (m/v). The system is submitted to controlled heating in a water
bath at 55 ºC for 40 min to promote the ammonia distillation to the headspace, thus favouring its
absorption into the acceptor solution (a 0.001% phenol red aqueous solution). The digital images
were acquired by a smartphone camera under controlled conditions (illumination, distance from the
camera to the Eppendorf, and region of interest) and the intensity of the reflected radiation was
converted to the RGB spectral space using a free app (PhotoMetrix®). As measurements were based
on the consumption of the acid form of the indicator (pink), values of the G channel were used as
analytical parameter because of the color complementarity. A linear response was achieved within
5.0-50.0 mg L-1 ammonium (r = 0.996), as confirmed by the lack-of-fit test, with coefficient of variation
(n = 10) and limit of detection (95% confidence level) estimated at 2.3% and 3 mg L -1, respectively.
The procedure requires only 3 μg of phenol red indicator, 100 mg of sodium hydroxide, and 500 μL
of sample digest and generates as low as 1.8 mL of residue per determination. With simultaneous
processing, a sample throughput of 12 h-1 was achieved. The procedure was successfully applied to
protein determination in milk, beans, and lentils with results in agreement with the micro-Kjeldahl
reference procedure at the 95% confidence level.
1 Brasil. Ministério da Agricultura, Pecuária e Abastecimento. MET POA/11/02/01.2013.
2 Soares, S.; Rocha, F.R.P. Talanta, 199, 2019, 285-289.
––––––––––––––––––––– Analytical chemistry & Energy –––––––––––––––––––––

390
EVALUATION OF GLASS MICROFIBER FILTER AS A SUPPORT FOR THE
ANALYSIS OF ORGANIC SOLVENTS BY LASER-INDUCED BREAKDOWN
SPECTROSCOPY
Layla P. Santos (PG)*, José Francielson Q. Pereira (R), Ivo M. Raimundo Jr (R)
State University of Campinas, Institute of Chemistry, PO Box 6154, 13083-970, Campinas (SP)
*e-mail: l234913@dac.unicamp.br
The direct determination of liquids in LIBS is usually difficult, encompassing experimental issues, such
as formation of splashes that can reach the lenses of the system1. Therefore, it is usual to transfer the
solution to a solid substrate, being the analysis performed after the evaporation of the solvent. This
strategy is quite straightforwardly for inorganic analysis, since the supports are organic materials, being
the paper the most common example2. However, this approach does not work for the Laser-Induced
Breakdown Spectroscopy (LIBS) for organic solvent analysis, fuels and biofuels3. Microfiber filters made
of borosilicate glass are commonly used in fast filtration, have the ability of retaining a large amount of
particles (0.5µm) and are resistant to most organic and inorganic solvents. In this work, glass microfiber
filters with a thickness of 25 mm and basis weight of 140 g/m² were used. LIBS measurements of organic
compounds were performed by employing a pulsed (5 ns) Nd:YAG laser (1064 nm), with pulse energy of
80 mJ, operating at 20 Hz. The laser pulse was focused on the filter by means of a lens with a focal
distance of 10 cm; the emitted radiation was collected with a 5-cm diameter lens, coupled to an optical
fibre with diameter of 105 µm, which guided the radiation to an echelle polychromator. An ICCD detector
(iStar DH 734, Andor Technology) was employed for spectral acquisition, with a gate delay of 2.0 µs and
gate width of 20 µs. The glass microfiber filters was placed on a rotating device, permitting to obtain a
spectrum with integration of 30 laser shots, focalised on different points. Figure 1 shows the spectral
regions of the dry borosilicate glass microfiber filters and after addition of 50 µL of deionized water. It is
notable the signal reduction after the deposition of water, producing an adequate baseline for solvent
measurement. The baseline is maintained constant up two set of shots, as the glass microfiber filters is
damaged during the third series of shots. Figure 1a-c shows the C I, C2, CH emissions obtained with the
addition of 30 µL of toluene on the filter, previously humidified with deionized water (50 µL). These results
indicate that glass microfiber filters is a useful support for LIBS analysis of organic compounds and
solvents.
Figure 1. Spectral signals provided by dry (----), humidified (····) glass filter, followed by the addition of toluene ( ̶ ̶ ̶ ).
1 Lazic V, Jovicevic S, Spectrochimica Acta Part B-Atomic Spectroscopy, 101, 2014, 288.
2 Zhang Y, Zhang TL, Li H, Spectrochimica Acta Part B-Atomic Spectroscopy, 181, 2021, 106218.
3 Moros J, Laserna J, Applied Spectroscopy, 73, 2019, 963.
[FAPESP, INCTAA, CNPq, FAPESP, CAPES]

391
ANALYSIS OF VOLATILE COMPOUNDS IN BORDER REGION FLIGHT
Luana E Knaul (PG)a*, Priscila M M Ramos (R)b , Aline T Toci (R)b, Andre L Rudiger (R)b,
Marcelo Kapp (R)b, Marcela Boroski (R)a,b
aFederal University of Latin American Integration (UNILA), PPGIES/ILATIT, Foz do Iguaçu, Paraná, Brazil, 85866-000
bFederal University of Latin American Integration (UNILA), ILACVN Foz do Iguaçu, Paraná, Brazil, 85866-000
*e-mail: luanaknaul16@gmail.com
Extra virgin olive oils (EVOO) are known for their aroma formed by numerous volatile organic
compounds (VOC), but EVOO are susceptible to adulteration with other oils and other substances.
This study aims to evaluate possible sources of tampering in EVOO through VOC analysis in
samples sold in the triple border region formed by the cities of Foz do Iguaçu (Brazil), Ciudad del
Este (Paraguay) and Puerto Iguazu (Argentina). 19 samples were collected between the years 2018-
2019 from Argentina (9), Portugal (5), Spain (3) and Chile (2). Previously, the analysis of the fatty
acid composition was carried out and 7 adulterated samples were detected. The analysis was
performed using the solid phase microextraction technique followed by gas chromatography coupled
to a mass detector (SPME/CG-MS) to detect volatile organic compounds and possible adulterating
compounds ¹ ². The presence of 100 volatile organic compounds was detected. The greatest
predominance were organic acids (19) followed by esters (17), aldehydes (14) and alcohols (14), in
smaller amounts ketones (8), ethers (6), hydrocarbons (6), phenols (6), organosulfur (2) and other
compounds (8). The compounds hexanal, E-2-hexenol, Z-3-hexenol and 1-hexanol characteristic of
high quality EVOO samples were identified in the samples, in which E-2-hexenol and 1-hexanol can
be used as a compound assist in determining the origin of EVOO, according to the literature ³ 4. In
the 7 adulterated samples, it was possible to evaluate the presence of 24 possibly adulterating
compounds, which were not present in the authentic EVOO samples. The composition of possible
adulterants was varied, corresponding to 41,7% esters, 12,5% aldehydes and 8,3% alcohol. Esters
are added to oils in order to impart significant aromas similar to EVOOs in order to camouflage
adulteration with other low quality oils. On the other hand, aldehydes such as 2-heptenal, E-2-
octenal, EE-2,4 decadienal and others are originated in vegetable oils as their oxidation occurs,
however an EVOO when adulterated with sunflower, soybean and corn oil, for example , will have
its highest percentages proportional to the levels of oxidation 5. The same applies to the high
percentage of alcohol, as the 1-octen-3-ol that is present in the adulterated samples, has its
concentration increased according to the level of oxidation of the oil. Through the analysis of VOCs,
it was possible to differentiate the samples according to the origin of the EVOOs, showing that this
can be used as a method of detecting adulterations.
1 Fernandez MLA, Assof M, Jofre V, Silva MF, Food Analytical Methods, 7, 2014, 2122.
2 Persurić Ž, Saftić L, Masek T, Pavelić SK, A case study, Lwt, 95, 2018, 326.
3 Cavaliere B, et al., Journal of Agricultural and Food Chemistry, 55, 2007, 1454.
4 Ozcan-Sinir G, Food Control, 188, 2020, 107433.
5 Guillen MD, Goicoechea E, Food Chemistry, 111, 2008, 157.
[PRPPG/UNILA), Fundação Araucária de Apoio ao Desenvolvimento Científico e

Tecnológico do Estado do Paraná, CAPES, SETI,LEIMAA ]

392
DESENVOLVIMENTO DE METODOLOGIA PARA DETERMINAÇÃO
QUANTITATIVA DE CHUMBO POR ESPECTROFOTOMETRIA UV-VIS.
Alexandre Muchon, Alex Almeida (R)

UNIFOR – MG, Centro Universitário de Formiga
A determinação de metais muitas vezes é realizada com o auxílio de equipamentos de ponta que
por sua vez detêm um alto preço de aquisição, manutenção e utilização, fatores que tornam-se um
empecilho ou barreira para pesquisadores iniciantes e pesquisas de base. Por estes motivos a
utilização de métodos voltados como por exemplo, para a espectrofotometria UV-Vis destacam-se
como uma opção viável para diversas pesquisas, tais como a determinação de metais pesados. O
chumbo por sua vez é um elemento que não apresenta nenhuma função essencial para o ser
humano, e a presença do mesmo torna-se motivo de grande preocupação, necessitando então de
que hajam formas de detecção do metal em alimentos e no ambiente. Diante desta necessidade e
em busca e formas alternativas na determinação de chumbo, é proposto neste trabalho a avaliação
do metal utilizando o reagente analítico 8-hidroxoquinolina.

393
DETERMINATION OF Ca, Mg, K, AND Mo IN PLANT MATERIALS BY LASER-
INDUCED BREAKDOWN SPECTROSCOPY
Sady L. Almeida (PG)a*, Aline de C. Elias (PG)a, Juliana Naozuka (R)b, Alex Virgilio (R)c,
Fábio R. P. Rocha (R)c, Cassiana S. Nomura (R)a
a University of São Paulo, Department of Fundamental Chemistry, Institute of Chemistry, São Paulo, SP, Brazil,
05508-220.
b Federal University of São Paulo, Department of Chemistry, Diadema, São Paulo, SP, 04021-001.
c Center for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brasil, 13416-000.
*e-mail: sadylobo@usp.br
Laser-induced breakdown spectroscopy (LIBS) is a versatile and promising analytical technique

which allows direct analysis of solid materials from different sources. LIBS also shows as
characteristics a multi-element capacity, fast response and in situ or remote analysis due to the
availability of portable instruments 1. However, the difficulties in performing quantitative analysis,
mainly due to matrix effects, are challenges that need to be overcome in order to be competitive with
other well-established techniques used for quantitative analysis of complex samples2. In this context,
the selection of appropriate calibration approaches is imperative. External calibration (EC) is the
most usual strategy, but the need for at least 3 standards covering the analyte mass range fractions
is a challenge. In the present work a new calibration approach known as slope ratio calibration
(SRC) was evaluated. SRC is based on the proportionality between the emission intensity of the
analytes and the amount of ablated mass for the sample using a single solid calibration standard
with a similar matrix3. The analytes mass fractions are determined by the slope ratio of a standard
and the test sample. The viability of SRC-LIBS was evaluated for Ca, Mg, K and Mo determination
in plant Certified Reference Materials (CRMs), NIST SRM 1515 Apple Leaves and NIST SRM 1547
Peach Leaves. Analyzed samples were pressed using an automatic hydraulic press (Xpress 3635,
Spex SamplePrep) by transferring 0.35 g of sample and 0.15 g of paraffin binder to a 13 mm die set
and applying 8 ton cm−2. Experiments were carried out with a LIBS J200 Tandem (Applied Spectra,
USA) composed of a Nd:YAG laser operating at 266 nm. LIBS instrumental parameters as laser
energy (20 mJ), delay time (0.50 µs) and laser spot size (65 µm) were optimized aiming the highest
emission signals and signal-to-background ratios. The SRC was carried out using 135, 203, 270,
337, 405 and 445 laser pulses. The accuracy of the SRC and EC methods were evaluated by
comparing the results obtained and the certified reference values. For the SRC linear correlation
coefficients higher than 0.98 were obtained for all the analytes and quantitative recoveries were Mg
(95%), Ca (113%), K (109%) and Mo (88%) were obtained. In contrast, the recoveries of EC method
were Mg (127%), Ca (175%), K (101%) and Mo wasn’t determined due to lack of linearity, probably
caused by matrix effects. Overall, the proposed SRC method demonstrated better accuracy and
simplicity when compared with EC.
1 Costa, V. C.; Augusto, A. S.; Castro, J. P.; Machado, R. C.; Andrade, D. F.; Babos, D. V; Sperança, M. A.; Gamela, R.
R.; Pereira-Filho, E. R. Química Nova, v. 42, 2019, 527.
2 David A. Cremers & Rosemarie C. Chinni (2009), Applied Spectroscopy Reviews, v. 44, 2009, 457.
3 Nunes, Lidiane C.; ROCHA, Fábio R. P.; KRUG, Francisco J. Journal of Analytical Atomic Spectrometry, v. 34, 2019,
2314.

394
DETERMINATION OF CALCIUM IN CRUDE OIL USING COLORIMETRY WITH
DIGITAL IMAGES
Luiza Baumann (PG)a, Kéllen F. Anschau (PG)a, Caroline M. Senger (PG)a, Jennefer M.
Queiroz (UG)a, Edson I. Müller (R)a*
*e-mail: edson_muller@yahoo.com.br
Crude oil extraction is usually realized together with emulsified water, which are formed due to the
turbulent flow in the pipelines. These emulsions contain some unwanted species, such as sediments,
high water content and salts.1 Among the salts associated with the aqueous phase of water/oil
emulsions, there are the calcium salts, which, when subjected to high temperatures, react with the
water vapor forming the hydrochloric acid. As a result, corrosion of the crude oil distillation equipment
occurs.2 For this reason the objective of this work was to develop a colorimetric method, based on
complexation reactions with calcium using calcein, murexide and calcon, for the determination of
calcium in petroleum. For that, a portable system was used based on the capture of digital images
by an endoscopic camera and the processing and treatment of the images was done using the
PhotoMetrix PRO app.3 At first, the range of pH and concentration of each complexing agent
necessary for the production of stable complexes with calcium were evaluated. After that, a three
variable Doehlert matrix (DOE), quadratic model, was also used to evaluate the influences of the
total volume of solution (1 to 2 mL), camera focal length (2 to 20 mm) and light intensity (30 at 180
lx) for digital image acquisition. In addition, the regions of interest for capturing digital images (32x32,
64x64 and 128x128 pixels) were optimized and the possible interference caused by the presence of
magnesium was evaluated. All images were processed in the PhotoMetrix PRO application using
the multivariate calibration model by the partial least squares (PLS) method. The sample preparation
method used was based on the ASTM D4807 standard to isolate the saline species contained in the
crude oil in a 0.45 μm membrane after filtration. To evaluate the predictive capacity of the PLS model,
5 samples of crude oil were selected, with calcium concentrations ranging from 5 to approximately
900 mg kg-1. The accuracy of the method was evaluated comparing the results with the values
obtained by microwave-assisted wet digestion using single reaction chamber (MAWD-SRC) with
determination by ICP OES. Determination coefficients greater than 0.997 were obtained for the
calibration models built for the three complexing agents, using 5 factors. The RMSECV values
obtained for the regression model for calcein, murexide and calcon were 0.26, 0.35 and 0.32 mg L-
1
, respectively. Besides, no significant differences were observed between the proposed method and
the reference methods (ANOVA, 95% confidence interval). The detection and quantification limits
were 0.62 and 2.05 mg kg-1, 0.12 and 0.40 mg kg-1 and 0.15 and 0.51 mg kg-1, for the complexing
agents calcein, murexide and calcon, respectively. No significant interference of magnesium was
observed in petroleum samples used in this work. Finally, the digital images with PhotoMetrix are
considered alternative method for the determination of Ca crude oil samples, due to its possibility of
miniaturization, low cost, operational ease and speed in obtaining and processing digital images.
1Umar AA, Saaid IBM, Sulaimon AA, Pilus RBM, Journal of Petroleum Science and Engineering, 165, 2018, 673.
2Gray MR, Eaton PE, Le T, Petroleum Science and Technology, 26, 2008, 1924.
3Costa ABD, Helfer GA, Barbosa JL, Teixeira ID, Santos RO, Santos RBD, Voss M, Schlessner SK, Barin JS, Journal of
the Brazilian Chemical Society, 32, 2021, 675.
[CNPq, CAPES, UFSM]

395
DETERMINATION OF INORGANIC CONTAMINANTS IN CACHAÇA BY ICP-OES
Cleidiane G. Lima (R)a*, Mairlane S. de Alencar (PG)b, Denilson G. Queiroz (R)a, Crisiana de
A. Nobre (R)a, Renata de O. Silva (R)a, Vitor P. de A. Silva (R)a
a Nucleus of Technology and Industrial Quality of Ceará-NUTEC, Instrumental Chemistry Laboratory-LQI, Fortaleza, CE,
Brazil, CEP 60.440-552
b Federal University of Ceará, Department of Chemical Engineering , Fortaleza, CE, Brazil, CEP 60.440-900
*e-mail: cleidiane.lima@nutec.ce.gov.br
The cachaça is a genuinely Brazilian drink with an alcohol content of 38% to 48%, being the second
most consumed alcoholic beverage in our country. It can be produced by hand in copper stills or by
industrial process in stainless steel column. It is very important to know the chemical composition of
cachaça, both the organic and inorganic parts, so that the injection of substances harmful to human
consumption can be controlled. This study aimed to determine the inorganic contaminants copper
and lead, volatile acidity and alcohol content in artisanal and industrial cachaças produced in the
state of Ceará and compare them with the limits required by Normative Instruction nº 13 of June 29,
2005, as well as with the European legislation. 5 artisanal samples and 5 industrial samples were
analyzed. Copper and lead contaminants were quantified by Inductively Coupled Plasma Optical
Emission Spectrometry (ICP-OES). The alcohol content and volatile acidity through the methodology
described in the analytical Norms of the Instituto Adolfo Lutz (2008). Among the analyzed samples,
seven showed compliance with the limits required by Brazilian legislation. An artisanal cachaça had
a copper content of 6.184 mg/L, with a maximum limit of 5 mg/L. Comparing with the European
legislation, only 3 samples were within the established limits, being two industrial and one artisanal.
The others had a copper content of 3.590 mg/L; 4.340 mg/L; 2.443 mg/L; 4.809 mg/L; 6.184 mg/L;
2.295 mg/L and 3.853 mg/L, where the maximum allowed limit is 2 mgL. Two samples of industrial
cachaça had alcohol content of 31.8% and 30.8%, values lower than the minimum required limit,
which is 38%. The high copper content is mainly due to the distillation of artisanal cachaça being
done in copper stills, where the main contamination by this metal occurs.
1 Castro JK, Vieira LP, Batista SD, Quimica Nova, 21, 2007, 16
2 Lajunen LHJ, Spectrochemical Analysis by Atomic Absorption and Emission, The Royal Society of Chemistry, 1991.
3 Aquino. F. W; Nascimento, R. F.; Rodrigues, s. ; Cassimiro, a. R. S. . Determinação de marcadores de envelhecimento
em cachaças. Ciência e Tecnologia de Alimentos, Campinas, v. 26, p. 145-149, 2006.
4 Assunção DA, Pinto BG, Leal AC, Augusti R, Melo JOF, Ramos AS, Silva MR, Capobiango M. Research, Society and
Development, v. 9, 2020, e55491110054
[FUNCAP, SECITECE, UFC, NUTEC]

396
DETERMINATION OF POLYCYCLIC AROMATIC NITROGEN HETEROCYCLES
IN FINE AIRBORNE PARTICLES USING A GREEN MINIATURIZED
EXTRACTION PROCEDURE
*Pedro Victor Bomfim Bahia (PG)a,b,c, Valdiria Almeida de Oliveira (UG)a, Madson Moreira
Nascimento (R)b,c, Leila Oliveira Santos (R)d, Gisele Olimpio da Rocha (R)a,b,c, Jailson
Bittencourt de Andrade (R)b,c,e, Maria Elisabete Machado (R)a,b,c
a Universidade Federal da Bahia, Instituto de Química, Programa de Pós-Graduação em Química;
b Centro Interdisciplinar de Energia e Ambiente – CIEnAm;
c Instituto Nacional de Ciência e Tecnologia em Energia e Ambiente - INCT E&A
d Universidade Federal do Sul da Bahia, Centro de Formação em Tecnociencias e Inovação –CFTCI
e Centro Universitário SENAI-CIMATEC, Salvador, BA.
*e-mail: pedrovictorbahia@hotmail.com
Polycyclic aromatic nitrogen heterocycles (PANHs) are formed and released into the environment by
incomplete combustion of fossil fuels, in spills or effluents of several industrial activities, oil drilling, refining,
and storage and coal tar distillation1,2. The exposure human to PANHs through the inhalation of fine particulate
matter (PM2.5) can cause several health problems from chronic respiratory or cardiovascular diseases due to
their toxic, mutagenic, carcinogenic, and teratogenic effects3. PANHs are found in the environment at low
concentrations, thus is necessary to apply sensitive analytical methods capable of quantifying these
compounds at atmospheric levels. In the literature, studies that determine these N-compounds in particulate
matter usually includes two or more steps: extraction of analytes from the matrix, followed by their purification
and pre-concentration. However, large volumes of solvent, long extraction times and intensive sample
manipulation are usually required2. Although in recent years the introduction of green sample preparation
methods in environmental matrices has increased, studies related to PANHs in PM 2.5 are very scarce. The
main objective of the present study was to develop and validate a green method based on miniaturized solid-
liquid extraction combined with ultrasound-assisted for the determination of twenty PANH in PM2.5 samples by
gas chromatography-mass spectrometry (GC-MS). A simplex-centroid mixture design was applied to optimize
the extraction solvent composition (acetone, hexane and dichloromethane) and a full factorial design was used
to optimize the extraction conditions, where the extraction time (10-30 minutes), sample size (2.8-5.3 cm2) and
solvent volume (400-900 µL) were the studied variables. Through the response surface and Pareto chart
obtained from the experimental design were found the best solvent mixture (hexane:acetone (60:40,v/v), and
since the solvent volume is a significant parameter for the experiment, was optimized using a univariate
approach, presenting optimal condition using 450 µL. The methodology was validated in terms of matrix-
matched calibration and the LOD and LOQ values obtained ranged from 0.05 ng m  -3 (1,4,8-trimethylcarbazole)
to 0.11 ng m -3 (3-methylindole) and from 0.1-0.37 ng m -3 for the same compounds, respectively. Determination
coefficients (R2) ≥0.99 were obtained for all compounds within the linear region (0.5-40 ng mL-1). The mean
recoveries for PANHs ranged from 75% to 119%, with intraday and interday relative standard deviation (RSD)
<20%. Evaluation of the microscale method using Analytical Greenness (AGREE) assessment metric
demonstrated the green potential of the developed method. Finally, the proposed method was successfully
applied to samples collected in three turns in a bus station, in the city of Salvador, State of Bahia, Northeastern
Brazil. The sampling was performed at a semi-enclosed underground floor of this bus station, where buses
were in an idling position during commuting. The carbazole, indole and quinoline classes were detected with
concentrations in the range of 0.29 ng m3 (2-methylcarbazole) to 6.08 ng m 3 (carbazole). The highest
concentrations among the studied PANHs were observed in the afternoon period for carbazole and 2,4-
dimethylquinoline these compounds are found in diesel emissions and incomplete combustion process4. The
microscale green procedure was found to be precise, accurate and suitable to employed for the determination
of PANHs in airborne particle samples.
1 Lintelmann, J. et al. Journal of Chromatography A, 1217(10), 2010, 1636–1646.
2 Alves, C. A., et al. Science of The Total Environment, 595, 2017, 494–504.
3 Chibwe, L. et al. Environmental Science & Technology 53, 2019, 2981-2989.
4 Claxton, L. Mutation Research/Reviews in Mutation Research, 763, 2015, 30-85.
[CNPq, FAPESB, FINEP]

397
DETERMINATION OF TOTAL CONCENTRATION AND BIOACCESSIBILITY OF
ESSENTIAL AND NON-ESSENTIAL ELEMENTS IN DIETARY SUPPLEMENTS BY
ICP-OES
Thaís dos S. Berón (PG)(a), Gustavo R. Bitencourt (PG)(a), Aline L. H. Müller (R)(a), Rodrigo C.
Bolzan (R)(a), Paola de A. Mello (R)(a)
aDepartamento de Química, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil, 97015-900
Dietary supplements are defined as products to supplement the diet and provide essential nutrients for
the humans. May contain vitamins, minerals, herbal or botanical products, amino acids, or any
combination of these1.The large consumption of dietary supplements can be associated with several
factors, as awareness of the people with health care, higher consumption by athletes and bodybuilding
exercises practitioners for better performance and gain of muscle mass, among others 1. Among the
nutrients provided by dietary supplements, there are the essential and non-essential elements and their
bioaccessibility is crucial. Bioaccessibility is defined as the fraction of a nutrient that is released from its
matrix in the gastrointestinal tract, making it available for efficient absorption by intestine 1. Different
methodologies can be applied to determine the bioaccessibility of elements, and the in vitro methods
have been widely used, mainly due to their lower cost and easier application 1. The Bioaccessibility
Research Group of Europe (BARGE) developed a unified bioaccessibility method (UBM) in order to
produce a validated and standardized in vitro procedure1. Another bioaccessibility in vitro method was
also established in a consensus by the COST (European Copperation in Science and Technology) action
INFOGEST international network2. Thus, in this work the bioaccessibility of some essential (Ca, Co, Cu,
Fe, K, Mg, Mn, Mo, Na, Ni, Zn, V) and non-essential elements (Al, As, Ba, Be, Bi, Cd, Cr, La, Pb, Sr) was
determined by using the unified BARGE method and INFOGEST method and subsequent determination
by inductively coupled plasma optical emission spectrometry (ICP-OES). For this, five samples of dietary
supplements of different classes (vitamins and minerals, minerals, and botanicals) were used. Total
concentration of the essential and non-essential elements was determined by ICP-OES after microwave-
assisted wet digestion. The BARGE procedure consisted of a three-stage process, simulating the
digestive processes of the mouth, stomach, and small intestine, where the first stage simulates the gastric
phase (saliva + gastric fluid) and the second stage simulates the gastrointestinal phase (duodenal fluid +
bile). On the other hand, the INFOGEST method comprises the gastric and the gastrointestinal phase in
an only stage (saliva + gastric fluid + intestinal fluid). All extractions were performed at 37°C as suggested
by both protocols, because the enzyme activity and chemical characteristics, such as solubility, are
affected by the temperature. The bioaccessible fractions of the elements using the BARGE method in the
gastric phase was similar to that after the gastrointestinal phase for the dietary supplements for some
analytes whereas others were lower after the second step. Bioaccessible fractions using BARGE method
were low for some elements as Al, Cr, Fe, and Zn, up to 28% depending on the sample, or medium to
high as for Ba, Cu, Mg, Mn, Mo, and Sr, all in the range up to 45-100%. For same elements, as Ag, Be,
Cd, La, Na, Ni, and V, the concentrations in extracts were below the limit of quantification (LOQ). The
bioaccessible fraction using the INFOGEST method was about 1% for Al, 12 to 23% for Ba, 1% to Ca,
24% to 132% for Cu, 5 to 17% for Cr, 1 to 2% for Fe, 60 to 91% for K, 0.13 to 50% for Mn, 61 to 126%
for Mo, and 12 to 40% for Sr. Using this method, for Ag, As, Be, Cd, Co, La, Na, Ni, Pb, and V,
concentrations in extracts were below the LOQ. Investigated protocols demonstrated to be not in
agreement for those investigated analytes in dietary supplements and this in an important point to
consider when dealing with protocols for bioaccessibility.
1. Wragg J, Cave M, et al. Science of the Total Environment, 409, 2011, 4016.
2. Silva E, Farias L, Cadore S. Journal of Food and Analysis, 68, 2018, 65
[CNPq, CAPES, UFSM]

398
DEVELOPMENT OF A FIELD TRIAL FOR DETERMINATION OF
ADULTERATION OF FUEL ETHANOL BY METHANOL
Vanessa Gomes Furtado da Cruz 1, Cristiane Gimenes de Souza 1, Rafael Cavalcante dos
Santos 1, Renan de Oliveira Muniz 1, Débora França de Andrade 1, Luiz Antonio d'Avila 1
Universidade Federal do Rio de Janeiro
Ethanol is the most used biofuel in Brazil. However, this fuel has been adulterated with methanol, of
fossil origin, which, in addition to environmental and tax impacts, can cause damage to consumers'
health due to its toxicity. The National Agency of Petroleum, Natural Gas and Biofuels (ANP), as of
Resolution No. 19 of 20151, establishes that the maximum content of methanol in fuel ethanol must
not exceed 0.5% by volume. With the enactment of ANP Resolution No. 696, of 2017/08/31,
amended on 2017/10/032, it became mandatory to analyze the methanol content in fuel ethanol by
suppliers of this fuel and liquid fuel distributors, in order to curb the use of methanol as an ethanol
adulterant. The reference method for the determination of methanol in ethanol is based on the gas
chromatography technique, NBR 16041/2015 (ANP, 2015). An alternative method, based on high
performance liquid chromatography, has been developed3 but both require expensive equipment
only feasible to be carried in laboratories. Thus, the main objective of this work was to develop a
simple, fast and low-cost test, capable of being carried out in the field, using Nile blue chloride dye
and processing digital images of colorimetric reactions for determine adulteration of fuel ethanol with
methanol. An experimental plan was constructed with the objective of identifying the best
concentration of this dye and relating it to the colorimetric responses in ethanol and methanol
solvents. Concomitantly, the influence of different cameras and smartphones were evaluated, and
the results were obtained in the RGB system (Red, Green, and Blue), using the ImageJ software,
and compared to select the channel with the highest sensitivity. As a result, the combination of a
portable camera and a 64-megapixel smartphone allowed the identification of a greater difference in
the colorimetric response between ethanol and methanol in terms of RGB values. An increase in the
difference in the colorimetric response between methanol and ethanol was observed when working
with low concentrations of dye. The linearity of the method was determined in the ranges of 0 to
100%, 0 to 20% and 0 to 1%, by volume of methanol in ethanol. The linearity was verified by the
determination coefficients (R2) whose values were greater than 0.90. The Red Channel showed a
more sensitive response compared to the other channels (Green and Blue). The developed method
is promising, requiring the optimization of experimental conditions to be used in the identification of
adulteration of fuel ethanol by methanol in the field, making the monitoring of fuel ethanol more
accessible and, therefore, improving its quality.

399
DEVELOPMENT OF METHODOLOGY FOR THE DETERMINATION OF
CANNABINOIDS IN CANNABIS-BASED PRODUCTS FOR MEDICINAL
PURPOSES
Mychelle A. Monteiroa*, Anna S. C. L. Dantasa, Matheus N. de Souzaa, Patrícia C. de Limaa,
Maria E. D. Limaa, Daniela S. Santanaa, Ricardo L. N. Maranhoa, André Colonesea, Soraya M.
Ochsa
a Fundação Oswaldo Cruz, Departamento de Química/Instituto Nacional de Controle de Qualidade em Saúde, Rio de
Janeiro, Rio de Janeiro, Brazil, 21040-900
*e-mail: mychelle.monteiro@incqs.fiocruz.br
The use of medicinal products based on the Cannabis sativa plant has been increasingly studied.
Products derived from this plant are available in Brazil, among them, one drug, cannabis products (CP)
authorized by Anvisa, individual import products and those provided by patient associations.1 Once these
products are used for therapeutic purposes, quality control of these products is essential to ensure that
they meet the requirements of quality, purity, and safety. So, the importance of methods for analysing the
content of cannabidiol (CBD) and tetrahydrocannabinol (THC) active compounds and their related
substances, CBDA, THCA, CBN and CBG is evidenced (Figure 1). The importance of controlling THC is
also highlighted, since it is a psychoactive substance.2 Despite the growing use, there is no monograph
for the evaluation of cannabinoids in Brazilian Pharmacopoeia (BP) or in other compendia accepted in
Brazil, according to the legislation RDC nº 511/2021.3 Therefore, this work aimed to develop a quick and
easy method to quantify cannabinoids in products for medicinal purposes. The method was developed
by HPLC-UV, with C18 stationary phase (150 x 4.6mm, 5µm), gradient elution of the mobile phase
composed by 0.1% formic acid in acetonitrile and water, flow rate of 0.6 mL.min-1 and 220nm wavelength
detection, in which it was possible to resolve the peaks of CBD, THC, cannabidiolic acid and cannabinol.
Sample preparation was carried out using ethanol:water (95:5) as extraction solvent and inversion, vortex
and ultrasound agitation steps, in addition to centrifugation. The developed method was applied to two
products and met the established requirements of precision, accuracy, linearity and selectivity4,5.
Considering the importance of a pharmacopeial method to establish minimum quality requirements for
medicines and products for medicinal purposes and considering the quantity of these products available
in Brazil, it was found that is important to evaluate the implementation of other products in the method
developed for a future validation, in order to insert the method in BP.
Figure 1. Molecular structure of CBD, THC, CBN, CBDA, THCA and CBG.
1 Agência Nacional de Vigilância Sanitária. Autorização Sanitária de produtos de Cannabis, Brasília, 09 ago. 2021.
https://www.gov.br/anvisa/pt-br/assuntos/educacaoepesquisa/webinar/medicamentos/medicamentos.
2 Lewis MM; Yang Y; Wasilewski E; Clarke, HA; Kotra, LP; ACS Omega 2017, 2, 6091
3 BRASIL. Resolução da Diretoria Colegiada - RDC No 511, de 27 de maio de 2021. Dispõe sobre a admissibilidade de códigos
farmacêuticos estrangeiros. Diário Oficial da União: seção 1, Brasília, DF, n 101, p. 146, 31 mai. 2021
4BRASIL. Resolução da Diretoria Colegiada - RDC Nº 166, de 24 de julho de 2017. Dispõe sobre a validação de métodos analíticos e
dá outras providências. Diário Oficial da União: seção 1, Brasília, DF, n 141, p. 87, 25 jul. 2017
5 AOAC INTERNATIONAL. Guidelines for Standard Method Performance Requirements (Appendix F). Rockville: AOAC International,
2016.
[INCQS, FIOCRUZ, CNPq]

400
EVALUATION OF 2-HYDROXYETHYLAMMONIUM-BASED PROTIC IONIC
LIQUIDS AS ELECTROLYTES FOR ENERGY PRODUCTION
José F. de Macedo (PG)a,*, Maria E. C. V. Nascimento (UC)a, Erica P. Fernandes (PG)a, José
C. dos Santos Jr (PG)a, Gerlon de A. R. Oliveira (PG)b, Luciano M. Lião (R)b, Eliana M.
Sussuchi (R)a
a Research Group of Nanomaterials and Electrochemical Sensors - SEnM, Graduate School of
Chemistry, Federal University of Sergipe, 49.100-000, São Cristóvão, SE, Brazil
b Laboratory of Nuclear Magnetic Resonance – LabRMN, Institute of Chemistry, Federal University of Goiás, 74.690-900,
Goiânia, GO, Brazil

*e-mail: macedojf92@gmail.com
The development of new electrolytes based on less toxic and more secure materials that promote
sustainable chemical processes is of great interest to reduce environmental pollution and modernize
electrochemical energy storage devices1. Protic ionic liquids (PIL) serve as electrolytes and feature
a “free” proton facilitating proton transport under non-aqueous conditions. PILs, also called “design
solvents”, have adjustable nature and physicochemical properties such as low flammability, low
volatility, high conductivity and high thermal, electrochemical and ionic stability. This potential set of
properties makes it possible to application PIL in different systems for energy generation2. In this
work, ionic liquids based on 2-hydroxyethylammonium and different anions - acetate (HEAA),
ascorbate (HEAs), citrate (HEC) and oleate (HEAOl) - will be studied through electrochemical
techniques for application as electrolytes for energy generation. The PILs were characterized by
infrared spectroscopy (FTIR), shown in Fig. 1, and nuclear magnetic resonance (NMR). The low
toxicity of ionic liquids was determined by the lethal concentration (LC50%) through bioassay with
Artemia salina. Studies on its thermogravimetric, rheological, conductivity and electrochemical
properties are in progress. Initial tests by cyclic voltammetry (CV) using the three-electrode system,
with the glassy carbon electrode (GC) as the working electrode in contact with the electrolytes
prepared by dissolving 1.0 mol L-1 of PIL in 10.0 mL, demonstrated a good electrochemical window
for ionic liquids (Fig. 2).
Figure 1. FTIR spectra of protic ionic liquids. Figure 2. Cyclic voltammograms of the glassy carbon
electrode in 1.0 mol L-1 solutions of PIL at 100 mV s-1, 25ºC.
1 Pradhan, S.; Kumar Sahu, P.; Priyadarshini, S.; Puhan, S. Mater. Today Proc. 2022.
Stettner, T.; Balducci, A. Energy Storage Mater. 2021, 40, 402–414.
[PPGQ/UFSE, CNPq, SEnM, LCM, CLQM, SergiTec, LabRMN IQ/UFG, ASCi, CAPES, CNPq,
FAPITEC/SE]

401
EVALUATION OF EXTRACTION CONDITIONS FOR SALTS AND SEDIMENTS
FROM THE PRE-SALT OIL AND CHEMICAL CHARACTERIZATION
Caroline M. Sengera (PG), Kéllen F. Anschaua (PG), Luiza Baumanna (PG), Denise P. Oliveiraa
(UG), Aline L. H. Müllera(R), Edson I. Müllera (R)*
aFederal University of Santa Maria, Department of Chemistry, Santa Maria, RS, Brazil, 97105-900
*e-mail: edson_muller@yahoo.com.br
Petroleum consists of a natural mixture of hydrocarbons, normally present in the liquid state, and to a
lesser extent by compounds of sulfur, nitrogen, oxygen, metals and other elements in low concentrations.
Among other constituents, there may be the presence of water or sediment in the oil that cause problems
in the refinery, such as corrosion of equipment, malfunctions in the distillation unit, in heat exchangers
and affect the quality of the product. These problems may also be related with the presence of inorganic
materials such as salts and sand associated with the aqueous phase.1 For the characterization of the
saline species present in the sediments, the ASTM D4807 standard was used, which consists of the
solubilization of the sample with toluene and subsequent vacuum filtration.2 The 0.45 µm porosity
membrane is weighed before and after filtration to calculate the amount of sediment present in the oil.
However, this methodology employed by the standard can induce the solubilization or crystallization of
salts present in the oil. In order to verify if the change in the nature of the solvent affects the shape and
composition of the crystals in the saline species retained on the surface of the nylon membrane, in this
work, the solvent used for filtration was replaced by different proportions of the mixture of toluene +
ethanol (10% + 90%, 20% + 80% and 30% + 70%). Other changes were also evaluated in the filtration
stage, such as the use of membranes with different porosities (0.11, 0.22 and 0.45 µm), application of
magnetic stirring, vacuum levels (-40 to 80 KPa) and temperature (70, 80 and 90 °C). Scanning electron
microscopy (FEG-SEM) technique (Carl Zeiss) with a secondary electron (SE) detector and
compositional analysis using an energy dispersive X-ray detector were used to evaluate the crystal
morphology of species saline. For chemical characterization, under all conditions evaluated, the solids
retained in the nylon membrane were solubilized with water and then Ca and Na were determined in an
inductively coupled plasma optical emission spectrometer (Spectro Cirus CCD ®, Spectro Analytical
Instruments) and the determination of Cl- and SO42- was performed using ion chromatography (Thermo
Scientific). Three oil samples named A, B and C were used. First, an initial characterization was carried
out, evaluating the contents of °API, water and sediment. Sample A had the highest sediment content
(1.33%) and high water content (5.55%) and was used for all ASTM D4807 assessments. To determine
the total concentration of Ca and Na in the three samples, digestion of 0.5 g of sample with 6 mL of HNO3
was applied under heating by microwave radiation for later determination by ICP OES. However, for the
determination of SO42- and total Cl-, an extraction method assisted by microwave radiation was used with
10 g of oil and 20 mL of water. The results obtained from changes in the ASTM D4807 standard, showed
that there was no influence in the conditions of the temperature, agitation and vacuum level on the amount
of solids retained in the membrane. However, use of the toluene and ethanol mixture produced a
significant influence on the amount and form of solids retained on the membrane surface compared to
ASTM D4807 using toluene. The greatest reduction in the amount of solids retained in the 0.45 µm nylon
membrane was observed for the ethanol + toluene mixture (30% + 70%). The modification of the
morphology and size of the crystals retained on the membrane surface was also observed, with a
predominance of cubes formed by Na, Cl and Ca. The use of 0.22 and 0.11 µm membranes showed that
some solvent mixtures promoted the crystallization of solids smaller than 0.45 µm that were not retained
on the membrane surface recommended by ASTM D4807. The elemental analysis of the solids retained
on the surface of the membranes showed a selective solubilization of some salts, with emphasis on a
greater solubilization of Ca salts in relation to salts containing Na.
1 Speight JG, The Chemistry and Technology of Petroleum, 2006.

2 ASTM, Standard Test Method for Sediment in Crude Oil by Membrane Filtration (D 4807-05): 2005.
[UFSM, CNPq, CAPES]

402
EVALUATION OF LA-ICP-MS FOR ELEMENT DETERMINATION AND
ELEMENT DISTRIBUTION IN MULTIVITAMINS
Flávia B. Hanzel (PG)a*, Ana B. Viana (PG)a, Arthur F. Burg (UG)a, Viviane L. Garcia (UG)a,
Valderi L. Dressler (R)a
aFederal University of Santa Maria 1, chemistry department, Santa Maria, Rio Grande do Sul, Brazil
*e-mail: flaviabritohanzel@gmail.com
Multivitamin supplements present several essential vitamins and minerals and as the purpose to fill
the deficit that may be related to deficiency in the fixation or synthesis of these species.1 A growing
part of the population is consuming vitamins and minerals in the form of supplements, and in some
cases, the supplement represents the main source of ingestion of such substances. 1 Regardless of
the reason, multivitamin supplements gain every day more space in commerce. While humans need
several essential elements to survive, they must be present at certain levels. On the other hand,
several elements are not essential and can be present in supplements as contaminants, often at
toxic levels.1 Inductively coupled plasma mass spectrometry (ICP-MS) combined with laser ablation
(LA-ICP-MS) allows analysis of samples without the need for time-consuming sample preparation
and little sample handling.2 The purpose of the present work aims to develop a method for
quantitative analysis of multivitamins and investigate element distribution (mapping). A calibration
procedure with hydroxyapatite was investigated for the determination of Mn, Sr, Cd, Ba, Li, Al, Cr,
Co, Ni, Cu, Zn, Mg and Fe in multivitamins by LA-ICP-MS. Hydroxyapatite enriched with reference
solutions was used as material for matrix matching to construct calibration curves. Calcium was used
as internal standard (IS) as it is present in samples and hydroxyapatite, and it is relatively
homogeneously distributed. The sample preparation consisted only of grinding and pressing to make
tablets by using a hydraulic press. The effect of carrier gas flow rate, laser energy, and laser beam
diameter were evaluated on the signal intensity of each element and normalized signals with 43Ca.
The homogeneity of the samples and the effect of milling on the distribution of elements in the tablet
were also evaluated. As example, the signals for Sr before and after grinding are shown in Figure 1.
In addition, an image of Sr distribution in a sample surface without grinding is shown. Similar results
were obtained for the other elements. Limits of detection in the order of ng g -1 to mg g-1 and good
precision (RSD below 20%) were achieved. In conclusion, the proposed LA-ICP-MS method proved
to be suitable for the determination of the elements in multivitamins.
Figure 1. A) Sample signal for 87Sr
without grinding; B) Sample signal
for 87Sr after grinding; C) Image of
the distribution of 87Sr in the sample.
1 Damodaram, S.; et al. Química dos Alimentos de Fennema. 4ª edição, São Paulo. ARTMED, 900 p, 2010.
2 Nunes, M. A.G. Avaliação da calibração em LA-ICP-MS usando papel filtro para análise de plantas. Tese (doutorado) -
Universidade Federal de Santa Maria, Santa Maria, 2015.
[UFSM, CAPES, CNPq]

403
EVALUATION OF MGCL AND CACL AS TARGET MOLECULES FOR CHLORIDE
DETERMINATION IN ETHANOL FUEL SAMPLES USING HR-CS GF MAS
Leandro Kolling (PG)a, Júlia de Lima Cesar Fernandes (UG)a, Maria Goreti R. Vale (R)a,b,
Márcia Messias da Silva(R)a,b*
a Universidade Federal do Rio Grande do Sul, Instituto de Química, Porto Alegre, RS, Brazil, 91501-970,
b Universidade Federal da Bahia, Instituto Nacional de Ciência e Tecnologia do CNPq - INCT de Energia e Ambiente,
40170-115, Salvador, BA, Brazil
*e-mail: mmsilva@iq.ufrgs.br
Nonmetal determination in a variety of samples has been simplified by using the high-resolution
molecular absorption spectrometry and the direct introduction of samples. Even so, the
determination of trace elements (such as nonmetals) in fuel samples is still a challenge.1–3 However,
this determination is mandatory due to environmental concern and regulatory issues. In order to fulfil
these specified needs, a reliable method for chloride determination in ethanol fuel samples via HR-
CS GF MAS was developed and presented in this work. For the chloride monitoring, two target
molecules were evaluated: the diatomic molecules CaCl and MgCl. The method consisted of
introducing the liquid fuel samples directly into the graphite furnace, followed by the forming reagent
(Ca or Mg) in the vaporizer. For the optimization of the proposed method, parameters such as:
pyrolysis and vaporization temperatures, the working wavelengths, and the nature and amount of
forming reagents in different media (aqueous and organic) were investigated. For calibration, the
use of standard solutions of three different sources of chloride in aqueous (NaCl and HCl) and
ethanolic media (LiCl) were evaluated. All the obtained curves presented R² greater than 0.991 and
a linear range between 0.5 and 7.3 mg kg-1 of chloride. This working range was suitable as it covered
the maximum chloride content established in the Brazilian legislation (ANP Resolution No. 7, 2015).4
By using the proposed method (for both CaCl and MgCl) in the analysis of fortified samples,
satisfactory results were obtained with recoveries between 80% and 115%. For seven real samples
analyzed, the chloride concentrations were between 0.6 and 2.0 mg kg-1 (as shown in Table 1).
These values were in agreement with the results obtained using a reference method (NBR 10985
modified by Avelar and Barbeira, 2007).5 Besides, the results for both CaCl and MgCl were
statistically equivalents at 95% confidence level (paired t-test).
Table 1. Determination of chloride (in mg/kg) in real ethanol fuel samples. Results expressed as mean ± standard
deviation. The t-values are presented in parentheses (n=3; t-crit.: 4.30).
Sample Reference Method* CaCl MgCl

E1 2.06±0.09 2.0±0.2 (0.52) 2.0±0.3 (0.35)
E2 0.66±0.02 0.6±0.1 (1.04) <0.8 (LOQ)
E3 0.92±0.09 0.7±0.1 (3.81) 0.9±0.4 (0.09)
E4 0.93±0.10 0.8±0.1 (2.25) 0.9±0.1 (0.52)
E5 0.64±0.07 <0.5 (LOQ) <0.8 (LOQ)
E6 1.10±0.10 1.1±0.1 (1.73) 1.3±0.2 (1.73)
E7** <0.45 (LOD) <0.2 (LOD) <0.3 (LOD)
5;
* NBR 10895 modified by Avelar and Barbeira (2007) ** Anhydrous ethanol fuel sample
1. L. S. G. Teixeira, T. J. Chaves, P. R. B. Guimarães, L. A. M. Pontes e J. S. R. Teixeira, Anal. Chim. Acta, 2009, 640,
29–32.
2. F. V. Nakadi, A. L. Carcinoni Soares e M. A. Mesquita Silva Da Veiga, J. Anal. At. Spectrom., 2014, 29, 1871–1879.
3. L. Kolling, M. B. Dessuy, M. G. Rodrigues Vale e M. Messias da Silva, Talanta, 2022, 238, 122998.
4. ANP - Agência Nacional do Petróleogás natural e Biocombustíveis, RESOLUÇÃO ANP No 7,DE 21.2.2013 - DOU
22.2.2013, .
5. H. de Magalhães Avelar e P. J. S. Barbeira, Fuel, 2007, 86, 299–302.
[CAPES, CNPq]

404
FABRICATION OF A TAILORED "PLUG AND PLAY" 3D PRINTED MIP-BASED
STRAW FOR SCOPOLAMINE REMOVAL IN ALCOHOLIC BEVERAGES
Lara F. M. Oliveira (UG)*, Weida R. Silva (PG)a, João Flávio S. Petruci (R)a
aFederal University of Uberlândia, Institute of Chemistry, Uberlândia, Minas Gerais, Brazil, 38 408 - 100
*e-mail: laramagalhaes1800@gmail.com
The ingestion of abuse drugs – such as scopolamine (SCP) – causes inhibition of the senses,
temporary memory loss and reduction of the motor function. Such substances are surreptitiously
mixed with alcoholic beverages to potentialize their psychoactive effects enabling assault,
kidnapping, extortion, robbery, or rape of the victim1. This sort of crime is commonly known as “Good
Night Cinderella”. Some strategies have been developed to prevent these types of crimes, such as
drinks tests chips or other devices that change color whether the beverage is contaminated.
However, this practice is not so feasible to be implemented in a real situation. In this context, the
present study proposes the fabrication of a “plug-and-play” device containing a molecularly imprinted
polymer (MIP) for efficient removal of scopolamine in alcoholic beverages, avoiding the potential
victim’s ingestion. The MIP was previously synthetized as follows: SCP (template) was mixed with
acrylamide (functional monomer) with the ratio of 1:5 (template/monomer). Ethylenedimethacryl
(EDGMA) was used as a cross-linking reagent, 1,1′-Azobis (cyclohexanecarbonitrile) (ABCN) as
radical initiator and methanol as porogenic solvent2. After the synthesis procedure, the polymers
were initially washed with methanol. Then, the template was removed using a Soxhlet system and
the polymers were oven-dried for 24 hours2. Two devices were fabricated to accommodate the MIP
for scopolamine removal. The device 1 consisted of a polyethylene tube with length of 4 cm and
diameter of 0.6 cm. Both ends were sealed with glass wool and luer lock connectors. The device 2
was fabricated using a 3D printer with length of 4 cm and diameter of 2 cm. Both ends were sealed
with porous filter and glass wool. A peristaltic pump was used to circulate the samples through the
devices. Initially, 5 mL of a 50 µM scopolamine solution was passed through device 1 containing 10
mg of MIP using flows ranging from 0.5 to 3 mL min-1. The amount of scopolamine that passed
through the device was quantified using Capillary Electrophoresis. The maximum amount (30%) of
SCP retained in the device 1 was obtained with 2 mL min-1. Next, the combination of two columns of
the device 1 were evaluated to increase the SCP removal at the optimized flow. In this condition,
48% of SCP was retained. The same procedure was evaluated with an alcoholic sample spiked with
50 µM of SCP, which revealed 41% of trapping efficiency. Next, the device 2 was prepared using 40
mg of the MIP. At a flow of 0.5 mL min-1, an efficiency of 56% was obtained for SCP extraction in a
spiked sample. Such promising results revealed that the proposed strategy may be feasible for the
efficient removal of SCP in alcoholic beverages. Additional experiments have been performed to
increase the amount of scopolamine adsorbed to ensure the complete removal of this unwanted
substance from alcoholic beverages.
a) b)
Figure 1. Devices to accommodate the MIP (a) device 1 and (b) device 2.
1. Dufayet, L.; Alcaraz, E.; Dorol, J.; Rey-Salmon, C.; Alvarez, J. C.; Forensic Toxicologyl. 38, 2020, 264.
2. Silva, W. R.; Soté, W. O.;Petruci, J. F. S.; Batista, A. D.; Comar Junior, M.; European Polymer J..166, 2022, 111024.
3. Ribeiro, M. M. A. C.; Barreto, D. N.; Flávio da S. Petruci, J.; Richter, E. M.; Microchem. J. 72,2022,106985.
[IQ-UFU, FAPEMIG, CAPES, CNPq]

405
FRACTIONATION AND PRE-CONCENTRATION OF AGNPS USING HUMIC ACID
Aline M. de Andrade (PG)a,b*, Marco A. Z. Arruda (R) a,b
a Spectrometry, Sample Preparation and Mechanization Group - GEPAM, Institute of Chemistry, University of Campinas
– Unicamp, P.O. Box 6154, Campinas, SP, Brazil, 13083-970
P.O. Box 6154, Campinas, SP, Brazil, 13083-970

*e-mail: alinemdandrade@outlook.com
The applications of nanoparticles (NPs) to different materials and products have been growing, whilst
also increasing the concerns regarding their environmental exposure. NPs can reach environmental
matrices through different pathways, depending on their life cycle1. Silver NPs (AgNPs) have special
attention due to their antimicrobial characteristic, and their use has been reported in different
industrial products and goods. However, one of the main difficulties found in tracking those NPs
relies on the fact that their expected concentration in the environment is in the range of ng L-1.2 Single
particle inductively coupled plasma mass spectrometry (spICP-MS) is a technique that provides
information not only on mass concentration but also on number concentration, mass, size
distribution, and elemental composition of spherical NPs at concentrations of ng L-1. The compromise
between the low dwell time, number concentration in the suspension, and total acquisition time
makes feasible the NPs characterization, for instance, in the 104 NPs mL-1 concentration for
millisecond of dwell time. The commitment allows counting a certain quantity of NPs to ensure a
sufficient sampling of NPs for further statistics data treatment. 3 Once the NPs are found in the
environment at ng L-1, then, their number concentration is expected to be low. An alternative can be
designed through a pre-concentration method, which can be considered a challenge not only
because of the concentration range, but also the instability nature of NPs. As reported in the
literature, the humic substances demonstrate interaction with NPs and stabilization effects
depending on NPs coating surface, through electrostatic or steric interactions1. This work aims to
pre-concentrate AgNPs through a fractionation method employing humic acid (HA) due to its
interactions with NPs. Preliminary qualitative experiments demonstrate the NPs characterization
suitability from the proposed strategy considering the obtained profiles shown in the Figure 1. The
fractionation procedure allowed to obtain an expected profile as a log-normal distribution (d), around
80 nm, when compared with the suspension containing only 3000 NPs mL-1 in (a) 10 mg L-1 of (HAs)
and in (b) in water.
Figure 1: 3000 NPs mL-1 without fractionation procedure in (a) 10 mg L-1 HAs, and (b) in water. Resuspended fractionation
after procedure (c) blank, and (d) sample containing 80 nm NPs.
Therefore, the proposed method can be considered as a new strategy to improve the NPs
identification and characterization process even at lower expected concentrations in the
environmental water matrices.
1 Bundschuh M, Filser J, Lüderwald S, McKee MS, Metreveli G, Schaumann GE, Schulz R, Wagner S, Environmental
Sciences Europe, 30:6, 2018, 1
2 Wimmer A, Urstoeger A, Funck NC, Adler, FP, Lenz L, Doeblinger M, Schuster M, Water Research, 171, 2020, 115399.
3 Mozhayeva D, Engelhard C, Journal of Analytical Atomic Spectrometry, 35, 2020, 1740.

406
LIQUID-LIQUID MICROEXTRACTION AND SMARTPHONE-BASED
COLORIMETRY FOR DETERMINATION OF FLAVONOIDS IN BERRIES
Luís C. Martinsa*, Samara Soaresa, Fábio R. P. Rochaa
aCenter for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil, 13416-000
*e-mail: luis.claudio.martins@usp.br
Flavonoids are polyphenol compounds widely distributed in plants and fungi that exhibit a wide range
of applications, including antioxidant and anti-inflammatory properties. Flavonoids may form metal
chelates due to the carbonyl and hydroxyl groups in their chemical structure, which is effective to
prevent free radical generation and thus damage of biomolecules by metal-induced oxidative stress.
Metal chelation is also exploited for screening of the total flavonoid content (TFC) in food, beverages,
and medicinal plants by molecular UV-vis spectrophotometry, e.g. complexation with 2% to 10%
Al(III) and analytical measurements performed in 410-430 nm (acid medium) or 510 nm (alkaline
medium)1. However, these methods are not specific and final results may be affected by e.g.
anthocyanins and phenolic acids. In this context, this work aimed to develop a simple and cost-
effective alternative procedure for preconcentration of flavonoids from berries (blackberry and
blueberry), through liquid-liquid microextraction with low-temperature partitioning and smartphone-
based digital-image colorimetry. The procedure consists of the ultrasound-assisted extraction of
flavonoids using acetonitrile (ACN), partitioning of their aluminum complexes to the aqueous phase
under low temperatures (-20 °C), and direct image acquisition at the bottom of the tubes. Photometric
measurements were based on the intensity of reflected radiation measured under controlled
illumination, distance from camera to the object, and region of interest, then converted to the RGB
color system using a free mobile app (Color Grab, Loomatix®). Channel B values were taken as
analytical parameter due to the complementarity with the color of the complex formed. The results
were expressed in quercetin equivalent concentration (mg L-1 QE) with TFC determined either by
standard addition (SA) or external calibration (EC). Extraction with ACN avoided interferences from
anthocyanins, whereas complex formation in acetate buffer pH 3.5 hindered interferences of
phenolic acids, mainly chlorogenic acid. Under the optimized conditions, the procedure showed
linear response up to 10 mg L-1 QE (33 µmol L-1), r > 0.998, with limits of detection and quantification
of 0.5 and 1.7 mg L-1, respectively. Coefficient of variation of 1.3% (n = 10) was achieved. The slopes
of analytical curves obtained in the presence and absence of the sample extract and poor recoveries
achieved using EC (e.g. 34% and 57% for blueberry and blackberry, respectively) indicated matrix
effects, which were successfully circumvented by SA (recoveries of 101% and 121% for blueberry
and blackberry, respectively). The accuracy was assessed in relation to the spectrophotometric
reference procedure2, with results in agreement at the 95% confidence level. These analytical
features demonstrate that the proposed procedure is a simple and cost-effective approach for the
determination of flavonoid content in berries, with low reagent consumption and waste generation.
1 Pękal A, Pyrzynska K. Food Analytical Methods, 7, 2014, 1776.
2 Papoti VT, Xystouris S, Papagianni G, Tsimidou MZ. Italian Journal of Food Science, 23, 2011, 252.

407
MULTIVARIATE OPTIMIZATION OF A HS-SPME+GC×GC-MS PROCEDURE TO
DETERMINE FINGERPRINTS OF THE VOLATILE FRACTION OF LAGER BEERS
Isabela C. M. Cunha* and Fabio Augusto *
Institute of Chemistry and National Institute of Science and Technology in Bioanalytics (INCTBio), University of
Campinas (Unicamp) – CP 6154, 13083-970 Campinas SP, BRAZIL
*e-mail: faugusto@unicamp.br
Beer is obtained from the fermentation of malted cereals with of additives such as hops. Its volatile
fraction is extremely complex, consisting mainly on aliphatic alcohols and esters, heterocyclic
compounds, aldehydes and essential-oil related substances from hops. Due to the complexity of the
composition of their volatile fraction, analytical procedures to screen these samples should combine
high sensitivity, capability of resolution and fast isolation of the target compounds. This could be
achieved by Headspace Solid Phase Microextraction (HS-SPME) coupled to Comprehensive Two-
dimensional Gas Chromatography with Mass Spectrometric Detection (GC×GC-MS), to obtain a
broad fingerprint of the volatile fraction. However, to achieve the full power of this combination,
careful optimization of both HS-SPME and GC×GC is necessary operational parameters to produce
useful volatile fingerprints. In this work this was performed using a combination of univariate and
simultaneous multivariate approaches. Standard canned lager beer from the same brand were used
as test samples. Before use samples were decarbonated by a freeze procedure designed to
minimize loss of volatile compounds and efficient elimination of CO2. After opening, the content of
previously refrigerated (-4 °C) beer cans was transferred to appropriate vials, fast frozen by
immersion on LN2 for 5 min, submitted to ultrasound until complete tawing and decarbonation and
immediately used. Preliminary tests were performed to select the best HS-SPME fiber: 65 m
polydimethylsiloxane / divinylbenzene (PDMS/DVB) was selected. Also, the GC×GC cryogenic
modulation conditions were optimized by visual inspection of chromatograms. The best use of the
separation space and suitable peak resolution was possible with PM = 6 s, 10 psi pressure for the
cold focusing jet and demobilization jet at 250 °C. The optimization of extraction parameters was
performed by a multivariate approach using a Doehlert matrix. An aliquot of Vam mL of sample was
transferred to a septum-sealed vial; 2,0 mL of saturated aqueous NaCl solution was added. The vial
contents was thermostated at Text °C and a PDMS/DVB fiber exposed to the stirred vial headspace
for text min. The optimized response was the total integrated peak area. The results of the multivariate
optimization pointed that the extraction efficiency was maximum with Vam = 7,00 mL, Text = 50 °C
and text = 30 min. Using the optimized extraction conditions, it was possible to resolve and detect
~240 to ~270 chromatographic peaks on the volatile fraction of the samples. These results point out
that HS-SPME combined to GC×GC-MS can be a powerful tool to obtain qualitative and quantitative
information regarding the composition of beers.
1. Cunha, J.; Andrade, D. E.; De, A.; Para, Q.; De, C.; Bolini, M. A., Universidade Estadual de Campinas, Faculdade De
Engenharia De Alimentos.
2. Silva, G. A. da; Augusto, F.; Poppi, R. J.; Food Chem. 2008, 111, 1057.
3. Pedroso, M. P.; Antonio, L.; Godoy, F. De; Henrique, C.; Fidélis, D. V.; Correa, E.; Jesus, R.; Augusto, F. 2009, 32,
421.
4. Teófilo, R. F.; Ferreira, M. M.; Quim. Nova, 2006, 29, 338.

408
RAMAN SPECTROSCOPY-BASED IMMUNOASSAY FOR IN SITU MONITORING
OF HUMAN STRESS LEVELS
Javier E.L. Villa (R)a,c,*, Maria D.P.T. Sotomayor (R)b, Luis M. Liz-Marzán (R)c
aInstituto de Química, Universidade Estadual de Campinas (UNICAMP),13083-970, Campinas-SP, Brazil
bInstituto de Química, Universidade Estadual Paulista (UNESP), 14800-060, Araraquara-SP, Brazil
cCIC biomaGUNE, Paseo de Miramón 182, 20014, San Sebastián-País Vasco, Spain
*e-mail: jelv@unicamp.br
Cortisol is a steroid hormone that plays a key role for regulating anti-stress responses of human
body. For this reason, cortisol is considered a reliable stress biomarker and also called the ‘stress-
hormone’. Even though cortisol response to stress is essential for survival, its abnormal levels
represent serious health risks (e.g. Cushing’s and Adison’s disease for higher and lower cortisol
levels, respectively).1 Since cortisol concentrations in biological fluids can serve as a biomarker for
stress and various diseases, its determination represents a crucial task for human health care.1
Additionally, as a consequence of the increasing levels of physiological stress (because of
globalization, living style and competition), the development of reliable, sensitive (concentrations in
the order of ng/mL) and selective (biological complex matrices) methods capable to quantify cortisol
at physiological relevant concentrations are urgently required.2 In this work, we have developed a
novel cortisol biosensor by combining a highly sensitive analytical technique (surface-enhanced
Raman spectroscopy, SERS) and a highly specific assay (competitive immunoassay), see inset
figure. To fabricate the gold nanotags, three
types of nanoparticle shapes were tested
(spheres, rods and stars) and compared in
terms of sensitivity using three different
lasers (532, 633 and 785 nm). The Raman
reporter used was 4-mercaptobenzoic acid
combined with SH-PEG-COOH (Mv=5000)
to modify the nanosurface and to obtain
stable nanoparticles for further surface
modification. BSA bound to cortisol was
covalently bound onto the nanoparticles via
EDC/NHS reaction with the -COOH groups. Since stars shaped nanotags showed the greatest
sensitivity, they were the ones used in all the immunoassay experiments. Regarding to the capture
agent, magnetic beads (MBs) of ~800 nm diameter, with –COOH groups on the surface were used
to allow a simple and fast separation. Cortisol monoclonal antibody was covalently bound to the MBs
beads via EDC/NHS reaction with the -COOH groups, thereby promoting well-oriented linkage of the
antibodies. The experimental conditions to carry out the immunoassay probe were optimized to
maximize the sensitivity and the reproducibility. Under such optimized conditions, the proposed
method was applied to quantify cortisol in biological fluids, particularly urine and serum. The percent
of cortisol recoveries in these samples were ranged between 85 and 94% and there was no
significant difference with the cortisol concentration reported in a serum certified reference material,
thus demonstrating an excellent accuracy. Additionally, there were no significant differences
between these recovery values and those obtained using the well-developed analytical technique
high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). Nevertheless,
the proposed immunoassay exhibited some advantages over HPLC-MS, such as: (1) lower limit of
detection, (2) minimal sample preparation, (3) portability, and (4) lower duration of sample analysis.
Therefore, the proposed method is suitable for monitoring human stress levels by means of cortisol
measurements and presents a great potential for a wide variety of point of care applications.
1 Kufe, D. W.; Pollock, R. E.; Weichselbaum, et al. Cancer Medicine, 6th ed.; BC Decker: Hamilton (ON), Canada, 2003.
2 Kaushik, A.; Vasudev, A.; Arya, S.K.; et al. Biosens. Bioelectron. 2014, 53, 499-512.
[FAPESP, European Research Council, Spanish State Research Agency]

409
SMARTPHONE-BASED DIGITAL-IMAGE PHOTOMETRIC PROCEDURE FOR
LACTOSE DETERMINATION IN BOVINE MILK
Rita C. M. Alves*a, Luís C. Martinsa, Fábio R. P. Rochaa
aCenter for Nuclear Energy in Agriculture, University of São Paulo, 13416-000, Piracicaba, SP, Brazil,
*e-mail: rita.malho@usp.br
Milk is a highly nutritive food, source of carbohydrates, vitamins, proteins, calcium, and other
nutrients needed in all stages of human and animal development. The main carbohydrate in milk is
lactose, corresponding to 40 to 50 g L-1 of the composition in bovine milk1. It can cause health
problems for consumers who have low levels of the lactase enzyme in the body (named as lactose
intolerance). Lactose determination has been performed mainly by High-Performance Liquid
Chromatography (HPLC) and titrimetric methods, such as the Lane-Eynon method, recommended
by the Brazilian Ministry of Agriculture, Livestock, and Supply (MAPA)2. These procedures are time-
consuming and require laborious sample preparation, being incompatible with the high analytical
demand. In this way, development of alternative analytical methods for lactose determination in milk
and dairy products is important. This work aimed at the development of a simple cost-effective, and
environmentally friendly analytical method, exploiting smartphone-based digital-image photometry
for lactose determination in milk and dairy products. It is based on a modification of Benedict's
method3, involving formation of Cu(I)/2,2'-biquinoline-4,4'-dicarboxylic acid (BCA) complex4, instead
of the copper(I) oxide precipitate, aiming at higher sensitivity and better precision. A smartphone
camera was used for images acquisition under controlled conditions, which were converted to the
RGB color system. The analytical signal was based on the intensity of the reflected radiation
measured at channel G, which corresponds to the complementary color of the complex. The sample
preparation in whole, semi-skimmed, and skimmed milk involved precipitation of protein in ethanolic
medium. The procedure consists of the addition of 250 µL of sample together with 250 µL of a 10-
fold diluted Benedict's reagent, and 125 µL of 0,01 mmol L-1 BCA and subsequent heating in a water
bath for 10 min at 90 °C. Under the optimized conditions, the proposed procedure yielded a linear
response within 1.0 – 30.0 mg L-1 (r = 0.994), with limits of detection and quantification of 0.3 mg L -
1
and 0.9 mg L-1, respectively, and coefficient of variation of 3.2% (n = 12). The slopes of the
analytical curves obtained with and without the addition of milk samples were in agreement, with
differences lower than 5%, indicating the absence of matrix effects. The procedure takes less than
10 min and consumes as low as 160 µg Cu(I) and 430 µg BCA per determination. The achieved
sensitivity and limit of detection estimated is compatible with determination of lactose in milk and
dairy products categorized with low content of the sugar.
1 Brito ABN, Giulietti M, Crystal Research and Technology, 42, 2007, 6.
2 Brasil, Ministério de Agricultura, Pecuária e Abastecimento, Instrução normativa n°68, 2006, 49.
3 Benedict SR, J. Biol. Chem. 227, 1907, 101.
4 Soares, S.; Rocha, F.R.P. Microchemical Journal. 162, 2021, 6.
[CNPq, CAPES, FAPESP, INCTAA]

410
SPECIATION OF VOLATILE FATTY ACIDS AS A TOOL FOR ANAEROBIC DIGESTION
PROCESS MANAGEMENT TO INCREASE THE BIOMETHANE PRODUCTION
Fabiane G. Antesa*(R), Deisi C. Tapparob(PG), Rafael Favrettoc(PG), Ricardo L. R.
Steinmetza(R), Airton Kunza(R)
aEmbrapa Suínos e Aves, Concórdia, SC, Brazil, 89715-899
b Faped, Sete Lagoas, MG, Brazil, 35700-039
C UDESC, Lages, SC, Brazil 88519-000
*e-mail: fabiane.antes@embrapa.br
Anaerobic digestion (AD) is a well-recognized approach to recover energy from organic wastes into
a circular economy concept since this technology allies the mitigation of environmental impacts and
economic value addition to residues. Basically, it consists of a biological process where a consortium
of microorganisms uses organic materials as substrate producing mainly carbon dioxide and
methane1. The composition of the residues submitted to AD has a great influence on the process
performance and to establish the best biodigester operation the use of some analytical tools are an
important concern. In this sense, the speciation of volatile fatty acids (VFA, C2-C7) is an indicator of
suitable biodigester efficiency since these molecules are intermediaries during the AD process. The
titration VFA/total alkalinity (VFA/TA) method provides an indirect result of AD process stability.1 The
result of VFA/TA is a relation based on buffer capacity and VFA production and can indicate an
initially inhibition on process. On the other hand, the quantification of each VFA enables more
assertive process management. Therefore in this work a headspace-gas chromatography (HS-GC)
method was validated for determination of VFA on samples collected from a laboratory scale
Continuous Stirred Tank Reactor (CSTR) treating cattle manure and submitted to increasing organic
loading rate (OLR). For this, a GC equipment (Nexis 2030, Shimadzu) equipped with an HS auto-
sampler (HS-20, Shimadzu) and flame ionization detector (FID) was used. Sample pre-treatment
(centrifugation, pH adjustment, dilution) was established to minimize matrix effect on analytical
results to enabling calibration using internal standard (IS) method (IS: octanoic acid). Linear range,
limits of detection and quantification (LOD and LOQ) were determined for the following VFA: acetic,
propionic, butyric, i-butyric, i-valeric, valeric, n-valeric, i-caproic, hexanoic and heptanoic. Values of
LOD and LOQ were on the range of 1.5 and 5 mg L-1, respectively which was suitable for VFA
determination. Accuracy was evaluated by recovery tests (standard addition on samples, n = 3) and
results were between 90 and 105% for all analysed VFA. Results for acetic and propionic acid, the
main VFA detected on analysed samples, compared to VFA/TA (obtained by titration following
standard method) are shown on Figure 1.
Figure 1. Concentration of VFA compared to VFA/TA on samples collected during CSTR operation treating cattle
manure at increasing OLR. VS: Volatile solids.
As can be observed on Figure 1, samples with higher concentration of VFA also presented higher
VFA/TA. As acetic acid is the precursor of biomethane, its accumulation on OLR up to 3.5 is not
necessarily an operation problem. However, the accumulation of propionic acid (OLR > 6.0) indicates
biodigester over-feeding and highlights the importance of VFA speciation by the proposed HS-GC
method.
1 Kunz A, Steinmetz RLR, Amaral AC, Fundamentals of anaerobic digestion, biogas purification, use and treatment of
digestate. SBERA, 2022

411
ULTRASOUND-ASSISTED EXTRACTION METHOD FOR METALS
DETERMINATION IN LIGNOCELLULOSIC BIOMASS
Karen Giacobe (PG)a*, Débora P. Almeida (PG)a, Vanessa R. do Nascimento (PG)a, Cezar A.
Bizzi (R)a
a Federal University of Santa Maria, Department of chemistry, Santa Maria, Rio Grande do Sul, Brazil, 97105-900
*e-mail: karengiacobe@gmail.com
Biomasses are wastes coming from animals, vegetables, or microorganisms, which can be used by
the chemical industry for the production of biofuels (generation second ethanol, bio-oil, biochar,
among others), chemical platforms, or energy generation.1 Biomass from agro-industrial or forest
waste is formed mainly from cellulose, hemicellulose, lignin, extractives, and inorganic constituents.
Those inorganic constituents, which are normally undesired for the industrial process, can cause
problems with the equipment and reactions while biomass is converted.2 The extraction of the
mineral fraction can be conventionally performed by means of mechanic stirring, leaching, Soxhlet,
or extraction. Alternatively, other methods can be proposed aiming the reduction of time, reagents
and energy delivered into system.3 In this work ultrasound-assisted extraction (UAE) was proposed
to increase the efficiency of extraction procedure when compared with conventional extraction
protocols. The proposed methods was investigated for metals (Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Na,
P, Sr e Zn) determination by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES,
Optima 4300V DV, Perkin Elmer, EUA) in energy cane, sugar cane straw, bagasse, pine residue
and eucalyptus residue. The parameters evaluated for the UAE, optimized with sugar-cane bagasse,
were the volume and reagent for the extractant solution, frequency and delivered energy of the
ultrasound (US), extraction temperature and time, sample mass of biomass of cane bagasse, and
the effect mechanic stirring (replacing the US), as well as sample mass (from0,250 up to 25 g,
keeping the same ration sample: solvent). The optimal conditions found for UAE were: US bath
operating at 45 kHz and 70% amplitude (74,71 W/dm3 as delivered energy), 30 min extraction at 50
°C, sample mass of250 mg for 20 mL of 1 mol L-1 of H2SO4(with the same efficiency for 25 g for 2 L
of 1 mol L-1 of H2SO4). The elements determined by ICP OES (Al:200 ± 12; Ba: 4.65 ± 0.63; Ca:288
± 45; Cu: 3.84 ± 0.11; Fe:290 ± 22; K:1248 ± 70; Mg:314 ± 24; Mn:19.2 ± 0.9; Na: 11, 74 ± 0.22;
P:103 ± 7; Sr:3.64 ± 0.08; and Zn:5.53 ± 1, 00 µg g-1), were totally extracted from the biomass when
UAE was used (from 95 to 100% for most elements, except for Al, Fe and Cu, that extracted below
60%). However, while the sample mass was increased, it was possible to observe a decrease in the
extraction efficiency of conventional extraction method (extraction efficiency of 90% extraction, in
average, using the mechanical stirring and heating). When UAE was used for up to 25 g of sample,
a relatively high extraction efficiency was achieved (104% extraction, in average). In a kinetic study,
it was possible to observe an efficient extraction using UAE since the first minute (93 ± 5%, in
average), which was much more efficient when compared with conventional protocol (58 ± 6%, in
average for mechanic stirring, and 70 ± 6%, in average for just heating). Thus, it is possible to
observe that even for a relatively short period of extraction (1 min), the proposed UAE method was
efficient. In addition, UAE methods bring some advantages as the short extraction time (from 1 to 30
min), the possibility of using diluted H2SO4 solution (1 mol L-1), being in agreement with green
analytical chemistry recommendations.
1 Cortez,L. A. B. et al. Biomassa para energia, 773, 2008.

2 Eom, I-Y. et al. Characterization of primary thermal degradation features of lignocellulosic biomass after removal of
inorganic metals by diverse solvents. Bioresource technology, 102, 1991, 3437.
3 Jiang, L. et al. Influence of different demineralization treatments on physicochemical structure and thermal degradation
of biomass. Bioresource technology, 146, 2013, 254.
[CAPES, CNPq, FAPERGS]

412
USB MICROSCOPE CAMERA AS TOOL FOR DEVELOPMENT OF SOLID PHASE
DIGITAL COLORIMETRY METHOD
Marcos D. S. Pereiraa* (PG), Leonardo S. G. Teixeiraa (R), Rodolfo M. M. Santanaa (R)
aInstituto de Química, Universidade de Federal da Bahia, Salvador, BA, Brasil
*e-mail: marcosdiogopereira@gmail.com
Solid phase spectrophotometric methods were reported as good alternatives for improving the
analytical sensitivity of colorimetric analysis. Nevertheless, the solid support and measurement cell
design characteristics, especially the thickness, should be carefully handled to avoid light beam
scattering and attenuation, that cause difficulties in detection1. In this context, image analysis could
be an strategy to overcome these hindrances due to needless radiation-solid phase interaction to
enable the analytical measurement. The present work aims to evaluate the colorimetric
determination of total iron in tap water using 1-(2-tiazolylazo)-2-naphthol (TAN) immobilized on C18
bonded silica with a digital USB microscope camera. The images were video recorded through a 50x
- 500x 2mp Digital Microscope connected to a smartphone and analyzed by the ImageJ software
exploiting RGB, HSV and grayscale. All solutions were handling using a multicommutated flow
analysis system (Figure 1). The studied physical and chemical parameters were (i) pH (Britton-
Robinson buffer 5.0 to 7.5) (ii) eluent volume and concentration HCl (150 to 400 μL and 10 to 250
mmol, respectively) (iii) sample volume (100 to 300 μL), and (iv) flow rate (0.8 to 2.0 mL min -1).
Optimization step established (i) pH = 6.5, (ii) 350 μL of HCl 100 mmol L-1, (iii) 250 μL of sample,
and (iv) carrier flow rate 2.0 mL min-1 as the recommended operational conditions. The USB
microscope camera allowed the solid phase support reduction (ca. 3.5 ± 0.1 mg), which meant a 10-
fold reduction in comparison to solid phase colorimetric methods reported. Additionally, a flow
impedance reduction was also observed, which led to an analytical throughput of 45 determinations
per hour. Under recommended conditions, sensitivity gain and broader linear range were observed
using greyscale regard of RGB. Those observed effects using grayscale occurred because of the Fe
concentration increasing that led to the color changing associated to 1:1 Fe-TAN complex (purple
color) formation instead 1:2 complex (brown color) that affect the analysis in RGB color space. The
proposed method presented a linear range with superior limit equal to 70 mg L-1 (R2= 0.996), LOD
and LOQ were estimated as 1.2 and 4.1 mg L-1, respectively. Furthermore, the method showed good
precision (RSD ≤ 5.5%, n = 6, 1 mg L-1) and accuracy (recovery values from 100 to 109%). The
analyzed tap waters presented an average of (8.1 ± 0.6) mg L-1 Fe content.
Figure 1. Flow multicommutated system with USB microscope camera for solid phase digital colorimetry method
development
1 Rocha, F. R. P., Raimundo, I. M., Teixeira, L. S. G. Direct Solid-Phase Optical Measurements in Flow Systems: A
Review. Analytical Letters.vol.44, 528-559, 2011
[CNPq, FAPESB, CAPES, UFBA]

413
CHEMOMETRICS
414
A CHEMOMETRIC STUDY FOR OPTIMIZING THE IN-SITU SYNTHESIS OF
LASER-INDUCED CU NANOPARTICLES ON 3D-PRINTED ELECTRODES
Fernando Amaral F. (PG)a*, William B. Veloso (PG)a, Gabriel N. Meloni (R)a and Thiago R. L.
C. Paixãoa
a Universidade de São Paulo - USP, Instituto de Química – IQ/USP, São Paulo, SP, Brazil, 05508-000
*e-mail: fernandoaf@usp.br
Chemometrics can be used to improve and develop new experimental designs by applying statistical
tools such as dimensionality reduction, classification and regression, linking (and screening)
experimental variables to experimental outcomes, and possibly highlighting otherwise hidden
relationships between the studied variables1. The use of multivariate experiments allows the
rationalization of the impact of different experimental variables into the results, allowing fast
screening of experimental parameters. Here we report the application of the response surface
methodology (RSM, Figure 1) for optimizing the in-situ synthesis of Cu nanoparticles on 3D-printed
electrodes. The fabrication method relies on using a CO2 laser, which is scanned over the electrode
surface in a raster pattern, to reduce copper nitrate, which is deposited on the electrode surface via
dropcasting. Multivariate experiments were performed exploring the following synthesis parameter
space: laser power, focal distance, laser raster velocity, raster line separation (lateral resolution),
concentration and number of additions of copper nitrate solution. All electrodes were tested towards
the direct electrochemical oxidation of glucose2 and the anodic peak current values were used as
the modelled response for the RSM study. All experimental design, data processing and visualization
was done in Python, a general purpose and open source programming language3. The optimization
highlighted the most relevant laser parameters impacting the Cu nanoparticle synthesis, hence the
electrode behavior towards direct glucose oxidation, providing insights into parameters somewhat
overseen in the literature, that are actually critical in the nanoparticle synthesis. Our study provides
a fresh perspective into laser-induced/assisted electrode modifications, which can help to improve
or develop new electrochemical sensing surfaces.
Figure 1. (A) Anodic peak currents at 0.8 V of varying experimental conditions from cyclic voltammograms (B) of 4 mmol
L-1 glucose in 0.1 mol L-1 NaOH and (C) response curve obtained by RSM in Python. Voltammetric scan rate of 20 mV s-1
1. Sergio L.C. Ferreira, Valfredo A. Lemos et al. Microchemical Journal 140 (2018) 176–182.
2. Kumar, KP Akshay, et al. Electrochem. Commun. 120, 2020, 106827.
3. Shigeaki Morita. Analytical Sciences 36 (2020) 107-112.
[USP, CNPq, CAPES, FAPESP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

415
A GREEN SOLUTION FOR AUTHENTICATION OF ROSEWOOD ESSENTIAL OIL
BY HANDHELD NIR SPECTROMETER AND DD-SIMCA CLASSIFICATION
Tereza C. M. Pastore (R) a*, Lilian R. Braga(R)b, Daniele C. G. da C. Kunze(R)b, Liz F.
Soares(R)b, Floriano Pastore Jr (R)b, Alessandro C. de O. Moreira(R)a, Priscila V. dos Anjos
(R)b, Caroline S. Lara (PG)c, Vera T. R. Coradin (R)a, Jez W. B. Braga (R)b,d.
aLaboratório de Produtos Florestais, Serviço Florestal Brasileiro, Brasília, DF, Brasil, CEP 70818-900
b Institutode Química (IQ), Universidade de Brasília (UnB), Brasília, Distrito Federal, Brasil, 70910900
cInstituto Nacional de Pesquisa da Amazônia, R. Bem-Ti-VI, s/n - Petrópolis, Manaus - AM, 69060-001
dInstituto Nacional de Ciência e Tecnologia em Bioanalítica, 13083-970, Campinas, SP, Brasil, CEP 13083-970
*e-mail: lilianrodribraga@gmail.com
Rosewood essential oil is extracted from the species Aniba rosiodora Ducke, a native of the Amazon
rainforest, and is highly valued by the cosmetics and fragrance industries due to its unique aroma
and rich linalool content (78 - 93% v/v).1-3 Due to its commercial relevance and the extensive
exploration of A. rosiodora, there is a growing concern with requirements of sustainable production
by national and international regulatory environmental agencies, as well as the quality conditions to
meet the demands of industries for pure and authentic oil. In this context, this work proposes a
simple, fast and direct method to verify purity and authenticity using a portable NIR spectrometer
and DD-SIMCA classification model. For the study, 68 batches of rosewood oil (130 samples/bottles)
were purchased from different origins and production processes (production plants, local markets,
community associations, street markets, and internet providers). Eleven lots (21 samples/ bottles)
were donated by the company Magaldi Agro Comercial e Industrial LTDA, which were considered a
reference standard by its precedence and purity confirmed by GC-MS analysis. Another 12
independent batches from different origins (26 samples/bottles) were considered compatible with
the standard rosewood oil. The NIR spectra of all samples were obtained in triplicate, directly from
the commercially available oil using the portable spectrometer MicroPHAZIR™ RX Analyzer (1595
– 2396 nm and 8 nm resolution). Data analysis was performed in the MATLAB® program using
Toolbox DD-SIMCA4. Initially, the data from the 47 samples/ bottles of 11 different brands were
organized into a single matrix and the Duplex algorithm was applied to obtain the training and
validation sets in a 2:1 ratio, totaling 31 and 16 samples, respectively. Then, to increase the
homogeneity and representativeness of these samples, mixtures were prepared to be added to the
training and validation sets. A total of 33 training mixtures and 28 validation ones were made with
the authentic samples separated for each purpose. All mixtures were prepared using calibrated
micropipettes and the total volume of each mixture was set at 1.50 mL. Then, the triplicate NIR
spectra of the mixtures were acquired and included in their respective training or validation sets.
Despite the wide color, viscosity, and aroma variation of the acquired samples, the ones that showed
chemical profiles compatible with authentic and unadulterated samples in the GC-MS analysis also
showed good agreement in the results by NIRS. The vast majority (95.4%) of the commercial
samples purchased on the internet or in local markets were considered unauthentic, presenting
different chemical profiles or adulterations. The method is fast, relatively low cost, and highly efficient
(98.9% accuracy with authentic and false samples). In addition, the analysis does not require pre-
treatment, uses only 0.2 mL per sample, is non-destructive, and does not generate chemical
residues. Due to that, it is considered a “green solution” to be used in quality control or for inspection
purposes to evaluate if the samples meet the criteria established by CITES.
1 May PH, Barata LES, Econ. Bot. 58, 2004, 265.
2 Maia JGS, Andrade EHA, Couto HAR, Silva ACM, Marx F, Henke C, Quimica Nova, 30, 2007, 1906.
3 Fidelis CHV, Augusto F, Sampaio PTB, Krainovic PM., Barata LES, J. Essent. Oil Res. 24, 2012, 245.
4 Zontov YV, Rodionova OYe, Kucheryavskiy SV, Pomerantsev AL, Chemom. Intell. Lab. Syst., 167, 2017, 23.
[Program CTSP CITES, OTCA, INCTBio]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

416
ANALYSIS OF INDUSTRIALIZED FOOD LABELS AND THEIR HEALTH RISKS
THROUGH PRINCIPAL COMPONENT ANALYSIS (PCA).
Fernanda Martins Gomes (UG)a, Deangelis Damasceno (R)a, Monise Cristina Ribeiro Casanova (R)a*
aInstitutoFederal de Educação, Ciência e Tecnologia de Goiás, Campus Senador Canedo, Senador Canedo, Goiás,
Brazil, 75250-000
*e-mail: monise.coltro@ifg.edu.br
The nutritional profile is multifactorial and it is characterized by sequential changes in the diet pattern
resulting from social, economic, technological and cultural changes that directly affect the lifestyle
and health profile of the population. The main changes have occurred in the last two decades with
the adoption of a dietary pattern with a high content of saturated fat and sugar, in addition to foods
with low fiber content. The technological possibilities of large-scale food production and its
conservation for a long time have been causing the spatial and temporal disruption of production
and access to food and increasing the consumption of industrialized foods. According to the 2017-
2018 Family Budget Survey, compared to the same survey carried out in 2002-2003, the evolution
of food consumption at home indicates an increase in the proportion of processed foods. A diet rich
in saturated fats increases serum cholesterol levels and the incidence of cardiovascular diseases,
such as myocardial infarction1. From the last, the objective of this study was to collect nutritional data
on the amount of total fat, saturated fat, protein and sodium contained in labels of different
industrialized foods such as heavy cream, powdered milk, margarine, sandwich cookies, microwave
popcorn and cheese bread from different brands and different origins and through principal
component analysis (PCA) to evaluate possible positive correlations between type of food, brands,
origin and establish a grouping between processed foods and their degree of harm to health.
Through the PCA of the different brands of margarine, it was found that PC1 and PC2 contribute
with 87.56% of the total variance and comparing the loadings and scores graph it is noted that the
margarines from Chile are richer in sodium while those from Brazil, Argentina, Canada and Spain
are richer in total and saturated fats – Chile is a country with an extremely mountainous region,
whose geological characteristics of the place make its soils and consequently its waters rich in
minerals, especially Ca, Mg and At. In the manufacture of margarine, an emulsion, in the aqueous
phase, this mineral-rich water is used, which may explain the higher concentration of sodium in
margarines from Chile. Excess of sodium increases blood volume contributing to heart attacks,
strokes, stomach cancer and even kidney problems. The other countries have margarine richer in
total and saturated fat – and studies show that high levels of saturated fat increase the risk of cancer,
Alzheimer's disease and Parkinson's disease1. The cause may be linked to the way it is produced
by hydrogenation, which considerably increases the saturated fat content2. When the milk samples
are analyzed, analyzing the loading graphs and PC1 scores (48.48%) vs. PC3 (21.25%) – it is noted
that milk samples from Canada and Argentina are the ones with the highest concentration of total
and saturated fats compared to the others. This may be correlated with the season/climate prevailing
in countries that are closer to the extremes of the globe and the concentration of fat in milk. The
colder the climate, the lower the induction of heat stress in the animal, causing an increase in rumen
pH, an increase in the acetic/propionic acid ratio and consequent increase in the fat content in milk 3.
Animal nutrition also influences results, since a low accumulation of trans fatty acid in the animal diet
leads to a higher fat content in milk.
1 The Global Burden of Diseases, Injuries, and Risk Factors Study, The Lancet, v. 396, n. 10258, p. 1129-1306, 2020.
2 FREIRE, L. As novidades no mercado de margarinas. Rev. Óleos & Gorduras, São Paulo, Ano 3. Ed. 16, p. 31-37, 2017.
3 NORO, G. Fatores ambientais que afetam a produção e a qualidade do leite em rebanhos ligados a cooperativas
gaúchas. Dissertação de mestrado. Porto Alegre, RS, 2004.
[CNPq]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

417
ARTIFICIAL NEURAL NETWORKS FOR COFFEE ASSESSMENT BY HIGH-
RESOLUTION MASS SPECTROMETRY
Victor Cardosoa, Julia Balogb, Tamas Karancsib, Guilherme Sabina,c, Leandro Hantaoa
a University of Campinas, Campinas, Brazil
b Waters Research Center, Budapest, Hungary
c OpenScience, Campinas, Brazil
*e-mail: victor.cardoso@iqm.unicamp.br
Coffee is one of the most consumed beverages worldwide1,2. This product presents an important
role in the global economy, mainly in Brazil which is the top producer 1,3. The large consumption of
coffee requires efficient quality control to ensure its quality, meeting the wide consumer demands1-
3
. Thus, business intelligence is mandatory in this process, providing coffee assessment through
quick analysis and interpretation of large amount of data. To achieve this objective, a sampler
prototype based on laser ablation rapid evaporative mass spectrometry (LA-REIMS) was used to
analyse coffee powder by high resolution mass spectrometry (HRMS). This prototype allowed to
perform direct analysis of a sample per minute in the same well plate with minimal sample
preparation, seeking to supply the ascending demands of coffee industries. For this study, 31 types
of espresso coffee samples were used, with different sensory properties and processing parameters,
such as bitterness, body, acidity, intensity, and roasting level. The dataset contains 54 spectra of
each coffee type, being 1674 spectra in total. After suitable pre-processing, the samples were divided
into training, validation, and test sets for the modelling process. As an alternative to PLS-DA, artificial
neural networks (ANN) in tandem with Bayesian optimization were applied to model this dataset.
This approach presented an automated hyperparameter selection based on probabilistic estimations
and classification errors, aiming to find the global optimal solution5. Also, ANN can easily deal with
outliers, numerous classes and non-correlated variables, and massive data sets6, which are issues
found in this data set. ANN presented better results to discriminate the coffee types, achieving 95%
of accuracy against up 90% in partial least square discriminant analysis (PLS-DA). Regarding outlier
samples, they only affected PLS-DA model since the high values of T2 and Q residuals were
measured in this algorithm5, while ANN is capable to deal with non-homogeneous classes. Also,
ANN was evaluated to estimate sensory properties of those coffee capsules such as acidity,
bitterness, body, intensity, and roasting level, achieving high values of accuracy for those models.
This study proved that the LA-REIMS prototype has provided reliable coffee analysis. The developed
method provided quick, clean, and high-throughput analysis compatible with coffee industry
demands. A challenge is the data treatment due to the large volume of data, but ANN provided
excellent results to identify the type of coffee capsules and to estimate sensory properties. We
believe that in a near future, this method can be adapted and optimized to larger scales and as a
routine analysis in coffee industries.
1 V. Cardoso, G. Sabin, L. Hantao, BrJAC. 8 (2021) 91-106.
2 M. Ayseli, H. Kelebek, S. Selli, Food Chem. 338 (2021) 127821.
3 M. Baqueta, A. Coqueiro, P. Valderrama, J. Food Sci. 84 (2019) 1247-1255.
45 V. Cardoso, G. Sabin, L. Hantao, Anal. Methods. 14 (2022) 1540-1546.
5 V. Cardoso, R. Poppi, Microchem. J. 164 (2021) 106052.
6 F. Marini, R. Bucci, A. Magrí, A. Magrí, Microchem. J. 88 (2008) 178-185.
[CAPES, CNPq, Fapesp, OpenScience, and Waters Corporation]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

418
ASSESSING COFFEA ARABICA AND COFFEA CANEPHORA BLENDS BY GAS
CHROMATOGRAPHY-MASS SPECTROMETRY AND CHEMOMETRICS
Marcos V. V. Lyrio (PG)a*, Bruno Q. Araújo (R)a, Pedro H. P. da Cunha (PG)a, Danieli G.
Debona (PG)a, Roberta Q. Frinhani (PG)a, Bárbara Z. Agnoletti (PG)a, Paulo R. Filgueiras (R)a,
Lucas L. Pereira (R)b Eustáquio V. R. de Castro (R)a
a Federal University of Espírito Santo/UFES, Laboratory of Research and Development of Methodologies for Analysis of
Petroleum (LabPetro), Department of Chemistry, Campus Goiabeiras Vitória, Espírito Santo, Brazil, CEP 29075-910
b Federal Institute of Espírito Santo, Department of Food Science and Technology, Venda Nova do Imigrante, Espírito
Santo, Brazil, CEP 29375-000

*e-mail: mrvaleriovieira@gmail.com
Authentication is essential for food quality control, especially for commercial products with a high
market value or geographical indication. Although more than 100 coffee species exist, the Coffea
arabica (arabica) and Coffea canephora (conilon) are still the most representative in a national and
international scenario. Due to their chemical and sensory characteristics, including the price
difference between both species, coffee blends are commercially available to combine single
sensory attributes and reduce the production cost. Given the commercial importance of coffee
beans, analytical techniques have been applied to quality control (e.g., FTIR, MS).1-3
In this sense, the present work proposes to develop a new approach based on volatile compounds
assessment by gas chromatography-mass spectrometry with static headspace sampler (HS-GC-
MS) and chemometrics. The coffee samples were analyzed on a GC-MS system (QP2010 Ultra,
Shimadzu) equipped with VB-wax capillary column (polyethylene glycol, 30 m × 0.32 mm i.d. × 0.25
µm film; ValcoBond, VICI Metronics). The peak area of the individual compounds from total ion
chromatograms (TIC) and extracted ion chromatograms (EIC) were acquired from LabSolutions
software (Data Analysis v. 2.72). The data (TIC and EIC) were treated through partial least squares
regression and variable selection by uninformative variables elimination (UVE-PLS) for the
evaluation and quantification of 72 blends containing 24 mixing levels (0 to 100 wt% conilon:
arabica). In addition, univariate models were developed based on peak area ratio (i.e., [A1/(A1+A2)])
calculated from TIC and EIC data. The calibration set consisted of 48 selected samples and the 24
remaining samples were used in the external validation. A total de 54 volatile compounds were
identified by HS-GC-MS which 1-hydroxy-propan-2-one, acetic acid, 5-methyl-2-
furancarboxaldehyde, formic acid, propanoic acid, butyrolactone, 3-methyl-butanoic acid, and 2-
methoxy-4-vinylphenol were the most relevant compounds for the regression models. TIC-UVE-PLS
model presented root mean squared error of prediction (RMSEP), coefficient of determination (R2p),
and limit of quantification (LOQ) of 4.7%, 0.98, and 8.7%, respectively. On the other hand, EIC-UVE-
PLS presented RMSEP and R2p of 3.3% and 0.99, respectively, and higher values of LOQ (11%)
when compared to TIC-UVE-PLS model. The randomized test for accuracy indicated that these
models are statistically equal at a significance level of 5%. EIC univariate model based on 4-ethyl-
2-methoxyphenol and cyclopent-4-ene-1,3-dione ratio resulted in RMSEP, R2p, and LOQ of 5.0%,
0.99, and 8.7%, respectively. Meanwhile, the TIC univariate model with 2-methoxyphenol and 2-
methyl-1H-pyrroleshowed RMSEP, R2p, and LOQ of 6.5%, 0.96, and 15%, respectively. According
to the randomized test, there is no significant difference between univariate models. Moreover, all
univariates and multivariates models were statistically similar at a significance level of 5%. The
multivariate and univariate models developed with coffee volatile chemical data are useful for blends
authentication and may be used an additional tool to coffee quality control.
1 Cagliani LR, Pellegrino G, Giugno G, Consonni RQ, Talanta, 106, 2013.
2 Abubakar Y, Gemasih T, Muzaifa M, Hasni D, Sulaiman MI,. IOP Conf Ser Earth Environ Sci., 425, 2020.
3 Toci AT, Farah A, Pezza HR, Pezza L, Crit. Rev. Anal. Chem, 46, 2016.
[LabPetro, EMBRAPA, CNPq, FAPES, CAPES]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

419
AUTHENTICATION OF FRESH QUAIL EGGS USING A PORTABLE NIR
SPECTROMETER AND DD-SIMCA
Taynná K.L. Araújoa, David D.S. Fernandesb, Wellington S. Lyrab , Edilene D.T. Moreirac,
Paulo H.G.D. Diniza,d*
a UEPB,
PPGQ, Campina Grande, Paraíba, Brazil, CEP 58429-500
b UFPB,Department of Chemistry, João Pessoa, Paraíba, Brazil, CEP 58051-900
c UFPB, Department of Chemistry and Physics, Areia, Paraíba, Brazil, CEP 58397-000
d UFOB, POSQUIPA, Barreiras, Bahia, Brazil, CEP 47808-021
*e-mail: paulo.diniz@ufob.edu.br
In Brazil, the consumption of quail (Nothura maculosa) eggs has increased in recent years due to
their good sensory properties, besides having a nutritional value higher than chicken eggs (in terms
of amounts of protein, antioxidants, calcium, iron, and phosphorus). In 2019, Brazil was the second
largest producer in the world with a production of about 3786.95 billion eggs. After oviposition,
inadequate storage conditions affect egg quality for consumption due to deterioration of the internal
quality, and higher rate transfer of CO2 through the shell.1 Thus, in this work we developed an
analytical methodology using a portable NIR spectrometer (Texas Instruments®) and DD-SIMCA2
to authenticate quail eggs with real freshness for guaranteeing consumers’ safety.
For this, a total of 90 samples were studied: 50 fresh eggs stored at room temperature for up to 7
days and another 40 eggs stored under different simulated conditions: UV for 4 h (10 samples); 40
°C for 24 h (10 samples); 40 °C for 48 h (10 samples); UV for 2 h + 40 °C for 24 h (5 samples); and
UV for 2 h + 40 °C for 24 h (5 samples). Storage by temperature was carried out in an oven (Quimis®)
with temperature control at 40.0 ± 0.1 ºC, while the incidence of ultraviolet radiation was performed
in a Fotolight UV Exhibitor MD2-A4 (Carimbos Medeiros®). The spectra were pre-processed with
offset correction (OFF), standard normal variation (SNV), 1st derivative Savitzsky-Golay with 2nd
order polynomial and 11-points window (SG) e multiplicative scattering correction (MSC). The target
class (i.e., fresh quail eggs) were divided into training (40 samples) and test (10 samples) using the
Kennard-Stone algorithm. Training samples were used to construct the model, and the other 10
samples were jointed with the degraded samples to test the robustness of the constructed model in
terms of sensitivity, specificity, and efficiency.
Table 1. Results obtained for DD-SIMCA in the authentication of fresh quail eggs.
OFF SNV SG MSC
DD-SIMCA (α = 0.05)
Model Test Model Test Model Test Model Test
Principal Components 6 13 14 7
Sensitivity (%) 95.0 100 100 40.0 97.5 50.0 95.0 90.0
Specificity (%) - 100 - 100 - 100 - 100
Efficiency (%) 98.3 73.7 78.7 94.9
It is important to highlight that, regardless of the pre-processing technique used, all models were
able to correctly identify all degraded samples (i.e., a 100% of specificity), which is very
advantageous from the point of view of the consumer, who will not purchase or ingesting eggs under
inadequate storage conditions. Comparatively, the DD-SIMCA/OFF model used only 6 principal
components to obtain the highest efficiency of 98.3%, with only two samples assigned as extremes
in the target class. The proposed analytical methodology demonstrates that it is possible to
authenticate fresh quail eggs under appropriate conditions of consumption from those ones that were
stored improperly. Here, the main advantage is the rapid acquisition of analytical information using
a low-cost, portable equipment, without the need for sample preparation. Consequently, this method
can be applied in the in-situ monitoring of quail eggs directly at the production farm or even on
supermarket shelves. Furthermore, as it does not invade and destroy the sample, besides using no
reagents and generating waste, the proposed methodology also stands out for being in accordance
with the principles of Green Analytical Chemistry.
1 Brasil YL, Cruz-Tirado J.P., Barbin DF, Food Control, 131, 2022, 108418.
2 Zontov YV, Rodionova OY, Kucheryavskiy SV, Pomerantsev AL, Chemom. Intell. Lab. Systems, 167, 2017, 23.
[UEPB, UFPB, UFOB]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

420
AUTOMATIC DATA PROCESSING OF TLC PLATES FOR ANALYSIS OF
PHYTOCANNABINOIDS IN CANNABIS SAMPLES
Rayssa Rocha Zeidan dos Santos (PG)a*, Jez Willian B. Braga (R)a, Luciano C. Arantes (R)b
aUniversidade de Brasília (UnB), Instituto de Química (IQ), Brasília, Distrito Federal, Brazil, 70910900
bPolícia Civil do Distrito Federal (PCDF), Instituto de Criminalística (IC), Brasília, Distrito Federal, Brazil, 70610907
*e-mail: rayssa.rocha@aluno.unb.br
Despite the pharmacological potential of phytocannabinoids, the illicit use of Cannabis is still their
main destination. Thus, the analysis of seized plant-related material is one of the main demands of
forensic laboratories. Thin layer chromatography (TLC), which consists of separating the
components of a mixture through differential migration over a thin layer of adsorbent retained on a
flat surface after the application of an appropriate solvent, configures a practical, low cost and
effective solution for separation, identification and even quantification. However, the separation
capacity, sensitivity and the subjective interpretation of the results are challenges for quantification
purposes, which leads to studies for the improvement of the method to identify and quantify the main
phytocannabinoids present in Cannabis products.1 The present work aims to develop a software to
automatically process images of TLC plates and enable the identification and quantification of the
main phytocannabinoids contained in seized samples. In a 10x15 cm Silica gel 60 TLC plate, a total
of 30 applications for independent TLC runs were made at the respective origin points.
Subsequently, the plate was placed inside a TLC chamber to elude for ~15 minutes with toluene as
the mobile phase.2 Then, images were acquired in a common scanner and imported to Matlab®.
After noise reduction and baseline correction, chromatograms of all 30 runs were obtained with an
automatic and simple program; data were divided into seven layers (red, green, blue, hue, saturation,
luminosity, and grayscale), allowing for calculations of area, height, and migration distance of the
peaks of interest. The separation of the main phytocannabinoids was confirmed based on TLC runs
containing analytical standards of Tetrahydrocannabinol (THC), Cannabinol (CBN), Cannabidiol
(CBD), and Cannabichromene (CBC). Preliminary results for quantification were obtained by ten
consecutive dilutions of a seized sample analyzed in triplicate in a single TLC plate. Overall, the blue
and saturation image layers presented the highest sensitivity, and the peaks showed greater
intensity and sharpness, consisting of results with an appropriate signal/noise ratio for identification
and quantification purposes. The software presented a promising alternative to visual data analysis
of TLC plates, bringing evident improvements in sensitivity and resolution, besides providing a more
objective and accurate comparison of migration distances and peak integration. A better separation
was obtained for most phytocannabinoids in relation to other references. Furthermore, the integration
is performed in the whole stain (3D integration) with automatic detection of the stain size, position,
background correction, and PCA noise filtering. Despite an initial background observed for the peaks
in the first dilutions, the results for THC presented areas with relative standard deviations of ~10%
and a clear linear pattern according to the dilution factors. Linear regressions presented a very good
coefficient of determinations (R2), 0.94 and 0.98 for the blue and saturation layers, respectively. The
method presented good selectivity and sensitivity for application in seized cannabis samples.
Despite the variation caused by the manual application of the samples in TLC plates, the initial
quantitative results showed adequate precision and accuracy, representing a promising alternative
for the visual analysis of TLC plates.
1 Degani, A. L. G.; Cass, Q. B.; Vieira, P. C. Cromatografia um breve ensaio. Quím. Nova na Escola. Cromatografia. No.
7, Maio, 2018.
2 Moffat, A. C.; Osselton, M D.; Widdop, B.; Watts, J. Clarke’s Analysis of Drugs and Poisons in pharmaceuticals, body
fluids and postmortem material. 4th Ed., London, Pharmaceutical Press, 2011.
[CAPES, FAPDF, PCDF, UnB]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

421
CHALLENGES IN QUANTITATIVE ANALYSIS USING NEAR-INFRARED
HYPERSPECTRAL IMAGING (HSI-NIR): COFFEE PURITY QUALITY CONTROL
CASE STUDY
Cristiane Vidal (PG)a*, Gina Cardozo (R)b, Celio Pasquini (R)a
aUniversity of Campinas (UNICAMP), Department of Analytical Chemistry, Institute of Chemistry, Campinas, SP, Brazil,
13083-681
bInstituto de Tecnologia de Alimentos (ITAL), Campinas, SP, Brazil,
*e-mail: crisvdl@unicamp.br
A proposal method for coffee purity quality control using near-infrared hyperspectral imaging and
chemometric classification model (PLS-DA, Partial Least Square Discriminant Analysis) is
described. The objective is the development of a fast method for determining the maximum of
1% w/w of impurities (husks) in coffee, condition to grant the brand a Purity Stamp (Selo de Pureza1),
which is a Brazilian Coffee Quality Control Program. The official method² is based in degreasing the
coffee with chloroform, filtering, drying, sieving, subsampling, visual inspection and manual sorting
of impurities under a stereoscope. The sample is weighted at many steps, and the impurity mass
percentage is calculated. This procedure is laborious, time-consuming, and it has many analytical
error sources. The last step of visual sorting is subjective and requires a very specialized and trained
analyst. Thus, the objective of this work is perform direct analysis of the coffee (i.e. without any
sample preparation) and asses the quantity of impurities based on an index of the ratio between the
image pixels classified as impurities and the total image pixels (PPI index, Percentage of Pixels of
Impurity). Four PLS-DA classification models for coffee and impurities classes were developed
varying the training/validation set and the spectral range. The sensitivity and specificity were close
to 100% for both classes for all models. Mixtures containing 1 % w/w of impurities in coffee were
prepared and analysed using a Sisuchema hyperspectral system (156 x 156 µm pixel size) operating
at 928-2500 nm. The pixel class prediction using the PLS-DA models were used to build the chemical
images of these mixtures, and to the PPI calculation (Figure 1). The PPI values varied between 0.04
and 1.25 for 27 mixtures containing 1% (w/w) of impurities (Figure 1), with high standard deviation.
The coffee granulometric range was one of the causes of this high PPI fluctuation, demonstrated
when coffee samples were sieved, and the images were analyzed by size fraction. Thus, it was
assessed that a reliable relationship between the PPI value and the maximum allowable impurity
content (1 % w/w) by carrying out direct measurements of the coffee was not established yet. Finally,
the results of this work allowed us to identify limitations in relation to the use of images in the
quantitative detection of impurities caused by the granulometry of the sample and the presentation
of the sample to the imaging system.
Figure 1. Chemical maps of the samples by PLS-DA model predictions (colorbar: coffee in blue and impurities in yellow)
for four coffee samples (M01, 07, M18, M27) containing 1 % w/w of impurities.
1 ABIC - Associação Brasileira da Indústria de Café. Programa Permanente de Controle Da Pureza Do Café (PPCPC).
[https://www.abic.com.br/certificacoes/pureza/]
2 Menezes Jr, J. B. F.; Bicudo, B. A. A. Revista Adolfo Lutz, v. 11, 1958, pp 13–47
[INCTAA, FAPESP, CNPq, CAPES, EDUCORP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

422
CHEMIMETRY APPLIED FOR DISCRIMINATION OF COFFEE REGARDING THE
TYPE OF ROASTER
Danieli G. Debona (PG)a, Marcos V. V. Lyrio (PG)a, Bárbara Z. Agnoletti (PG)a, Elivelton O.
Robrigues (PG)a, Paulo R. Filgueiras (R)a, Emanuele C. S. Oliveira(R)b, Roberta Q.
Frinhani(PG)a, Taís R. Moreira (PG) c, Lucas L. Pereira(R)b, Eustáquio V. R. de Castro(R)*a
a Federal University of Espírito Santo/UFES, Department of Chemistry, Campus Goiabeiras Vitoria, Espírito Santo,
Brazil, CEP 29075-910
b Federal Institute of Espírito Santo/IFES, Department of Food Science and Technology, Venda Nova do Imigrante,
Espírito Santo, Brazil, CEP 29375-000

c Federal University of Espirito Santo/UFES, Department of forest and wood sciences, Center of Agrarian Sciences and
Engineering, Jerônimo Monteiro, Espirito Santo, Brazil, CEP: 29550-000

*e-mail: eustaquiovinicius@uol.com.br
Chemometrics describes the statistical and mathematical approaches used to optimize experiments
and extract information from complex data sets. Among the areas of chemometrics, partial least
squares discriminant analysis (PLS-DA) maximizes the separation between predefined classes while
capturing most of the information1. PLS-DA is widely used in food studies, for coffee it has been
applied in the discrimination between species2, in the authentication of roasted coffee3, and
classification according to the genotype4. In a complementary way, Hydrogen Nuclear Magnetic
Resonance (1H NMR) can be used together with PLS-DA, since it is capable of providing chemical
information about the molecular level.In this perspective, the objective of this work was to
discriminate coffee samples submitted to different roasters using 1H NMR and PLS-DA, as well as
to identify the chemical markers responsible for this discrimination. The roasting experiment was
carried out in two roasters: Ikawa Pro by Ikawa®, in which heat transfer occurs by convection, and
Probatino by Probat®, which roasts by conduction and convection. 64 samples were analyzed for
each roaster, totaling 128 samples. The 1H NMR spectra were acquired in a spectrometer model
VNMRS-400 MHz, with a magnetic field of 9.4 Tesla and a 5 mm probe 1H/19F/X, at 25 °C, according
to the methodology established by Briosch Júnior et al.5. The 128 samples were separated into
training (70%) and test (30%) sets, using the Kennard-Stones algorithm, for the construction of the
PLS-DA model. The spectra were autoscaled, the number of latent variables in the model was
determined by 5-fold cross-validation. The models were evaluated by the parameters: sensitivity,
specificity and class accuracy. The integrals in regions of interest of chemical compounds described
in the literature for coffee5 were determined from the spectra obtained. The model was built using 3
latent variables. Quantitatively, 98% of the training set samples were correctly classified. While for
the test set, all samples were correctly classified (100%). The accuracy and hit rate results were
greater than 98%. The sensitivity and specificity of the model were 98% for the training set, whereas
for the test these parameters had a value of 100%. The results indicate great potential of the
association 1H NMR and PLS-DA is demonstrated in the discrimination of coffees submitted to
roasters with different operating principles. With regard to chemical markers, sugars, chlorogenic
acids, caffeine, trigonelline, and hydroxymethylfurfural were responsible for the discrimination of
coffees.
1 Ballabio, D.; Consonni, V. Analytical methods, 5, 2013, 3790.
2 Agnoletti, BZ.; Oliveira, ECS; Pinheiros, PF; Saraiva, SH. Revista Virtual de Química, 11, 2019, 785.
3 Harahap, RH.; Hakim, L; Djuwendah, E; Karyani, T. Journal of Physics: Conference Series, 1341, 2019, 022006.
4 Marquetti, I, Link, JV, Lemes, ALG, dos Santos Scholz, MB, Valderrama, P, Bona, E. Computers and Electronics in
Agriculture, 121, 2016, 313.
5 Brioschi Junior D, Guarçoni RC, da Silva MCS, Veloso TGR, Kasuya MCM, Oliveira ECS, da Luz JMR, Moreira TR,
Debona DG, Pereira LL, Food Chemistry, 342, 2021, 128296.
[Sicoob, UFES, PPGQUI, LabPetro, Coffee Design, CNPq, FAPES, CAPES]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

423
CHEMOMETRICS WEB APPS PART 1: DATA HANDLING APP
Bernardo Cardeal Darzé*, Licarion Pinto*, Aderval S. Luna*
a Rio de Janeiro State University, Institute of Chemistry, Department of Analytical Chemistry, Rio de Janeiro, RJ, Brazil,
20550-900
*e-mail: bernardocardeal@outlook.com
The development of graphical interfaces helps to spread the use of chemometrics tools by non-
chemometricians. Therefore, developing interfaces that make algorithms easier and reduce the
probability of misuse by anyone is vital to increasing chemometrics relevance. It is essential to
highlight that the great majority of these interfaces were developed by or with the contribution of a
chemist, once some adjustments should be made in the algorithm to extract information that is more
relevant for chemical data. Several interfaces were developed in MATLAB [1-4], but a few were
developed in free languages such as R [5-6]. This work reports the release of the data handling app,
an R application, to perform an initial evaluation and treatment of the data. This application accepts
data directly from .txt, e .csv or .xlsx file extensions. It allows using the data directly from highly used
file types and, in some instances, directly from the instrument, so it does not require a deep
knowledge of R software to use the application. The data handling app allows removing specific
variables or samples, selecting a region of interesting variables, performing basic statistical analysis,
descriptive analysis, several types of data imputations, spectral transformation such as smoothing,
baseline and scatter correction, peak alignment, variable preprocessing, and normality inducing
transformations. Three datasets with some common features of chemical data were pretreated to
highlight the app's usability. The results were discussed using figures obtained directly from the data
handling app to highlight the main functionalities of the application. So, the main idea of this
application is to allow users to perform all the basic data pre-treatment properly and perform an initial
analysis of the data even with no knowledge of R programming. Besides, the data handling app is
an open-access code available to be used on RStudio free environment at the computer or on cloud
computing using computers, tablets, cell phones, or similar devices, even without installing any
software for usage. The code can be obtained by emailing the corresponding authoror by accessing
it through the following webpage https://licarionpinto.shinyapps.io/Data_Handling_app/. All the app
functionalities can be used on this webpage even from a smartphone without installing the R software
or any package on the used device, even if there is an idle time limitation.
1 Eigenvector Research, PLS Toolbox, (2022).
2 A.C. Olivieri, H.C. Goicoechea, F.A. Iñón, MVC1: an integrated MatLab toolbox for first-order multivariate calibration,
Chemom. Intell. Lab. Syst. 73 (2004) 189–197. https://doi.org/10.1016/j.chemolab.2004.03.004.
3 D. Ballabio, A MATLAB toolbox for Principal Component Analysis and unsupervised exploration of data structure,
Chemom. Intell. Lab. Syst. 149 (2015) 1–9. https://doi.org/10.1016/j.chemolab.2015.10.003.
4 D. Ballabio, V. Consonni, Classification tools in chemistry. Part 1: linear models. PLS-DA, Anal. Methods. 5 (2013) 3790.
https://doi.org/10.1039/c3ay40582f.
5 C. Troein, S. Siregar, M. Op De Beeck, C. Peterson, A. Tunlid, P. Persson, OCTAVVS: A graphical toolbox for high-
throughput preprocessing and analysis of vibrational spectroscopy imaging data, BioRxiv. (2019) 2019.12.17.879387.
https://doi.org/10.1101/2019.12.17.879387.
6 T.M. Antonelli, A.C. Olivieri, Developing and Implementing an R Shiny Application to Introduce Multivariate Calibration
to Advanced Undergraduate Students, J. Chem. Educ. 97 (2020) 1176–1180.
https://doi.org/10.1021/acs.jchemed.9b00850.
[CNPq, FAPERJ, UERJ (Programa Pró-Ciência), FAPERJ, CNPq]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

424
CHEMOMETRICS-ASSISTED RAMAN SPECTROSCOPY FOR ASSESSING
WATER CONTENT IN BIODIESEL FROM TWO SYNTHETIC ROUTES
Maycom Cezar Valerianoa*, Antônio Carlos Ferreira Batistab, Mónica Benicia Mamián-Lópeza
aFederal University of ABC, Human and Natural Sciences Center. Santo André, São Paulo, Brazil, 09210-580.
b Federal University of Uberlândia. Pontal Institute of Exact and Natural Sciences. Chemistry Department, Ituiutaba,
Minas Gerais, Brazil, 38304-402
*e-mail: maycom.cezar@ufabc.edu.br
consumer, consequently
Several undesirable processes can alter the biodiesel quality from production until the final
changing its stability parameters. Its susceptibility to go under degradation processes is caused
mainly due to exposure to humidity, temperature rise, contact with metals, light, and oxygen from
the air, making necessary regular monitoring of its stability parameters.1 Due to the chemical nature
of its composition and starting compounds for the synthesis, vibrational spectroscopy techniques are
an attractive option for providing structural information, allow a non-destructive analysis for most
samples, require minimum preparation and amount of sample, and in situ analysis.2 In particular,
Raman spectroscopy measures the inelastic scattering of electromagnetic radiation when interacting
with a sample, providing information-rich signals related to the structure and amount of specific
functional groups.3 This characteristic, along with the ability of spectrometers to generate a large
volume of data in short periods, demand the use of computational methods to model information
that is not observable by separate analysis of specific bands, thus optimising industrial and research
processes. This work combines Raman spectroscopy with chemometric tools for classifying the
biodiesel samples according to their synthetic route and water content.
Figure 1: (a) Pre-processed Raman spectra (b) PLS-DA classification model.
Table 1: PLS-DA classification of biodiesel in compliance to ANP n° 45/2014 resolution for water content.
Compliant Non-Compliant Accuracy Sensibility Specificity
Calibration Set 16 6 1 1 1
Validation Set 7 3 0.7 1 1
1. Knothe, G. Some aspects of biodiesel oxidative stability. Fuel Processing Technology 88, 669–677 (2007).
2. Lôbo, I. P., Ferreira, S. L. C. & Cruz, R. S. da. Biodiesel: parâmetros de qualidade e métodos analíticos. Química
Nova 32, 1596–1608 (2009).
3. Sala, Oswaldo. Fundamentos da espectroscopia RAMAN e no infravermelho. Unesp, 2008.
[PRH-ANP, UFABC, FAPESP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

425
CLASSIFICATION OF COMMERCIAL CACHAÇAS AGED IN DIFFERENT
WOODEN BARRELS USING SERS AND CHEMOMETRIC METHODS
Carlos A. Riosa,*(PG), Márcia C. Breitkreitza (R)

aInstitute of Chemistry, State University of Campinas, Campinas-SP, Brazil, 13084-971
*e-mail: c228209@dac.unicamp.br
The SERS (Surface Enhanced Raman Scattering)1 effect allows to significantly increase the Raman
signal to detect low concentrations of compounds of interest. In Analytical Chemistry, several
applications have been reported in the literature using SERS for this purpose, for example, detection
of pesticides/herbicides/insecticides in food2, toxic substances in processed foods3 and analysis of
different compounds in beverages.4 In the beverage market, Brazil stands out worldwide for the
production and export of cachaça. The aging of cachaça in wooden barrels is a very important step,
since the drink recently distilled has aggressive sensory characteristics and a strong alcoholic flavor,
therefore it is not pleasant for consumption.5 Oak barrels are preferred due to the significant sensory
improvement, however, it can be expensive, since this is not a typical Brazilian wood.6 In this context,
counterfeiting using native and cheaper woods, such as umburana, jequitibá, balsam and peanuts 7
are found. The objective of the study was to develop classification models to discriminate samples
of cachaça aged in oak barrels from samples aged using other types of wood, using Raman spectra
with the aid of the SERS effect to detect minor components. SERS substrates were constructed by
covering an A4 sheet with a layer of wax except for small circumferences, on paper, with a diameter
of 4 mm. On the hydrophilic region (paper), 15 µL of the concentrated solution (25x) of nanoparticles
was added. The synthesis of gold nanoparticles was based on the modified procedure of the Lee-
Meisel method.8 After drying for 24h, the same volume of sample was added. In total, 33 samples of
cachaça of different brands were obtained and aged in 7 types of barrels: peanut, balsam, oak,
stainless steel, jatobá, jequitibá and umburana. The spectra were collected using a laser
spectrometer at 785 nm and a CCD detector cooled at -50 °C. The samples were exposed to the
laser for 10 s in a total of 8 accumulations. The spectrum region used was from 350 to 1700 cm-1
with a resolution of 2 cm-1. The spectra were processed in Matlab (v. 8.3) using the PLS-Toolbox
package (v. 8.6.2). The spectra were preprocessed by correcting the baseline by the WLS method
(weighted least squares) with a sixth order polynomial, normalization by unit length vector, smoothing
using the Savitsky-Golay filter, with a first order polynomial and nine-point window and finally, were
centered on the mean. For the construction of the classification model, the partial least squares
discriminant analysis method (PLS-DA) was used. The Kennard-Stone9 algorithm was used to
separate samples in the calibration and validation set. The PLS-DA model was built using 8 latent
variables with 224 spectra in the calibration set and 96 in the validation set. In cross-validation, the
“randow subsets” method was used, with 12 data splits and 20 interactions. The explained variance
in matrix X was 93.44% and in vector y was 73.17%. The classification error for validation set was
6%. For the validation set, the values for sensitivity, specificity and accuracy were 86.7, 98.5 and
94.8%, respectively.
1 Moskovits, M. J., Chern. Phys., vol. 69, 1978, p. 1
2 Fan, Y., Lai, K., Rasco, B.A., Huang, Y., Food Sci. Tech., vol. 60, 2015, p. 352
3 Lin, M., He, L., Awika, J., Yang, L., Ledoux, D.R., Li, H., Mustapha, A.J., Food Sci., vol. 73, 2008, p. 129
4 Ilaslan, K., Boyaci, I.H., Topcu, A., Food Control, vol. 48, 2015, p. 56
5 Decreto Nº 6871, de 4 de julho de 2009. Capítulo VII, Seção IV: Das bebidas alcoólicas destiladas. Brasilia, 2009.
Disponível em: http://www.planalto.gov.br/ccivil_03/_Ato2007-2010/2009/Decreto/D6871.htm. Acesso em: 10/jul/2022
6 Miranda, M.B., Souza, M., Nilo, G., Souza, B., Eduardo, A., Horri, J.A., Ricardo, A., Food Sci. Tech., vol. 27, 2007, p.
897
7 Bernardes, C.D.; Barbeira, P.J.S., Food Anal. Methods, vol. 9, 2016, p. 1053
8 Lee, P.C., Meisel, D., Phys. J. phy. Chem., vol. 86, 1982, p. 3391
9 Kennard, R.W., Stone, L.A., Technometrics, 1969, p.1
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

426
CLASSIFICATION OF THE VARIETY OF GREEK HONEY USING MACRO-
ELEMENTS, PHYSICOCHEMICAL PARAMETERS AND CHEMOMETRIC
Erklaylle G. C. da Silva (PG)a*, Maria-Anthi Priakou (PG)b, Marios Kostakis (R)b, Loukas
Gialouris (PG)b, Carolina S. Silva (R)c , Marilena Dasenaki (R)b, Maria Fernanda Pimentel (R)a
aDepartment of Fundamental Chemistry, Federal University of Pernambuco, Recife, PE, Brazil
b Laboratoryof Food Chemistry, Department of Chemistry, National and Kapodistrian University of Athens, Zografou,
Greece, GR-15771
cFaculty of food sciences and Nutrition and Centre of Biomedical Cybernetics, University of Malta, Msida, Malta
*e-mail: erklaylle.gabriely@hotmail.com
Honey products, beneficial for human health, consist of a complex mixture of sugars, minerals,
vitamins, enzymes, volatiles and phenolic compounds. Both botanical and geographical origin affect
its composition, sensory attributes, and the market price. In the absence of standardized
regularization of such products, they have become a major target for falsification,[1] making it
important to understand how the composition of honey is related to its origin. Most studies of the
botanical origin of honey are carried out by chromatography and physicochemical analysis, together
with chemometrics, especially principal components analysis (PCA) and discriminant analysis
(DA)[1]. DA approaches, however, are not suitable for food authenticity, being necessary to use of
class modelling approaches [1]. The present work aimed to classify different varieties of monofloral
Greek honey based on their elemental composition and physicochemical properties using soft
independent modelling by class analogy (SIMCA) and data-driven SIMCA (DD-SIMCA)[2]. For this
work, 187 samples of 8 varieties were studied - 3 honeydew (fir, oak and pine) and 5 honey blossom
(arbutus, chestnut, heather, orange and thyme). These were analysed with different methodologies:
(i) Microwave Plasma Atomic Emission Spectrometry (MP-AES) to quantify macro-elements (Na,
Ca, Mg, K) and (ii) the standardised methods proposed by International Honey Commission to
determine the physicochemical parameters (pH, acidity, electric conductivity and moisture). The 5-
components PCA model, explained 91.04% of accumulated variance, showing a difference between
honeydew and blossom honeys. The blossom samples had higher concentrations of Na and
moisture, compared to honeydew samples, which had higher concentrations of Mg, K, pH, acidity
and conductivity. The chestnut variety (blossom honey) showed similar scores values when
compared to the honeydew. When Class-modelling models were built, it was possible to obtain more
than 80% true negative for fir, oak, orange, pine and thyme. In general, the best models showed that
the false positive honey samples were those of the same type (honeydew/blossom honey) This
preliminary study shows that the authentication of Greek honeys remains challenging due to the
variety of chemical profiles within and among the botanical origins. However, some of the studied
varieties have shown promising results and new variables, such as trace elements, should be further
included.
Table 1. Results of classification of different varieties of honey.
SIMCA CDD-SIMCA (classic) RDD-SIMCA (Robust)
True positive True Negative True positive True Negative True positive True Negative
Arbutus 100% 43% 88% 58% 88% 45%
Chestnut 100% 32% 71% 55% 86% 38%
Fir 88% 90% 63% 97% 50% 97%
Heather 100% 42% 75% 69% 75% 69%
Oak 88% 82% 100% 86% 100% 85%
Orange 100% 82% 100% 84% 100% 85%
Pine 83% 77% 50% 88% 83% 86%
Thyme 100% 84% 100% 90% 83% 89%
1 Tsagkaris, A. et al. Royal Society of Chemistry, 11, 2021, 11273.
2 Pomerantsev, A. L., Rodionova, O. Y., Journal of Chemometrics, 28, 2014, 438.
[FACEPE, CNPq. INCTAA, NUQAAPE, CAPES]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

427
COMPARISON BETWEEN VARIABLE SELECTION ALGORITHMS IN PLS
REGRESSION FOR PREDICTION OF Fe and Ti IN SOIL
Giovanna Abrantesa(PG), Mirelly Gomesa(PG), Angelo J. Maiab(PG), Rennan C. Nascimento
b
(R), Yuri J. A. B. Silvac(R), Germano Verasa*(R)
aState University of Paraíba, Chemistry Department, Campina Grande, Paraíba, Brazil, 58.429-500
bFederal Rural University of Pernambuco, Agronomy Department, Recife, Pernambuco, 52.171-900
cFederal University of Piauí, Agronomy Department, Bom Jesus, Piauí, 64.900-000
*e-mail: germano@servidor.uepb.edu.br
The determination of metals is important to identify potentially toxic elements or others with levels
above or below desirable levels for the intended purposes for the soil. The most commonly used
analyzes are spectrometry (AAS,AES) and DRX for mineralogical characterization. However, these
analyze are relatively time-consuming and expensive. Some works have already been developed
on simultaneous multicomponent analysis of metals using DR-NIR. Allied to the use of analytical
techniques, methods can be associated for processing chemometric data that allow analyzes with
an acceptable degree of confidence, making the separation of analytes unnecessary and, therefore,
obtaining an environmentally friendly process. The chemometric tools should allow the obtaining of
parsimonious and robust models due to the complexity of the matrix under study. A useful approach
to building models in high-dimensional and chemically complex datasets is the selection of variables
to eliminate or, at least, reduce the influence of uninformative variables. This study aimed to compare
different PLSR models for predicting Fe(9255-68950mg/kg) and Ti(1602-10450mg/kg) in soil
samples. The regression algorithms chosen were PLSR using all variables(full PLSR), and those
with selection of variables, by intervals(iPLS) and by intervals selected by the Successive Projections
Algorithm(iSPA-PLS) and firefly bioinspired algorithm[1] (FFiPLS). For the models created from
algorithms with variable selection, 20 intervals selected. For the models built by FFiPLS, population
conditions of 100 fireflies in 100 life cycles were stipulated. The samples (101) used in this work
were obtained from soils from the Ipojuca River basin, located in the state of Pernambuco between
the parallels 8°09'50” and 8°40'20” of south latitude and the meridians 34°57'52” and 37°02'48”
longitude west of Greenwich. The reference analysis of the metals was by ICP-OES, PerkinElmer,
model Optima DV7000 with coupled cyclonic chamber system after extraction by acid digestion.
Subsequently, the samples were dried in an oven at 50°C for 48 hours and were measured in the
region from 1000-2500 nm using a PerkinElmer FT-IR/NIR spectrometer, Frontier model, coupled
with a diffuse reflectance accessory. The samples were divided into calibration (76 samples) and
prediction (25 samples) sets using the SPXy algorithm. The chemometric models were built on raw
data and pre-processed by MSC, SNV, mean centering, baseline adjust, autoscaling, mean
reduction and Savitzky-Golay smoothing and derivation (second-degree polynomial and 17-point
window). All data were treated in Matlab environment. To define the most suitable model, the figures
of merit R²pred, RMSEP(mg/Kg), biaspred(mg/Kg), REP and SDV were evaluated. The FFiPLS proved
to be more suitable for Fe using SG preprocessing because it has lower RMSEP(4.73e+3),
biaspred(248.72), REP(16.82), R²pred(0.77) and SDV(4.85e+3), other models presented higher R²pred,
however they used a greater number of variables(>11) and overfitting of the models. For Ti, the best
model obtained was from the FFiPLS preprocessed by baseline correction, as it also presents low
RMSEP(680.84), biaspred(208.48), REP(17.95) and SDV(786.55) and R²pred(0.79) from a bit of
number of latent variables. Based on the results, it was possible to observe that the FFiPLS obtained
more parsimonious models for predicting Fe and Ti in soil by presenting more adequate parameters,
which confirms its viability.
1 OLIVEIRA, DLB et al Bio-inspired algorithm for variable selection in i-PLSR to determine physical properties, thorium
and rare earth elements in soils from Brazilian semiarid region. Microchemical Journal, v. 160, Part A, 2021, p. 1-7.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

428
COMPARISON OF MULTIVARIATE CALIBRATION MODELS BUILT WITH
DIFFERENT MASS SPECTRA RESOLUTIONS
Jussara V. Roquea* (R), Marcella F. Rodriguesa (PG), Michel M. F. Vieiraa (PG),
Gabriel Henry M. Dufrayera (UG), Gabriel F. dos Santosa (R), Boniek G. Vaza (R)
aFederal University of Goiás, Chemistry Institute, Goiânia, Goiás, Brazil, 74690-900
*e-mail: jussara_roque@ufg.br
The constant advances in analytical instrumentation in the last decades allow the quantitative and
qualitative study of different types of samples. In this context, high-resolution analytical
instrumentation, such as mass spectrometry has been widely used and typically generates a large
data amount1-3. In this way, data analysis becomes an essential task to give interpretable and valid
results. However, this task can become time-consuming and requires a powerful computer for data
processing. Thus, this work aimed to evaluate different mass spectra resolutions used for building
multivariate quantitative models. Thirty-six samples of a mixture of naphthenic acids (NAs) were
prepared in a range of 0.5 to 180 μg mL-1. The NAs used were: cyclopentanecarboxylic, benzoic,
cyclohexanebutyric, 1-naphthoic, 9-anthracenecarboxylic, pentadecanoic, 2-methyloctadecanoic,
decanoic, and 3,5-dimethyladamantane-1-carboxylic acids. Mass spectra data were obtained using
a High-Performance Liquid Chromatography (Agilent Technologies) coupled with Q Exactive hybrid
Quadrupole-Orbitrap mass spectrometer (Thermo Fisher). From the original data acquired (77187
variables with 0.001 resolution ranging from 100 to 400 m/z), the other three datasets were obtained
by decreasing the resolution until reached the nominal mass. Briefly, the mass values were rounded
to decrease the decimal places and the intensities of all equal results were summed to give the new
intensity value. Partial least squares (PLS)4 regression was used to build multivariate models with
mass spectra and NAs concentration by using random cross-validation. Samples were split into
calibration (24) and prediction (12) sets a thousand times to achieve the root mean square error of
calibration (RMSEC) and prediction (RMSEP). These parameters were pairwise compared by the
Tukey test (α = 0.05). Ten latent variables were used to build PLS models with the normalized mass
spectra and mean-centered NAs concentrations. Figure 1 shows the dataset mass spectra and
RMSEs histograms.
Figure 1. Mass spectra of each dataset (1-4) in different resolutions and the respective root mean square error histograms
of calibration (RMSEC) and prediction (RMSEP) in μg mL-1 unit.
In Figure 1, the mass spectra of datasets 1-3 was one hundred times more intensity that dataset 4.
This fact is due the sum of the intensities in step of building the datasets. Additionally, we observed
the increase of RMSEs with the increase in datasets resolution, with the frequency maxima
increasing for RMSEC and decreasing for RMSEP. Tukey test shows that the RMSEs of datasets 1-
3 are significantly different (p-value = 3.77×10-9) from dataset 4. These results show that a high-
resolution spectrum is not necessary to provide an accurate quantitative model, reducing the data
processing time. Thus, for this quantitative analysis, the mass spectra with the nominal mass
resolution were enough to build an efficient PLS model to predict accurately NAs concentrations.
1 Mattoli L, Gianni, M, Burico M, Castro JK, Vieira LP, Batista SD, Mass Spectrometry Reviews, 2022.
2 Donohoo KB, Wang J, Goli M, Yu A, Peng W, Hakim MA, Mechref Y, Electrophoresis, 43, 2022.
3 Pereira I, Aguiar DVA, Vasconselos G, Vaz, BG, Fundamentals and Applications of FT Mass Spectrometry, 2019.
4 Roque JV, Cardoso W, Peternelli, LA, Teófilo, RF, Analytica Chimica Acta, 1075, 2019.
[Petrobras, LaCEM]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

429
COMPARISON OF PCA AND ASCA CHEMOMETRIC APPROACHES FOR
EXPLORING GC×GC-MS DATA OF ROASTED COFFEE BEANS
Victor H. C. Ferreiraa*, Leandro W. Hantaoa, Ronei J. Poppia
aUniversity of Campinas, Institute of Chemistry, Campinas, São Paulo, Brazil, 13083-970
*e-mail: vhcferreira@outlook.com
Coffee is one of the most consumed beverages in the world and Brazil is one of its most important
producers and consumer markets. Although many different traits affect the products’ flavor and
aroma, consumers tend to consider aspects such as roasting level and bean type as indicators of
coffee quality. Therefore, the analysis of the volatile compounds of different types of roasting of
varied coffee beans may help find compounds related to better consumer evaluations. A design of
experiments (DoE) was conducted to evaluate the roasting process of raw coffee beans in the lab
considering two main factors: different roasting procedures; and distinct subgroups of green Coffea
arabica beans. Roasting procedures were tested in three levels, while beans’ subgroups had five
levels related to the five subgroups of green beans obtained from the Regional Cooperative of Coffee
Growers in Guaxupé (Cooxupé). Beans were classified according to coffee cupping done by
Cooxupé professionals and codified in letters from A to E in descending order, with A being the best
and E the worst bean out of the five. All roasted samples were submitted to headspace SPME and
then analyzed by comprehensive two-dimensional gas chromatography (GC×GC-MS). Two
chemometric tools were then used to explore the chromatographic data. The first one was principal
components analysis (PCA), which could only find patterns related to the roasting process. No PC
was able to properly distinguish any bean subgroup. The second tool was ANOVA-simultaneous
component analysis (ASCA), which considers the DoE in its multivariate analysis and generates a
different model for each of the factors and interactions. In this work, however, only the factors were
deemed statistically significant. The model related to the roasting factor had a score plot that was
like the pattern found with PCA, whereas the model associated with the bean subgroup easily
distinguished bean A from the others. A comparison of PCA and ASCA results concerning bean
subgroups is shown in Figure 1. From the loadings of the ASCA model, some compounds were
identified as important markers of bean A, such as 2,6-dimethylpyrazine, 1-hydroxy-2-butanone, and
2-furanmethanol, which are related to roasted, caramel, and coffee odors, as well as nutty and sweet
flavors. Thus, ASCA proved to be better suited to the dataset as it provided information from both
aspects sought by coffee consumers.
Figure 1. Score plots of samples identified according to the five bean subgroups, coded from A to E. PCA model (a) and
ASCA model for the bean subgroup factor (b).
[CNPq, CAPES, FAPESP, Nova Analitica, and Cooxupé]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

430
DETERMINATION OF ACETYLSALICYLIC ACID BY RAMAN
SPECTYTROSCOPY USING UNIVARIATE ANALISYS
Walace M Moreira (PG)a*, Benedito R Alvarenga Junior (PG)b, Luis G Q Durango (PG)b ,Moacir R Forim (R)b,
Renato L Carneiro (R)b
a Federal Institute of Maranhão, Departament of higher Education, Açailândia, MA, Brazil, 65930.000
bFederal University of São Carlos-UFSCar, Departament of Chemistry, São Carlos, SP, Brazil, 13560-905
*e-mail: walace.moreira@ifma.edu.br
The acetylsalicylic acid (ASA) is a nonsteroidal anti-inflammatory drug with antipyretic properties
commonly used as analgesic throughout the word[1,2]. The quality control in pharmaceutical industry
is performed by using chromatographic techniques, such as HPLC. However, this technique
presents disadvantages such as high cost, time and consume of organic solvents. Raman
spectroscopy associated with chemometrics tools is an alternative method in the ASA analysis. This
approach presents huge analytical versality, presenting uncounted advantages and versatility, such
as no invasive analysis, high analytical frequency, low cost and minimal preparation of samples.
ASA presents high solubility in organic solvents such as ethanol, chloroform and acetone[3]. The use
of these solvents brings some drawbacks in the Raman analysis, since organic solvents often
present peaks overlapping the signal of the analyte. A strategy to increase the ASA water solubility
is promote its ionization using a strong base. This work aims to use Raman spectroscopy for
quantification of acetylsalicylic acid in commercial tablets employing sodium hydroxide solution (0.3
mol.L-1) as a strategy to increase drug solubility and without the use of chemometric tools, using only
a conventional calibration curve. In addition, Principal Components Analysis (PCA) was applied to
evaluate the ASA stability in solution at different pH’s during kinect monitoring studies. For
multivariate kinect monitoring, four standard solutions of ASA were prepared in the concentration of
3 % m/v at pH ranged from 6 to 12. In the first two hours of monitoring, Raman spectra were collected
each 30 min, and after 2 h, spectra were collected every hour up to 8 h. For quantitative analysis,
acetylsalicylic acid standard in NaOH solution were prepared from 1 % to 5 % (m/v). Solutions were
prepared adding different volumes of NaOH according to mass of acetylsalicylic acid used. This
procedure had as goal to obtain the same pH for all solutions (pH 7) in order to avoid ASA
degradation and maximize the ASA solubility. Moreover, two brands of commercial tablets containing
500 mg of ASA were analyzed and compared to the reference method (HPLC). The degradation
kinect monitoring using Raman and PCA allowed to verify the stability of ASA in different pHs over
time. The scores of PCA shows that pH 6 presented promote the better compromise between
stability and water solubility for ASA. Moreover, it was possible to observed the kinect degradation
by variation of PCA score along the time. The method proposed for the univariate quantification give
rise to a calibration curve coefficient of determination of 0.991. The method was capable to quantify
acetylsalicylic acid in commercial tablets presenting a limit of detection (LOD) and quantification
(LOQ) of 0.205 % and 0.623 % m/v, respectively. The relative deviation of this method was 0.63%
for brand A and 0.88% for brand B, when compared to reference method. The proposed method
presented several advantages compared to standard method such as low cost, fast determination,
and simpler sample preparation.
1 Korolkovas A, Burckhalter, JH, Química Farmacêutica, Guanabara Koogan, 1ª ed., 1982, Brasil.
2 Sallum LF et.al., Spectrochim. Acta - Part A Mol. Biomol. Spectrosc. 133, 2014, 107.
3 Maia GD, Giulietti M, J. Chem. Eng. Data, 53,2008, 256.
[IFMA, PPGQ/UFSCar,Departamento de Química/UFSCar]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

431
DETERMINATION OF API GRAVITY AND TOTAL SULFUR CONTENT IN
CRUDE OIL SAMPLES USING MACHINE LEARNING ALGORITHMS
Henrique Faccinª* (UG), Vitoria H. Cauduroª (PG), Edson I. Müllerª (R)

a Federal University of Santa Maria, Department of Chemistry, Santa Maria, RS, 97105-900, Brazil
*e-mail: henrique.faccin@ufsm.br
Crude oil is a complex matrix constituted by a large variety of hydrocarbons and other organic
molecules containing sulfur, nitrogen and oxygen.1 The feasibility of extracting crude oil and its
subsequent refining is evaluated, among other logistical and economic factors, from the study of the
elemental composition and physicochemical properties of petroleum samples from exploration
wells.2 For example, a high sulfur content is strongly associated with a higher incidence of corrosive
processes in refineries and in the oil derivatives industry. A high sulfur content is also generally
associated with lower API gravity oils. Determination of such properties is based on international
standards and is usually extremely expensive, laborious and not eco-friendly. Thus, this study aimed
to use attenuated total reflectance mid-infrared spectroscopy (FT-MIR/ATR), a rapid instrumental
analysis that requires a small sample volume and no prior sample preparation. To estimate the API
gravity and the total sulfur content of crude oil samples, machine learning algorithms such as Partial
Least Squares Regression (PLS-R) and Support Vector Regression (SVR) were used. A total of 116
oil samples were evaluated, presenting API gravity between 19 and 41° and total sulfur content
between 660 and 8,938 µg·g-1. Spectra were obtained in triplicate using FT-MIR/ATR, in a region
between 4,000 and 650 cm-1 (3,351 variables), with resolution of 4 cm-1. For each proposed model,
the samples were splitted into two groups through the Duplex algorithm, so that 70% of the
observations were selected as the training set and 30% as test set. Outliers were evaluated and
detected using robust estimators.3 A full-cross validation was performed to evaluate the final models.
The best PLS-R model was selected according to the number of latent variables associated with the
lowest root mean square error in cross validation (RMSECV) obtained. To calculate the SVR, the
parameters of cost (C), margin (ε) and gamma (γ) were optimized for a radial kernel aiming at the
smallest mean square error (MSE). Referring to SVR, the dimensionality of the data was previously
reduced by Principal Component Analysis, considering the number of principal components
necessary to reach an explained variance of 99%. The computational calculations were carried out
using the R Statistical Software (version 4.2.0), with prospectr, mdatools4 and e1071 packages. The
main models obtained are summarized in Table 1. It is important to emphasize that the performance
of the SVR algorithm considering the full IR spectrum is comparable with the PLS-R algorithm with
previous variable selection by the VIP (Variable Importance on Projection) criteria greater than or
equal to 1.
Table 1. Best models calculated using the evaluated machine learning algorithms.
Variable Algorithm Pretreatment Spectra region R² (C) RMSEC RMSECV RMSEP
API gravity PLS-R MC, MSC Full spectra 0.914 1.20° API 1.47° API 1.61° API
API gravity PLS-R MC, MSC VIP ≥ 1 0.920 1.24° API 1.49° API 1.38° API
API gravity SVR radial MC, MSC Full spectra 0.911 1.27° API 1.44° API 1.42° API
Total sulfur PLS-R MC, SNV Full spectra 0.901 327 µg·g-1 568 µg·g-1 1,011 µg·g-1
Total sulfur PLS-R MC, SNV VIP ≥ 1 0.970 173 µg·g-1 404 µg·g-1 798 µg·g-1
Total sulfur SVR radial MC, SG Full spectra 0.948 308 µg·g -1 524 µg·g -1 725 µg·g-1
R² (C): coefficient of determination for calibration; RMSEC: root mean square error of calibration; RMSECV:
root mean square error of cross validation; RMSEP: root mean square error of prediction. Finally, by means
of the SVR algorithm with radial kernel, root mean square errors of prediction (RMSEP) were obtained in the
order of 1.4° for the API gravity and 725 µg·g-1 for the total sulfur content.
1 Mohammadia M, Khorramia MK, Vatanib A, Ghasemzadeha H, Vatanparastc H, Bahramiand A, Fallah A, Spectrochimica

Acta Part A: Molecular and Biomolecular Spectroscopy, 232, 2020, 118157.
2 Speight JG, The Chemistry and Technology of Petroleum, CRC Press, 5th ed., 2014.
3 Rodionova OY, Pomerantsev AL, Analytical Chemistry, 92, 2020, 2656.
4 Kucheryavskiy S, Chemometrics and Intelligent Laboratory Systems, 198, 2020, 103937.
[UFSM]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

432
DETERMINATION OF QUALITY PARAMETERS OF PULP OF Spondias mombin
L. USING NEAR INFRARED SPECTROSCOPY WITH PARTIAL LEAST SQUARE
AND VARIABLES SELECTION ALGORITHM
Mirelly Gomes (PG), Lêda Figueiredo (PG), Germano Veras*(R)
State University of Paraíba, Chemistry Department, Campina Grande, Paraíba, Brazil, 58.429-500
*e-mail: germano@servidor.uepb.edu.br
Brazil occupies the third place in production and commercialization of tropical fruits consumed, after
China and India. Besides the known fruits there are also a great variety of species that are little
known, even by the Brazilians themselves, but that have great potential for commercialization in
natura or as pulps, for example. In terms of Brazilian regions, the Northeast has tropical fruits that
are still little explored, among which the fruit of Spondias mombin L., known as cajá or taperebá in
the North, stands out. Besides the outstanding flavor of the fruit, the cajá contains antioxidant
substances, flavonoids, phenolic acids, vitamins A and C, anthocyanins and carotenoids in
considerable amounts. However, when the fruit is processed these subtances may be lost, affecting
brix, total acidity and pH, for example. In this sense is essential to guarantee that the commercialized
product meets the characteristics that the consumer wishes in terms of nutritional and functional
properties. However, the recommended analysis are expensive and time consuming that difficult the
quality control of the products into account that most of the production is conducted by small
companies with local or regional commercialization. Therefore, the objective of this work was to
evaluate the quality parameters of S. mombin pulp evaluating total titratable acidity (TA), pH and
Total Soluble Solids (TSS), using Near Infrared Spectroscopy (NIRS) associated with Partial Least
Squares Regression algorithms: iPLS, iSPA-PLS and FF-iPLSR[1] on raw and pre-processed data.
The data were processed using Standard Normal Variate (SNV), Multiplicative Scatter Correction
(MSC) and Savitzky-Golay derivation (with combinations of first and second derivatives, fist and
second degree polynomial and 17, 21 and 25-point window). The data were obtained using 36
samples with spectral range of 900 to 1400 nm. The results were evaluated using the predictive
ability in terms of the Elliptical Confidence Region (ECR), Root Mean Square Error of Calibration
(RMSEC) and Prediction (RMSEP), Coefficient of Determination of calibration (R2cal) and prediction
(R2pred) and bias. The calibration (27 samples) and prediction (9 samples) sets were selected by
SPXy algorithm. In terms of the results obtained, the determination of TSS with iSPA-PLS using SNV
and MSC preprocessing obtained the best results. In contrast, the chemometric models to determine
pH and TA the best results presented R2cal and R2pred distant from unity, not consolidating an efficient
model for determining these properties. Thus, the algorithm used iSPA-PLS can be used to build
effective and parsimonious chemometric models, capable of associating the responses of each
variable and obtaining the best answers to the proposed problem and in a much less expensive way
in terms of cost and time.
1 Oliveira, D. L. B.,Pereira, L. H.S, Schneider, M.P., Silva, Y. J. A. B., Nascimento, C. W. A., Straaten, P. V., Silva, A. B.,
Gomes, A. A., Neto, G. V. Bio-inspired algorithm for variable selection in i-PLSR to determine physical properties, thorium
and rare earth elements in soils from Brazilian semiarid region. Microchemical Journal, 2020.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

433
DIFFERENTIATION BETWEEN SPECIAL AND TRADITIONAL GREEN COFFEE
BEANS USING SPECTROPHOTOMETRIC AND CHEMOMETRIC ANALYSIS
Winston P. C. Gomesa* (PG), Wanessa R. Melchertb (R)

aCenter for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil
bCollege of Agriculture "Luiz de Queiroz", University of São Paulo, Piracicaba, SP, Brazil
*e-mail: winstonpcg@usp.br
The green coffee bean is the dried seed (endosperm) of the coffee fruit, peeled and before going
through the roasting process1. The objective of this work was to determine compounds present in
green coffee beans using spectrophotometric analysis and to find the differentiation of the beans, in
special or traditional ones, through chemometrics. For this, the contents of total phenolic compounds
(TPC), reducing (RS), non-reducing (NRS), and total (TS) sugars were quantified of 10 samples of
special coffees and 8 of traditional coffees. The extraction used was according to the cup test
protocol2. The results obtained were submitted to Shapiro-Wilk, Levene, and Student's t analyses,
for means, at a confidence level of 95% (α = 0.05) and Principal component analysis (PCA) was
employed, following the autoscaling of the data (Figure 1).
Figure 1. Biplot of the first two main components for the samples of special green coffee (E1 to E10, orange color) and
traditional (T1 to T8, green color) referring to the spectrophotometric analysis (a) and the altitude correlated with the
spectrophotometric analysis (b).
In Figure 1a, it generated the PCA referring only to the compounds determined by
spectrophotometry, while in Figure 1b, the altitudes of the cultivars of the coffee samples were
correlated with the spectrophotometric analyzes. Statistically, the levels of TPC, NRS, and TS
showed significant differences, with the special coffee having the highest content of these
compounds. Species responsible for being aromatic precursors due to Maillard and Strecker's
reactions and final acidity, being factors that will influence the final characteristics of the drink 3. In
addition, it was possible to classify the green coffee beans into special and traditional, both using
only spectrophotometric analyzes and when correlating the altitudes of the samples, since there was
the formation of distinct clusters. In this way, there are advantages that allow classification without
the use of a trained and experienced evaluator, since their previous experience can influence the
results, due to their expertise in a particular type of coffee. In addition to being a simple, fast and low
cost methodology.
1 Gomes WPC, Pires JA, Teixeira NN, Bortoleto GG, Gutierrez EMR, Melchert WR, Journal of Food Measurement and
Characterization, 16, 2022, p. 1.
2 Specialty Coffee Association of America – SCAA, SCAA Protocols, 2009. 1.
3 Pereira LL, Moreira TR, Quality determinants in coffee production. Springer, 2021.
[CAPES, FEALQ FAPESP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

434
DIFFERENTIATION OF GREEN COFFEE BEANS BY ICP OES AND
CHEMOMETRIC TOOLS
Gustavo M. Alves (UG)a*, Winston P. C. Gomes (PG)b, Wanessa R. Melchert (R)a

a College of Agriculture "Luiz de Queiroz, University of São Paulo, Piracicaba, SP, Brazil
bCenter for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil
*e-mail: gustavomaximiano@usp.br
Green coffee bean is the endosperm of the coffee fruit peeled and before going through the roasting
process1. The objective of this work was to determine minerals present in green coffee beans using
elemental analysis of coupled plasma atomic emission spectroscopy inductively (ICP OES). For this,
the minerals Na, Mg, K, P, Ca, Mn, Fe, Cu, and Zn were quantified using ICP OES of 10 samples of
specialty coffees and 8 of traditional coffees. Dry digestion2 was performed in two ways: i) natura or
ii) extract of the samples obtained by the cup test protocol3. The results obtained were submitted to
ANOVA and Tukey analyses with a confidence level of 95% (α = 0.05) and Principal component
analysis (PCA) was used in the differentiation, following the data autoscaling (Figure 1).
a) b)
Figure 1. Biplot of the first two main components for the samples of special green coffee (S1 to S10, orange color) and
traditional (T1 to T8, green color) referring to the spectroscopy analyses: (a) in natura and (b) the extract.
Statistically, the content of Mg, K, F, Ca, Mn, Fe, and Cu have significant differences when the
analysis is conducted directly to green coffee beans (Figure 1a), whereas the samples of specialty
coffees presented higher concentrations of Mn and Cu. The other minerals presented higher
concentrations on traditional coffee ones. For the analysis of the extracts (Figure 1b), the levels of
Mg, Cu and Zn were higher on specialty coffees and these minerals presented significant difference.
In Fig. 1a, sample S10 has a tendency to cluster with traditional samples, justified by the fact that it
has a content close to the content found in traditional samples for mineral K, which is the main
mineral that influences PC1. This influence may be since the coffee bean is processed by the dry
process, consequently presenting K levels like robusta coffees4, which are usually traditional coffees,
in addition to being an indication that the S10 sample may have gone through the dry process. Fig.
1b, sample T2, despite not being close to the grouping of special samples, it ended up being
influenced by PC1, which has great influence by the mineral Cu. Cu is reported as a good
discriminator of species5, so the sample T2 may be of the arabica species, therefore causing this
sample to have the influence of PC1. Therefore, special and traditional green coffee beans can be
differentiated implementing spectroscopy analyses linking their mineral concentrations, both in
natura and in extract.
1 Gomes WPC, Pires JA, Teixeira NN, Bortoleto GG, Gutierrez EMR, Melchert WR, Journal of Food Measurement and
Characterization, 16, 2022, p. 1.
2 Instituto Adolfo Lutz, Métodos químicos e físicos para análise de alimentos, 4ª Edição, 2008, Brasil.
3 Specialty Coffee Association of America – SCAA, SCAA Protocols, 2009. 1.
4 Preedy V, Processing and impact on active components in food. Academic Press, 2015.
[CNPq, CAPES, FEALQ, FAPESP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

435
DIGITAL IMAGE COMBINED TO INFRARED SPECTRA IN ACAI
AUTHENTICATION AGAINST ADULTERATION BY SEQUENTIALLY
ORTHOGONALIZED ONE CLASS PARTIAL LEAST SQUARES
Larissa O. Onça a* (PG), Alisson S. Araújo ª (PG), Mateus P. Schneider a (PG), Paulo H. G. D.
Diniz b (R), Adriano G. Araújo a
a Universidade Federal do Rio Grande do Sul, Institute of Chemistry. Porto Alegre, Rio Grande do Sul, Brazil, 91501-900
bUniversidade Federal do Oeste da Bahia, Center of exact sciences and technologies. Barreiras, Bahia, Brazil, 47808-
021
*e-mail: lariolion@gmail.com
Acai is the berry of a palm scientific named Euterpe oleracea, which is endemic to the Amazon
region. Mixed with water, its edible berries undergo maceration and sieving. Acai has a noteworthy
nutritional composition, due to its content of lipids, fiber, proteins and polyphenols, among which are
flavonoids and anthocyanins providing high antioxidant proprieties. Once acai’s consumption is
wildly spread through the country and abroad, its quality control become necessary to ensure
protection against intentional and unintentional fraud, guarantying economical credibility and food
safety.1,2 As counterfeiting may happen in different steps of the production process or even by mixing
new substances, for instance, cassava flour, beet pulp or maize starch, analytical methods for fraud
detection is a challenge. Therefore, the present work aims to the acai’s pulp authentication while
applying analysis of combined Fourier transformed near infrared spectroscopy (FTIR) and digital
imaging data. The approach proposed here is to access acai’s authentication by sequential
orthogonalized one class partial least squares regression (SO – OCPLS). Different sources built a
180 group of samples, among of which 60 were the pure ones (target class) and 120 were
adulterated the adulterated ones (untargeted class). Acai sample spectra were registered through
the IR wavenumber range of 3600 and 600 cm-1. Afterwards, data underwent Savitzky-Golay
smoothing with first derivate and second-degree polynomial ith window-size 15 followed by a
multiplicative scattering correction (MSC). A total of 180 acai pulp samples were used to build the
model, 60 unadulterated samples (class 1) and 120 adulterated samples (60 adulterated with
cassava flour and 60 with added wheat flour) associated with class 2. Sample partitioning was
carried by Kenard-Stone algorithm in different steps, regarding the signal source’s block and sample
class. Samples among Class 1 on Block 1 (digital images) were partitioned into 35 to the training
group, 10 to the test and 15 to the validation. Samples from Class 1 but on Block 2 (FTIR) underwent
the same partition ratio. Half of untargeted samples were starch adulterated meanwhile the other
half had been mixed with cassava flour. First, a principal component analysis showed the best
spectra ranges for sample segregation. Then, digital imaging data (block 1) and FTIR (block 2) were
separately modeled by OC – PLS. After that, both data blocks were modeled together, side by side,
or in other words in an augmented column-wise structure, also by OC – PLS and then with SO –
OCPLS. With significance level (α) of 0.1, the best results presented efficiency of 70 %. The outcome
results show that augmented column-wise matrices, composed by blocks of different spectra
sources, and processed by sequential orthogonalization may be an alternative to the acai’s pulp
authentication. Moreover, the presented methodology should represent a low budget analysis and
fits with green chemistry requirements, for its respectively no use or production of chemical reagents
and residues.
1 PALA, D.; BARBOSA, P.O.; SILVA, C. T.; SOUZA, M. O. de; FREITAS, F. R.; VOLP, A. C. P. & FREITAS, R. N. Clinical
Nutrition, v. 37 (2), p. 618-623, 2018.
2 Araújo, A.; Marinho, W. & Gomes, A. A. Food Analytical Methods, 2017.
3 Lobato, K. B. De S.; Alamar, P. D.; Caramês, E. T.D. S. & Pallone, J. A. L. Journal of Food Engineering, v. 224, p. 105-
111, 2018.
4 Gomes, A. A.; Khvalbota, L.; Onça, L.; Machyñáková, A.; Spánik, I. Food Chemistry, v. 382, 132271, 2022.
[CNPq, UFRGS, PPGQ-UFRGS, NSP-Governo Eslovaco]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

436
DIRECT AND SIMULTANEOUS DETERMINATION OF NITROGEN AND
CARBON IN SOYBEAN LEAF BY NEAR INFRARED SPECTROSCOPY
Isadora C. M. Silvaa(UG*), Roberta O. Santosb (PG), Jocelene Soaresc(PG), Gilson A. Helfera

(R), Adilson B. da Costab,c(R)
aUniv.de Santa Cruz do Sul, Depto. de Eng., Arq. e Comp., Santa Cruz do Sul, Rio Grande do Sul, Brazil, 96815-900
bUniv.de Santa Cruz do Sul, Prog. de Pós-Grad. em Sist. e Proc. Industriais, Santa Cruz do Sul, RS, Brazil, 96815-900
cUniv. de Santa Cruz do Sul, Prog. de Pós-Grad. em Tec. Ambiental, Santa Cruz do Sul, RS, Brazil, 96815-900
*e-mail: isadoracristina@mx2.unisc.br
The determination of macro and micronutrients in plant tissue is increasingly used in agriculture, as
it allows the assessment of the nutritional status of plants, as well as verifying the effective absorption
of nutrients present in the soil. Due to the great demands of these analyses, near-infrared
spectroscopy (NIRS) associated with multivariate calibration methods has stood out for its precision,
speed, low cost, and little sample handling1. In this context, the objective of this work was to develop
an analytical methodology for direct and simultaneous determination of nitrogen and carbon
elemental in soybean leaves, and to compare the performance between conventional and portable
equipment. The soybean leaves were dried and ground to a thickness of 1 mm for determination of
carbon and nitrogen. These concentrations were measured for reference method, using an
elemental analyzer (Flash EA1112, Thermo Electron
Corporation, Milan, Italy). A conventional infrared
spectrometer (Spectrum 400, Perkin Elmer) was used
with accessory of near infrared reflectance (NIRA), in
region 1000 - 2500 nm and a portable near infrared
spectrometer (DLP NIRscan Nano EVM, Texas
Instruments), in region 900 - 1700 nm, was specially
developed (Figure 1). All spectra were obtained in
absorbance mode, with 16 and 6 scans for sample
using conventional and portable spectrometer,
respectively. As a pre-processing method was
applied normalization followed by multiplicative
scatter correction (MSC) and mean center. The Figure 1. Portable near infrared spectrometer.
partial least squares regression (PLS) calibration
models were developed, using SOLO+MIA software
(Eigenvector Research, Inc), with approximately 30 soybean leaf samples collected in April 2021
and applied to the simultaneity determination of carbon and nitrogen in 15 set samples collected in
April 2022. The best results were obtained using the conventional spectrometer data pre-processing
by normalization followed by multiplicative scatter correction (MSC) and mean center. The best
results for coefficient of determination (R2), and square root of the average error of calibration
(RMSEC) and prediction (RMSEP) were obtained by the conventional spectrometer (Table 1). The
methodology developed for the portable spectrophotometer can certainly be applied as an analytical
alternative (screening method) for rapid and low-cost determination of elemental nitrogen.
Table 1. Results of the PLS models.
Spectrum 400, P.E. DLP NIRscan Nano
N, % C, % N, % C, %
R2 cal 0.976 0.764 0.883 0.553
RMSEC 0.113 0.298 0.252 0.404
RMSEP 0.342 0.691 0.781 1.501
Range 2.8 – 5.5 42.5 – 45.2 2.8 – 5.5 42.5 – 45.2
1 Baumann, L, Librelotto, M, Pappis, C, et al. Journal of Chemometrics. 34, 2020; 3281.
[FAPERGS, CNPq, CAPES]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

437
DISCRIMINATION OF COFFEE FROM DIFFERENT ALTITUDES USING FT-IR
ASSOCIATED WITH MAIN COMPONENTS ANALYSIS
Cristhiane A. Filetea, Sillas R. Marianoc, Ana C.M. Guerrac, Alice D.C. Calimana, Emanuele CS
Oliveiraa, Aristodemos P. Hassemb, Ediézio V. Carvalhob,Eldelon O. Pereirab, Gabriel GG
Rosab, José MS Oliveirab, Túlio LB Limab, Lucas L. Pereiraa
aFederal Institute of Espírito Santo, Venda Nova do Imigrante Campus. Coffee Design, Venda Nova do Imigrante,
Espírito Santo, Brazil.Brazil, 29375-000.
bInstituto Capixaba de Pesquisa Technical Assistance and Rural Extension, Vitória , Espírito Santo, Brazil.Brazil, 29052-
010.
cInstituto Federal do Espírito Santo, Rodovia ES-482 (Cachoeiro-Alegre) Km 47 CEP: 29500-000 Rive, Alegre - Espírito
Santo, Brasil.
*email:lucas.pereira@ifes.edu.br , lucaslozada@hotmail.com
The quality of specialty coffees is influenced by multiple factors, including altitude, since in higher
regions fruit maturation occurs more slowly, allowing the accumulation of the main chemical
precursors associated with the quality of the beverage (Bertrand et al., 2012). Furthermore, post-
harvest techniques have been improved so that it is possible to modify the sensory and chemical
profile of the beverage. In this sense, analytical techniques have been used to better understand
these changes, for example, spectroscopy in the infrared region. This technique has been used in
studies to evaluate the sensory quality of coffee and relate it to the chemical composition (Ribeiro;
Ferreira; Salva, 2011) and to evaluate fermentation methods in robusta coffee (Oliveira et al., 2020).
The study aimed to evaluate the influence of altitude on the chemical profile of coffees produced in
Caparaó Capixaba, through wet processing. For this, an experimental plot of 120Kg of arabica
coffee, from 3 properties with altitudes from 795m to 1250m, was processed and later analyzed by
mid-infrared spectroscopy in the 630 FTIR spectrometer (Agilent) in order to assess whether there
are differences between the treatments and different altitudes. From the infrared spectra obtained,
principal component analysis (PCA) was applied in order to visualize discrimination between the
coffees. The first main component in the Scores chart held 68.8% of the explained variance, while
the third main component explained 5.3%, so there is more than 70% of the information contained
in the two main components. Thus, there was a discrimination between the region of higher altitude
and the region with lower altitude along PC1. And through the Loading chart, it was possible to
identify the variables with greater weight that contributed to this separation. It was observed that the
regions in the ranges 3000-2900 cm-¹ 1700 cm-¹, and 1400-1000 cm-¹ were more significant. The
region 3000-2900 cm-¹ can be associated with classes of alkane compounds, that characterize
carbon chains that form the compounds present in coffee. The range of 1700 cm-¹ has mainly C=O
bonds referring to carbonyls, which are associated with some functions such as carboxylic acids, in
addition to being associated with the presence of ester, confirming the presence of lipids. And finally,
the region 1400-1000 cm-¹ characterizes bands of C–O bonds present in alcohols present in sugars
and C–N bonds present in amines, which can be related to amino acids and proteins that constitute
coffee. These results demonstrate that the chemical composition of coffee is influenced by altitude
and these changes observed in the chemical classes indicate the complexity of controlling
edaphoclimatic factors, results also found by Joët et al. (2010), where the authors noted differences
in polysaccharide bases and chlorogenic acids in arabica coffee samples as a function of different
altitudes.
1 Bertrand, B., et al. Climatic factors directly impact the volatile organic compound fingerprint in green Arabica coffee bean
as well as coffee beverage quality. Food chemistry, v. 135, no. 4, p. 2575-2583, 2012.
2 Joët, Thierry, et al. Influence of environmental factors, wet processing and their interactions on the biochemical
composition of green Arabica coffee beans. Food chemistry, v. 118, no. 3, p. 693-701, 2010.
3 Ribeiro, JS; FERREIRA, MMC; SALVA, TJG Chemometric models for the quantitative descriptive sensory analysis of
Arabica coffee beverages using near infrared spectroscopy. Talanta, v. 83, no. 5, p. 1352–1358, 2011.
4 Oliveira, EC DA S. et al. Infrared spectroscopy for the study of fermented conilon coffee. Brazilian Journal of
Development, v. 6, no. 4, p. 19248–19259, 2020.
[Research Projects Bank (SEAG), FAPES].
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

438
DISTINGUISHING JATROPHA mollissima SAPS ACCORDING TO THEIR
GROWING LOCATION USING NIR AND PATTERN RECOGNITION TECHNIQUE
Caroline L. Fernandesa, José F. B. Rodriguesb, Tiago S. Silvaa ,Isabel C. V. dos Santosa,

Caike R. de Lima a, Solomon K. S. Amoahb, Djair A. Fialhob, Luanna A. D. M. de Medeirosb,
Rosana A. Rosendob, Simone da S. Simões *a
a Universidade Estadual da Paraíba, Prog. de Pós-graduação em Química, Campina Grande, Paraiba, Brazil, 58429-500
b Universidade Federal de Campina Grande, Dep. de Engenharia de Materiais, Campina Grande, Paraiba, 58109-970.
*e-mail: simonesimoes@servidor.uepb.edu.br
Jatropha mollissima is a medicinal plant endemic to the Brazilian Northeast. It is popularly used as
an antiophidic and anti-inflammatory agent. Pharmacological studies demonstrated correlations
between secondary metabolites and bioactivity in plants of the genus Jatropha. Among these
secondary metabolites are flavonoids, phenols, tannins, and saponins which may vary depending
on planting location, sap collection time, weather conditions, and others. The handling of Jatropha
mollissima, as well as other medicinal plants, is a challenge when it refers to the quality control of
their material since plants cultivated in different regions can produce distinct metabolites and present
variations in their concentration. This fact may be directly related to seasonality [1]. For this reason,
analytical methods must be applied to evaluate the quality of the material collected at each location.
Chromatography is a technique commonly applied to identify plant metabolites composition and thus
provide information on quality characteristics that can be related to the geographic origin of plants
[2]. However, this method requires the use of chemical reagents and is time-consuming. Given the
above, the objective of this work was to discriminate the Jatropha molíssima sap from three distinct
locations in the state of Paraiba. For this, plant saps were collected from the three locations for 12
months and their meteorological data were registered. The saps were lyophilized and stored properly
until the time of spectra recording. The spectra of all samples (108) were recorded in the near infrared
region (8000-4000 cm-1) and pattern recognition methods (unsupervised and supervised) were
applied. The principal components analysis (PCA) with 5 PCs (principal components) explained
72.96% of the data variance and suggested that the determining factor for the separation of the three
classes would be the soil composition and climatic conditions of the cultivation site geographical
origin (Figure 1). The supervised pattern recognition model (PLS-DA) showed good predictive ability
with sensitivity and selectivity values close to 1 and low error rates in the prediction step for the three
classes (3.8%, 4.5% and 0%). Thus, because of the positive results obtained, it is concluded that
NIR spectroscopy combined with pattern recognition methods is a technique capable of
distinguishing differences between Jatropha molissima saps cultivated in three distinct locations in
state of Paraiba.
Figure 1: Scores plot for a) growing location and b) seasonality
1. Ferreira, W. S. J., Ladio, A. H.; Albuquerque, U. P. Journal of Ethnopharmacology, p. 238–252, 2011.

2. Wang, C.-Y.; Tang, L.; Jiang, T.; Zhou, Q.; Li, J.; Wang, Y.-Z.; Kong, C.-H. Industrial crops and products, v. 160, p.
113090, 2021.
[CAPES, CNPq, FAPESP-PB]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

439
ELEMENTARY ANALYSIS AS A TOOL IN DIFFERENTIATING BETWEEN
CIGARETTE BRANDS MARKETED IN BRAZIL
Lucas S Rodrigues (PG)*, José M. O Godoya (R)
aPontifícia Universidade Católica, Departamento de Química, Rio de Janeiro, RJ, Brazil.
b Pontifícia Universidade Católica, Departamento de Química, Rio de Janeiro, RJ Brazil.
*e-mail: lucassrodrigues31.12.2014@gmail.com
The presence of trace metals in cigarettes offers potential interest from a forensic and criminological
point of view, which deals with the determination and classification of tobacco brands, with the aim
of differentiating legal brands from smuggled brands [1]. Therefore, this work aimed to develop and
validate methods for determining the metallic content present in cigarettes, in order to assess the
feasibility of discrimination between different cigarettes brands, analyzing cigarettes after the
process of calcination and acid digestion. These methods include chemical analysis by spectrometric
techniques (CV-AAS, ICP-OES and ICP-MS) and subsequent data interpretation by Principal
Component Analysis and Hierarchical Cluster Analysis. The distribution of metals and semimetals
(As, Ba, B, Cd, Co, Cu, Fe, Mn, Hg Ni, Sr and Zn) of 10 cigarettes samples of brands were
determined using ten replicas for each of the brands studied. Arsenic in cigarettes was determined
using ICP-MS, Mercury was determined using CV-AAS and the others elements was determined
using ICP-OES. These brands were acquired in tobacconists and local stores, featuring as the most
common consumer brands available to consumers in Brazil. The chosen brands were Lucky Strike
blue blend, Rothmans blue, Dunhill Carlton Blend (both from Souza Cruz), Chesterfield Terras
Brasileiras, Marlboro Red Selection (both from Phillips Morris), Camel Blue, Winston Classic (both
from Japan Tobacco International) Gift, Eight King Size and San Marino King Size (both Paraguayan
brands Tabacaria Tabesa). The analysis of PCA indicated the presence of four principal
components that together explained more than 80% of the variability of the sample data. These
results presents proved the existence of significant differences between tobacco brands in their
elementary content, allowing the differentiation between illegal brands and legalized brands and the
grouping of legal brands according to the tobacco companies to which they belong, being
corroborated by the Cluster Analysis Hierarchies (HCA). In this case, HCA analysis by UPGMA
method was performed using the scores of each brand studied against the four principals
components. HCA used for the interpretation of possible similarities between the analyzed metals
and semimetals was performed by Ward's method, and interpreted together with Pearson's linear
correlation test indicated similarities between the elements Ba, Sr, in addition to indicating the
proximity between the Mn and the Cd. Clusters between Fe and Zn with Sr and Co with Mn were
also detected, and the elements As, B, Cu, Ni and Hg were considered entropy elements of the
clusters formed.
1 Bernal, J.P; Amigo, J.M; Fernández-Torres, R; Bello, M.A; Callejon-Mochon, M. Trace-metal distribution of cigarette
ashes as marker of tobacco brands. Forensic Science Internacional, 2010.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

440
EMPLOYING AUTO-MACHINE LEARNING ALGORITHMS FOR PREDICTING
THE PHYSICAL PROPERTIES OF BIODIESEL AND ITS BLENDS USING
INFRARED SPECTROSCOPY DATA
Aderval S. Lunaa*, Alexandre R. Torresb, Camilla L. Cunhab, Igor C. A. Limaa, Luis G. Nonatoc
aUniversidadedo Estado do Rio de Janeiro, PPG-EQ, Rio de Janeiro, RJ, Brazil, 20550-013
b Universidadedo Estado do Rio de Janeiro, Faculdade de Tecnologia, Resende, RJ, Brazil, CEP 27537-000
cUniversidade de São Paulo, Instituto de Ciências Matemáticas e da Computação, São Carlos, SP, CEP 13566-590
*e-mail: asluna@uerj.br
This work aims to develop an auto-machine learning method to determine the cold filter plugging
point and kinematic viscosity at 400C of biodiesel, diesel, and mixtures samples using vibrational
(middle-IR) spectroscopy data. The biodiesel was obtained by the transesterification reaction and
later purified. The first dataset was composed of 108 blends (biodiesel obtained from different
biomass such as soy, corn, sunflower, and canola) with binary, ternary and quaternary mixtures. The
second dataset was composed of 227 blends of diesel-biodiesel and diesel-biodiesel-ethanol,
respectively. The physical properties of the samples were obtained according to ANP Resolution
45/2014 and ABNT NBR 10441. The middle-infrared spectra were acquired from 7,800 to 450 cm-1,
with a 4 cm-1 resolution and 20 scans. The spectra' baseline alignment was carried out using the
asymmetric least squares method. A Savitzky–Golay filter was applied to a set of digital data points
to smooth the data. This work used a first-order polynomial and a zero derivative function to smooth
the spectra. The dataset was split into training and test sets using the function CreateDataPartition
from the caret package. It was adopted 70% for training and 30% for test sets. In this work, the model
training process was carried out using the open-source Python library LazyPredict. The LazyPredict
returns the trained models and their performance metrics. The kinematic viscosity at 400C of the
biodiesel samples and their blends could be modeled using the middle-infrared spectra dataset by
different auto-machine learning algorithms with an RMSEP (≤ 0.02 mm2. s-1) similar to the
experimental error obtained after log transformation. The cold filter plugging point of the biodiesel
samples and their blends could be modeled using the middle-infrared spectra dataset by different
auto-machine learning algorithms with an RMSEP (≤ 1.60C) similar to the experimental error
obtained by traditional methodology. Based on the lower computational time and the same
performance observed by the RMSEP and R2 values from different algorithms, it is recommended to
use Ridge or Ridge Cross-Validation Regression models for both physical properties using middle-
infrared spectral data [1].
1 Luna, AS. Employing auto-machine learning algorithms for predicting the physical properties of biodiesel and its blends
using vibrational spectroscopy data, Trabalho final do MBA em Ciência de Dados, USP, São Carlos, 2022.
[CNPq, FAPERJ, CAPES, UERJ]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

441
EVALUATION OF Al, Na, Ca AND Mg LEACHING IN DIFFERENT COFFEE
SAMPLES
Charlie G. Gomes (PG)*a, Daísa H. Bonemann (PG)a, Sabrina H. Scherdien (PG)a, Miguel P.
Soares (UG)a,
Anderson S. Ribeiro (R)a, Mariana A. Vieira (R)a
Federal University of Pelotas, Chemistry Postgraduate Program, Pelotas, Rio Grande do Sul, Brazil, 96010-900
*e-mail: charlieggomesii@gmail.com
Coffee consumption is always a subject discussed in various spheres of society today and over time.
Since its discovery in Africa of the years of mass dissemination, the fruit of coffee has always had
its clashes, with claims of improvement in intellectual and physical performances to the risks of its
consumption, such as addiction, for example. What is indisputable is its growing consumption and
all the economy in its surroundings, which through various products and innovations never fall into
forgetting, becoming one of the most popular beverages in the world, second only to water1. In view
of this, it is evident that a more detailed control of its composition becomes inevitable, especially in
an elementary character. Thus, the objective of this work was to present results for the determination
of metals in samples of conventional coffee and espresso at their different fractions (total coffee
powder, infusion, and infusion powder). The determinations were made using the microwave-
induced plasma optical emission spectrometry (MIP OES) technique. For the preparation of solid
fractions (powder and sludge), a digester block with reflux system was used. The best decomposition
conditions were optimized from a factorial planning: 750 mg of sample with the addition of 2.0 mL
and 5.0 mL of H2SO4 and HNO3, respectively, at 300°C for 3 h. For the liquid fraction (infusion), 0.2
mL of HNO3 and 3.0 mL of H2O2 at 120 °C in digester block were used for 10 mL of samples3. The
accuracy was verified by a mass balance between the concentrations of the fractions of the samples
(total powder, infusion powder and infusion) and by the analyte addition tests. Both presented
recoveries between 81 to 119%, for Al, Ca, Na and Mg, considered satisfactory and proving the
accuracy of the method. The method allowed the determination of Al, Na, Ca and Mg in 22 different
coffee samples, with 3 different types of conventional infusion and 3 by espresso coffee machines.
It was possible to perceive the percolation behavior of the elements from the solid fraction to the
liquid fraction of coffee, with the percentage varying from 5% to 55% for Al; from 12% to 45% for Ca;
from 30% to 80% for Mg and from 30% to 98% for Na approximately. The results obtained for the
concentration of Al, Na, Ca and Mg were also used to verify the agreement of behavior in the
elemental composition in the solid fraction and in the liquid fraction, in relation to the type of coffee
infusion and its origin (brand). Thus, multivariate statistical tools were applied to the results, showing
through the PCA analysis, clusters of similarities between the samples. For the solid fraction, an
approximation in the composition of the samples was observed in agreement with their respective
marks for some cases. As for the liquid fraction, there was a loss of this information, indicating that
the concentration at this stage is more influenced by the interaction of the analyte with the infusion
water or with the way it is linked to the sample matrix, than with the origin. of coffee. Through these
methods and the multivariate statistical tools, the similarity in the elemental composition of the
samples in their solid fraction was observed. Thus, the proposed method was able to reliably
determine these 4 elements and extract as much information as possible from the samples obtained.
1 Arruda, AC, Minim VPR, Ferreira MAM, Minim LA, Silva MN, Soares CF. Ciência e Tecnologia de Alimentos, 29, 2009,
750.
2 Bezerra MA, Ferreira SLC, Novaes CG, Santos AMP, Valasques GS, Cerqueira UMFM, Alves JPS. Talanta, 194,941-
959, 2019.
3 Rainelde ACJ, Chagas AVB, Felix CSA, Souza RC, Silva LA, Lemos VA, Ferreira, SLC. Talanta, 191, 530, 2019.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

442
EXPLORATORY ANALYSIS BASED ON FTIR DATA OF BIO-OILS OBTAINED
FROM THERMAL LIQUEFACTION IN A BATCH SYSTEM
Flávio M. Andrade (PG)a*, Reinaldo A. S. Junior (PG)b, Raquel S. Silva (PG)b, Pedro N. A.
Neto (UG)b, Diogo A. Simões (R)c, Maria Fernanda Pimentel (R)b
a Federal University of Pernambuco, Department of Fundamental Chemistry, Recife, Pernambuco, Brazil, 50.740-560
b FederalUniversity of Pernambuco, Department of Chemical Engineer, Recife, Pernambuco, Brazil, 50.740-590
c Federal University of Pernambuco, Department of Biochemistry, Recife, Pernambuco, Brazil, 50.670-420
*e-mail: flvandrade@gmail.com
The use of thermochemical methods to convert biomass into drop-in fuels has been reported in the
literature. Most are based on pyrolysis or gasification processes, for which a preliminary drying step
is mandatory. On the other hand, Thermal Liquefaction (TL) has often been reported as a promising
technique for biomass conversion with the advantage of using wet biomass. A variety of polar
solvents can be used in this approach, including water in near or supercritical conditions. The major
product of this process is a bio-oil that can be further processed to produce biofuels, which can
replace fossil fuels such as gasoline and diesel1. In the present work, middle infrared (MIR) spectra
were acquired from bio-oil samples produced using different raw materials and process conditions.
An exploratory analysis based on Principal Component Analysis was performed. Sixty-six (66)
samples were produced in batch experiments carried out in a lab-scale reactor (model 4576, Parr
Instrument Co.) according to the conditions fixed in four experimental designs. The variables
considered were: temperature (280-350 °C), catalyst content (0-0.5 mol.L-1 of K2CO3 for water tests),
and heating time (0-30 minutes). Water and ethanol were used as solvents; bagasse and sugarcane
straw were employed as biomasses. The MIR spectra were obtained in the wavenumber range of
4000 to 550 cm-1 with resolution of 4 cm-1, using a benchtop FTIR spectrometer (Spectrum 400,
Perkin-Elmer) with the Attenuated Total Reflectance (ATR) accessory. For each sample, an average
of 32 scans were performed.
Figure 1. PCA performed with IR spectra of bio-oil samples.
The scores and loadings of the first two principal components (explaining 81,62% of variance) are
presented in Fig. 1. Regarding PC1, the most positive loadings are in the regions around 1734 cm-1 and
1044 cm-1, which might refer to (C=O) and (C-O) stretching vibrations. On the other hand, as indicated
by the positive loadings of PC2, the region around 3382 cm -1 (O-H bond) is principally responsible for
explaining the variability among the bio-oils obtained from sugarcane bagasse. The negative loadings of
PC2 highlighted the regions between 2919-2849 cm-1 (C-H, bond) and around 1715 cm-1 (C=O vibrations)
as being crucial to describe the bio-oils produced from sugarcane straw, which may indicate the presence
of oxygenated aliphatic components. PCA performed on the infrared data set showed clear differences
between the variabilities of the bio-oil samples, which were mainly due to the type of solvent (PC1) and
biomasses (PC2).
1Arturi KR, Kucheryavskiy S, Søgaard Eg, Fuel Processing Technology, 150, 2016, 94.
[NUQAAPE, INCTAA, FACEPE, CNPq, CAPES, PETROBRAS, SUZANO, KLABIN, EMBRAER]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

443
EXPLORATORY ANALYSIS OF THE CHANGE IN THE CHEMICAL
COMPOSITION OF COFFEE HARVESTED AT DIFFERENT TIMES
Marinalva M.B. Simmer*, Emanuele C.S. Oliveiraa, Lucas L. Pereiraa, Aldemar P.Morelia
aInstituto Federal do Espírito Santo, Group Coffee Design, Venda Nova do Imigrante,ES, Brazil, 29375-000
*e-mail:mariabratz13@gmail.com
The phenological coffee cycle is an object of great interest to coffee growers and Coffea arabica,
above all, presents variations in maturation cycles, and this phenomenon has been relativized due
to soil and climate conditions. In this study, we present in an unprecedented way how the chemical
composition of coffees subjected to different processing is influenced according to the harvest
period. Thus, a portion of coffee was harvested in September (2020) and another in November
(2020) and subjected to four processes: yeast fermentation, Washed, Semidry and Control. The
fermentation processes were delimited by a period of 36h. These coffees were then, extracted by
HS-SPME-GC/MS. After the pre-processing of the generated infrared spectra, statistical analysis
was performed, using the multivariate technique of principal component analysis (PCA), which
allowed the visualization of the discrimination of the samples collected at the two times in the
treatments yeast fermentation, Washed, and Semidry. For the Natural treatment, it was not possible
to observe the separation. The changes observed in these treatments in relation to the chemical
composition of the coffee between different times may be related to the environmental conditions.
Studies show that low temperatures, for example, are characterized by delaying the ripening of
grains, which, in turn, leads to a greater accumulation of chemical compounds related to beverage
quality (Bertrand et al., 2012). As the infrared spectroscopy showed differences in the characteristics
of the coffees from September and November, the analysis of volatile compounds also allowed to
observe a formation of groups between the harvest periods as well as between the processings. The
complex chain of aromas and flavors formation in coffees is mainly attributed to volatile compounds.
In this study, a total of 41 volatile compounds were identified in coffees and the main ones
responsible for the separation were cited. For Washed coffee in the first harvest, it was observed
that some of the main volatile compounds identified here and reported as precursors in quality and
which were responsible for separating it from the Washed treatment of the second harvest were:
Methylpyrazine, 2- Furanmethanol/Furfuryl alcohol, Isovaleric acid, Texanol and 1-Furfuryl-2- formyl
pyrrole. Another group formed by the Washed (second harvest), Semidry (first and second harvest),
yeast fermentation (first and second harvest), and Natural (first harvest) treatments showed some
similarities regarding the identified volatile compounds. It was not significant to discriminate between
them. And finally, the last and third group was formed by natural coffee in the second harvest. The
compounds that best represented it and separated it from the other treatments were: 2-Acetylfuran,
Furfural, Isomaltol, 2,3-Butanediol, Acetol acetate, 5-methylfurfural and 2-Methyltetrahydrofuran-3-
one. These results reinforce that coffees harvested at different times tend to show changes in
chemical characteristics since the Washed treatments in the first harvest showed different
compounds compared to the second harvest and the Natural treatment that also discriminated the
compounds between September and November. These changes can be related to numerous
extrinsic and intrinsic factors. However, one must take into account, above all, the climatic changes
during the development and maturation of the fruits.
1 Bertrand, B. et al. Climatic factors directly impact the volatile organic compound fingerprint in green Arabica coffee bean
as well as coffee beverage quality. Food Chemistry, v. 135, n. 4, p. 2575–2583, 2012.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

444
FEASIBILITY OF PORTABLE NIR SPECTROPHOTOMETER AND
MULTIVARIATE ANALYSIS FOR QUALITY OF WHEY PROTEIN
SUPPLEMENTS
Jennifer A. Cavalcante (PG)a*, Jarbas J. R. Rohwedder (R)a
a Universidade Estadual de Campinas, Departamento de Química Analítica/Instituto de Química, Campinas, São Paulo,
Brazil, 13083-970
*e-mail: jennifer_cavalcante@yahoo.com
The consumption of dietary supplements has grown significantly in recent years in Brazil, reaching
a peak of 21% growth per year in 2021[1]. Whey Protein supplements are one of the most consumed
in diets designed for physical activity practitioners[2]. Therefore, it is extremely important that the
nutritional table of these products corresponds to the levels present in the protein formulations. In
total, 60 samples of powdered food supplements in the range of 25 to 89% (m/m) of protein
concentration described on the label were purchased in physical stores and e-commerce. Near
Infrared Spectroscopy (NIRS) in diffuse reflectance mode was used to acquire the spectra of
samples without any pre-treatment. A portable spectrometer, DLP NIRscan Nano EVM (Texas
Instruments Incorporated, Dallas, USA) was used in all experiments. The spectra were obtained in
the Hadamard transform mode and the spectral range from 900 nm to 1700 nm wavelength. Each
spectrum obtained represents the average of 25 scans. Teflon powder (Supreme Lubrificantes,
Santa Catarina, Brazil) was used as blank sample. Independent triplicates were performed and a
total of nine spectra were obtained from each sample, totaling 540 spectra. The Unscrambler® X
v10.4 program (AspenTech, Massachusetts, USA) was used for the pre-processing of the spectra,
using the Multiplicative Signal Correction (MSC) in conjunction with the First Derivative with a four-
point window. The program was also used in Principal Component Analysis (PCA) and in the
construction of the multivariate calibration model using Partial Least Squares Regression (PLS). A
set of 40 samples selected by the Kennard-Stone method had the protein content values determined
by the Dumas method and, from these, the PLS model was developed. 26 samples were used for
the calibration set and 14 for the external validation set. The results found are described in Table 1.
After validation of the method, it was used to predict the protein content in 20 new samples, obtaining
relative error values in the range of -13.26 to 16.78% in relation to the value labeled. These values
are within the limits recommended by ANVISA, where a tolerance of more or less 20% is allowed in
relation to the values declared on the label[3].
Table 1. Results found for the calibration and external validation models
RMSE
R-Square
(%protein, m/m)
Calibration 1,91 0,97
Validation 2,07 0,96
Prediction 2,67 0,92
The proposed method has advantages over other well-established methods such as the Kjeldahl
and Dumas method as it is a method of direct, fast, low-cost analysis and direct application in the
field. The results found demonstrate that the proposed method can be used to predict protein in food
supplements with an error of 2%, ten times smaller than the value accepted by ANVISA.
1 Associação Brasileira de Empresas de Produtos Nutricionais. http://www.abenutri.org/artigos/. Accessed july 11, 2022.
2 Associação Brasileira de Empresas de Produtos Nutricionais. http://www.abenutri.org/proteste-teste-whey-protein/.
published 2018. Accessed july 11, 2022.
3 ANVISA. Resolução da Diretoria Colegiada - RDC nº 360, de 23 de dezembro de 2003. Diário Oficial da União.
https://www.gov.br/agricultura/pt-br/assuntos/inspecao/produtos-vegetal/legislacao-1/biblioteca-de-normas-vinhos-e-
bebidas/resolucao-rdc-no-360-de-23-de-dezembro-de-2003.pdf/view. Published 2003. Accessed july 11, 2022.
[INCTAA, FAPESP, CAPES]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

445
FROM THE STUDIO TO THE CANVAS: DISCRIMINATING ARTWORKS OF A
BRAZILIAN CONCRETE ARTIST BASED ON CHEMOMETRIC MODELLING
Vitor P. Amaral (PG)a, Marcelo M. Sena (R)a*, Alessandra Rosado (R)b, Luiz A. C. Souza (R)b
a UFMG, Chemistry Departament, Belo Horizonte, Minas Gerais, Brazil, 31270-010
b UFMG, CECOR/ School of Fine Arts, Belo Horizonte, Minas Gerais, Brazil, 31270-010
*e-mail: marcsen@ufmg.br
The most comprehensive knowledge about materiality in artworks has been explored with the use
of analytical techniques in the multidisciplinary field of cultural heritage, with the aim of investigating
on authenticity, dating, conservation, restoration of artworks and elucidating the artist's creative
process. There is a growing use of chemometrics in cultural heritage due to the complexity of the
information obtained by the employed analytical techniques, with the aim of extracting as much
relevant information from the raw analytical data as possible.1 The proposed study analysed 13
artworks of the “Objeto Ativo” series by the Brazilian artist Willys de Castro, exhibiting two of them
proven originality. These artworks are dated in the early 1960s in wood on canvas, the shapes of
the objects are rectangular with two sizes, whose approximate dimensions are 70x2x11 cm and
35x4x8 cm. The analytical techniques FITR/ATR, Raman spectroscopy and gas chromatography
(GC-MS) were applied aiming to understand and compare material composition of these artworks.
Through FTIR spectra and a PCA (principal components analysis) model, it was possible to observe
the differentiation of the binders from the paint samples, acrylic in red, and oil in green, on PC1
scores (Fig. 1). This discrimination is mainly due to the following PC1 loadings: spectral peaks at
1723, 1158, 1358, 760 and 681 cm-1 present negative loadings characteristic of the acrylic binder
components, while peaks at approximately 1750, 1199, 1124, and 1083 cm-1 present positive
loadings associated to the oil binder composition.
Figure 1. A) Scores plot of PC1 x PC2; B) Loadings plot of PC1.
Raman analysis was used for the determination of pigments and fillers in the artworks, comparing
their spectra with the respective standards by KREMER®. The results identified titanium white and
lithopone as white pigments; blue and green phthalocyanines, as blue and green pigments; and red
azo and iron oxide as red pigments. Calcite, rutile and talc were the fillers found. In conclusion, a
PCA model applied to FTIR/ATR spectra proved to be a good tool for the discrimination of acrylic
and oil binders in paint samples. This allowed a global visualization of all the studied samples,
confirming the initial assignments verified by GC-MS and thus contributing to data interpretation.
Also, Raman spectroscopy identified pigments and fillers, allowing the characterization of all three
basic compounds on these paints: binder, pigment, and filler.
1 Madariaga JM, Analytical chemistry in the field of cultural heritage, Analytical Methods, 7, 2015, 4848.
[CAPES; Getty Conservation Institute (GCI)]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

446
GENERATION OF SECOND-ORDER ELECTROCHEMICAL DATA FOR
INTERFERENCE-FREE CALIBRATION OF GALLIC ACID IN RED WINES
Montemerlo Ab,c , Azcarate S a,b Camiña J a,b, Messina G
a Facultad Ciencias Exactas y Naturales, Universidad Nacional de La Pampa, Av.Uruguay 151, (6300) Santa Rosa, La
Pampa, Argentina. Tel: 02954-245220 int 7433
b Instituto de Ciencias de la Tierra y Ambientales de la Pampa (INCITAP-CONICET-UNLPam), Santa Rosa, La Pampa,
Argentina.
c Instituto de Química de San Luis, “Dr. Roberto Antonio Olsina” (INQUISAL-CONICET-UNSL), San Luis, Argentina.
*e-mail:amontemerlo@gmail.com
Wine consists of a solution of ethanol in water, which also contains a variety of organic compounds,
such as organic acids, amino acids, sugars, volatile compounds and polyphenols in different
concentrations. In particular, the interest in the latter is continuously growing due to its bioactive
nature and its close relationship with the final qualities of the resulting product. Specifically, gallic
acid is one of the most widely used phenolic compounds as a marker for the characterization of wine
products. The use of sulfur dioxide is a common practice in oenology that is currently considered
essential for the correct finish of wines. However, there is a growing concern about the reduction of
its levels in the final product, which makes it necessary to search for complementary agents. Among
the possible alternatives is ascorbic acid or vitamin C. In recent years, its use as an additive for
winemaking has been reported due to its antioxidant properties, its reducing nature and because it
is a healthy compound. It is important to note that ascorbic acid will never replace the use of sulfites,
but it can be an excellent complement to it. The rise of multidirectional calibration enabled the
quantification of numerous analytes in complex samples without interference, even in the presence
of components not modeled in the calibration, commonly referred to as a second-order advantage.
However, classical chemometric methodologies assume that the data comply with the property of
bilinearity, which is sometimes not possible to achieve with all instrumental methods. Among them,
electrochemistry is capable of providing information with high potential to be used in the development
of novel second-order calibration methodologies, although they may not meet the aforementioned
conditions. This work raises the possibility of developing a simple and versatile methodology for the
interference-free quantification of gallic acid in the presence of ascorbic acid, based on chemometric
modeling of electrochemical data. Second-order data were generated by cyclic voltammetry (CV) at
different scanning speeds and various modeling strategies were implemented to treat the data
obtained. The quantification of gallic acid in the presence of ascorbic acid was possible using a
model based on the resolution of multivariate curves-alternating least squares (MCR-ALS). It was
found that the optimal number of estimated components for calibration is equal to one. While the
analytical figures of merit found, REP% 9.4, LOD 6.094x10-5 mol L-1, show the possibility of using
this methodology for the quantification of gallic acid in complex samples. The studies carried out
open the doors for wine quality analysis through new and innovative methodologies based on the
potential of electrochemical data, combined with chemometric techniques, which make it possible to
obtain results in a simple, fast and reliable way for quality control in the wine industry.
1 F. Tekos et al., “Assessment of antioxidant and antimutagenic properties of red and white wine extracts in vitro,”
Metabolites, vol. 11, no. 7, 2021, doi: 10.3390/metabo11070436.
2 F. A. Chiappini, F. Gutierrez, H. C. Goicoechea, and A. C. Olivieri, “Achieving the analytical second-order advantage
with non-bilinear second-order data,” Anal. Chim. Acta, vol. 1181, p. 338911, 2021, doi: 10.1016/j.aca.2021.338911.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

447
GRAPE JUICE CLASSIFICATION WITH RESPECT AGRICULTURAL PRODUCTION
SYSTEM BY MEANS OF VISIBLE SPECTROSCOPY CHEMOMETRICS ASSISTED
Carlos H. Junges (PG)a*, Natalia A. P. C. Reis (PG)b, Fernando S. Diogo (UG)a, Adriano A.
Gomes (R)a, Celito C. Guerra (R)c, Marco F. Ferrão (R)a,d
a Universidade Federal do Rio Grande do Sul, Instituto de Química, Porto Alegre, RS, Brasil, CEP 91501-970
b Universidade Federal do Rio Grande do Sul, Faculdade de Farmácia, Porto Alegre, RS, Brasil, CEP 90610-000
c Empresa Brasileira de Pesquisa Agropecuária, Unidade Uva e Vinho, Bento Gonçalves, RS, Brasil, CEP 95701-008
d Instituto Nacional de Ciência e Tecnologia, Bioanalítica, Campinas, SP, Brasil, CEP 13083-970
*e-mail: carlosjunges88@gmail.com
Grape juice is a food rich in nutrients and a natural

source of numerous chemical compounds, especially
phenolic compounds. The chemical constitution of
grape juice depends on several factors, such as the
grape variety used, climate, harvest, geographic
origin, state of maturation and cultivation process1.
Considering the planting of grapes for juice production,
two mechanisms are established: the conventional
one, defined by the use of artificial pesticides and
fertilizers, and the organic one, in which chemical
products are not applied. It is important to develop an
analytical method that enables for the differentiation of Figure 1. Flowchart of the steps involved in the
study.
the two commercial forms of natural grape juice (organic or conventional) in order to protect the
consumers from frauds in its commercialization, especially the organic juice with the highest market
value2. In this context, we evaluated the fusion of data obtained by visible spectroscopy at different
pHs, assisted with chemometric methods to discriminate Brazilian grape juices by conventional and
organic production systems. In the study, the LDA method (linear discriminant analysis) was applied
combined with a tool genetic algorithm (GA) to optimize the performance of training and test sets
generating more reliable models and to select the most
significant variables from the data set. MATLAB® software
version 7.1.0 (The MathWorks Inc., USA) was used for the
chemometric procedures. According to the flowchart in Figure
1, then, 46 whole grape juices from 11 different varietals were
used as samples. The preparation consisted of first
homogenizing the grape juice using a blender and then the juice
was filtered. The samples were prepared by diluting 450 µL of
filtered grape juice with 15 mL of each buffer solution, in order
to acquire solutions with different pH values (2, 4, 6, 8, 10 and
12), evaluated using a calibrated portable pH meter (Metrohm©
827). The samples obtained were analysed at room temperature Figure 2. Result of the discriminant
function.
in quartz cuvettes with a 1 cm optical path in a Varian Cary® 50 UV-Vis spectrophotometer (Agilent
Tech. Inc., USA). The results (Figure 2) showed that the more alkaline pHs presented greater selection
of variables with a hit rate of up to 100% and 92% for the training and test sets, respectively. The select
absorption wavelengths represent phenolic compounds profile present in grape juice and the relative
amount of these compounds changes according to the agricultural system applied3. PLS-DA
methodology was used in the data set in order to contrast with the LDA models and the results revealed
a lower rate of correct classification. Therefore, the proposed method proves to be relevant as an
option in quality control, standing out for being accessible and easy to perform.
1 Sen I, Tokatli F, J. Food Compos. Anal., 45, 2016, 101.
2 Chang JD, Zheng H, Mantri N, Xu L, Jiang Z, Zhang J, Songs Z, Lu H, Int. J. Food Sci. Tech., 51, 2016, 2474.
3 Grinder-Pedersen L, Rasmussen SE, Bügel S, Jørgensen LV, Dragsted LO, Gundersen V, Sandström B, J. Agr. Food
Chem., 51, 2003, 5671.
[CAPES, CNPq, INCT-Bio, EMBRAPA]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

448
IDENTIFICATION OF BLOODSTAINS OF DIFFERENT AGES DEPOSITED ON
SUBSTRATES USING HANDHELD NIR SPECTROMETER
Aline C. S. Fonseca (PG)a*, Maria Fernanda Pimentel (R)b, Ricardo S. Honorato (R)c, Rasmus
Bro (R)d, José Francielson Q. Pereira (R)e
a Federal University of Pernambuco, Fundamental Chemistry Department, Recife, Pernambuco, Brazil, 50740-560
b Federal University of Pernambuco, Chemical Engineering Department, Recife, Pernambuco, Brazil, 50740-550
c Federal Police, Recife, Pernambuco, Brazil, 50030-230
d University of Copenhagen, Department of Food Science, Frederiksberg, Denmark
e State University of Campinas, Chemistry Institute, Campinas, São Paulo, Brazil, 13083-970
*e-mail: aline.csfonseca@ufpe.com.br
The identification of blood traces is essential in cases of violent crimes. Recent studies show the
usefulness of portable equipment to identify bloodstains at crime scenes [1,2 No published studies
exist using handheld near-infrared equipment to identify bloodstains deposited at different times.
The present work is an extension of the study developed to identify bloodstains on nine different
types of floor tiles (5 ceramic and 4 porcelain tiles) using hierarchical classification models (HCM) [2].
HCMs were originally built using blood samples deposited for 6 days. Two decision rules were
established for the development of this model. The first rule used as decision criterion the Q-Residual
obtained through a PCA model built using only samples of human blood (HB) and animal blood (AB).
This rule separates common false positives (CFP) from blood samples. The 2nd rule discriminates
between HB and AB samples using the Q-Residual obtained from a PCA model built using only HB
samples. To evaluate the effect of aging in the classification results, spectra were collected from
bloodstains of 31 volunteers deposited at different times (0, 1, 9, 20, 30 and 162 days) using a
handheld near-infrared spectrophotometer (MicroNir 1700). This provided 124 bloodstain samples.
These were then used in the prediction set of the HCM. As shown in Table 1, for the original model
(built with 6 days-old samples) the correct classification rate of HB samples was above 80% for
samples deposited between 0 and 30 days. Samples aged 162 days had a correct classification of
66%, due to degradation of bloodstains over time. Some HB samples were classified as AB; 6% of
the samples were misclassified as CFP. To improve the classification results, 0 day- and 1 day-old
samples were separately added to the training set and two other HCMs were built. This addition of
fresh samples to the training set increased the rate of correct classification to values above 92% for
samples deposited between 0 and 30 days. Samples aged 162 days also improved in classification
percentage; however, there were still samples classified as CFP. The correct classification
significantly increased when 0 day samples were introduced into the training set, especially those
from the 1st day (and 100% for the first hours). Therefore, for forensic purposes which usually deal
with fresher samples, the 2nd HCM model (0 days and 6 days) represents the best application.
Table 1. Results of classification of HB samples with different deposit times in hierarchical classification models.
Training sets: samples with 6
samples with 6 and 0 days samples with 6 and 1 day
days
Deposit Deposit Deposit
%HB %AB %CFP %HB %AB %CFP %HB %AB %CFP
(days) (days) (days)
0 82% 15% 3% 0 100% 0% 0% 0 84% 16% 0%
1 81% 17% 2% 1 97% 3% 0% 1 97% 3% 0%
9 85% 14% 1% 9 98% 2% 0% 9 99% 1% 0%
20 87% 10% 3% 20 90% 8% 2% 20 92% 6% 2%
30 87% 11% 2% 30 93% 6% 1% 30 94% 4% 2%
162 66% 28% 6% 162 73% 21% 6% 162 83% 14% 3%
1 Pereira, J. F. Q., Silva, C.S., Vieira, M. J. L., Pimentel, M. F., Braz, A., Honorato, R. S., Microchemical Journal, 133,
2017, 561.
2 Fonseca, A. C. S., Pereira, J. F. Q., Honorato, R. S., Bro, R., & Pimentel, M. F, Spectrochimica Acta - Part A: Molecular
and Biomolecular Spectroscopy, 267, 2021, 120533.
[CNPq/INCTAA, NUQAAPE, CAPES]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

449
IDENTIFICATION OF CANNABIS SATIVA L. IN SEIZED DRUG SAMPLES BY
ATR-FTIR OBTAINED FROM ETHANOLIC EXTRACTS AND PLS-DA
Rafaela Dornelas Vieira (UG)a, Fabiana Casarin (R)a, Laiz de Oliveira Magalhães (R)a, Ícaro
Noé Caixêta (UG)a, Luciano Chaves Arantes (R)a,b, Jez Willian Batista Braga (R)*b
aUniversidade de Brasília (UnB), Instituto de Química (IQ), Brasília, Distrito Federal, Brazil, 70910900
bPolícia Civil do Distrito Federal (PCDF), Instituto de Criminalística (IC), Brasília, Distrito Federal, Brazil, 70610907
*e-mail: jez@unb.br
Illegal drugs are substances administered without medical supervision to obtain a recreational
psychoactive effect that can cause dependence. According to UNODC1, in 2019 Cannabis sativa L.
was the most consumed drug of abuse in the world, being still proscribed in most countries. In Brazil,
the use of Cannabis and its commercialization is prohibited by law nº 11.343 of August 23, 2006. 2
However, the use of marijuana or its derivative products has been studied for medicinal treatments
against glaucoma, cancer, in HIV positive patients, autism, etc.3 Considering its illicit use, there is a
wide variety of forms of marijuana presentation, among the most common, are the beck, hashish,
skunk, in natura plant, pressed plant material, oil, and dry macerated plant.4 Therefore, many
seizures contain plant material that can be easily confused with other species of licit trade, requiring
an efficient screening method to differentiate Cannabis sativa L. from other plants of similar
appearance. The main screening method is based on the colorimetric test performed by the
extraction of phytocannabinoids, mainly tetrahydrocannabinol (THC), in a basic medium in an
organic phase and complexation with the Fast Blue B salt. The test can be easily and quickly
performed, with little solvent and low cost. Despite the advantages, reports of false positive errors
are found with this test due to the interference of other substances, such as chamomile, guarana,
and nutmeg.4 In the attempt to minimize these problems and propose a confirmatory method for
forensic analysis, the present work evaluated the use of Attenuated Total Reflection Fourier
Transform Infrared Spectroscopy (ATR-FTIR) and the Partial Least Squares for Discriminant
Analysis (PLS-DA) model to differentiate the drug-containing Cannabis sativa L. from other plants
known to interfere in the colorimetric method. Ethanol 99.8% was used for ethanolic extraction.
Approximately 27.5 mg of the sample (dry ground vegetable) were weighed and 200 µL of ethanol
was added, the solutions were mixed in a vortex for approximately 30 seconds and centrifuged for 2
minutes. Then, aliquots of 20.0 µL were taken to be analyzed in the ATR-FTIR using a heated ATR
accessory (Agilent Technologies). The spectra were acquired from 360 marijuana samples seized
by the Civil Police of the Federal District (PCDF) and 135 samples from other vegetables of legal
use. The spectra were analyzed by Principal Component Analysis (PCA) and PLS-DA to perform
exploratory and discriminant analysis, respectively. The PCA models showed well-defined groups
for Cannabis and the other vegetables, being that the region between 1550 and 1650 cm-1 presented
better results for pattern recognition and discrimination. Both models, developed with the entire
spectra and between 1550 and 1650 cm-1 presented low false positive and negative results along
with efficiency rates higher than 92%. The method was considered fast, and efficient, using just a
small amount of sample and without generation of chemical waste, which may assist the routine
analysis of Cannabis sativa L. in forensic laboratories.
1 UNODC. Executive Summary: Conclusions and policy implications. World Drug Report. 2019.
2 Brazil, law nº 11.343 of August 23, 2006. http://www.planalto.gov.br/ccivil_03/_Ato2004 - 2006/2006/Lei/L11343.htm.
Accessed at: 26/01/2022.
3 Santos NA, Souza LM, Domingos E, França HS, LacerdaJr V, Beatriz A, Vaz BG, Rodrigues RRT, Carvalho VV, Merlo
BB, Kuster RM, Romão W, Forensic Chemistry, 1, 2016, 13.
4 Custódio MF, Magalhães LO, Arantes LC, Braga, JWB, Journal of the Brazilian Chemical Society, 32, 2021, 513.
[CNPq, CAPES, FAPDF, Agilent]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

450
MACHINE LEARNING TO THE PREDICTION OF TOTAL ACID NUMBER IN
PETROLEUM BY ESI(-)-ORBITRAP HRMS
Gutierri Ricardo dos Santos Gonçalves Salgueiro (PG)a*, Thamara Andrade Barra (PG)a,
André Ferreira Young (R)b, Débora de Almeida Azevedo (R)a
aUniversidade Federal do Rio de Janeiro, LAGOA-LADETEC/IQ, Rio de Janeiro, RJ, Brazil, 21941-598
b Universidade Federal Fluminense, Department of Chemical and Petroleum Engineering, Niteroi, RJ, Brazil, 24210-240
*e-mail: gutierri.salgueiro@iq.ufrj.br
The heteroatomic compounds present in petroleum, reported as polar compounds, are constituted
by species containing N, S and O with aromatic and naphthenic systems. The presence of some of
these compounds, as the organic acids, can generate problems that affect the entire oil chain, e.g,
corrosion of equipment in refineries (pipelines, pipes and tanks)1. The present work aims to use the
electrospray ionization in the negative mode with Orbitrap high resolution mass spectrometry (ESI(-
)Orbitrap HRMS) and machine learning to predict a property related to heteroatomic compounds
present in petroleum, the total acid number (TAN). Thirty-four crude oil samples, with TAN values
ranging from 0.03 to 4.96 mg KOH g-1, were measured by ESI(-)-Orbitrap HRMS. Then, predictive
models were applied by multivariate calibration of partial least squares (PLS) and support vector
machine (SVM) types, using Python language to the absolute intensity values of each peak in the
high resolution mass spectra with molecular formula assigned after processing. The samples were
separated in calibration and validation groups using Shuffle&Split method at the proportions of 75 %
and 25 %, respectively. The validation group was used to evaluate the model for relative mean
square error of prediction (RMSEP). This procedure was repeated 10 times in which, several values
of the parameters of each model were tested. Table 1 summarizes the optimized models in
comparison to references2,3.
Table 1. Prediction error (RMSEP) of each optimized model and a comparison with the RMSEP from
others works.
Model RMSEP RMSEP from other works Technique
2
0.77 FT-ICR MS
PLS 0.22
0.16 3 ATR-FTIR
SVM 0.13 0.68 2 FT-ICR MS
The results obtained were compared with those from other studies and showed good RMSEP values.
The F test was performed to compare the prediction ability between the models and showed both
models have the same prediction effectiveness. This is the first study to evaluate the
physicochemical properties of Brazilian crude oils for the TAN using ESI(-)-Orbitrap HRMS and
Python language with PLS and SVM.
1 Terra, LA; Filgueiras, PR; Tose, LV; Romão, W; Souza, DD; Castro, EVR; Oliveira, LMSL; Dias, JCM; Vaz, BG; Poppi,
RJ. Petroleomics by electrospray ionization FT-ICR mass spectrometry coupled to partial least squares with variable
selection methods: Prediction of the total acid number of crude oils. Analyst, 139, 2014.
2 Vaz, BG; Abdelnur, OV; Rocha, WFC; Gomes, AO; Pereira, RCL. Predictive petroleomics: Measurement of the total acid
number by electrospray fourier transform mass spectrometry and chemometric analysis. Energy and Fuels, 27, 2013.
3 Rivera-Barrera, D; Rueda-Chacón, H; Molina, VD. Prediction of the total acid number (TAN) of Colombian crude oils via
ATR–FTIR spectroscopy and chemometric methods. Talanta, 206, 2020.
[CNPq, FAPERJ, CAPES, Petrobras]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

451
METABOLOMICS ASSAYS FOR DIFFERENTIAL DIAGNOSIS OF INFERTILITY
IN MEN WITH VARICOCELE
Hélder V. Carneiro (PG)ac, Filipe T. L. Neto (PG)b, Ricardo O. Silva (R)c, Ronmilson Marques
(PG)c Licarion Pinto (R) cd*
a University
of Delaware, Department of Chemistry and Biochemistry, Newark, Delaware, USA, 19717
b Universidade
Federal de Pernambuco, Departamento de Cirurgia, Recife, PE, Brazil
c Programa de pós graduação em Química, Universidade Federal de Pernambuco, Recife, PE, Brazil,
d Universidade do Estado do Rio de Janeiro, Departamento de Química Analítica, Rio de Janeiro, RJ, Brazil,
*e-mail: jose.licarion.neto@uerj.br
Despite being considered major cause of male infertility, the relationship between varicocele and the
incapacity of fathering children is not well defined1. The disease can be described as an abnormal
dilatation of the spermatic veins which drain the venous blood from the testes2. The traditional
diagnosis of infertility in men with varicocele can take up one year3. In this scenario, is it possible to
find a less time-consuming way of detecting infertility? This question guided the goal of this work,
which was to perform metabolomics analysis of men’s seminal fluid that were diagnosed with
varicocele using HPLC-DAD and chemometrics. The metabolomics study used here had three
principal steps: 1) Sample preparation, 2) Instrumental Analysis and 3) Machine Learning models.
The sample preparation was made through DLLME. The Instrumental Analysis was optimised using
a HPLC-DAD (Shimadzu UFLC - 13327) with a C18 column (Luna, 00G-4252-E0, H20-209701, 250
x 4,6 mm, 100 Å, 5 µm), water and methanol were used as mobile phase. The HPLC-DAD presents
itself as an affordable instrumentation for medium and small hospitals and clinics, once it is easy to
operate, robust, reproducible and present a low operational cost. The samples were submitted to
exploratory multivariate analysis and data classification with the MATLAB software. The samples
were divided in three groups: C – Control (fertile men without varicocele); VF – fertile men with
varicocele, and VI – infertile men with varicocele. Robust PCA analysis did not indicate anomalous
samples presence, but there wasn’t a clear separation between classes with this analysis. PLS
analysis promoted better discrimination between the classes, with an accuracy of 95% and 75% for
global triage of C, VF and VI group. The LDA analysis with variable selection using genetic algorithm
showed an accuracy of 100% when it is built a model to identify the fertility in men with varicocele.
Figure 1 shows the scores results obtained for classification(a), loadings of the selected variables
(b), selected variables in the chromatogram calculated as a mean of all VI samples (in red) and all
VF samples (in green) (c). With this methodology it is expected to help the early indicative of infertility
in men with varicocele. This is an initial study, that will be increased by the analysis of more samples.
Figure 1. Scores for the discriminant function(a), loadings(b) and the selected variables (c).
1 Fonseca RP, Macedo LC Saúde e Pesquisa, v. 8, n. 1, p. 167, 2015.

2 Baigorri BF, Dixon, RG, Thiememedical Publishers. Seminars in interventional radiology. [S.l.], 2016. v. 33, n. 3, p. 170.
3 Medicine, P. C. of the American Society for R. et al. Fertility and sterility, Elsevier, v. 103, n. 3, p. e18, 2015.
[DQF/UFPE, HC/UFPE, CAPES, CNPq, FACEPE, UERJ]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

452
METABONOMIC MODELS FOR SCREENING ANALYSIS OF PERIPORTAL
FIBROSIS STAGING OF BY MANSONI SCHISTOSOMIASIS USING HPLC-DAD
Julieth G. Herrera (PG)a*, Milena L. Rodrigues (PG)b, Edmundo P. Lopes (R)bc, Ana Lúcia C.
Domingues (R)bc, Ricardo O. Silva (R)a, Licarion Pinto (R)d
a Universidade Federal de Pernambuco, Departamento de Química Fundamental, Recife, PE, Brazil, 50740-560
b
Universidade Federal de Pernambuco, Centro de Ciências Medicas, Recife, PE, Brazil, 50670-901
c
Universidade Federal de Pernambuco, Hospital das Clinicas, Recife, PE, Brazil, 50670-901
d Universidade do Estado do Rio de Janeiro, Departamento de Química Analítica, Rio de Janeiro, RJ, Brazil, 20550-013
*e-mail: julieth.gonzalezherrera@ufpe.br
Schistosomiasis is a tropical disease that has an acute and chronic course. Periportal fibrosis (PPF)
is delimited to peripheral intrahepatic branches, without promoting destruction of hepatocytes,
however, inducing portal hypertension. Schistosomiasis is one of the most prevalent diseases
among those transmitted to water collections, being mainly a risk in populations in rural areas and
urban peripheries. The eggs of infected patients may develop in the liver granulomatosis lesions that
will form the PPF. The intensity of this fibrosis, depends on the intensity of the infection and the
immune response, and will determine the morbidity of the disease. In Brazil, it is estimated that about
1.5 million people live in areas with high risk of contracting Mansoni Schistosomiasis 1. Due to the
difficulty in accessing hospitals, especially in rural populations, other strategies have been used that
can accurately detect PPF, using a single serum sample. These studies are emerging in an attempt
to develop methods that are capable of identifying and evaluating PPF through biological markers
present in the blood 2. So, this work aimed to develop and evaluate screening models based on
metabolic analysis for staging PPF in patients with Mansoni Schistosomiasis using liquid
chromatography and chemometrics, employing ultrasound (US) as gold standard method. The study
was carried out with 94 serum samples, 19 from patients with a mild stage of PPF, 30 intermediate
and 45 advanced. The analysis conditions of the samples were previously optimized. In sample
preparation, DLLME and QuEChERS methods were tested. For the mobile phase of the
chromatographic analysis, different gradients of acetonitrile and methanol were tested. The
optimization revealed that DLLME and mobile phase gradient with methanol lead to higher number
of visible peaks in the elution. With the defined working conditions, the 94 samples were analyzed
using an HPLC-DAD. The data obtained were pre-processed to correct retention time shifts and
baseline, and later submitted to pattern recognition multivariate analysis using the MATLAB
software. ROBPCA analysis did not indicate the presence of outlier. Among the metabonomic
models built, the DD-SIMCA stands out, which obtained 100% sensitivity, 64% specificity and 68%
accuracy for the mild stage of the disease. 92% sensitivity, 63% specificity and 75% accuracy for
the advanced stage of PPF. Models with high sensitivity values are more interesting for the present
screening analysis since a false negative prejudice the early treatment. So, the present work
proposes a model that can be used for PPF diagnosis triage and for earlier control of the disease.
The integration of analytical techniques and metabonomic models in clinical practice may provide a
tool to help the triage for staging PPF in patients with Mansoni Schistosomiasis. In addition, it is
possible to monitoring the disease with suitable frequency, since serum samples would be collected
repeatedly.
1 Ministerio da Saude, Vigilância da Esquistossomose Mansoni, 4a edição, 2014, Brasil.
2 Barreto, A. V. M. S. Arquivos de Gastroenterologia, 56, 2017, 108.
[DQF/UFPE, CENAPESQ/UFRPE, HC/UFPE, CAPES, CNPq, FACEPE 14/2019 - INOVA IAM]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

453
META-HEURISTIC VARIABLES SELECTION METHOD TO IDENTIFY
ADDITION OF BOVINE IN GOAT MILK
Vitor D. Alves (PG)a*, Elainy V. S. Pereira (R)b, Germano Veras (R)b,
David D. S. Fernandes (R)c, Paulo H. G. D. Diniz (R)d.
aUFPE, Department of Fundamental Chemistry, Recife, Pernambuco, Brazil, 50670901
bUEPB, Department of Chemistry, Campina Grande, Paraiba, Brazil, 58429500
cUFPB, Department of Chemistry, João Pessoa, Paraiba, Brazil, 58429500
dUFOB, Pure and Applied Chemistry Post-Graduate Program, Barreiras, Bahia, 47810059
*e-mail: vitordantas.alves@ufpe.br
Goat milk consume has been increasing since beginning of XX century, from helping on feeding and
nutrition of rural families to high production of its derivative. This milk a rich source of proteins and
carbohydrates, its nutritional proprieties, highlighting great digestibility, high alkalinity, also use as
an alternative to lactose allergic and/or intolerant, comparing to bovine milk. These benefits are from
its sui generis chemical composition, like interesting fat percentage to produce derivatives and β-
casein presence assisting lactose digestion, presenting as great cost-benefit alternative to other
species milk. Europe and Asia rules production and market of goat milk, followed by African an
American continents [1]. Most of american products are made for export to countries with higher
consume, as happens in Brazil. Dairy goats’ production is concentrated in five states, Minas Gerais,
Bahia, Pernambuco, Paraíba and Rio Grande do Norte, producing together around of 280 thousand
litres per year. Exportation and economic value increasing put a target on goat milk to potential
adulterations using cheaper products to increase volume [2]. From water, common added in liquid
products, to milk from other species as an adulterant making harder to identify adulteration using
conventional methods of analysis. Most of these techniques are destructive, inaccurate and produce
residue, even considering that milk is perishable, so it’s required a rapid, accurate, non-destructive
and cheaper technique. In this sense, Near Infrared Spectroscopy has been showing interesting
uses to identify food adulterations, also in analysis in situ using portable devices. Portable
instruments are suitable in these studies because of their low cost compared to a benchtop, allowing
remote inspections. Near-infrared Spectroscopy (NIR), in association with chemometrics provides
interpretation of information that allows distinguish authentic and adulterated samples. From this
view, this work proposes the goat milk added with cow milk identification using portable NIR
spectrometer along with chemometric methods. For that it was associated metaheuristic algorithms
and discriminant analysis. Variable selection methods chosen wavelengths associated to chemical
information of milk to represent dataset. Thus, data were divided by Kennard-Stone Algorithm,
training set were built LDA models with 8 variables by Genetic Algorithm (GA) and 7 variables by
Ant Colony Optimization (ACO). Test set used in GA-LDA and ACO-LDA showed efficiency of
models with 100% of accuracy, sensitivity and specificity to discriminate goat milk authentic samples
from adulterated, even with only 5% of cow milk added. Peaks selected by GA and ACO matched to
C-H, N-H and O-H stretching bonds, correspondents, respectively, to carbohydrates, proteins and
water, major components of both milks. The difference between percentage of these constituents
were the key to assure adulteration of goat milk by bovine. Therefore, the developed models
recognized goat milk samples adulterated by cow milk using fast, non-destructive and accurate
methods.
1 FAOSTAT, Food and Agriculture Organization of The United Nations Statistics Division. 2021.
2 Pereira EVS, Fernandes DDS, Araújo MCU, Diniz PHGD, Maciel, MIS, LWT – Food Science and Technology, 127, 2020.
[NUPEA/UEPB, LAQA/UFPB]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

454
MULTIVARIATE CALIBRATION MODEL FOR ANALYSIS OF MIXTURES OF
LACUSTRINE AND MARINE OILS USING APPI (+) FT-ICR MS
Leonardo M. Ferreiraa (UG), Danielle M.M. Francoa (PG), Jussara V. Roquea (R), Mário D.
Rangela (R), Boniek G. Vaza* (R)
aFederal University of Goiás, Chemistry Institute, Goiânia, Goiás, Brazil, 74690-900
*e-mail: boniek@ufg.br
In several sedimentary basins of the Brazilian continental margin, there is more than one type of
source rock, i.e., oils of different origins are generated1. Identifying the contribution of each type of
crude oil in each accumulation is an important strategy for exploring such areas 2. This study's
objective was to build a multivariate calibration model using the partial least squares (PLS)
regression method3,4 to predict the percentage proportion of lacustrine oil in samples from one
Brazilian sedimentary basin. Thirty-seven samples were prepared from the mixture, in different
proportions (from 0 to 100%), of two endmembers: one of lacustrine origin and the other of marine
origin, both from the same basin. All prepared samples were analyzed using a Fourier transform ion
cyclotron resonance mass spectrometry (FT-ICR MS) having atmospheric pressure photoionization
(APPI) in positive mode ion as the ionization source. It stands out that four latent variables were
chosen, and the values of the model quality statistical parameters are: root mean square error of
calibration (RMSEC) of 4.6686 and prediction (RMSEP) of 6.2575. Furthermore, in Figure 1 (A), it’s
possible to verify the linear relationship between the actual percentage proportion of lacustrine oil in
the mixtures and the predicted percentage proportion of lacustrine oil. It can be observed that the
model can predict the content of the lacustrine oil with acceptable error because the points in the
graph are close to the diagonal line, indicating that the predicted values are close to the real ones.
A better idea of this prediction error can be seen in Figure 1 (B). That’s because it’s possible to
analyze the residual value of each mixture, i.e., the residual value varies approximately from -12%
to +15%.
Figure 1. Real and predicted values for the model calibration and prediction set (A) and residue from mixtures (B).
Thus, it’s inferred that the model created could predict the lacustrine oil content in the mixtures
prepared with acceptable error values, considering the complexity of the samples, due to the ultra-
high resolution of FT-ICR MS combined with chemometric tools.
1 Trindade LAF, Brassell SC, Organic Geochemistry, 19, 1992, 13.
2 Neto EVS, Hayes JM, AAPG Bulletin, 83 (3), 1999, 496.
3 Ferreira MMC, Antunes AM, Melgo MS, Volpe PL, Química Nova, 22 (5), 1999, 724.
4 Geladi P, Kowalski BR, Analytica Chimica Acta, 185, 1986, 1.
[Petrobras, CNPq, LaCEM]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

455
MULTIVARIATE DIGITAL IMAGE ANALYSIS FOR NON-DESTRUCTIVE
PHENOTYPIC DISCRIMINATION OF SESAME SEEDS
Wilson B. N. Filho*a, Mirla J. A. Cidadeb, Francisco dos Santos Panerob, Oscar J. Smiderlec
a Federal University of Roraima, Agrotechnical School, Boa Vista, Roraima, Brazil, Campus Murupu.
b Federal University of Roraima, Chemistry department, Boa Vista, Roraima, Brazil, 69310-000
c Brazilian Agricultural Research Corporation, Boa Vista, Roraima, Brazil, 69301-970
*e-mail: wilson.botelho@ufrr.br
The use of digital images as a platform to perform chemical, clinical, nutritional analysis, in quality
control and to determine properties in substances has been explored in recent years, which allows
the characterization of compounds and their physicochemical properties through a set of variables
related to color that a given sample presentes 1,2. Digital Image Processing and Analysis (PADI) was
used to extract attributes from digitized images of sesame seeds. With the aid of a microtiter plate,
approximately 20 seeds of each sample were placed in each microwell, to be later scanned using
an HP officejet 7610 scanner. The images were saved in JPEG (Joint Photographics Experts Group
format) with a resolution of 300 dpi 3,4. To perform the classification, Principal Component Analysis
was used, PCA having as an attribute for its discrimination, the color of the seeds. For the
development of the work, the software for processing and digital analysis of images, ImageJ, was
used, which made it possible to extract the RGB channels and colorimetric profile of the digitized
images. Readplate puggling was used allowing the extraction of RGB values from all wells
simultaneously. For chemometric analysis, the ChemoFace software was used, because in addition
to being free software, it has a friendly graphical interface. With the application of PADI, attributes
were extracted from the digitized images, which could be used for their discrimination through PCA.
The digital processing and analysis of the images made it possible to efficiently discriminate the
sesame samples, when paired with the standard (GC), obtaining an accumulated variance greater
than 95%, which makes the use of PCA highly relevant. When the behavior of the cultivars and the
lineage without the presence of the pattern (GC) was evaluated, it was observed that the PCA was
quite efficient in the discrimination of the samples, obtaining PC1 values for the pairings between
the cultivars and the lineage between 90.63 to 99.06% of the total variance of the data. By grouping
all the samples, PC1 obtained values of 96.21% with the presence of commercial sesame and
96.79% with its absence. Several works have demonstrated the use of this technique combined with
chemometrics to determine several parameters in varied samples such as Damasceno et al, (2015)5
in the determination of pH in drinking water, Colzani et al, (2017)3 in the determination of manganese
in batteries and Dominguez; Centuríon, (2015)4 in the discrimination of the color of honeys from
different regions.The results obtained in this work indicate that the proposed methodology can be a
simple analytical alternative for the phenotypic discrimination of seeds to perform their classification,
having color as an attribute.
1 Neto GFS, Fonseca A, Braga JWB, Química Nova, 39, 2016, 876.
2 Gomes MC, Trevizan LC, Nóbrega JÁ, Kamogawa, MY, Química Nova, 31, 2008, 1577.
3 Colzani H, Rodrigues QEAG, Fogaça C, Gelinski JMLN, Pereira-Filho ER, Borges EM, Química Nova, 40, 2017, 833.
4 Domínguez MA, Centuríon ME, Microchemical Journal, 118, 2015,110.
5 Damasceno D, Toleto TG, Godinho MS, Silva CP, Oliveira SB, Oliveira AE, Química Nova, 38, 2015, 836.
[UFRR, CNPq, Embrapa]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

456
MULTIVARIATE OPTIMIZATION FOR THE EXTRACTION OF PHENOLIC
COMPOUNDS IN FRUITS AND NUTS FROM THE BRAZILIAN CERRADO
Darlisson S N Silva (PG)*a, Mikael K A Mendes (PG)a, Carlos A Galinaro (PG)b, Tiago L S
Coelho (PG)a, Ronaldo C Coelho (PG)a, Hudson de C Silva (UG)a, Matheus S Silva (UG)a, Ana
R A Nogueira (R)c, Emanuel Carrilho (R)b, Cícero A Lopes Júnior (R)a, Edivan C Vieira (R)a.
aUniversidade Federal do Piauí, Departamento de Química, Teresina, Piauí, Brasil, CEP: 64049-550
bInstituto de Química de São Carlos, Departamento de Química, São Carlos, São Paulo, Brasil, CEP: 13560-970
cEmbrapa Pecuária Sudeste, São Carlos, São Paulo, Brasil, CEP: 13560-970
*e-mail: darlisson.ufpi@hotmail.com
The Brazilian Cerrado is one of the largest savannas with a great biodiversity. Some species studied
such as Pequi (Caryocar brasiliense), Buriti (Mauritia flexuosa), Tuturubá (Pouteria macrophylla),
Murici (Byrsonima crassifolia), Tucum (Astrocaryum vulgare), Guabiraba (Campomanesia
xanthocarpa), Cashew nut (Anacardium occidentale) and Brazil nut (Bertholletia excelsa) have
revealed a high antioxidant potential. This biological activity has been associated with phenolic
compounds. A wide variety of solvent combinations have been used to improve polyphenols
extraction in plant matrices. So, the objective of this study was to develop a simple, efficient, and
reliable extraction method for extracting phenolic compounds from fruits and nuts samples from the
Brazilian Cerrado. After freeze-dried and lipids extraction with petroleum ether:ethyl ether (1:1: v/v),
equal portions of each sample were mixed to obtain a blend that was used for the optimization runs.
The simplex-centroid mixture design was used to optimize the extraction of phenolics from the matrix
blend by evaluating three solvents: water (w), ethanol (e), methanol (m) and their mixtures1. Briefly,
ca. 15 mg of blend sample was extracted with 1.5 mL of extraction solution proposed by centroid-
simplex mixture design. The mixture was stirred at 800 rpm for 2 h and centrifuged at 10,000 rpm
for 30 min. Total phenolic content (TPC) was based on Folin-Ciocalteu method and the
Figure 1. Contour surface for the quadratic
measurements using microplate reader2. An external
model. TPC is predicted as a function of the calibration curve was prepared with gallic acid (A =
ratio of water, ethanol, and methanol solvents. 0.0045c + 0.0693, r = 0.9972). The runs of experimental
design were carried out in triplicate and randomly. The
highest TPC was obtained using the binary mixtures
(50/50) of water:ethanol (692 ± 51 mg GAE/100 g) and
water:methanol (663 ± 3 mg GAE/100 g). The quadratic
model is well-fitted, reliable (Fcal (68.70) > Ftab (3.69)) and
high predictive describing ca. 97.72% of data variance.
Fig. 1 shows the surface contour plotted applying Eq. TPC
(mg GAE/100 g) = + 617.31w + 251.87e + 470.65m +
1075.50we + 506.81wm - 373.68em, which illustrates the
effect of extractor solution composition on TPC. The
numerical analysis showed that the best solvent mixture
for the extraction of phenolics from blend matrix is 66% of
water and 34% ethanol, with a predicted of TPC 734 mg
GAE/100g (desirability of 0.9999). TPC found applying the
optimal extraction condition was 745 ± 16 mg GAE/100g,
which is not significantly different from the predicted value (p<0.05). The optimal extraction mixture
was applied to the samples and the TPC in mg GAE/100g found was 465 ± 20 for the pequi, 635 ±
30 buriti, 1688 ± 46 tuturubá, 382 ± 27 murici, 981 ± 95 tucum, 1486 ± 71 guabiraba, 120 ± 17
cashew nut, and 428 ± 35 Brazil nut. The results revealed that using mixtures to extract phenolics is
more effective when compared to using pure solvents. A future step will be to evaluate the phenolic
extracts using LC-MS/MS as well as investigate their biological activities.
1 Mendes, MKA, Oliveira CBS, Veras MDA, Araújo BQ, Dantas C, Chaves MH, Lopes Júnior CA, Vieira EC. Talanta, 205,
2019, 120100.
2 Bobo-García G, Davidov-Pardo G, Arroqui C, Vírseda P, Marín-Arroyo MR, Navarro M. Journal of the Science of Food
and Agriculture, 95, 2015, 204–209.
[CNPq]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

457
NEW STRATEGY TO ASSESS FOOD INTEGRITY BASED ON ONE CLASS
CLASSIFICATION OF MULTIBLOCK DATA
Larisa Onçaa,b , Liudmyla Khvalbotab, Andrea Machyňákováb and Ivan Špánikb, Adriano A.
Gomesa,b*
aFederal University of Rio Grande do Sul, Institute of Chemistry, Porto Alegre, RS, Brazil, 91501970
bSlovak University of Technology in Bratislava, Institute of Analytical Chemistry, Faculty of Chemical and Food
Technology, Bratislava, Slovakia, 81237
*e-mail: araujo.gomes@ufrgs.br
In this work, a new approach to deal properly with food integrity1 by modelling methods based on
data recorded from different sources is proposed. This new method called sequentially
orthogonalized2 one class partial least square3 (SO-OC-PLS), combines an orthogonalization step
between the different data sets (data blocks) to eliminate redundant information followed by definition
of an acceptance area for a target class by OC-PLS3. The proposed method was evaluated in two
case studies. The first one was controlled scenario with simulated data. Simulated data set was
designed to show the benefits of the proposed method. In the first step, both blocks 1 and 2 were
modelled by OC-PLS followed by reorganization to side by side row-wise augmented structure. The
prepared dataset underwent OC-PLS treatment and finally was processed by SO-OC-PLS. In the
second case study, the approach was applied on UV-VIS and IR data, in order to differentiate Slovak
Tokaj Selection wines of high quality from other lower market value wines from Slovak Tokaj wine
region. Varietal Tokaj wines, Tokaj cuvée, different Tokaj “putňa” selections and Tokaj essence of
vintages (1959–2017) were included in this study. All 58 samples were stored in bottles at 4 °C. After
opening, the wine aliquot was transferred into 20 mL vial and equilibrated at 20 °C before
measurements. The samples were obtained directly from producers. All calculation was carried out
in MatLab 2010a environment. The training set contained 30 wine samples form target group of 40
wines selected by by Kernnard-Stone algorithm. Additionally, test set contains 10 target and 18 non-
target wine samples. The proposed method was evaluated in relation of individual data blocks and
row-wise augmented data considering the sensitivity (SEN), selectivity (SPC) and efficiency (Eff).
Individual blocks did not achieve high classification efficiency even if data were combined in a row-
wise augmented matrix. This finding reinforces the idea that multiblocks should be treated together
and not as a single set of data. In general, the proposed method achieved results in compliance with
the level of overlap defined in data setup for all α values. Efficiency was practically similar compared
to 0.01 and 0.05 level but showed a decrease if α equals to 0.1. It was expected, that
orthogonalization process would reduce the number of latent variables when comparing the model
that cross both blocks of data. In both cases, the proposed method shows improvements in the
results in relation to the demise of the disregarding the structure of multiblocks. It was proved that at
least in discussed case studies; properly treated multiblocks can significantly improve food
classification. In the proposed method, the orthogonalization of a block in relation to the others is
used to discard the common and retain only distinct information. This allowed estimating an
acceptance area with high efficiency in the prediction step.
1 Montgomery, H, Haughey, SA, Elliott, CT. Global Food Security, 26, 2020, 100447.
2 Biancolillo, A, Måge, I, & Nᴂs, T. Chemom. and Intell. Lab.Syst., 141, 2015, 58.
3 Xu, L., Goodarzi, M., Shi, W., Cai, C.-B., Jiang, J.-H. Chemom. Intelli. Lab. Syst. 139, 2014, 58-6.
[CNPq, PPGQ/UFRGS, STUBA, NSP-Slovakia, Marie Sklodowska Curie Actions-SASPRO2]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

458
PHYTOCHEMICAL PROSPECTION OF MAYTENUS ILICIFOLIA (ESPINHEIRA
SANTA), BY 2D TLC AND MACHINE LEARNING.
Thiago Silva Dias 1, Rodolfo Santos Barboza 1, Daniella Lopez Vale 1

1 UFRJ - Universidade Federal do Rio de Janeiro
Among the therapeutic plants related in RENISUS (Relação Nacional de Plantas Medicinais de
Interesse do Sistema Único de Saúde), list of medicinal plants recognized by the brazilian health
program - SUS (Sistema Único de Saúde), there is the Maytenus ilicifolia, popularly known as
“espinheira santa”. The related proven pharmacological properties1 as well as its intense popular
use as herbal medicine guide to the need to develop a quality control method for the commercial
phytoterapics products based in Maytenus ilicifolia leaves, because it becomes a health public safety
problem. The aim of this work was the development of a phytochemical prospection method
combining two-dimensional thin-layer chromatography (2D TLC), computerized densitometry and
machine learning analysis. For this, leaves of Maytenus ilicifolia samples of commercial herbal
medicines were dried and crushed (≤ 0,50 mm), extracted with MeOH/H2O (3:1) (v/v) under
ultrasound (15 min). The extract was partitioned with n-hexane (5 × 5 mL). The defatted extracts
were analyzed by 2D TLC in mobile phase and development conditions to monitor polyphenolic
substances2 (flavonoids, phenolic acids, among others - stationary phase: silica gel; mobile phase:
AcOEt/HCOOH/AcOH/H2O 100:11:11:27. Thus, using image analysis tools, such as computerized
densitometry to determine the relative percentage of substances present in the leaves of the studied
plant samples and the machine learning technique for hierarchical grouping 3 of images, it was
possible to suggest the formation of 2 groups of plants, being the plants "probably therapeutic" -
when a similar composition was observed with the reference Maytenus ilicifolia plant - and the plants
"probably non-therapeutic" - when the chemical profile was different from the reference plant. The
results obtained confirmed the need for a rigorous quality control of plants marketed as herbal
medicines, since the phytochemical prospection showed that most of the samples studied did not
present the pharmacological active principles of the species Maytenus ilicifolia.
1 BERSANI‑AMADO, C. A. et al. An International Journal Devoted to Pharmacological and Toxicological Evaluation of

Natural Product Derivatives, v. 14, n. 7, p. 543-545, 2000. 2 WAGNER, H.; BLADT, S. Springer Science & Business
Media, 1996. 3 BOŽIČEVIĆ, A. et al. Analytical chemistry, v. 89, n. 23, p. 12682-12689, 2017.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

459
PLS PREDICTION OF ACID VALUE IN ANDIROBA OILS BASED ON NIR
SPECTROSCOPY
Rayanna Falcão da Rocha Reis (UG)a*, Neirivaldo Cavalcante da Silva (R)a
aFederal University of Pará, Faculty of Chemistry, Belém, Pará, Brazil, 66075110
*e-mail: rayanna.reis@icen.ufpa.br
Andiroba seed oil is a non-wood forest product (NWFP) extracted from andiroba plant species
Carapa guianensis Aubl. and Carapa procera D.C, mainly found in the Amazon rainforest. It has a
high commercial value and plays an important role for the economic development of local
communities with wide use by cosmetic industries and a series of medicinal properties1. As
established by ANVISA, vegetable oils need to meet minimum quality characteristics. Among them,
the acid value (AV) is an indicator of the state of conservation of oils. The reference analysis for
determination of the AV in vegetable oils, according to the recommendations of the Association of
Official Analytical Chemistry and American Oil Chemists' Society, involves a time consuming wet
analytical method that uses significant amounts of chemicals. On the other hand, the use of near
infrared spectroscopy (NIRS) together with chemometrics represents a high potential analytical
alternative for assessing AV in andiroba oils. NIRS is directly influenced by vibrations of O-H, N-H
and C-H chemical bonds, which are present in the major chemical components of the composition
of andiroba oil. When combined with chemometrics, NIRS can align with policies focused on the
development of more sustainable and environmentally responsible methodologies, as indicated by
UN Sustainable Development Goals. This work investigates the potential of a NIR based PLS
regression model to predict AV in andiroba oils. This application has not been described in the
literature so far. Thirty-nine andiroba oil samples were acquired from a 6-year historical bank of a
company located in Pará state. AV analyses were carried out as in the reference method described
by Adolfo Lutz Institute and expressed as the number of mg of potassium hydroxide needed to
neutralize 1 g of the sample2. Absorbance spectra were acquired in a FTNIR spectrometer (MB3600,
ABB) in the range from 13000 to 4000 cm-1, with 16 scans and absorbance values obtained every 4
cm-1 and using a 1mm optical length quartz cuvette. Absorption bands are mainly due to
combinations (4,500 and 4,000 cm-1) and first overtones (6,000 to 5,500 cm-1) of stretching of C-H
bonds from typical compounds present in andiroba oil. Spectra were preprocessed using first
derivative with a second-order polynomial and Savitzky-Golay filter with a 21-points smoothing
window. A PLS model was developed to determine AV (5,5 to 81,2 calibration range) in the andiroba
oils, with 29 calibration samples and 10 external validation samples, selected using the SPXY
algorithm. The model predictive performance was evaluated using the figures of merit RMSE, bias
and R2 for cross-validation and external validation. Six latent variables were considered as the ideal
number, with basis on venetian blinds cross-validation. The PLS loadings are mainly related to the
region of C-H combination bands, around 4300 cm-1. R2 values were 0.99, 0.96 and 0.96; and RMSE
values were 2.3, 3.8 and 3.1, for calibration, cross validation and external validation, respectively.
No significant bias was observed, according to a t-test at a confidence level of 95%. The results
obtained in this work demonstrate the NIR spectroscopy as a feasible alternative to determine AV in
andiroba oil samples. Additionally, the proposed approach can also be applied to investigate
methodologies for determining other quality characteristic of andiroba and other vegetable oils
typically found in Amazon rainforest.
1 Amaral, LFG.; Fierro, IM, Brazilian Journal of Pharmacognosy, 23, 2013, 716.
2 Odair Zenebon, Neus Sadocco Pascuet and Paulo Tiglea - São Paulo: Instituto Adolfo Lutz, 2008.
[FAPESPA, CNPq and Amazon Oil Company]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

460
PRACTICAL ASPECTS OF THE USE OF PORTABLE NIR SPECTROMETERS IN
THE SUGARCANE PRODUCTION
Bruno H. S. Melo (PG)a*, Rafaella F. Sales (R)b, Jorge S. P. Silva (R)b, Deborah M. C. Peixoto
(R)b, Lourival S. B. Filho (R)b, Aluska G. C. A. Sousa (R) b, M. Fernanda Pimentel (R)b.
a Federal University of Pernambuco, Department of Fundamental Chemistry, Recife, Pernambuco, Brazil, 50740-560
b FederalUniversity of Pernambuco, Department of Chemical Engineering, Recife, Pernambuco, Brazil, 50740-590
*e-mail: bruno.hsmelo@ufpe.br
In a previous work¹, we demonstrated the feasibility of using a handheld near infrared spectrometer
(MicroNIRTM) to monitor important quality parameters, brix and pol, in different stages of an industrial
sugar production process. In this study, less concentrated samples (brix up to 45%w/w) were measured
by transmittance, while samples with brix greater than 45%w/w were analyzed using diffuse reflectance.
PLS (partial least squares) and SVM (support vector machine) were used to build models. It was possible
to identify the presence of non-linearities in the dataset, mainly for the more concentrated samples. The
current work aims to continue this previous study by evaluating 2 approaches: 1. The ability of the models
built with data from previous seasons (2019/2020 and 2020/2021), using diffuse reflectance, to predict
brix and pol of a new dataset from the current season (2021/2022). 2. Spectral acquisition using
transflectance, with minimal sample preparation, to build brix and pol models, using 2 portable
spectrometers operating in different wavelength ranges, MicroNIRTM (950 to 1650 nm) and NeoSpectraTM
(1350 to 2500 nm). In both approaches, spectra were pre-processed using first derivative (2nd order
polynomial function and 5-point window) and SVM was employed as regression technique. F-tests (at
95% confidence) were used to compare the RMSEPs values. For the first approach, the collected
samples (138 for brix and 107 for pol) were predicted in the pre-existing models. For brix, the RMSEP
(1.63%w/w) was similar to that obtained previously (1.44%w/w), indicating a satisfactory performance of
the model (F-test, 95% confidence). For pol, the RMSEP (3.1%w/w) was higher than that of the previous
study (2.44%w/w), suggesting a poor performance of the model in predicting pol in the new samples.
These results may be influenced by the variation in the content of the reducing sugars (RA) due to climatic
changes between seasons, that interfere in pol determination. The Table below summarizes the results
for the 2nd approach. For brix, NeoSpectraTM data presented similar results to those obtained in the
previous study (0.59 and 1.44%w/w). For MicroNIRTM data, it was possible to construct a global model
with RMSEP of 0.92%w/w. This value is relatively high for very diluted samples (brix around 1.7%w/w),
but it can be satisfactory to facilitate the industrial routine, using only one model.
Instrument Quality parameter Range %(w/w) ncal npred RMSEP

1.7 - 44.5 53 23 0.52
Brix
45.2 - 93.6 107 45 1.37
NeoSpectraTM
1.7 - 16.9 35 13 1.04
Pol
34.8 - 88.1 73 32 2.17
Brix 1.7 - 93.6 175 74 0.92
MicroNIRTM 1.5 - 16.9 36 14 0.45
Pol
34.8 - 88.1 82 33 3.33
For pol, the high variability of the data did not allow building global models. For the models with more
concentrated samples, there was no significant improvement in the RMSEP values in relation to the
previous work. The RA concentration may also have interfered in these results. For models with less
concentrated samples, the MicroNIRTM result showed a significant improvement in the RMSEP value,
compared to that obtained previously. Finally, despite the improvements obtained in some cases in
relation to the previously tested conditions, from this preliminary study, it is possible to notice that there
is not a single instrument nor mode of spectra acquisition that would be better for all industrial samples
and brix and pol determination.
1 Melo BHS et al., Food Chemistry, 369, 2022, 130919.
[NUQAAPE; INCTAA, CNPq, CAPES, FACEPE]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

461
PREDICTING TOKAJ WINE AGE BY MEANS OF GC×GC IMAGE AND
CHEMOMETRIC TOOLS
Nemanja Koljančića, Olga Vyviurskaa, Larissa Onçaa,b, Adriano A. Gomesa,b and Ivan Špánikb*
aSlovak University of Technology in Bratislava, Institute of Analytical Chemistry, Faculty of Chemical and Food
Technology, Bratislava, Slovakia, 81237
bFederal University of Rio Grande do Sul, Institute of Chemistry, Porto Alegre, RS, Brazil, 91501970
*e-mail: ivan.spanik@stuba.sk
Wine age is a parameter of great significance in wine quality1. However, there is no way to access this
information directly. A wine’s vintage can be predicted based on the presence and content of volatile
organic compounds (VOCs)2,3. One of the most efficacious methods for analyzing VOCs is
comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry
(GC×GC-TOF-MS). However, the extensive information generated by this analytical technique can make
data processing challenging and time consuming. Therefore, we proposed an age prediction method
based on GC×GC-TOF-MS digital images and multivariate calibration. In total, 71 wine samples were
taken from the Tokaj wine region, which is located between Slovakia and Hungary. The samples include
three groups of wines: botrytized wines, varietal wines as well as wine essence, which were obtained
from four wineries. The analyzis was performed on a GC×GC-TOF-MS with a cryogenic modulator. VOCs
from wine samples were extracted by solid phase microextraction (SPME) using PDMS/CAR/DVB SPME
fiber. The GC column setup consists of DB-FFAP in the first dimension and BPX-50 in the second. Helium
was used as a carrier gas with flow rate 1 mL min−1. The primary oven program was: 40 °C (15 min) to
220 °C (5 min) at 2 °C min−1. The cryogenic modulator operated with a modulation period of 10 s. For
the temperature program in the second dimension a temperature offset of 5 °C was used.. The signal
acquisition rate was 100 spectra/s in the m/z range 29-550. The final images were converted into color
histograms generating a 71×1792 matrix including greyscale, red-green-blue (RGB) and Hue-Saturation-
Intensity (HSI) color channels.The data set was split into a calibration (51 samples) and an external
prediction set (20 samples). Partial Least Square (PLS) model,variable selection by interval approach
(iPLS) and successive projection algorithm (iSPA-PLS) were performed. For all cases the optimized
number of latente variable (LV) was established by a minimum value of the RMSECV in the cross-
assessment step. Interval-based models were computed for partitions of 5, 10, 15, 20 and 25 equidistant
intervals. All calculations were performed in a MatLab environment. For the PLS global model best results
were achieved using 2 variable latent (VL): RMSECV equal to 1.06 year and REPcv 5.2%; RMSEP 0.92
year REPpred equal to 4.4%). In turn, interval-based iSPL and iSPA-PLS models showed the following
results respectively: 2 VL – 10 intervals RMSECV = 1.05; REPcv = 5.1% and RSEMP = 0.93; REPpred
= 4.5% and 2 VL – 10 intervals RMSECV = 0.90 year; REPcv = 4.4%, RSMEP = 0.81 year and REPpred
= 3.9%. It is important to highlight that for all cases significant bias was absent with 95% of statistical
confidence. Therefore, exploring the composition of volatiles via GC×GC-MS through image data with
low computational demand proved to be feasible and efficient to provide good age estimates of Tokaj
wines.
1 Khvalbota L, Machyňáková A, Čuchorová J, Furdíková K, Špánika I, Journal of Food Composition and Analysis, 96,
103719, 2021
2 Marti MP, Busto O, Guasch J, Journal of Chromatography A, 1057, 2004, 211
3 Bailly S, Jerković V, Meurée A, Timmermans A, Collin S, Journal of Agricultural and Food Chemistry, 57, 2009, 8557
4 Vyviurska O, Koljančić N, Gomesa AA, Špánika I, Journal of Chromatography A, 1675, 463189, 2022
[Agency of the Ministry of Education of Slovak Republic, Slovak University of Technology

in Bratislava, UFRGS, CNPq, Marie Sklodowska Curie Actions-SASPRO2, National
Scholarship Programme of the Slovak Republic]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

462
PREDICTION OF CHALLENGING COFFEE PROPERTIES USING GC-FT-
ORBITRAP MS AND CHEMOMETRICS
Andre C. Paivaa, Carlos A. Teixeiraa, Leandro W. Hantaoa, *
a University of Campinas, Department of Analytical Chemistry, Campinas, São Paulo, Brasil, 13083-970
*e-mail: wang@unicamp.br
Coffee is one of the most consumed beverages worldwide. In 2019, this market generated
approximately 30 billion dollars and Brazil was ranked as the leader in coffee beans exportation 1.
Among the reasons that promote coffee intake are taste, cultural, and visual appeal 2. In this context,
the five human senses play a major role when it comes to the introduction of a new variety of coffee
to consumers, with smell being one of the most important senses explored during a beverage
sensory evaluation. This sense is stimulated by volatile and semi-volatile organic compounds (VOC
and SVOC, respectively) present in the beverage’s aroma. VOC and SVOC characterization have
been explored a lot by researchers. More than 900 different compounds were already identified in
roasted coffee beans 3, which guarantees a range of possibilities that could be explored through
sensory analysis to predict coffee properties. In fact, coffee producers and panelists have been using
sensory analysis to predict specific sensory attributes for decades. Yet, although this practice is
commonly adopted in this field, with protocols and specific procedures, gaps still exist and may
prejudice the quality of the results for sensory and coffee properties prediction, since the sensory
analyst's conduct will always bias the data. Therefore, the development and establishment of
standardized and complementary sensory-chemistry protocols remain an unmatched demand that
should be addressed. Aiming to explore this field, this study investigated the development of
chemometric models able to accurately predict sensory information about coffee samples using their
chemical information, which would be a prediction approach free from human bias. In this context,
solid-phase microextraction in headspace mode extracted the compounds from coffee matrices,
which were then separated and identified using gas chromatography coupled to a high-accuracy and
high-resolution mass spectrometer with Fourier transform. A new automated SCRIPT, which corrects
RESHAPE for non-integer MS acquisition rates, was explored to import the MS-based
chromatographic data 4. Second-order data for 118 samples were joined in a single three-
dimensional array (118×8820×351) and then reshaped to a matrix (118×3095820). A filtering step
was investigated to remove low-intensity variables unrelated to the study goal, which decreased
them from 3,095,820 to 124,890. The use of chemometric tools to investigate and identify insights
and trends in it prevents relevant information from being overlooked and discarded. Since
classification models were built for a multiclass problem in this study, another challenge faced during
data processing was dealing with the effect of unbalanced classes. There is a relative difference
between the number of samples belonging to a majorly represented class and the number of
samples belonging to a less represented class, which leads to a scenario of unbalanced classes
when samples are split between training and validation subsets and influences the model accuracy.
Finally, finding indices that assess the global classification performance of each tested model, and
not each specific class, was another concern in this study. Metrics such as accuracy, extended
Matthew correlation coefficient, and averages precision and sensitivity were used 5. For example,
built models exhibited average sensitivities between 0.86 and 0.95.
1 (2022) OEC - Coffe - Product Trade, Exporters and Importers. https://oec.world/en/profile/hs92/coffee
2 Seninde DR, Chambers E, Beverages, 6, 2020, 1.
3 Mussatto SI, Machado EMS, Martins S, Teixeira JA, Food Bioprocess Technol, 4, 2011, 661.
4 Hantao LW; Teixeira CA; Ferreira VHC, Harvard Dataverse, V4, 2022.
5 Ballabio D, Grisoni F, Todeschini R, Chemometrics and Intelligent Laboratory Systems, 174, 2018, 33.
[UNICAMP, CNPq, CAPES, Petrobras, FAPESP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

463
PURITY OF ALMOND FLOUR USING HANDHELD NEAR INFRARED
INSTRUMENTS AND PLS REGRESSION
José Marcelino Netto (PG)a, Fernanda Honorato (R)b*, Maria Fernanda Pimentel (R)b
aUFPE, Departamento de Química Fundamental, Recife, PE, Brazil, 50740-560
b UFPE, Departamento de Engenharia Química, Recife, PE, Brazil, 50740-590
*e-mail: fernanda.honorato@ufpe.br
The almond seed, commonly imported, is mainly found in powder form (flour) and very susceptible
to economically motivated fraud due to its high price. This adulteration is characterized by the
addition of low-quality ingredients, which negatively affect the nutritional performance [1]. In the
current study, methods using near-infrared spectroscopy (NIRS) were developed to determine the
purity of almond flours. Three different handheld NIR instruments (DLPR NIRscanTM Nano: 900 -
1700nm; MicroNIR™: 950 - 1650 nm; and NeoSpectra: 1350 - 2500 nm) were evaluated to verify
the purity. The results were compared with a benchtop FT-NIR (FT-IR Frontier - Perkin Elmer: 900 -
2500 nm). Fifty-four almond flours of different varieties and granulometries were acquired in local
markets and adulterated with low-cost flours widely consumed in Brazil, such as cassava flour, oat,
peanut and flour mixtures, totalling one hundred and twenty-four samples (eighty-six samples for the
calibration set and thirty-eight samples for the prediction set using the KSXY algorithm). Partial Least
Squares regression (PLSR) was employed to predict the purity of the sample using 60% to 99%
(m/m) purity as calibration range. The PLS models showed coefficients of determination (R²) greater
than 0.90 for all models, and root mean squares errors of prediction (RMSEP) between 3.3-4.8%
purity. The regression coefficients of the PLS models had significant values in the spectral regions
around 1200 and 1400 nm (for NanoNIR and MicroNIR), and around 1400-1600 for NeoSpectra,
regions, providing important information for content discrimination of the flours, such as lipid, starch
and protein [2]. The results of the calibration models indicate that the handheld NIR instruments are
an efficient tool for quantifying the purity of almond flour and, consequently, for identifying adulterated
samples. These portable instruments are advantageous as they allow in-situ analysis as well as
having lower cost when compared with benchtop FT-NIR instruments.
Figure 1. Graph of predicted x measured values for almond flour purity using 1st Derivative as pre-processing for
handheld NIR devices (a.b.c) and FT-NIR (d). Calibration samples in black color and external prediction samples in red
ones.
1Tibola CS, Silva SA, Dossa AA, Patrício DI, Journal of Food Science, 83, 2018, 2028
2 Kumar K, Resonance, 26, 2021, 429
[NUQAAPE, FACEPE, CAPES, CNPq, INCTAA]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

464
QUALITY ATTRIBUTES ANALYSIS AND AUTHENTICATION OF ARGENTINEAN WINE
VINEGARS BASED ON MULTIPLE SPECTRAL SOURCE FUSION STRATEGIES
Wagner, Marcelo*ab, Ortiz, Danielac, Zaldarriaga Heredia, Jorgelinaab, Montemerlo,

Antonellad, Azcarate, Silvanaab, Camiña, José ba Facultad Ciencias Exactas y Naturales,
Universidad Nacional de La Pampa, Santa Rosa, La Pampa, Argentina, 6300
b Instituto de Ciencias de la Tierra y Ambientales de la Pampa - CONICET, Santa Rosa, La Pampa, Argentina, 6300
c EEA Anguil, Instituto Nacional de Tecnología Agropecuaria, Anguil, La Pampa, Argentina, 6326
d Instituto de Química de San Luis, “Dr. Roberto A. Olsina” (INQUISAL-CONICET-UNSL), San Luis, Argentina, 5700
*e-mail: marcelowagner_14@hotmail.com
Wine vinegars are highly valued mainly regarding the implemented acetification process. From a
technological point of view, there are two production methods: submerged cultivation method, which
is used in the industry to quickly obtain low-quality vinegars, and traditional method, in which the
acetic fermentation is carried out inside wooden barrels, which are filled with wine to ferment, giving
rise to high-quality vinegars that are more appreciated for their greater organoleptic complexity1.
Particularly in Argentina, most wine vinegars are made through industrial fermentation processes
and few of them are produced in a traditional way but without added value2. Currently, the search
for fast, accurate and low-cost determinations makes UV-Vis and near-infrared (NIR) spectroscopies
one of the most popular alternatives to determine the concentrations of chemical compounds as well
as for authenticity studies3.The aim of this work was to develop an analytical methodology based on
UV-vis and NIR measurements in combination with multi-block analysis to quantify a multi-
parametric analysis for the quality control of wine vinegars and, additionally, to differentiate them
according to the implemented production method. Different determinations that are normally used
for the quality control of these products were carried out applying standard methods. For vinegars,
the following results were obtained: total acidity 3.70-6.82%, fixed acidity 0.11-1.08%, volatile acidity
3.63-6.70%, pH 2.49 - 3.07 and polyphenol concentration 0.1 1.98 g/L eq. Ac. Gallic. Spectra of each
sample were obtained from direct measurement for NIR (944-1670nm) and by dilution for UV-Vis
(178-890nm). With the information generated, predictive models were built for the quantification of
each parameter through partial least squares (PLS) regression analysis with each data set obtained
by each individual technique. Likewise, in order to exploit the particularities of the different data
blocks, different data fusion strategies (low and mid-level) were performed and compared. The
predictive capacity of each model was evaluated from the calibration regression coefficients (R2),
the mean square errors of calibration (RMSEC), cross-validation (RMSECV) and prediction
(RMSEP), as well as the percentage error of prediction (REP). The best results were achieved when
the mid-level data fusion strategy with SO-PLS was applied. All the parameters analyzed showed
REP values below 12%. From spectral data, a classification approach to differentiate and classify
vinegars produced by traditional and industrial systems was conducted. Principal component
analysis (PCA) and SO-PLS discriminant analysis (PLS-DA) were performed on the data set as
exploratory and classification analysis, respectively. From PCA, a clear differentiation of the samples
of wine vinegars produced industrially from those produced traditionally was observed, and the
classification analysis using SO-PLS-DA yielded a correct classification rate of 96%. The synergistic
combination of spectral information and chemometrics can be a powerful tool to perform a quality
control of wine vinegars and to verify the type of acetification carried out for its production. However,
future studies involving a larger number of samples are needed to validate the results and
conclusions presented here, in order to allow the implementation of this methodology in the
certification of these products.
1 Ríos-Reina, R., Azcarate, S.M., Camiña, J., Callejón, R.M. Food Chemistry, 2020, 323 , 126792..
2 Antonucci, F., Pallottino, F., Paglia, G., Palma, A., Aquino, S., & Menesatti, P. Food and Bioprocess Technology, 4(5),
2011, 809.
3 Azcarate, S. M., Ríos-Reina, R., Amigo, J.M.,& Goicoechea, H. C. (2021). TrAC Trends in Analytical Chemistry, 143,
116355.
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

465
SOIL ACIDITY STATUS FROM SEMI-ARID AND HUMID ENVIRONMENTS
USING EDXRF SPECTROSCOPY AND CHEMOMETRICS
Pedro G. C. Lucena (UG)a*, Ramon V. S. Aquino (PG)b, Juliet E. S. Sousa(R)c, Valdomiro S.
Souza Júnior (R)c, José G. A. Pacheco Filho (R)b, Claudete F. Pereira (R)a
a Universidade Federal de Pernambuco, Departamento de Química Fundamental, Recife, Pernambuco, Brazil, 50670-
901
b Universidade Federal de Pernambuco, Departamento de Engenharia Química, Recife, Pernambuco, Brazil, 50670-901
c Universidade Federal Rural de Pernambuco, Departamento de Agronomia, Recife, Pernambuco, Brazil, 52171-900
*e-mail: pedro.correialucena@ufpe.br
Soil acidification is a natural process, especially common in humid environments. This process leads
to a reduction of soil pH, base saturation percentage (BSP), and cation exchange capacity (CEC).
However, human activities can accelerate it, leading to productivity reduction and degradation of this
natural resource1,2. For large-size areas, obtaining information concerning the soil parameters can
be costly and time-consuming. Since the soil elemental composition may be affected by soil acidity,
spectroscopic techniques such as Energy-Dispersive X-ray Fluorescence (EDXRF) coupled with
chemometrics are valuable tools for soil acidity analysis. Nonetheless, the contributions focused on
the qualitative assessment of soil acidity are limited. Qualitative indicators are advantageous
because they are easier to interpret and do not require technical knowledge by non-professional
users to interpret the results3. Therefore, this study proposes a screening method for soil acidity
status using EDXRF spectral data and supervised pattern recognition analysis. For this, 176 soil
samples from humid and semi-arid tropical environments from the Pernambuco state (Northeast
Brazil) were analyzed by a benchtop EDXRF instrument (at 6.5 and 35 kV, under air atmosphere).
The chemometric analysis was carried out using the peak intensity of the elemental emission lines
employing R software. Classification models were developed using Partial Least Squares
Discriminant Analysis (PLS-DA) and Random Forest (RF) to classify samples into two classes
according to the following values: pH (< 5.0 or ≥ 5.0), CEC (< 15 cmolc kg-1 or ≥ 15 cmolc kg-1), and
BSP (< 50% or ≥ 50%). The samples were divided into training (70%) and test (30%) by the Naes
algorithm4. Cross-validation (k-fold = 5) was used for model validation. Twenty-seven emission lines
related to 22 elements (Mg, Al, Si, P, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Ga, As, Br, Rb, Sr, Zr,
and Nb) were employed to build the models. As shown in Table 1, RF models outperformed PLS-
DA ones. The results suggest that RF models can model the complex and non-linear relationships
of soil chemical composition and its acidity status. The variable importance analysis demonstrated
that Ca, Mn, As, and Zn were classification-relevant elements for pH. For CEC, Zn, Cu, Fe, and Zr
were the most important elements. For BSP, K, Ca, Ni, and Mn demonstrated higher importance.
Despite the importance of Mg and Al for chemical weathering and acidity, these elements were not
very important for the classifiers. Therefore, the results demonstrated the high accuracy (> 90%) of
the developed method to provide rapid qualitative assessment and monitoring of the acidity status
of soil in humid and semi-arid tropical environments.
Table 1. Classification accuracy of PLS-DA and RF models for pH, CEC, and BSP.
pH CEC BSP
Dataset
PLS-DA RF PLS-DA RF PLS-DA RF
Training 81.3% 100% 84.6% 100% 81.3% 100%
Test 77.4% 92.5% 86.8% 92.5% 83.0% 90.6%
1 Pincus, L. N.; Ryan, P. C.; Huertas, F. J.; Alvarado, G. E., Geoderma, Elsevier, 2017.
2 Ajiboye, G. A., Oyetunji, C. A., Mesele, S. A.; Talbot, J., Communications in Soil Science and Plant Analysis, Taylor &
Francis, 2019.
3 Lepak, N., Newingham, B. A., Kachergis, E., Toledo, D.; Moffitt, J., Rangelands. Elsevier, 2022.
4 Næs, T., Journal of Chemometrics, Willey, 1987.
[INTAA/CNPq/FAPESP, CAPES, NUQAAPE]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

466
SOME APPLICATIONS OF MULTIVARIATE CALIBRATION WITH MACHINE
LEARNING TECHNIQUES USING THE PYCARET LIBRARY
Fernando Schimidta*, Marcela H. Miyabaraa, Alexandre Pancottib, Mariângela F. Santiagoc

a Instituto Federal de Goiás, Mestrado prof. em Tecnologia, Gestão e Sustentabilidade, Goiânia -GO, Brazil, 74050-110
b Universidade Federal de Jataí, Central analítica, Jataí-GO, Brazil, 75801-615
c Universidade Federal de Goiás, Faculdade de Farmácia, Goiânia-GO, Brazil, 74605-170
*e-mail: schimidt99@gmail.com
In this work, we developed multivariate calibration models for three different sample sets (analysis
of caffeine in water, benzene in atmospheric air, and organic matter in soils) using an open-source
machine learning library with Python1 language. This library is easy to install and use, allowing the
construction and management of different models (continuous data regression and also discrete
data classification) with few lines of code and fast execution, not requiring advanced hardware. The
results of the multivariate calibration models obtained using the library are shown in Table 1 below
and consist of: (1) analysis of caffeine in water by pre-concentration of 56 samples in vegetable
fibers and determination by energy dispersive X-ray fluorescence spectroscopy2; (2) analysis of
organic matter in 108 soil samples by near-infrared spectroscopy3; (3) analysis of benzene in 820
atmospheric air samples collected using a multi-sensor gas device4. The last two were obtained from
public repositories and the first from a master science dissertation. All the three sets of samples
(data matrices) were pre-treated with Principal Component Analysis (PCA) and the scores obtained
were used as input data for the models. All spectra were previously mean centered and also treated
with second derivatives. The air sample data matrix were previously autoscaled. The PyCaret library
currently (version 2.3.10) has 25 regression models for multivariate calibration calculations. All the
installation and execution/calculations of the library were done on the Google Colab platform. The
selected training set is tested with all models in the library and classify them using k-fold cross-
validation method to evaluate the metrics. The output prints a score grid that shows for each
calibration model: mean absolute error (MAE), mean squared errors (MSE and RMSE), correlation
coefficient R2 (between real and predicted data), root mean square logarithm error (RMSLE), mean
absolute percentage error (MAPE) in subsets (fold), and training time (in seconds).
Table 1. Results obtained with three samples sets using PyCaret library.
Total Variance RMSE R2 RMSE R2
Samples Best model PCs
sam. total cal cal pred pred
Extra Trees
Caffeine/water 56 6 98.5% 8.108 0.6084 3.263 0.9670
Regressor
OM/soils Bayesian Ridge 108 10 89.7% 3.168 0.8502 3.437 0.8323
C6H6/air Bayesian Ridge 820 9 99.6% 0.563 0.9921 0.562 0.9935
The summarized results shown in table 1 above contain the best RMSE calibration and prediction
errors (test set) obtained by the best model found for each samples set used, selected from the
general model table of the library. The models allow us to optimize its fit parameters, store the optimal
settings obtained, and make predictions with other datasets.
1 https://pycaret.gitbook.io/docs/
2 Miyabara MH, Dissertação de mestrado, Mestrado profissional em Tecnologia de Processos Sustentáveis, Instituto
Federal de Goiás - IFG, Goiânia-GO, 2021.
3 Rinnan and Rinnan, Soil Biology & Biochemistry, 39, 2007, 1664.
4 De Vito S, Fattoruso G, Pardo M, Tortorella F, Di Francia G, IEEE Sensors Journal, 12(11), 2012, 3215.
[Google Inc, PyCaret, GitHub]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

467
SUPERVISED MACHINE LEARNING MULTI-PARAMETER PREDICTIVE
MODELING OF CRUDE OIL AND ITS FRACTIONS BY FT-ORBITRAP
Carlos A. Teixeira*, Amilton M. de Oliveira, Leandro W. Hantao
Institute of Chemistry, University of Campinas, Campinas, SP, Brazil, 13083-970
*e-mail: carlostx@unicamp.br
The oil and gas industry always presents challenges and issues in terms of data handling. It can
become even more critical when dealing with the analysis of petroleum distillates by high-resolution
mass spectrometry. This work tries to shed some light on the field of machine learning and artificial
intelligence to solve oil and gas industry problems. Namely, we worked on predicting American
Petroleum Institute (API) gravity, total sulfur and nitrogen, and basic nitrogen concentration.
Somehow all parameters are related to the oil quality, therefore, its commercial value. Roughly, oil
with API gravity of around 40 can be marketed at high values due to its longer carbon chains [1]. On
the other hand, sulfur, and nitrogen-containing components despite environmental concerns also
play an important role in poisoning catalysts in the refining process. In some cases, even traces of
these components can disrupt the entire process, thus their concentration can be used as
hydrotreating process effectiveness [1]. Therefore, gathering such parameters quickly and cleanly,
especially in crude oil, is not only technically interesting but important for the petrochemical industry.
Mass spectra were acquired in the Q Exactive FT-Orbitrap mass spectrometer (ThermoFisher
Scientific, Austin, TX, USA), and further modeled in MATLAB© Version 9.12 (2022a) with Parallel
Computing Toolbox Version 7.6. In short, a duplex algorithm for data splitting was applied to set
calibration and validation indices. Next, Artificial Neural Network models were built to predict the
contents of API gravity, total nitrogen and sulfur, and basic nitrogen by Bayesian Optimization and
assessed through the validation set mean squared error (MSE). To have the most representative
model the median MSE was used as a guideline for the final model, which was finally evaluated with
an external and independent test dataset. In general, the calibration models could predict all target
parameters with test predictions error between 0,03 – 8,49 % and overall linearities of 0.87 – 0.97.
Therefore, in conclusion, the proposed approach seems to be adequate to solve the given analytical
problem. Moreover, data compression plays an important role when Bayesian Optimization is used,
since the calibration-validation circle repetition can be computation-consuming when full MS spectra
are used.
1 Giles, H. N., and Mills, C. O., Crude Oils: Their Sampling, Analysis and Evaluation, MNL68; ASTM International: West
Conshohocken, PA, USA, 2010.
[Nova Analítica, ThermoFisher, CENPES/ Petrobras, CAPES, FAPESP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

468
SURFACE-ENHANCED INFRARED ABSORPTION SPECTROSCOPY (SEIRA)
WITH UNIVERSAL ATTENUATED TOTAL REFLECTANCE (UATR) ACCESSORY
Felipe C. S. Trindade (PG)*a, Izabel G. S. Sobrinha (PG)a, Giovannia A. L. Pereira (R)a,

Claudete F. Pereira (R)a
Universidade Federal de Pernambuco, Departamento de Química Fundamental, Recife, Pernambuco, Brazil, 50740560
*e-mail: felipe.cstrindade@ufpe.br
Infrared (IR) spectroscopy is a well-established technique for the unambiguous identification of

molecular species. Nonetheless, this technique is not sensitive enough to detect small quantities of
analytes. This drawback can be overcome by using surface enhancement strategy, which means
the employment of substrate surfaces, (irregular metals, semimetals, semiconductors, or polar
dielectric nanostructures) to enhance IR spectra (Surface-Enhanced Infrared Absorption
Spectroscopy - SEIRA). SEIRA is a promising technique enable more accurate studies involving
different types of substances and the monitoring of chemical and biochemical phenomena. In the
present work, silver selenide quantum dots (QDs), stabilized with mercaptopropionic acid
(Ag2Se/MPA), were used for amplifying the IR signature of methyl violet (MV) organic dye and
atrazine (ATZ) herbicide, which is used to control pests in corn and sugar cane plantations1. These
QDs are one-pot synthesized in an aqueous medium, thus reducing the production cost and
minimizing environmental damage. Infrared spectra were recorded using a FTIR Spectrum 400
spectrometer (Perkin Elmer) with a universal attenuated total reflectance (UATR) accessory
(diamond crystal), in the spectral region from 4000 to 650 cm−1, averaging 32 scans at a spectral
resolution of 4 cm-1. This UATR accessory has only one internal reflection, which becomes a
challenge for SEIRA studies because the signal intensification obtained by the technique is directly
proportional to the area of electromagnetic radiation interaction with the sample.2 The SEIRA spectra
was recorded using a deposited QDs layer on the ATR accessory, background. The SEIRA effect of
QDs was evaluated using experimental enhancement factors (EF), determined as the ratio of peak
height of SEIRA bands and corresponding band intensity acquired without QDs. Promising results
were obtained for the MV dye, with enhancements up to 4 times the original signal in conventional
IR spectroscopy, using Ag2Se/MPA with MV concentration of 75 µg/mL. Furthermore, for ATZ 150
µg/mL, enhancements of up to 15 times were obtained. Principal Component Analysis (PCA) was
also performed using IR spectra without and with Ag2Se QDs, as PCA is a suitable way to describe
the interpoint distance using as few dimensions as possible. For this, all spectra were preprocessed
via a baseline offset correction and mean centered. The scores plot carries the main information on
the enhancement effect promoted by Ag2Se QDs vs. the molecules. Thus, the enhancement effect
was determined by using the interpoint distances from each pair of samples (i.e., with and without
Ag2Se–MSA) along the PC1 and PC2 axes using the Euclidian distance. This strategy determines a
novel parameter we call the ‘Multivariate Enhancement Factor’ (MEF) 3. Results obtained in this work
were very promising even though only a single one-reflection ATR accessory was used, with results
comparable with literature. In addition, MEF is a more realistic way to quantify the enhancement
effect across the entire spectrum rather than at a single wavelength, which is more suitable for
optimizing the experimental conditions during surface-enhanced studies. It is also worth noting that
the MEF is considered a generic metric that can also be used to quantify the enhancement effect in
Surface-Enhancement Raman Spectroscopy (SERS).
1 Sun, H. Sun, C. Ding, X. Lu, H. Liu, M. Zhao, G. Journal of Hazardous Materials. v. 403. p. 123953. 2021.
2 Wang, H. You, E. Panneerselvam, R. Ding, S. Tian, Z. Light: Science & Applications. v. 10. p. 161. 2021.
3 PEREIRA, C. F.; Viegas, I. M. A.; Sobrinha, I. G. S.; Pereira, G.; Pereira, G. A. L.; Krebs, P.; Mizaikoff, B. Journal of
Materials Chemistry C. v. 8. p. 10448. 2020.
[CNPq, CAPES-PRINT/UFPE, CNPq/FAPESP/INCTAA, Serrapilheira Institute]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

469
USE OF DESIGN OF EXPERIMENTS (DOE) ON ASSESSMENT OF COMMERCIAL
INHIBITORS OF CALCIUM CARBONATE SCALE IN O&G PRODUCTION
Nayanna S. Passos (PG)a,*, Vinicius Kartnaller( R)a, Vinicius Gonçalves (R)a, João Cajaíba
(R)a
a Instituto de Química, Pólo de Xistoquímica, Universidade Federal do Rio de Janeiro (UFRJ), Brazil, 21941-614
*e-mail: nayannasouzap@gmail.com (correspondence author only)
Despite its vast application on soil improvement[1], self-healing, carbon dioxide capture, drug
delivery[2] and others, undesired calcium carbonate precipitation is a great concern on oil and gas
industry, since it is one of the most common oilfield scales[3], followed by barium, strontium, and
calcium sulfates. Scale can damage separators, heat exchangers[4], vessels, pumps, demanding
constant production shutdowns. To avoid this bottleneck, industry dedicates resources on research
aiming to mitigate or retard this process as much as possible. The development of chemical
compounds acting as inhibitors is the greatest cost-effective[5] preventive method of treating this
issue. However, to detect, predict and inhibit this process, it is crucial to understand the reasons
behind its occurrence. Scale (or inorganic fouling) is a physicochemical process caused by the
supersaturation, nucleation, crystal growth and accumulation/adherence of precipitates on pipelines
and surface facilities. Those ions are derived from the production water which is coproduced with
oil/gas, which changes its composition from well to well, turning the process of mitigation more
complex to be solved[6]. Therefore, when a chemical compound is synthesized, it must be tested by
compatibility and static methods but, even when approved, it should still go through a dynamic
method, the Tube Blocking Test (TBT). Several works have discussed about some variables
impacting scale formation such as temperature[7], scaling ions concentration, salinity[8], and other
variables, but as far as we know, no previous work have evaluated at a screening level the
importance hierarchy of those parameters when chemical inhibition occurs. The present study has
considered temperature, calcium, bicarbonate and inhibitor concentrations in the presence of salinity
in order to judge which is of greatest importance on scaling process, through a fractional factorial
design of experiments. The results led to the conclusion that temperature is the most impactful
variable on scaling process, determining whether an inhibitor may act on some specific scenario,
followed by bicarbonate, calcium concentration and salinity. The statistical significance from the
selected inhibitor (four compounds were studied) was a great indication that its thermal stability was
not convergent with the chosen temperature, proved by the Thermogravimetric Analysis (TGA). That
is an important indication that a compound is not suitable for a range of formation water without the
need of additional compatibility and static methods.
1 Miftah A., Tirkolaei H. K., Bilsel H. IOP Conf. Ser. Mater. Sci. Eng., vol. 800, 2020, p. 1.
2 Pérez H. F.; García, M. G. J. Appl. Res. Technol., vol. 18, 2020, p. 245.
3 Merdhah A. B.; Yassin, A. A. M. Mar. Sci. Technol. Semin. 2007.
4 Oliveira, M. C. K.; Gonçalvez, M. de A. L.; Marques, L. C. do C. Fundamentos da Garantia de Escoamento.
Editora Interciencia. 1a Edição: 2018. Brasil.
5 Mpelwa M.; Tang, S. F. Pet. Sci., vol. 16, 2019, p. 830.
6 Houston, S. J. Formation waters in petroleum reservoirs : their controls and applications. University of Leeds. 2007.
7 Dyer, S. J.; Graham, G. M. J. Pet. Sci. Eng., vol. 35, 2002, p. 95.
8 Coto B.; Martos, C.; Peña, J. L.; Rodríguez, R.; Pastor, G. Fluid Phase Equilib., vol. 324, 2012, p. 1.
[ANP, PRH, UFRJ]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

470
USE OF VIRTUAL SAMPLES TO DISCRIMINATE AMONG GASOLINES BASED
ON PHYSICOCHEMICAL PROPERTIES OR DISTILLATION CURVES
Luan C. Barbosa-Patrício (PG)a, Rafaella de F. Sales (R)b*, Neirivaldo C. da Silva (R)c,
Eduarda F. da Silva (UG)b, Lívia R. e Brito (R)d, M. Fernanda Pimentel (R)b
aFederal University of Pernambuco, Department of Fundamental Chemistry, Recife, Pernambuco, Brazil, 50740-521
bFederal University of Pernambuco, Department of Chemical Engineering, Recife, Pernambuco, Brazil, 50740-521
cInstitute of Exact and Natural Sciences, Federal University of Pará, 66075-110, Brazil.
dScientific Police of Pernambuco, Instituto de Criminalista Professor Armando Samico, Recife, Pernambuco, Brazil,
52031-080
*e-mail: rafaella.sales@ufpe.br
The police in the state of Pernambuco, Brazil, must deal with forensic cases of fuel theft and
adulteration that require identification of the origin of gasoline samples, to decide whether or not
samples seized at different locations came from the same fuel source. In this country, the National
Agency of Oil, Natural Gas and Biofuels (ANP) establishes specifications and quality control
requirements for commercial automotive gasoline. This includes specification limits for
physicochemical properties, such as anhydrous ethanol content, specific gravity and distillation
parameters. These physicochemical tests have been used in combination with multivariate pattern
recognition techniques to discriminate among gasoline samples for different purposes. For example,
to differentiate gasoline samples according to their origin, using principal component analysis (PCA),
hierarchical cluster analysis (HCA) and linear discriminant analysis (LDA)1, or to detect and quantify
different adulterants in gasoline, using partial least squares discriminant analysis (PLS-DA)2. To
address forensic problems requiring gasoline discrimination in pairs, this work evaluated the potential
of physicochemical properties and distillation curves to differentiate one individual sample from
another. To overcome the need for numerous repeated measurements, an approach to generate
virtual samples from real gasoline measurements was developed. Measurement uncertainties of the
methods were considered in this approach. The real and virtual samples were then evaluated by
pattern recognition techniques. Fifty-four gasoline samples were collected from 9 regions of the state
of Pernambuco. These were fractionated and destined for analysis by tests recommended by ANP,
obtaining physicochemical properties and distillation curves data sets. Considering the estimated
expanded uncertainties of each method, 49 virtual samples were randomly generated from real
gasoline measurements. The Euclidean distances for all 1,431 possible pairs of the 54 samples were
analyzed for physicochemical properties and distillation curves. The data sets were pre-processed
with autoscaling and pairs of samples compared. Each class contained the real sample and its
respective 49 virtual samples. PCA was applied for exploratory studies and soft independent
modelling of class analogy (SIMCA) was used for classification. Using SIMCA, samples were
considered to be completely distinguished when the false positive rate (FPR) assumed a null value.
For the physicochemical properties data set, about 96% of the pairs were differentiated. For the
distillation curves data set, this percentage increased to about 99%. To validate the strategy, a new
common gasoline sample was analysed in duplicate by the assays of this study, obtaining two data
sets of physicochemical properties and distillation curves and evaluated by the proposed approach.
For the physicochemical data set, a FPR of 66% was obtained for the SIMCA model (33 samples
from one class were assigned to the other class). For the distillation curves data set, a FPR of 54%
was obtained (27 samples from the test set were predicted as training class), suggesting that these
samples were the same. These results show that the proposed methodology could be useful for
investigations requiring this type of gasoline comparison as fast as possible. Moreover, the strategy
developed could also be extended to other applications that require discrimination of samples in
pairs.
1 Barbeira PJS, Pereira, RCC, Corgozinho, CNC, Energy & Fuels, 21, 2007, 2212
2 Mendes G, Barbeira PJS, Fuel, 112, 2013, 163.
[NUQAAPE, INCTAA, CNPq, CAPES, FACEPE]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

471
UV–VIS REFLECTANCE SPECTROELECTROCHEMISTRY AND MULTIWAY
CALIBRATION IN THE SIMULTANEOUS DETERMINATION OF
NEUROTRANSMITTERS
Levi G. Oliveira (PG), Matheus C. Barreto (PG), Wallace D. Fragoso (R), Sherlan G. Lemos (R)*
Grupo de Estudos Avançados em Química Analítica, Department of Chemistry, Federal University of Paraíba, João
Pessoa, PB, Brazil
*e-mail: sherlan@quimica.ufpb.br
Spectroelectrochemistry comprises a set of techniques that provide simultaneous electrochemical and

spectroscopic information about a chemical system. The most popular is based on UV-Vis absorption
spectroscopy, with the normal transmission configuration being the most used due to optically transparent
electrodes. On the other hand, the near-normal reflection configuration has gained attention because it
simplifies the experimental setup for routine analysis when associated with disposable electrodes. In
spectroelectrochemistry the spectra collected along a voltammetric scan are vectors, which grouped
generate a data matrix per experiment/sample called second-order data. The information provided by
second-order data, once adequately decomposed, can be uniquely ascribed to a single source of
variation (an analyte, for example). Such feature allows to exploit the second-order advantage, which is
the possibility of quantifying analytes in the presence of unknown and unmodeled interferents. In
consequence, calibration models obtained from simpler standards can be used directly on complex
samples. In this work the association between UV-Vis reflectance spectroelectrochemistry and second-
order calibration in the development of an analytical method was evaluated. As a case study the
simultaneous determination of dopamine and epinephrine, which present strongly overlapping peaks in
voltammetry (Fig. 2c and 2d). Fig. 1 shows the experimental setup used. Dropsens DRP-110 printed
electrodes were used, containing working and auxiliary carbon electrodes, and an Ag/AgCl reference
electrode. Spectra were acquired using a Starline AvaSpec 2048 CCD detector, an Avalight-DHS light
source, and an Avantes FCR-7UVIR200-2 probe. Voltammetric scans were performed by an Autolab
PGSTAT101 potentiostat. The devices were interconnected by Trigger TTL cable. The control and
synchronization of the measurements were performed using the Nova 2.1 software. The first spectrum
was used as a reference. Since spectroelectrochemical data are trilinear in nature1, the calibration models
were built using Parallel Factor Analysis (PARAFAC). In Fig. 2 the spectroelectrochemical and
voltammetric signals for 2 mM dopamine and epinephrine standards are illustrated. Two spectral bands
are observed around 307 nm and 470 nm for dopamine, and around 304 nm and 495 nm for epinephrine.
Figures 3a and 3b show the decomposed spectral and voltabsorptometric profiles, respectively
(Corcondia = 100%). The recovered spectra refer to the oxidation products of the analytes, while the
obtained voltabsorptometric profiles indicate an increase in the concentration of oxidation products
throughout the scan, starting at 0.2 V. Four samples containing dopamine and epinephrine at
concentrations ranging from 2 to 5 mM were analyzed, obtaining RMSEPs of 0.04 mM and 0.05 mM for
dopamine and epinephrine, respectively. The predictions were satisfactory considering the inherent
challenges of the system under study – strong mutual interference of the analytes in both electrochemical
and spectral modes.
Fig 1. Scheme of the experimental setup Fig 2. Spectroelectrochemical signals of (a) Fig 3. (a) Spectral and (b)
employed. 1. light source; 2. spectrometer; 3. dopamine and (b) epinephrine. Voltammograms of voltabsorptometric profiles obtained
potentiostat; 4. spectroelectrochemical cell. (c) dopamine and (d) epinephrine with PARAFAC.
1 Garoz-Ruiz J, Perales-Rondon JV, Heras A, Colina A. Electroanalysis 31, 2019, 1254.

[CNPq, CAPES, FINEP]
––––––––––––––––––––– Chemometrics –––––––––––––––––––––

472
MISCELLANEOUS
473
CHEMICAL CHARACTERIZATION AND QUANTIFICATION OF TITANIUM
DIOXIDE NANOPARTICLE (TiO2) IN SEAFOOD BY SINGLE-PARTICLE-ICP-MS
Lisia M. G. dos Santos (R)a*, Cristiane B. Silva (R)a, Santos A. V. Neto (R)a, Renata A. Pereira
(PG)a, Silvana C. Jacob (R)a, Josino C. Moreira (R)a
a Departamento de Química, Instituto Nacional de Controle de qualidade em Saúde (INCQS/FIOCRUZ), Rio de Janeiro,
RJ, Brasil, CEP 21040-900
*e-mail: Lisia.gobbo@incqs.fiocruz.br
Titanium dioxide (TiO2) nanoparticles are commonly used in sunscreens as part of the formulation
to block the sun’s harmful UV rays1. As the use of nanoparticles in consumer products has increased,
concern has risen as to the health and environmental effects of nanoparticles, especially into the
marine environment2. The aim of the present study was to apply the validated methodology single
particle inductively couple plasma mass spectrometry (spICP-MS) (Nexion 300D, Perkin Elmer)
according to the parameters as described in ISO 17025, because this technique ability to provide
information on nanoparticle size, size distribution, nanoparticle number-based concentration, as well
as the dissolved titanium in the sample. About 0.5 g of the sample was weighed in triplicate and each
triplicate analysed five times and placed in a polypropylene Falcon tube with 15 mL water. The
samples were then submitted to ultrasound treatment for 2 h, centrifugation at 5,000 rpm for 30 min
and filtration through 0.22 µm pore membranes. Furthermore, to assess the sonification time on
TiO2NP standards were added before this stage and no significant losses were observed as shown
by the recovery study (78%). After filtration, each sample were sonicated and vortexed for 1 min. A
blank solution containing only ultrapure water, with no added sample, was subjected to the same
extraction process4. The quantification of dissolved Ti (1.73 ± 0.37 mg kg-1) did not differ significantly
from de total Ti (1.74 ± 0.04 mg kg-1), this sample was digested with acid by microwave. The results
by spICP-MS were the most frequent size was 93 nm, mean diameter 148 nm, particle concentration
(3.5 x 107 Ps-Tot/g) and the most of the particles identified had a size greater than 100 nm (61%)
followed by particles in the range 81-100 nm (31% Ps-total) and 61-80 nm (7%). This work has
demonstrated the ability to measure TiO2 extracted from seafood, and that a nano-sized fraction is
present this samples. Moreover, this study contributes to strengthen the literature on the application
of nanotechnology in products of sanitary interest, as well as their possible environmental and (eco)
toxicological impacts.
1 Garcia EB, Machado TSC, Ferraris FK, Amendoeira FC, Revista Analytic,13, 2015, 45.
2 Mackevica A, Olsson ME, Hansen SF, Journal of Nanoparticle Research, 20, 2018, 1.
3 International Standard Organization; General Requirements for the Competence of Testing and Calibration Laboratories,
ISO/IEC 17025, 2017.
4 Santos, LMG, Barata-Silva, C, Neto SAV, Fonseca MA, Magalhães CD, Moreira JC, Jacob SC, Química Nova, 44, 2021,
760.
[FAPERJ, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

474
IDENTIFICATION AND QUANTIFICATION OF SILVER NANOPARTICLES IN
TEXTILE SUBSTRATES BY SINGLE PARTICLE ICP-MS
Renata A. Pereira (PG)a*, Santos A. V. Neto (R)a, Cristiane Barata-Silva (R)a, Fabio S. Bazilio
(R)a, Silvana C. Jacob (R)a, Josino C. Moreira (R)a, Lisia M. G. dos Santos (R)a
a Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (Fiocruz), Departamento de
Química/ Setor de Elementos Inorgânicos, Rio de Janeiro, Rio de Janeiro, Brasil, CEP: 21.040-900
*e-mail: renattaali@gmail.com
The COVID-19 pandemic and the most recent global emergency of monkeypox have made even
more evident the need for new technologies to be applied quickly, effectively and cost-effectively to
contain these diseases. In this scenario, the application of nanotechnology in textile fibers has the
potential to become a resource for combating these and other pandemics 1. In this field, fabrics
impregnated with silver nanoparticles (AgNP) must be mentioned, because they demonstrated
antimicrobial action against several viruses and bacteria, additionally, the literature also points out
the improbability that microorganisms develop resistance to these nanoparticles2. Currently, AgNPs
are already being added to hospital clothes and fabrics. However, the form and concentration of
AgNPs in these products are essential to guarantee their antimicrobial properties3. Thus, the
objective of this work was to optimize an analytical methodology using the “single particle” ICP-MS
technique4 to verify the compliance of AgNP in two textile substrates, one made with natural fiber
and other made with synthetic fiber, both impregnated with an AgNP solution through the pad-dry-
cure methodology. The working range varied from 1.5 – 20 µg L-1, the size detection limit (LODT),
dissolved silver detection limit (LODi) and particle concentration detection limit were respectively
17.5 nm, 0.74 µg L-1 and 3.1X10-4 particles mL-1. Thus, the results obtained suggests the possibility
to use this technique to determine the size and concentration of silver nanoparticles, in addition, it
was observed that the natural textile substrates have a better bound with AgNP than synthetic textile
substrate. Thus, research in the area of health surveillance plays an important role in responding to
health emergencies, especially those applicable in new products that can be used to combat
diseases and prevent future risks.
1 Iumah CI. The Journal of the Textile Institute, 112, 2021, 2056.
2 Galdiero S, Falanga A, Vitiello M, Cantisani M, Marra V, Galdiero M. Silver nanoparticles as potential antiviral agents.
Molecules, 16, 2011, 8894.
3 Barata-Silva C, Vicentini-Neto SA, Magalhães CD, Jacob SC, Moreira JC, Santos LMG. Avaliação da qualidade das
máscaras comercializadas no Brasil em tempos de pandemia da COVID-19 quanto à presença de prata e de
nanopartículas de prata. Vigilância Sanitária em Debate: Sociedade, Ciência & Tecnologia, 9, 2021, 29.
4 Bazilio FS, Silva CB, Santos LMG, Neto SAV, Jacob SC, Abrantes SMP, Química Nova, 44, 2021, 868.
[FAPERJ, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

475
SYNTHETIC PEPTIDES-BASED SPR BIOSENSORS TOWARDS VISCERAL
LEISHMANIASES DIAGNOSIS
Jaqueline Volpe (PG)a, Adriano S. Silva (PG)a, Floriatan S. Costa (PG)a, Angélica R. Faria ®b,
Hélida M. Andrade ®b, Dênio E. P. Souto ®a*
a Laboratório de Espectrometria, sensores e biossensores, Federal University of Paraná, Curitba, Paraná, Brazil,
81530-000
b Biological Sciences Institute, Federal University of Minas Gerais, Curitba, Paraná, Brazil, 81530-000
*e-mail: denio.souto@ufpr.br
Peptides are versatile molecules that can be explored in biosensors construction, being promising
due to their great stability, standard synthetic protocol, cost-effectiveness, and tunable physio-
chemical properties, all of which make it possible to synthetically produce them to substitute proteins
or crude antigens as bioreceptors in antibody screening.1 In turn, considering the transducing
element in biosensing, devices based on surface plasmon resonance (SPR) are powerful techniques
for biomolecular interaction investigations, allowing analysis at real-time, and without the necessity
of chemical or biological probes. SPR is an optical system that explores the excitation of surface
plasmons on metallic substrates, which allows highly sensitive analysis according to refractive index
changes in the close medium. In this context, peptides-based SPR biosensors play an important role
in biosensor development, associating the easy quantity fabrication and better stability of the artificial
peptides with the ability of a real-time, sensitive, and label-free analysis of SPR.2 In terms of
biosensor construction, other important factor to be considered is the biofunctionalization step, which
can intrinsically affect the biosensor’s performance, since the environment of the biocomponent and
the condition of its immobilization directly affects the capacity of interaction of the receptor with the
analyte. For this reason, covalently attaching the bioreceptor on the sensor surface by forming a
self-assembled monolayer (SAM) is one of the most used methods, allowing good control of the
receptor orientation, high surface homogeneity, and satisfactory stability.3 In this work, two synthetic
antigenic peptides were applied for the first time in SPR-based biosensors for serological diagnosis
of Visceral Leishmaniases (VL). VL is an endemic neglected disease that affects humans and dogs
around the world, affecting more than 200,000 people each year.4 Consequently, diagnostic methods
are essential to obtain better infection control. The sensors construction proposed in this study
consisted of a previous SPR gold disk functionalization, utilizing a mixture of mercaptoundecanoic
and mercaptopropionic acid, forming a mixed SAM to covalent binding of two different peptides
(PEP1 and PEP2) or the blend of them (PEP1:2). This biofunctionalization step (peptide
immobilization) was characterized at real-time via SPR. The biosensors developed were also
electrochemically characterized by cyclic voltammetry (CV) and electrochemical impedance
spectroscopy (EIS), in which due to interfacial shifts caused by each of the steps, changes in the
conductivity, resistivity, and capacitance were observed. The three bioreceptor conformations
(PEP1, PEP2 and PEP1:2) were evaluated accordingly to sensibility and stability, for the detection
of specific antibodies in rabbit serums. Additionally, the ability of detecting positive and negative
cases of VL in dog serums was evaluated for each of the arrangements. Summarily, this study
explored new and promising biostructures, contributing to a better understanding of their applicability
via SPR biosensors, consisting of an important screening tool in an urgent matter such as VL.
1 Liu Q, Wang J, Boyd BJ, Talanta, 136, 2015, 114.
2 Karimzadeh A, Hasanzadeh M, Shadjou N, de la Guardia M, TrAC-Trends in Analytical Chemistry, 107, 2018, 1.
3 Souto DEP, Volpe J, Gonçalves CC, Ramos CHI, Kubota LT, Talanta, 205, 2019, 120122.
4 Cruz CSS, Barbosa DS, Oliveira VC, Cardoso DT, Guimarães NS, Carneiro M, Parasitology, 148, 2021, 639.
[CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

476
PRINTING OF LUMINESCENT MOFS CONTAINING LANTHANIDE IONS
PROMISING IN THE ENCODING OF OFFICIAL DOCUMENTS AND PAPER
MONEY
Jakson Ney da Costa Reis (R)a, Érico Moura Neto (R)a, Jarley Fagner Silva do Nascimento
(R)a
a Instituto Federal de Educação, Ciência e Tecnologia do Rio Grande do Norte
*e-mail: jakson.reis@ifrn.edu.br
In this work, the process of synthesizing metal-organic networks containing lanthanide ions (LOFs),
accelerated via a precipitating agent, capable of accelerating the MOF crystal nucleation process, to
explore the optical properties of this precursor solution (SP_Gd/ EU-MOF), as a photoluminescent
print in the encoding of official documents and paper money. An alternative to materials already
reported in the literature was demonstrated. SP_Gd/Eu-MOF was deposited on paper substrates
using inkjet printers. SP_Gd/Eu-MOF was characterized by photoluminescence spectroscopy before
and after its deposition on substrates, and the spectra showed very similar profiles, showing that the
interaction with the substrate surface did not promote significant structural defects. The material,
after being deposited, was excited by UV radiation (λ = 254 nm), and emitted the red color
characteristic of the Eu3+ ion. The luminescent ink showed good thermal stability in the heating test
of the paper substrate after being deposited, since it maintained red radiation emission even after
being heated to 200 °C, demonstrating its potential to act as a proof of authenticity in the document
encoding and paper money, revealing itself as another very low-cost alternative for the deposition of
photoluminescent inks.
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

477
NON-CHROMATOGRAPHIC ARSENIC SPECIATION IN RICE AND SHRIMP
SAMPLES USING A NANOPROBE BASED ON Ag NANOPARTICLES
Francisco L. F. Silva (PG)a,b, Francisco E. H. Viana (PG)a, Pierre B. A. Fechine (R)c, Abdullah
Akhdhar (R)d, Gisele S. Lopes (R)a, Wladiana O. Matos (R)a*
a FederalUniversity of Ceará, Laboratory of Analytical Chemistry Applied, Analytical and Physical-Chemistry
Department, Fortaleza, Ceara, Brazil, CEP
b State University of Ceará, Education Faculty of Crateus, Crateus, Ceara, Brazil, CEP
c Federal University of Ceará, Group of Material Chemistry, Analytical and Physical-Chemistry Department, Fortaleza,
Ceara, Brazil, CEP

d University of Jeddah, Department of Chemistry, Jeddah, Saudi Arabia.
*e-mail: wladianamatos@ufc.br
The toxicity of arsenic (As) depends on its chemical form, being inorganic arsenic (iAs) species
considered the most toxic.1 Rice and seafood samples are the main sources of As for humans
through diet2. Thus, it is important to monitor iAs in these foods. In general, iAs analysis is performed
by HG-AAS or LC-ICP-MS. However, studies have been carried out to develop simpler, faster, low-
cost, and green analytical methods to quantify iAs. In this sense, nanoprobes that change their
optical characteristics when interacting with an iAs3 can be employed to develop analytical methods
that meet all or some of these items. In the present work, silver nanoparticles (AgNPs) functionalized
with polyethylene glycol (PEG200) and sodium citrate3 were studied for further application in the
analysis of iAs in seafood and rice samples by UV-Vis spectrophotometry. For the synthesis of
AgNPs, 50 mL of 50 mmol L-1 AgNO3 solution was mixed with 5mL of PEG and then this mixture was
heated in a hot plate until boiling. After this, 4 mL of 1% w w-1 citrate was added, and the formation
of a yellow solution indicated the AgNPs synthesis. The spectrophotometric measurements of the
AgNPs colloidal solution shows a plasmonic resonance (SPR) band at 412 nm that is reduced when
the AgNPs interact with iAs, while a band at 600 nm increases. This means that iAs provoke a
dislocation of the SPR band. The characterization of the nanomaterial using Zeta potential and
Dynamic Light Scattering suggests that the nanoparticles have a negatively charged surface and
their hydrated size varies between 80-100 nm. A linear relation between iAs concentration and the
ratio of absorbance at 412 and 600 nm was used to quantify iAs in rice and shrimp samples. The iAs
was extracted from 500 mg of samples using 10 mL 2% w v-1 HNO3 under heating in a block digester
at 90°C for 1h. The extract was filtered and diluted to 20 mL. For iAs analysis, 1 mL of the extract
diluted was added to 2 mL of AgNPs solution and the iAs detection was performed by UV-Vis
spectrophotometry. For the shrimp sample, the extract was diluted 10-fold before spectrophotometric
measurements due to the intense coloring of the solution. A NIST 1568b – Rice flour was used to
evaluate the accuracy of the method for rice and a spike/ recovery test was applied for rice and
shrimp samples. The SRM analyses presented a recovery of 99% for iAs and spike/recovery tests
the recovery was between 98-106%. The limits of detection and quantification of the method were
0.66 and 1.98 ng g-1 iAs, respectively. The iAs content in rice samples was between 55.3 ± 0.4 and
110.0 ± 17.5 ng g-1 (4 samples), and 64.7 ± 0.2 ng g-1 for the shrimp sample. The presence of cations
(Al, Cd, Cr, Fe, Mn, Se, and Zn) and phosphate in the sample did not interfere with the As signal.
The proposed method was effective for iAs speciation in vegetable and animal tissue matrices (rice
and shrimp). It can be a low-cost alternative to As speciation analysis compared with HPLC-ICP-MS
and selective hydride generation, besides that, it is a simple and greener methodology. Future works
are necessary to comprehend the mechanism of the interaction AgNPs-As and apply the
methodology for a larger number of samples.
1 Bissen M, Fimmel F, Acta Hydrochimica Hydrobiology, 31, 2003, 9.
2 Silva FLF, Andrade-Neto DM, Menezes FL, As IP, Higuera JM, Fechine PBA, Costa ARA, Lopes GS, Matos, WO, Food
Chemistry, 345, 2021, 128781
3 Boruah BS, Daimari NK, Biswas R, Results Physics, 12, 2019, 2061.
[CAPES, CNPq, UFC, INCTAA]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

478
QUANTIFICATION OF ALBUMIN IN COWPEA USING DIGITAL IMAGING
Michely da S. Sousa (PG)a*, Ronaldo C. Coelho (R)b, Everson T. S. G. da Silva (R)a, Cícero A.
Lopes Júnior (R)a
a Federal University of Piauí, Departament of Chemistry, Teresina, Piauí, Brazil, Zip Code 64049-550
b Federal Institute of Piauí, Department of Chemistry, Teresina, Piauí, Brazil, Zip Code 64000-040
*e-mail: michelysousacx@gmail.com.br
Proteins play extremely important roles in most biological processes, e.g. acting as enzymes1. The
development of methodologies for protein determination has become of fundamental relevance in
several areas of knowledge1. Digital image colorimetry (DIC) has been applied in the determination
of various analytes such as iron, ascorbic acid, potassium2. To our best knowledge, protein
determination in bean samples has not been performed using DIC. Therefore, this study aims to
develop a simple, safe and low cost method for protein determination using DIC. For comparison
purposes, the Bradford method (1976)3 was used. The biofortified cowpea samples were analyzed
by a mobile device, where image captures were made with stabilized lighting conditions to obtain
and acquire high quality images (see Fig. 1). The images were analyzed by Chemostat software for
data processing and obtaining results through the relationships between RGB channels and
concentrations. Using digital imaging data a concentration of 7.63 mg/g albumin was found, while
using UV-Vis the concentration was 7.02 mg/g. Thus, obtaining a relative error of 8.7%, indicating
good agreement with the measurements. The use of cell phones as analytical devices is a current
trend, as they combine fast response and simplicity of analysis with the possibility of real-time
communication with computers or other devices to obtain information in situ. The application of digital
images in food analysis is very promising, opening up the possibility of automation.
Figure 1. General methodology for digital image colorimetry on a smartphone.
1 Zaia DAM, ZAIA, CTBV, LICHTIG, J, Química Nova, 21, 1998, 787.
2 Yanjun F, Jiawei L, Yaping G, Lianwu X, Gui Z, Measurement, 171, 2021.
3 Fennema OR, Principles of Food Science; Marcel Dekker Inc, New York, 1976
[CAPES, FAPEMA, UFPI]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

479
EXPLORING BIOPHYSICAL STUDIES TO EVALUATE THE INTERACTION
BETWEEN AMINO ACID DERIVATIVES WITH THIOUREA NUCLEUS AND
UREASE: APPLICATION IN SOIL SAMPLES
Thamilla M.S. Maciel (UN)a,*, Angelo de Fátima (R)b, Josué C. C. Santos (R)a
a Instituto de Química e Biotecnologia, UFAL, Maceió, Alagoas, Brazil.
bDepartamento de Química, UFMG, Belo Horizonte, MG, Brazil.
*e-mail: thamillamaciel@hotmail.com
Amino acids are important biomolecules that have improved several properties in the development of
prodrugs, such as increased bioavailability and decreased toxicity of the original drug.1 Furthermore, the
association of amino acid derivatives with enzyme inhibitors has already been reported, including urease,
with promising results.2 Thiourea, in turn, is already well known as a classic urease inhibitor; therefore,
its derivatives are reported as potential inhibitors of this enzyme.3,4 This work aimed to evaluate the
inhibitory capacity of amino acid derivatives (Fig. 1), which have thiourea nucleus in their structures,
exploring biophysical studies to obtain potent inhibitors with low toxicity and potential agricultural
application.
Fig. 1. Structures of amino acid derivatives. Initially, the values of the inhibitory concentration
(IC50) in the presence and absence of the humic
substance were determined using the reference
method (Berthelot reaction) to evaluate the
potential of these compounds to inhibit the
activity of Jack bean urease (model), as well as
the influence of organic matter (OM).5 The IC50
values varied from 0.30 - 0.75 µM, and the
absence of effect associated with the OM was
observed.
In the interaction process evaluation, a decrease in the urease (2 µM) intrinsic fluorescence was observed
using excesses of amino acid derivatives, characterizing the occurrence of quenching (110-L, 110-D, 110
ac-L, 110 ac-D, 116: 10 to 250 μM, and 106 ac-L, 106 ac-D: 10 – 30 μM). The preferential quenching
mechanism was evaluated based on bimolecular diffusion rate constant (kq) values which ranged from
0.25 to 6.94×1012 M-1 s-1. Since kq > 2.0×1010 M-1 s-1, it was possible to infer that the quenching
mechanism is preferentially static with complex supramolecular formation.6 The binding constant values
(Kb) were obtained at different temperatures for each derivative and ranged from 102 to 106 M-1. In
general, the results suggest no influence of enantioselectivity in the interaction process since the Kb
values for each enantiomer pair were analyzed by the ANOVA (one-way) test. The thermodynamic
parameters indicated a spontaneous process, with preferential (non-exclusive) interactions such as van
der Waals forces, hydrogen bonds, and hydrophobic interactions. The 3D fluorescence and UV-vis
studies indicated changes in the secondary structure concerning urease due to the process of interaction
with ligands. The synchronized fluorescence study revealed that derivatives 110ac L-D and 116 bind to
the microregion close to the Tir residue (active site) and interact close to the enzyme's active site. These
studies corroborate the competition assay, in which amino acid derivatives competed for the active site
in the presence of classical inhibitors (hydroxyurea, thiourea, acetohydroxamic acid, and NBPT). Studies
for different soil samples (n = 4) showed that all compounds were more potent when compared to the
NBPT. Thus, it is possible to conclude that the association between amino acid derivatives and thiourea
is a strategy that can produce potent urease inhibitors with wide application, main to agriculture
application in fertilizers.
1 Vale N, Ferreira A, Matos J, Fresco P, Gouveia MJ, Molecules, 23, 2018, 2318.
2 Kobashi K, Takebe S, Terashima N, Hase J, Journal of Biochemistry, 77, 1975, 837.
3 Bano B, Kanwal, Khan KM, Lodhi A, Salar A, Begum F, Ali M, Taha M, Perveen S. Bioorganic Chemistry, 80, 2018, 129.
4 Li W, Xiao Q, Hu C, Liu B, Sun R, Geoderma, 354, 2019, 113877.
5 Tavares, MC, Oliveira KA, Fatima A, Coltro WKT, Santos JCC, Talanta, 230, 2021,122301.
6 Shu Y, Xue W, Xu X, Jia Z, Yao X, Liu S, Liu L, Food Chemistry, 173, 2015, 31.
[CNPq, FAPEAL, CAPES, UFAL, PPGQB, IQB]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

480
FIT-FOR-PURPOSE MIP OES METHOD TO MEET THE REQUIREMENTS OF
THE BRAZILIAN REGULATIONS FOR K AND Na IN COCONUT WATER, AND
NUTRITIONAL ASSESSMENT
Larissa O. Menezes (UG)a, Rodolfo Lorençatto (PG)b, Fábio V. Durão (UG)a, Mariana T. Silva
(UG)a, Rafaella R. A. Peixoto (R)a, Christiane Duyck (R)a, Anderson A. Rocha (R)a,c*
a UniversidadeFederal Fluminense, Departamento de Química Analítica, Niterói, Rio de Janeiro, Brasil, 24020-141
b Agilent Technologies Brasil, Barueri, São Paulo, Brasil, 06455-000
c Núcleo de Estudos em Biomassa e Gerenciamento de Águas (NAB / UFF, Niterói, Rio de Janeiro, Brasil, 24210-330
*e-mail: araujo20002000@yahoo.com.br
The Brazilian regulations for processed coconut water establish the range concentrations of Na
and K, being necessary high-throughput analytical methods, as microwave-induced plasma
optical emission spectrometry (MIP OES) for quality control. Coconut water is the liquid part of
the coconut (Cocos nucifera L.) endosperm, found in the hermetic inner cavity of the fruit. The
regular consumption of this natural drink can benefit the health, considering the high levels of
essential elements, vitamins, amino acids, antioxidants, and its low calorie and fat content. In
2020, the Ministry of Agriculture, Livestock, and Supply established a new classification and
quality standards for coconut water, and their respective analytical parameters, with the
publication of the normative instruction nº9 of January 30.1 The aim of this work is to evaluate
the suitability of MIP OES to be applied to Na and K determination in natural and processed
coconut water to verify if their contents are in accordance with the Brazilian regulations. With
the application of the method to analyze samples marketed in Brazil, a nutritious and risk
assessment was conducted considering the Recommended Daily Intake (RDI) of K and the
Maximum Recommended Intake (MRI) of Na. Samples of natural (green and mature) and
processed coconut water were acquired at the local markets of Niteroi (RJ, Brazil). In total, 21
coconut water samples were analyzed, 6 natural, and 15 processed. The samples were treated
by acid decomposition assisted by microwave radiation before their MIP OES analysis.
Recoveries for Na and K were 103 % and 95 %, whereas the RSDs were 7.6 % and 1.3 %,
respectively. The LOD was 1.77 mg kg-1 for K and 2.31 mg kg-1 for Na. Sodium and K were
determined in the ranges of, respectively, 27-490 mg L-1 and 1,600-3,500 mg L-1 in natural
samples, whereas in processed samples the ranges were 26-168 mg L-1 (Figure 1) and 814-
2,054 mg L-1. All the processed samples were in accordance with the established regulation,
except one dehydrated sample for which K content was below the accepted range. Natural
coconut water is an excellent source of K in the diet as its daily consumption can supply more
than 20 % of the RDI of K for adults and 30 % for children.
Figure 1. Comparison of the determined Na contents with the label’s declared contents, and the normative Na range
according to Brazilian regulations.
1 Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Normative instruction nº9, de janeiro de 2020, dispõe sobre
padrões de identidade e qualidade para a água de coco, Brazil.
[NAB-UFF]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

481
INTEGRATION OF PAPER-BASED ANALYTICAL DEVICES WITH DIGITAL
MICROFLUIDICS FOR COLORIMETRIC DETECTION OF CREATININE
Larissa G. Velasco (PG)a*, Danielly S. Rocha (PG)a, Wendell K. T. Coltro (R)a
a Universidade Federal de Goiás, Instituto de Química, Goiânia, Goiás, Brazil, 74690-900
*e-mail: larissa.garcia.velasco@discente.ufg.br
Digital microfluidics (DMF) is a platform for manipulating nano- to microliter-scale droplets in an array
of insulated electrodes. This technology enables fluidic operations of multiple droplets in parallel,
which is one of the main advantages of the automation of operation such as: dispensing from
reservoirs, merging, mixing, splitting, and directed movement. In addition, the DMF is applicable in
different areas due to the reconfigurable arrangement of its devices, which enables coupling with
different analytical detection methods1. These include digital image colorimetry (DIC), whose
fundamental principle is the analysis of the pixel intensity of images obtained after colorimetric tests
using specific software2. However, the images obtained can be influenced by lighting conditions, and
maintaining a homogeneous distribution of light emission can be difficult. Therefore, this study
proposes the use of a paper-based analytical device (PAD) coupled with a DMF chip to make
possible the image capture and analysis through a desktop scanner due to its efficient light control.
As a proof-of-concept, the detection of creatinine (CR), a useful biomarker for the assessment of
renal function as it is the metabolite waste product of muscle contraction excreted by the
kidneysperformed3, was explored as model. Colorimetric detection of CR was performed using the
Jaffé reaction, which consists of the reaction of alkaline picrate with creatinine to form an orange-
red product4. First, a polyester plate containing 96 square zones distributed in 8 rows and 12 columns
and a scanner were used for the optimization of picric acid (AP) and NaOH concentrations. The
images captured using the scanner were analyzed with Corel Photo-Paint software in the CMYK
color system exploiting the yellow channel, since it has a higher signal intensity when compared to
the RGB color channel. The range of AP concentration analyzed was from 0.5 to 10 mmol L-1,
keeping the NaOH concentration fixed at 250 mmol L-1, and the best result was achieved using 5
mmol L-1. The NaOH concentration was optimized raining from 50 to 450 mmol L-1, and the highest
color intensity was observed at 300 mmol L-1. These experiments were performed using CR solutions
prepared at concentrations of 8 and 32 mg dL-1. For sample handling using DMF, the best results
were achieved using the surfactant Tetronic 90R4 at 0.1% (v/v). Under the optimized conditions, the
integrated system involving DMF and PAD was tested for detecting CR. Preliminary results revealed
a linear behavior in the concentration range between 2 and 32 mg dL-1. Future efforts will be devoted
to the complete automation involving the whole analytical procedure, i.e., from sample preparation
to detection.
Figure 1. Experimental process for creatinine detection (A) DMF chip, where the automated operations for colorimetric
reaction take place, coupled to the PAD to collect the reddish-orange product (B) scan of the DMF chip coupled with the
PADs (C) Analysis of images in the CMYK color system in the yellow channel (D) The intensity of CR at different
concentrations.
1 Campos RPS, Rackus DG, Shih R, Zhao C, Liu X, Wheeler AR, Analytical Chemistry, 91, 2019, 2506.
2 Fan Y, Li J, Guo Y, Xie L, Zhang G, Measurement, 171, 2021, 108829.
3 Mathaweesansurn A, Thongrod S, Khongkaew P, Phechkrajang CM, Wilairat P, Choengchan N, Talanta, 210, 2020,
120675.
4 Rossini EL, Milani MI, Carillho E, Pezza L, Pezza HE, Analytical Chimica Acta, 997, 2018, 16.
[CAPES, UFG, GME, INCTBio]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

482
DEVELOPMENT OF A COLORIMETRIC METHOD FOR DETERMINATION OF
ACID PHOSPHATASE AIMING FOR FORENSIC APPLICATIONS
Beatriz do Nascimento Oliveira (PG)*, Josué Carinhanha C. Santos (R)
Institute of Chemistry and Biotechnology, Federal University of Alagoas, Maceió - AL, Brazil, 57072-900
*e-mail: beatriz.oliveira@iqb.ufal.br
Sexual crimes are a form of violence with the highest incidence against women. In this way,
identifying biological fluids such as blood, saliva, and semen constitutes one of the crucial steps in
the analysis of crime scenes or samples for forensics. In this sense, the presence of seminal fluid
(semen) on the victim or clothes, for example, may constitute criminal evidence. Semen has a
complex composition; however, the acid phosphatase (AP) enzyme can be used as a marker since
it has concentrations up to 400 times higher in the seminal fluid compared to other body fluids.1 The
objective is to develop a methodology with detection colorimetric test that uses a small amount of
sample and is capable of being used in the field, with a quick and selective response, thus helping
to confirm criminal evidence. The proposed method is based on the detection of AP using ascorbic
acid 2-phosphate (AAP) as a substrate, which, after enzyme action, releases inorganic phosphate
and reduced ascorbic acid (AAr) at pH 5, which is used to detect the enzyme in the medium. In this
case, AAr reacts with Fe(III) ions leading to the formation of Fe(II) and oxidized ascorbic acid (AAo)
(Fig. 1A). Therefore, the reaction of Fe(II) ions generated in situ with K3[Fe(CN)6] in acidic medium
leading to the formation of Turnbull's blue (TB, Fe3[Fe(CN)6]2), a blue colloid dark ( = 710 nm).
Using TB as a colorimetric probe with max > 500 nm avoids possible spectral interferences due to
the composition of complex samples.
Figure 1. (A) General reaction scheme for TB formation. Application of the method for qualitative evaluation in (B) solution,
where (i) reference and diluted semen at the proportion (ii) 1:10 and (iii) 1:2; (C) cloth (cotton) with traces of human semen
(50 L), where: (1) contaminated cloth (before the reaction), (2) non-contaminated cloth; (3) diluted sample (1:5) and (4)
undiluted sample (direct application in the cloth surface). The colorimetric reagents were applied only in the cloth systems
(2), (3), and (4).
Initially, the main reaction parameters were optimized using AAr (reaction product, Fig. 1A). Under
the best analytical performance conditions, Fe(III) 0.34 mM and K3Fe(CN)6 3.4 mM were used, both
in HNO3 0.4 M. The stability of the generated TB was improved using polyvinylpyrrolidone (PVP) at
0.20% (m/v). In order to simplify the method, it was verified that there is no statistical difference (95%
confidence) between the analytical curves for AAr (5 to 40 M) adding the reagents separately or as
a single mixture [Fe(III) + K3Fe(CN)6 + PVP]. Then, optimized conditions were applied to detect AP,
using AAP (1 mM) as substrate at pH 5 (HAc/NaAc, 20 mM). In these conditions, an analytical curve
for AP was obtained, with a linear range of 0.50 - 4 U L-1, corresponding to the equation A710 nm =
0.32×CAP + 0.07 (n = 6, r = 0.9867) and LOD = 0.20 U L- 1. As a proof of concept, the method was
successfully applied to a real semen sample (CAAE: 58121322.7.0000.5013) in a qualitative
evaluation (Fig. 1B-C) after dilution of the semen in solution and applied to the surface of cloth
(cotton). New assays are under development to expand the method's application. Finally, the
proposed method proved to be applicable in different situations, with a response of up to 20 min,
being an alternative to use in the field and contributing to quick decision-making when necessary.
1 Bull H, Murray PG, Thomas D, Fraser AM, Nelson PN, Molecular Pathology, 55, 2002, 65.
[UFAL, IQB, PPGQB, FAPEAL, CNPq, CAPES, PROCARD]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

483
FAST, LOW-COST, AND HIGHLY SPECIFIC COLORIMETRIC RT-LAMP ASSAYS
FOR INFERENCE OF THE BA.1 AND BA.2 NEW VARIANTS OF CONCERN
Paulo Felipe N. Estrela (PG)a*, Carlos A. dos Santos (PG)b, Paola C. Resende (R)c,
Patricia M. Lima (PG)c, Thauane S. C. da Silva (PG)c, Leonardo S. V. Matilde (PG)c,
Marilda M. Siqueira (R)c, Elisângela P. Silveira-Lacerda (R)b, Gabriela R. M. Duarte (R)a
a Universidade Federal de Goiás, Instituto de Química, Goiânia-GO, Brazil, 74690-900
b Universidade Federal de Goiás, Instituto de Ciências Biológicas, Goiânia-GO, Brazil, 74001-970
c Instituto Oswaldo Cruz FIOCRUZ, Rio de Janeiro-RJ, Brazil, 21040-360
*e-mail: paulofne@hotmail.com
COVID-19 is currently the world's leading disease, responsible for more than 6.4 million deaths in
the last three years, with constant renewal through the emergence of new variants. In this scenario,
the World Health Organization has classified the variants with the most significant potential to
increase the health crisis as Variants of Concern (VOC). The VOC Omicron was responsible for the
last wave of COVID-19 worldwide, with BA.1 and BA.2 lineages currently dominating the
epidemiological scenario.1 VOC surveillance is essential because the high viral circulation among
immunized people favours the emergence of new variants capable of evading immune responses,
rendering existing vaccines ineffective.2 Although there is a need for surveillance, African, Asian,
and South American countries sequence less than 1% of their total COVID-19 samples.3 In general,
methods for identification of variants need sophisticated instrumentation, specialized experimental
labour and high cost and time. In contrast, the reverse transcription Loop-mediated isothermal
amplification (RT-LAMP) have been proposed as a viable alternative to detect specific mutations to
help in the surveillance of VOCs, mainly in places with scarce resources, due to instrumental
simplicity required, low cost and time.2 Thus, this study aimed to develop a low-cost bioanalytical
method for inference of the main circulating VOCs (BA.1 and BA.2 lineages) based on the
colorimetric RT-LAMP. For this, we designed two sets of primers LAMP targeting specific mutations
in either the Omicron lineages BA.1 and BA.2. The mutations S: N211del and S: L212I were used to
design BA.1 primers, and S: Δ24-26 and S: A27S for BA.2. BA.1 RT-LAMP reaction contained 0.2
µM of external primers, 1.6 µM of internal primers, 0.8 µM of loop primers, 60 mM of guanidinium
chloride, 1X WarmStart Colorimetric LAMP Master Mix and 1.5 µL of RNA sample. While the BA.2
RT-LAMP reactions, 0.4 µM of external primers, 3.2 µM of internal primers, 1.6 µM of loop primers,
1X WarmStart Colorimetric LAMP Master Mix and 1.5 µL of RNA sample. The assays were
isothermally heated at 65 °C for 40 min for BA.1, and 30 min for BA.2. At the end of the incubation
time, all tubes were removed from the thermoblock and the results were interpreted based on the
color of the reaction, which changed from pink (negative) to yellow (positive). With the optimized
reaction conditions, we confirmed the high specificity of the primers designed in reactions with
different variants of SARS-CoV-2. The limit of detection with a confidence interval of 95% was 1,56
x103 RNA copies µL-1 (Cq 25.20) for BA.1 lineages and 7,46 x102 RNA copies µL-1 (Cq 26.30) for
BA.2 lineages, as estimated by probit regression analysis. A panel of 267 previous sequenced RNA
samples were blind tested in two independent RT-LAMP assays for B.1 and BA.2. The RT-LAMP
test for BA.1 and descendants showed 96.63% sensitivity, 100.00% specificity, 100.00% positive
predictive value, 98.37% negative predictive value and 98.89% accuracy. While the RT-LAMP test
for BA.2 and descendants showed 90.00% sensitivity, 98.85% specificity, 75.00% positive predictive
value, 99.61% negative predictive value, and 98.52% accuracy. The simplicity of the entire process
and the accuracy of over 98% demonstrates an excellent potential for colorimetric RT-LAMP assays
to be used as an inference test for genomic surveillance, mainly to rationalize sequencing in
countries with scarce resources.
1 World Health Organization, 2022.
2 Santos CA, Silva LC, Júnior MNS, Mendes GM, Estrela PFN, Oliveira KG, Curcio JS, Resende PC, Siqueira MM,
Pauvolid-Corrêa A, Duarte GRM, Silveira-Lacerda EP, Scientific Reports, 12, 11500, 2022.
3 Kalia K, Saberwal G, Sharma G, Nature Biotechnology, 39, 2021, 1058.
[CNPq, FAPEG, DECIT, MPT-GO]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

484
DIGITAL MICROFLUIDICS FOR BIOANALYTICAL APPLICATIONS: FROM
SAMPLE PREPARATION TO COLORIMETRIC ANALYSIS
Danielly S. Rocha (PG)a*, Richard P. S. de Campos (R)b, Habdias A. Silva-Neto (PG)a, Gerson
D. F. Júnior (R)a, Fethi Bedioui (R)c, Aaron R. Wheeler (R)d, Wendell K. T. Coltro (R)a,e
a Universidade Federal de Goiás, Instituto de Química, Goiânia, Goiás, Brazil, 74690-900
b National Research Council Canada, Edmonton, Alberta, Canada
c Chimie ParisTech, Institute of Chemistry for Life and Health Sciences i-CLeHS Paris, France, 75005
d University of Toronto, Department of Chemistry, Toronto, Ontario, Canada, M5S 3H6
e Instituto Nacional de Ciência e Tecnologia de Bioanalítica, Campinas, São Paulo, Brazil, 13084-971
*e-mail: daniellysr@outlook.com
Digital microfluidics (DMF) is a versatile lab-on-a-chip platform that allows integration with several
detectors, including colorimetric sensors1. However, some challenges need to be addressed when
integrating DMF with optical detection techniques, such as light homogeneity on the device and data
processing. Here, we propose, for the first time, a simple and affordable strategy for background
light control and colorimetric detection on DMF. In this sense, a mini studio combined with a LED
panel and webcam detector was designed to generate homogenous lighting for colorimetric
readouts. As a proof of concept, the proposed integrated tool was used for indirect bioanalysis of
nitrosocysteine (CySNO); an important biomarker for diseases such as Parkinson and Alzheimer.
The movement of the droplets was controlled by MicroDrop software (version 3.1.3) and assays
were performed using either real-time actuation mode or with semi-automated protocol. Droplet
driving potentials ranged from 90 and 120 V, frequency of 10 kHz and time intervals ranging from 1
to 5 s. To promote the sample preparation, the photolytic cleavage of CySNO to nitrite (NO2-) was
performed using a low-cost 3D printed holder. For realizing the decomposition of CySNO on the
DMF chip, an aliquot of standard solution or sample (9 µL) was positioned on reservoir region and
exposed to UV light for 15 minutes. Posteriorly, the samples volume of approximately 1 μL were
moved to the detection zones to be mixed with a drop of Griess reagent. Finally, the colorimetric
analysis was performed in two simple steps: (i) the region of interest from colorimetric zones was
captured by a print screen command and converted as PNG file format; (ii) the image colorimetric
zone was analyzed by using Corel® PHOTO-PAINT™ X7 software, converting the color scale CMYK
to magenta channel and collecting the pixel intensity. The proposed mini studio was characterized
and exhibited excellent correlation to data acquired using a desktop scanner. Considering the
analytical parameters, the obtained linear range and limit of detection of CySNO were 12.5 to 400
and 2.8 µmol L-1, respectively. A recovery assay to three concentration levels of CySNO in different
biological matrices was performed and values for DMF tests ranged from 78 to 104% and 80 to
100% for the control serum and blood plasma samples. In addition, the proposed method was
compared with UV-Vis spectrophotometry technique. Both responses did not differ statistically at a
95% confidence level. Finally, synthetic blood serum and plasma samples were successfully
analyzed highlighting that the combination of the mini studio and DMF system has potential for
enabling protocols for sample preparation and colorimetric detection of clinically relevant biomarkers.
Figure 1. (A) DMF system components. (B) Integration between 3D printed UV chamber and DMF platform. (C)
Decomposition of CySNO by UV radiation and mixture of the degradation product with the chromogen (Griess reagent).
(D) Digital image capture of the drops using a webcam.
1 Choi K, Ng AHC, Fobel R, Wheeler AR, Annual review of analytical chemistry, 5, 2012, 413.
[CNPq, INCTBio, UFG, GME, Wheeler Lab]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

485
DEVELOPMENT OF A PAPER-BASED OPTOELECTRONIC NOSE FOR IN-SITU
DISCRIMINATION OF ESSENTIAL OILS
Bárbara Cristina Dias (PG)a*, João Flávio da Silveira Petruci (R)a
a Universidade Federal de Uberlândia (UFU), Instituto de Química, Av. João Naves Ávila, 2121, Santa Mônica,
Uberlândia-MG, Brasil, 38400902.
*e-mail: babbidias72@gmail.com
Essential oil (EO) is a mixture of volatile compounds obtained from aromatic plants usually
prescribed for a variety of health conditions. The identification of EOs is important for the quality
control of the products and to avoid adulteration1, therefore, ensuring safety for the consumers. The
analytical methods commonly employed are usually time consuming and require separation
techniques with high costs. In addition, in-situ identification is preferable to provide fast information
concerning the quality of the products. In this scenario, the concept of an array of colorimetric
sensors is extremely valuable, due to the inherent discrimination ability, simplicity, ease of
fabrication, low cost and minimum use of reagents and samples2,3. In this study, an optoelectronic
nose – a.k.a colorimetric sensor array – consisting of nine colorimetric reagents impregnated in a
paper-based analytical device (PAD) was developed to discriminate five types of essential oils (i.e.
rosemary, cedar, citronella, eucalyptus and lavender). The paper-based devices with circular spots
with a diameter of 3mm were fabricated using the wax printing method. The colorimetric reagents
employed in the array were: (A1) Mn-5,10,15,20-Tetraphenylporphyrin (metalloporphyrin), (B5)
methyl orange, (B6) bromocresol green + AgNO3, (B7) methyl red, (B10) alizarin (pH indicators),
(C1) nile red, (C7) eriochromocyanin, (C9) dispersive orange (solvatochromic dyes) and (E1) 2,4-
Dinitrophenylhydrazine (specific reagent for ketones and aldehydes). 1.5 µL of each reagent was
added to the respective circular spot onto the PAD surface. Each opto-nose was exposed to the
volatiles from each type of EO for 15 minutes with an airflow of 200 mL min-1. The digital images
from before and after exposure were acquired with a scanner (HP ScanJet) and the color information
(RGB parameters) was extracted with ImageJ. By using the difference of the RGB parameters (∆R,
∆G and ∆B) differential color maps were built for each oil using graphic software (Inskcape). Figure
1 presents the color map for each EO.
Figure 1. Representative differential map of the optoelectronic noses after the exposure to different types of essential oils.
The obtained color maps resulted in a unique fingerprint for each sample, demonstrating the potential
of the optoelectronic nose to discriminate different types of essential oils. Chemometric techniques,
such as PCA and HCA, were applied to classify the oils according to the responses provided by the
responses. The first two principal components generated were responsible for 95.1% of the total
explained variance. In addition, the HCA dendrograms revealed well-defined clusters for each
sample type. As future perspectives, we suggest the possibility of using other multivariate
techniques, such as linear discriminant analysis (LDA), to provide unknown sample identification
and, therefore, providing adulteration identification.
1 Raut JS, Karuppayil SM, Industrial Crops and Products, 62 (2014) 250.
2 Conrado JAM, Sequinel R, Dias BC, Silvestre M, Batista AD, Petruci JFS, Food Chemistry, 350, 2021, 129243.
3 Dias BC, Batista AD, Petruci JFS, Analytica Chimica Acta, 1187 (2021) 339141.
[UFU, CNPq, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

486
DETERMINATION OF ADULTERATION IN EXTRA VIRGIN OLIVE OIL BY
DIFFERENTIAL SCANNING CALORIMETRY AND PARTIAL LEAST SQUARES
Lucas H. Pereira (PG)a*, Juliana Pereira (PG)a, Jerusa S. G. Trevisan (R)a, Marcello G.
Trevisan (R)a
a Federal University of Alfenas, Institute of Chemistry, Alfenas, Minas Gerais, Brazil, 37130-001
* e-mail: lucas22henrico@hotmail.com
Adulteration is an intentional process of altering the composition of products with high economic
value in order to obtain greater profit, such as beverages, medicines, foods, among others1. In
general, this process is made employing cheaper materials that are difficult to be noted by
consumers. Due to superior nutritional quality and high economic value, extra virgin olive oil (EVOO)
is usually adulterated by adding a low-quality and low-cost, food-grade vegetable oils such as corn,
soybean, sunflower and canola oils2,3. Many analytical techniques are used to determine and
quantify adulteration in products4. Among the thermoanalytical techniques, Differential Scanning
Calorimetry (DSC) stands out for being a simple and fast (without the need of an elaborated sample
preparation) that requires a very small sample amount. The aim of this work was to develop and
apply an analytical methodology for the adulterants determination in EVOO using DSC with
controlled cooling with partial least squares (PLS). For this, several binary mixtures were prepared
using sunflower, corn and soybean oils as adulterants. These materials were weighed and prepared
in proportions from 5 to 95% (m/m). The set of the samples were submitted to DSC analysis using
the following parameters: dynamic atmosphere of N2 (50 mL.min-1); temperature from 30 to -80ºC
and from -80 to 30ºC with heating/cooling rate of 5ºC.min-1; around 8 mg of sample in sealed
aluminum crucible. The construction of the PLS model was carried out by building two blocks X and
y using DSC data, comprising a set of X (21 x 13152) and y (21 x 1) for each adulterant. To eliminate
the influence on the sample's mass variation, the curves were normalized by the respective initial
masses. Data were pre-processed, normalized by the respective standard deviations and mean
centered. The multivariate results were also compared with an univariate calibration using Tonset data
(correspondent to oil crystallization event). The PLS model was successfully constructed to quantify
the adulteration level. DSC-PLS shows prediction errors of 2.54, 2.78 and 4.73% m/m for sunflower,
corn and soybean, respectively, with 4-5 latent variables. The univariate calibration curve
constructed demonstrates linearity obtained the following equations (y = -0.239x - 38.331, R2 =
0.962); (y = -0.182x - 41.059 R2 = 0.949); (y = -0.201x - 40.906, R2 = 0.936); and shown prediction
errors of 3.92, 1.26 and 1.22% m/m, for sunflower, corn and soybean oil, respectively. In this work,
a strategy based on DSC was successfully developed for the determination of adulteration in EVOO.
This is a very suitable approach to assessing the quality of EVOO related to adulteration evaluated.
Furthermore, the univariate model also is a fast and easy preliminary analysis for adulterants
detection in EVOO. When compared with others methodologies available, this work shows
satisfactory results with errors under 5%.
1 Santana FB, Neto, WB, Poppi RJ, Food Chemistry, 293, 2019, 323.
2 Meenu M, Cai Q, Xu B, Trend in Food Science and Technology, 91, 2019, 391.
3 Torrecilla JS, Cancilla JC, Matute G, Díaz-Rodríguez P, Flores AI, Journal of Food Engineering, 118, 2013, 400.
4 Ellis DI, Muhamadali H, Haughey SA, Elliott CT, Goodacre R, Analytical Methods, 7, 2015, 9401.
[UNIFAL-MG, FAPEMIG, CNPq, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

487
ATAZANAVIR SOLID DISPERSION: IN VITRO DISSOLUTION EVALUATION
AND SOLUBILITY
Juliana Pereira (PG)a*, Lucas H. Pereira (PG)a, Marcello G. Trevisan (R)a, Jerusa S. G.
Trevisan (R)a
a Universidade Federal de Alfenas, Instituto de Química, Alfenas, Minas Gerais, Brazil, 37130-001
* e-mail: julianapereiraqa@gmail.com
Atazanavir (ATV) is an antiretroviral used in combination with others in the treatment of the infection
by human immunodeficiency virus (HIV). The disease is characterized by the weakening of the
immune system, resulting from the decline in the number of lymphocytes T-CD4+, and by the
appearance of opportunistic infections1,2. Since the beginning of the pandemic, the highly active
antiretroviral therapy (HAART) has been applied worldwide giving infected patients the possibility of
living with the disease3,4. ATV is a protease inhibitor with high specificity against HIV-1 virus
categorized as class II drug by the Biopharmaceutics Classification System (BCS), which means
that it has poor aqueous solubility and high permeability, i.e. characteristics that contribute to low
oral bioavailability5. Considering its importance as well as the limitation about solubility, this work
aims to investigate solubility of ATV through solid dispersion (SD) in a hydrophilic carrier, mannitol
(MAN), and to evaluate its dissolution under physiological pH and temperature conditions. For that,
the SD-ATV was obtained by Kofler Method, thought the melt of physical mixture containing ATV
and MAN in a stoichiometric ratio of 1:4 (mol/mol), respectively, placed in an adiabatic oven at
controlled heating rates (10°C/min) under an inert atmosphere of nitrogen. The sample (40.0 mg)
was heated up to 209°C and left in an isotherm for 5 minutes. After melt, both components the
mixture were crystallized by cooling. The characterization of material was performed by X-ray
diffraction, infrared spectrometry, thermogravimetry and differential scanning calorimetry. The DSC
curves and ray-X diffraction patterns results suggests ATV amorphization. Solubility study was
carried out through solubility equilibrium (shake-flask) and in vitro dissolution test was performed
using apparatus II. All data obtained from both methodologies were quantified using the molecular
absorption spectrophotometry technique and the results demonstrate that SD-ATV was able to
increase aqueous solubility of ATV by approximately 6-fold at pH simulating the stomach
environment (pH 1.4), 55-fold at pH representing the duodenum pH conditions (4.5 and 6.8) and,
23-fold at simulating blood plasma, after 72h of sample incubation. Compared to Atazanavir Sulfate
(ATVs), the solubility improvement was mainly at pH 4.5, 6.8 and 7.5, by approximately 30, 10 and
36-fold, consecutively within the same period. Through the dissolution test performed by the USP II
method, SD-ATV exhibited a release rate that assures bioequivalence (>85% within 30 minutes)
compared to ATVs at a pH simulating the stomach environment. According to the results, the solid
dispersion obtained in this work could be an alternative to other solid forms available of ATV.
1 Sperk M, Domselaar RV; Neogi U, International Journal of Molecular Sciences, 19, 2018, 2000.
2 Desai SN, Landay AL, Current Opinion in HIV and AIDS, 13, 2018, 22.
3 Dey S, Patro SS, Babu NS, Murthy PN, Panda SK, Journal of Pharmaceutical Analysis, 7, 2017, 134.
4 Nascimento ALCS, Fernandes RP, Quijia C, Araujo VHS, Pereira J, Garcia JS, Trevisan MG, Chorilli M, Chemmedchem,
15, 2020, 1018.
5 Charalabidis A, Sfouni M, Bergström C, Macheras P, International Journal of Pharmaceutics, 566, 2019, 264.
[UNIFAL-MG, FAPEMIG, CNPq, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

488
SIMPLE SPECTROPHOTOMETRIC PROCEDURES FOR ETHANOL AND
METHANOL DETERMINATION IN ALCOHOL-BASED SANITIZERS
Vivian Maringolo (PG)*, Caio C. V. Macedo (PG), Diogo L. Rocha (R)
Federal University of ABC, Center for Natural Sciences and Humanities, Santo André, São Paulo, Brasil,
CEP 09210-580
*e-mail: vivian.maringolo@ufabc.edu.br
Quality assessment of alcohol-based sanitizers is important to ensure product efficiency and to avoid
contamination. Ethanol is the main constituent of these sanitizers, which inactivates microorganisms
present on surfaces, such as the virus that causes COVID-19. For maximum efficiency, the sanitizer
must show ethanol content between 60 and 80%[1]. Nevertheless, these products may contain toxic
compounds, such as methanol[2], which is commonly found in samples with high ethanol content.
Therefore, analytical procedures are necessary to quantify these components in sanitizers. In this
work, simple spectrophotometric procedures were proposed for the determination of ethanol and
methanol in commercial gel and liquid sanitizers.
Figure 1: Effect of sulphuric acid concentration and reaction Figure 2: Response surfaces obtained from the models
time (at 50ºC) on analytical (-□-) and blank signals (-○-) for to demonstrate the interactions acetylacetone/KMnO4
ethanol determination. concentrations (A) and pH/concentration of the buffer.
1 0.9
0.8
A 0.4
B 0.7
0.8 0.6
0.7
0.35
0.5
0.6
Absorbance
Absorbance
0.6
Absorbance
0.4
0.5 0.3
0.3
0.4
0.4 0.25 0.2
0.3
0.1
6.0
0.2 0.2 5.7
0.2 0
347
5.4
337
1000
976
952
326
928
5.1
904
7.9
316
880
856
0.1
305
832
7.0
808
295
784
4.8
760
pH
736
284
6.2
712
688
274
664
640
5.4 4.6
263
616
592
253
568
544
4.5
520
0.0 242
496
4.3
472
0
232
3.7
448
424
221
400
2.8 KMnO4 (mmol L-1)
211
8 10 12 14 16 18 20 22 Acetylacetone (mmol/L) 4.0

Ammonium Acetate (mmol L-1)
200
0 1 2 3 2.0
H2SO4 (mol L-1) Time (min)
Table 1: Evaluated parameters and selected values for Table 2: Evaluated parameters and selected values for
ethanol determination (univariate method). methanol determination (multivariate method).
Evaluated Selected Optimum

Parameter Parameter -1 0 1
range value value
KMnO4, µmol L-1 50-400 400 KMnO4, mmol L-1 200 275 350 200
H2SO4, mmol L-1 500-3000 1500 H3PO4, mmol L-1 300 500 700 300
Reaction time ActAct1, mmol L-1 2.0 5.0 8.0 8.0
0-30 20 NH4Ac2, mmol L-1 400 700 1000 1000
(50 ºC), min
pH 4.0 5.0 6.0 6.0
1) acetylacetone; 2) ammonium acetate
Both procedures were based on the oxidation of the analytes by KMnO4 in acidic medium. The
remaining reagent was directly related to the ethanol content. For methanol determination, the
yielded formaldehyde was quantified after reaction with acetylacetone in ammonium acetate
medium. After optimization of ethanol determination (Figure 1 and Table 1), the detection limit (n=15,
99.7% confidence level) and the coefficient of variation (n=15) were estimated, respectively, at 24
µmol L-1 and 1.7%. The most common concomitants in the samples did not cause significant
interference. Recoveries between 84 and 95% indicated adequate accuracy of the method. The
results of analyzed samples by the proposed and the reference procedures did not show significant
differences (95% confidence level). For methanol determination, after multivariate optimization
(Figure 2, Table 2), the detection limit (n=10, 99.7% confidence) and coefficient of variation (n= 10)
were estimated, respectively, at 18 µmol L-1 and 3.1 %. Recoveries obtained between 45 and 115%
and interference studies indicated that additional evaluations are needed to improve the performance
of the method. Other compounds with hydroxyl groups were oxidized by permanganate, such as
glycerol (which also yield formaldehyde). Therefore, micro-distillation using low-cost instrumentation
is in development for sample preparation prior to methanol determination.
1 Harrington C, Walker H, The New England Journal of Medicine 148, 1903, 548.
2 Yangzes S, Grewal S, Gailson T, Grewal SPS, JAMA Ophthalmology, 139, 2021, 362.
[CAPES, FAPESP]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

489
ARSENIC SPECIATION IN IN VITRO ASSAYS
Altevir R. Viana (PG)a, Graciela M. Heidrich (PG)b, Cristiano R. B. Rhoden (R)a, Érico M. M.
Flores (R)b, Valderi L. Dressler (R)b, Sergio R. Mortari (R)a*
a Universidade Franciscana, Ciências Tecnológicas, Santa Maria, RS, Brazil, 97010-032
b Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brazil, 97105-900
*e-mail: mortari@ufn.edu.br
Arsenic trioxide (ATO) is an efficient drug for Acute promyelocytic leukemia (APL) treatment, in the
induction of remission and consolidation in adult patients. The objective of this work was to evaluate
possible changes of the chemical species of arsenic, in vitro assay, in cell line NB4 (APL). The
technique used for speciation analysis of arsenic was IC-ICP-MS, with experimental conditions
according to Rosa et al., 20191. In short, the separation of As species was achieved by using a
Hamilton, PRP-X100, 250 x 4.1 mm column and a mobile phase of 10 mmol L-1 (NH4)2HPO4 at pH
6.00 solution. NB4 strain (100,000 cells) were placed in plates containing 96 wells in the presence
of 5 μg mL-1 ATO. The cells remained for 24 h and 72 h in a CO2 oven at 37 ºC in a humidified
atmosphere. After then, the supernatant was collected for As speciation analysis. The results (Figure
1) indicate a low conversion (0.5 % and 3.0 %) of As(III) to As(V) at exposure to NB4 cells, for 24 h
and 72 h, respectively. From the results obtained, it can be concluded that from 24 h to 72 h of
incubation there was an increment in the oxidation of arsenic species in the in vitro assays with the
NB4 cell line, as well as the appearance of a signal (next to the trivalent As signal), which leads us
to think about the formation of organic arsenic species.
Figure 1. A) Chromatographic signal of As2O3 and As2O5 standards (5 μg L-1), and B) Arsenic species after incubation in
cells NB4 (24 h and 72 h).
1 Rosa FC, Pardinho RB, Moreira MS, Souza LGT, Flores EMM, Mortari SR, Dressler, VL, Leukemia Research, 76, 2019,
11.
[CNPq, UFSM, UFN]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

490
COMPATIBILITY STUDY BETWEEN BISOPROLOL HEMIFUMARATE AND
SOME EXCIPIENTS
Lucas Y. Sato (UN)*, Larissa G. Velasco (PG), Maria Inês G. Leles (R)
LAMES - Laboratório de Métodos de Extração e Separação - Instituto de Química, Universidade Federal de Goiás, Av.
Esperança s/n. Campus Universitário, 74690-900, Goiânia, GO - Brasil
*e-mail: yugosato@discente.ufg.br
Bisoprolol Hemifumarate, commonly sold as oral tablets, is a β1-adrenergic that functions as a

competitive antagonist to the two catecholamines responsible for increasing cardiac activity,
epinephrine and norepinephrine. Being β1 selective the Active Pharmaceutical Ingredient (API) has
an edge over other similar medication that treat cardiovascular diseases (Congestive Heart failure,
arrhythmia, etc) since β1 receptors are only found in the heart, kidneys and fatty tissues, and will
mainly affect these areas of the body1,2. Pre-formulation is fundamental in the process of coming up
with new stable, safe and efficient drugs. One of the goals of pre-formulation is to understand how
the API is going to behave in the presence of the excipients3. In the present work, the thermal stability
of Bisoprolol Hemifumarate was studied in the presence of the following excipients: Magnesium
Stearate, Crospovidone and Dibasic Calcium Phosphate Dihydrate (DCPD). The samples, supplied
by GeoLab Indústria Farmacêutica Ltda., were analyzed using TGA-DTG and DSC techniques. The
TGA-DTG curves of the API and excipients were obtained using the model TG/SDTA 851e (Mettler
Toledo) in an α-Al2O3 pan and heated from 25 ºC to 1000 ºC. The DSC curves of the API, excipients
and the binary mixtures 1:1 (w/w) were obtained using the model DSC 822e (Mettler Toledo) in a
closed aluminum pan with a pinhole and heated from 25 ºC to 300 ºC. Both techniques were
performed under a N2 atmosphere, as a purge gas, with the flow rate of 50 mL min-1 and at a heating
rate of 20 ºC min-1. The DSC curve of Bisoprolol Hemifumarate showed three endothermic events,
the first one in the temperature range of 25-90 ºC, related to its dehydration; a second event with a
peak at 100 ºC indicating the melting; and the last one in the range of 180-260 ºC related to its
thermal decomposition. Four events were observed in the DSC curve of Magnesium Stearate where
the first three, between 25-150 ºC, are associated with the dehydration and the last one, at around
200 ºC, the melting. The DSC curve of Crospovidone only showed one endothermic event, in the
range of 25-150 ºC, related to its dehydration. In the DSC curve of DCPD, there are two consecutive
endothermic events, from 100 ºC to 180 ºC and then from 180 ºC to 200 ºC, attributed to the loss of
water. All of these mass losses caused by dehydration and thermal decomposition can be observed
in their respective TGA-DTG curves. Finally, the behavior of the binary mixtures (API + Magnesium
stearate, API + Crospovidone, API + DCPD) were analyzed by DSC. All of the events described
previously with each compound still occur, in the same temperature range, in the DSC curves of the
binary mixtures, thus concluding that there are no incompatibilities between them.
1 Rosenberg J, Gustafsson F, Expert Opinion on Pharmacotherapy, 9, 2008, 293.
2 Bazroon AA, Alrashidi NF, Bisoprolol. In StatPearls. StatPearls Publishing, 2022.
3 Bharate SS, Bharate SB, Bajaj AN, Journal of Excipients and Food Chemicals, 1, 2010, 3.
[LAMES/UFG, GeoLab Indústria Farmacêutica Ltda, FUNAPE]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

491
DETERMINATION OF TOTAL BIOGENIC AMINES IN KOMBUCHA USING
DIGITAL IMAGES
Ferdynanda M. Silva (PG)*a, Wanessa R. Melchert (R)b
a Center for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil, CEP 13400-970
b College of Agriculture "Luiz de Queiroz", University of São Paulo, Piracicaba, SP, Brazi, CEP 13418-900
*e-mail: ferdynanda.silva@usp.br
Kombucha is a fermented drink obtained by infusing tea, sugars and a symbiotic culture of bacteria
and yeast known as SCOBY (Symbiotic Culture of Bacteria and Yeast.1,2 The microbiological and
chemical composition of Kombucha varies according to fermentation and can contain organic acids,
polyphenols, vitamins, proteins, amino acids, minerals and biogenic amines.3 Biogenic amines are
molecules of nitrogenous organic bases derived from the decarboxylation of the amino acid
precursor, indicative of food quality and ingestion of significant amounts can cause poisoning and
damage to the nervous, digestive, respiratory and circulatory.4 In this work, a simple procedure for
extraction and spectrophotometric determination of biogenic amines in kombucha was proposed.
The procedure was based on the employment of a colorimetric reagent, after pre-concentration
assisted by salting-out. The procedure was based on the reaction of biogenic amines with chloride
of 4-(dimethylamine) Azobenzene-4’-sulfonyl (Dabsyl) in an alkaline medium, forming an orange
compound. A camera from a cell phone and an app was used to register the images and convert the
images for RGB values. The effects of reagent concentrations and physical conditions (speed and
time of agitation and centrifugation) were evaluated by using a factorial experimental design and the
optimized condition consisted of vortex speed 9, stirring time 30 seconds, bath time 20 minutes, bath
temperature 70 ºC, reagent concentration 22 mmolL-1 and buffer solution concentration 0.95 mol L-
1
. A calibration curve was obtained with a linear range of 0,5 e 5,0 mg L-1 of histamine, described by
equation Analytical Signal= 0,1613 + 0,7474 C (mg L-1), R = 0,9989, under optimized conditions.
The coefficient of variation and the limit of detection was estimated at 0.7% (n=11, histamine 2,5 mg
L-1) and 0,3 mg L-1 of histamine, respectively. Were consumed by the proposed procedure only 300
μL of sample and 600 μL dabsyl and generated 1 mL of residue per determination.
1 Brasil, Ministério da Agricultura Pecuária e Abastecimento. Instrução Normativa n° 268 41/2019, de 17 de setembro de
2019.
2 Coelho RMD, International Journal of Gastronomy and Food Science, 22, 2020, 100272.
3 Morales D, Trends in Food Science & Technology, 105, 2020, 323.
4 Alvarez MA, Moreno-Arribas MV, Trends in Food Science & Technology, 39, 2014, 146.
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

492
MODIFICATION ON PAPER SURFACE FOR COLORIMETRIC UREASE
DETECTION EXPLORING ACID-BASE INDICATOR AND pH VARIATION
Maria Célia Tavares (PG)*,a,b Wendell K. T. Coltro (R)c, Ângelo de Fátima (R)d, Josué
Carinhanha. C. Santos (R)b
a Instituto Federal de Alagoas – IFAL Campus Batalha, Batalha, AL, Brazil.
b Instituto de Química e Biotecnologia, Universidade Federal de Alagoas – IQB/UFAL, Maceió, AL, Brazil.
c Instituto de Química, UFG, Campus Samambaia, GO, Brazil.
d Departamento de Química, UFMG, Belo Horizonte, MG, Brazil.
*e-mail: celia.tavares@ifal.edu.br
Ureases are enzymes produced by fungi, plants, and bacteria, associated with agricultural and
clinical problems. The urea hydrolysis in NH3 and CO2 leads to the loss of N-urea fertilizers in
soils and, besides, changes the human stomach microenvironment, favoring the colonization of
the H. pylori. In this sense, the enzyme activity determination using a simple, fast, and
economical way is a scientific and commercial emergency to mitigate the effects of its action. 1
For this, paper analytical devices that can be used as a rapid test have a differential due to their
ease of use, low sample volume, portability, and the integration possibility with a wide variety of
methods and techniques, such as colorimetric detection of response qualitative (yes/no) or
quantitative using scanner and/or smartphone. The change in pH caused by the hydrolysis
reaction of urea catalyzed by urease can be visually observed using acid-base indicators. Thus,
this work aims to develop a surface-modified paper colorimetric device for urease determination
by exploring different acid-base indicators. The flower-shaped devices were produced using an
Eva 16 mm craft punch Flower Helix Blister (Make Co., Ltd, China)2, three types of paper [filter
(P1), Whatman CHR n° 1 (P2), and CHR 3MM (P3)] without and with surface modification using
chitosan (0.1 to 4 mg mL-1), polyvinylpyrrolidone (PVP, 0.1 to 10% m/v), tetraethoxysilane
(TEOS, 4:1 TEOS:water) and sodium polysulfonate (PSS 1 and 10% v/v). The determination of
urease (0 - 4 U mL-1) was performed using urea (20 mM), and the indicators 0.05% (m/v)
(bromocresol and m-cresol purple, bromothymol blue, phenol, and neutral red) in 0.5 mM sodium
phosphate buffer (initial pH evaluated). A model system for each indicator and the results of
some parameters are shown in Figure 1.
Figure 1. A) pH evaluation; B) Substrate paper evaluation; C) Analytical curve for BTB indicator and PSS conditions; D)
Evaluation modification of the surface for BTB indicator; E) Device for comparative conditions modification of the surface
for BTB indicator. Conditions: urease 0-4 UmL-1, urea 20 mM, 0.05% (m/v) indicators, scanner /Corel Drawn X8 analysis.
The results indicated that the CMYK color channel and the Whatman CHR 3MM paper for all
acid-base indicators showed better linearity and sensitivity. Initial pH, paper type, and surface
modification influence the sensitivity and color intensity of the acid-base indicator since the initial
pH is dependent on the indicator pKa. The PSS and PVP modifiers are more effective, combining
simple preparation, low toxicity, greater color intensity, and sensitivity. Thus, we developed a
paper device with simple and low-cost fabrication using acid-base indicators to determine
urease.
1 Celik C, Sezgin GC, Kocabas UG, Gursoy S, Ildiz N, Tan W, Ocsoy I, Analytical Chemistry, 93, 2021, 6246.
2 Su M, Johnson MA, RSC Advances, 5, 2015, 55633.
[LINQA, PPGQB, IQB, UFAL, IFAL, UFG, UFMG, FAPEAL, CAPES, CNPq]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

493
SIMPLIFYING NITRATE DETERMINATION BY UV-VIS
SPECTROPHOTOMETRY IN THE PRESENCE OF INTERFERENTS
Marcela F. Silva (R)a*, Luana B. de Lima (UG) b, Claudia Telles Benatti (R)a
a Universidade Estadual de Maringá, Civil Engineering Departament, Maringá, PR, Brazil, 87020-900
b Universidade Estadual de Maringá, Chemistry Departament, Maringá, PR, Brazil, 87020-900
*e-mail: mfsilva2@uem.br
The presence of nitrate (NO3) in groundwater and surface water at concentrations above those
considered suitable for human consumption has become an increasingly frequent problem. One
of the most used methods is spectrophotometry, both using commercial kits and by direct
reading of the absorbance relative to nitrate at wavelengths in the 195 to 280 nm range, 1-3 as
the method 4500-NO3-B of Standard Methods for the Examination of Water and Wastewater 8
(SM-4500 method) based on reading absorbance at 220 nm (A220, due to NO3) corrected by
twice absorbance at 275 nm (due to organic matter, A275) and Adolfo Lutz Institute method 9 (ALI
method), based on reading the absorbance at 205 nm. These methods use a range of
wavelenght where interferences may occur due to the presence of organic matter (OM) and
chloride (Cl)4-6. Certain saline constituents (such as hydroxides and carbonates) at high
concentrations can also cause interference in the absorbance relative to NO3, which in turn uses
acidification with HCl to minimize such problems7. However, following the same thought, it can
be assumed that for waters with low concentrations of hydroxides and carbonate, the
acidification step in spectrophotometric methods may not be necessary. Thus, in this study, the
simplification (without the use of acidification) of SM-4500 and ALI methods in the presence of
Cl and OM interferents was verified using factorial planning 24. The evaluated concentrations of
NO3 (5; 10 mg L-1 NO3-N), OM (0; 20 mg L-1) and Cl (0; 50 mg L-1) were based on known
concentrations of these substances in real groundwater and surface water samples from
Maringá-PR region and on the limits of Brazilian regulations 10-13. Regarding the factorial
planning, it was found that for the ALI method, the concentration of NO3 (5 and 10 mg L-1 NO3-
N) is not a significant factor in the absorbance. This shows that this method does not obey Beer's
law for these concentration ranges, so it should not be applied to its intended purpose. The SM-
4500 method had as significant factors the NO3, Cl and MO concentrations and acidification of
the sample. On the other hand, applying its standardized form (with acidification) resulted in
correction absorbances (2A275) higher than 10 % of A220 for samples with OM and NO3
concentrations up to 20 mg L-1 and 10 mg L-1 NO3-N, respectively, which indicated that this
method should not be applied for this range. However, the method can be applied in its simplified
form, without acidification.
1 Birkmann J, Pasel C, Luckas M, Bathen D, Water Practice and Technology, 13, 2018, 879.
2 Collos Y, Mornet F, Sciandra A, Waser N, Larson A, Harrison PJ, Journal of Applied Phycology, 11, 1999, 179.
3 Jonhson KS, Coletti LJ, Deep Sea Research Part I: Oceanographic Research Papers, 49, 2002, 1291.
4 Albrektienė R, Rimeika M, Zalieckienė E, Šaulys V, Zagorskis A, Journal of Environmental Engineering and Landscape
Management, 20, 2012, 163.
5 Korshin G, Chow CWK, Fabris R, Drikas M, Water Research, 43, 2009, 1541.
6 Souto MAM, Okada M, Okada IA, Dovidauskas S, Revista do Instituto Adolfo Lutz, 65, 2006, 66.
7 Gross A, Boyd CE, Seo J, Journal of the World Aquaculture Society, 30, 1999, 388.
8 APHA/AWWA/WEF, Standard Methods for the Examination of Water and Wastewater, American Public Health
Association, American Water Works Association, Water Environment Federation, 23 rd ed, 2017.
9 Zenebon O, Pascuet NS, Tiglea P, Métodos físico-químicos para análise de alimentos. Instituto Adolfo Lutz, 2005.
10 Brasil. Ministério da Saúde, Portaria GM/MS n° 888, de 4 de maio de 2021. Altera o Anexo XX da Portaria de
Consolidação GM/MS no 5, de 28 de setembro de 2017, 2021.
11 Mantovani D, Batista, AT, Bezerra CO, Cusioli LF, Rezende D, Bergamasco R, Soares, PF, Journal of Exact Sciences,
18, 2018, 5.
12 Rezende D, Valim Junior NC, Nishi L, Mantovani D, Vieira AMS, Bergamasco R, Blucher Chemical Engineering
Proceedings, 2, 2015, 1954.
13 Rezende D, Nishi L, Coldebella PF, Mantovani D, Soares PF, Valim Junior NC, Vieira AMS, Bergamasco B,
Desalination and Water Treatment, 161, 2019, 203.
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

494
ANALYSIS OF THE CHELATING CAPACITY OF Fe2+ AND Cu2+ IN ROASTED
AND IN NATURA YERBA MATE INFUSIONS
Gabrielly Rodrigues Campos Cruz (UG)a, Helder Araujo de Oliveira (UG)a, Samuel Lopes
Arruda (UG)a, Claudemir Adriano Borgo (R)a, Marcos Canto Machado (R)a*
a Indaiatubana Foundation for Education and Culture, Technological Education Center, Indaiatuba, São Paulo, Brazil,
13349-003
*e-mail: marcos.machado@fiec.edu.br
The association of transition metals (e.g. iron and copper) as intensifiers of pathologies has been a
target of interest in the scientific community, aiming to prevent or remedy the high absorption of
these metals by the human body. In this context, the use of chelating agents has been increasingly
used to reduce the availability of metals in organisms. Although there are several chelating agents
available in therapy (e.g. EDTA, DTPA, penicillamine, deferoxamine, etc.) there are some
inconvenient side effects of their administration. However, the so-called medicinal herbs can also be
used as chelating agents, which are given in the form of extracts, infusions, and even in the form of
teas. Yerba mate (Ilex paraguarienses A. St.-Hill.), for example, is already used as a chelating agent
in the absorption of iron in patients with hemochromatosis and its use can be attributed to this
function, due to the large number of phenolic compounds derived from coumarin, tannins and
flavonoids in its composition1,2. Thus, the purpose of this work was to analyze the complexing
capacity of infusions against the free ions of Fe2+ and Cu2+ and to evaluate the ability of roasted
yerba mate teas (mate tea) and in natura herb (“chimarrão”) to verify if the process of roasting
influenced its chelating capacity. The preparation of the infusions was carried out similarly to its use
in the daily life of the population and following the instructions of the yerba mate supplier (roasted or
in natura). The analytical methodology was optimized based on the work of Granato3, using solutions
of FeSO4.4H2O (0.30mM), phenanthroline (0.80mM), EDTA (2mM), acetate buffer pH 6 (50mM) and
CuSO4.5H2O (0.1 mg/ml). The analysis of the samples (n = 10) containing the infusions, the negative
controls, and the analytical blank solutions was performed using 300 µL of these in a UV-VIS
microplate reader using wavelengths of 490 nm and 405 nm respectively for the analyzes of Fe 2+
and Cu2+. The complexed Fe2+ contents 64.6±2.4 % and 65.7±1.4 % respectively for the “chimarrão”
mate and mate tea samples, showing no statistical difference (p > 0.05) when using a T-test (CI =
95%). The complexed Cu2+ contents 57.3±5.0 % and 71.6±0.9 % respectively for the mate tea and
mate tea and “chimarrão” mate samples, showing a statistical difference (p < 0.05) when using a T-
test (CI = 95 %). The results shown were validated using the comparison of intermediate precision,
indicating that the methodology reproduces its results when performed by different analysts on
different days of analysis according to RDC No. 166/20174. It can be concluded that satisfactory
analytical results were obtained for the proposed objectives, and the methodologies used were able
to demonstrate the ability of the infusions to form complexes and the difference that the treatment of
the herb causes in its complexing activity.
1 Bastos HM, Silva AF, Torres E, Nutrire, 3, 2003.
2 Leopoldine M, Russo N, Chiodo S, Toscano M, Journal of Agricultural and Food Chemistry, 54, 2006.
3 Granato D, Análises químicas, propriedades funcionais e controle da qualidade de alimentos e bebidas, LTC, 2016.
4 ANVISA. Resolução nº 166. Dispõe sobre a validação de métodos analíticos, Ministério da Saúde, 2017.
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

495
RAPID AND LOW-COST COLORIMETRIC DETECTION OF SARS-COV-2 USING
FUNCTIONALIZED COTTON SWABS
André Lopes Ferreira (R)a*, Lucas Felipe de Lima (PG)a, William Reis de Araujo (R)a
a Portable Chemical Sensors Lab, Department of Analytical Chemistry, Institute of Chemistry, State University of
Campinas – UNICAMP, P.O. Box 6154, 13083-970, Campinas, SP, Brazil.
*e-mail: drelopes20@gmail.com
The COVID-19 pandemic has exacerbated our society’s health equity gap. Disadvantaged
populations have been disproportionally affected by COVID-19, lacking access to affordable testing,
a known effective tool for preventing viral spread, hospitalizations, and deaths. High-frequency
testing represents an excellent tool for outbreak and viral spread prevention that can only be realized
with tests that are accessible, low-cost, and rapid. However, the most widely used tests today, such
as reverse transcription polymerase chain reaction (RT-PCR), are slow, relatively expensive, and
require highly skilled workers and appropriate lab infrastructure. Colorimetric biosensors display
properties that may help overcome these issues. They are easy to prepare and use, rapid, portable,
and can utilize metallic nanoparticles whose optical properties have been well established and
characterized1. Such tests hold potential for deployment in remote locations and are amenable for
use both at the point of care and at home without the need for sophisticated equipment. However,
more affordable technologies using accessible equipment that does not require skilled personnel to
operate are needed to enable mass testing. Smartphones have been used both as a detector and
processor of portable biochemical assays, revolutionizing the field of mobile healthcare by
empowering citizens to perform rapid screening tests while obviating the need for expensive
equipment or high levels of expertise2. Here, we report the development of an accessible and
sensitive colorimetric biosensor fabricated on cotton swabs using human angiotensin-converting
enzyme 2 (ACE2), a natural receptor for SARS-CoV-23, and functionalized gold nanoparticles
(AuNPs) for COVID-19 diagnosis. The colorimetric biosensor leverages ACE2 immobilized directly
onto cotton to ensure selective and rapid recognition of SARS-CoV-2 spike protein (SP) present in
the nasopharyngeal/oropharyngeal (NP/OP) biofluids. The AuNPs modified with ACE2 enable a
visual readout of the results through the formation of a molecular sandwich with SARS-CoV-2, dyeing
the cotton surface for a positive COVID-19 test. Our biosensor enables COVID-19 diagnosis through
a simple, minute-time scale color shift that can be detected by the naked eye and whose quantitative
results may be obtained using a smartphone camera and a free RGB app. Under optimized
conditions, an analytical curve was built by incubating (3 min) the cotton swabs into solutions
containing increasing concentrations of SP (10–12 to 10–6 g mL–1), which provided a visually purple
gradient color from 10–12 to 10–8 g mL–1 of SP and resulted in a limit of detection of 0.154 pg mL−1
SARS-CoV-2 SP. The performance of the proposed method was assessed using 100
nasopharyngeal/oropharyngeal (NP/OP) clinical samples, yielding sensitivity, specificity, and
accuracy values of 96%, 84%, and 90%, respectively. Additionally, the color intensity correlated with
the cycle threshold (Ct) values obtained using the RT-PCR method. Collectively, our diagnostic test
can be easily assembled and costs only 15¢ per assay, and presents rapid (5 min) and sensitive
detection of SARS-CoV-2, thus providing a solution to enable high-frequency testing, particularly in
low-resource communities.
1 Tang L, Li J, ACS Sensors, 2, 2017, 857.
2 Fan Y, Li J, Guo Y, Xie L, Zhang G, Meas. Journal of the International Measurement Confederation, 171, 2021, 108829.
3 Zhang H, Penninger JM, Li Y, Zhong N, Slutsky AS, Intensive Care Medicine, 2020, 46, 586–590.
[CNPq, CAPES, FAPESP, University of Pennsylvania, Cesar de La Fuente-Nunez]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

496
EVALUATION OF PROPANIL AND 3,4-DCA PHYTOREMEDIATION
BY PISTIA STRATIOTES
Emerson J. S. Silva (R)a, Maria L. V. Schmitt (PG)b, Bruna P. Ramborger (PG)c, Rafael
Roehrs (R)d*
a Federal University of Pampa - UNIPAMPA, Natural Science, Uruguaiana, RS, Brazil, 97501-970
b Federal University of Pampa - UNIPAMPA, Biochemistry Postgraduation Program, LAQAT, Uruguaiana, RS, Brazil,
97501-970
c Federal University of Pampa - UNIPAMPA, LAQAT, Uruguaiana, RS, Brazil, 97501-970
d Federal University of Pampa - UNIPAMPA, LAQAT, Bagé, RS, Brazil, 96413-172
*e-mail: rafaelroehrs@unipampa.edu.br
Propanil is a commonly used pesticide in agriculture. In few days it degrades to 3,4-

dichloroaniline (3,4-DCA), which is more persistent and has a higher toxicity.1 Therefore, it is
necessary to find ways to remove it in order to combat environmental contamination. Most of
removal techniques have high cost and often end up generating highly toxic by-products. Yet,
phytoremediation comes as a low-cost technique for this removal and some phytoremediation
plants are already known, the species Pistia stratiotes is one of them.2 The objective of this study
was to evaluate the phytoremediation of propanil and 3,4-DCA by P. Stratiotes. To find out the
lethal dose for the plant, a sensitivity curve was constructed. The plants were exposed to
concentrations of 2.5; 5; 10 and 50 mg L-1 of propanil and 1.8; 3.7; 7.4 and 37.1 mg L-1 of 3,4-
DCA. Only the maximum concentration of each group caused death of the seedlings in 14 days
of treatment. To analyze the presence these compounds in the collected samples, High
Performance Liquid Chromatography (HPLC) was used in a reversed-phase, with C18 column,
an isocratic mobile phase of acetonitrile, methanol and water (30:24:46 v/v/v) in a pH 3 (aqueous
phosphoric acid solution (1:1, v/v)), the flow rate was 0.9 ml.min-1 and the injection volume 20
μL in a 220 nm wavelength performed at room temperature. A YL9100 chromatography system
(Young Lin, South Korea) equipped with a YL90 vacuum degasser YL9110 quaternary pump,
YL9150 auto-injector, YL9131 column oven, YL9160 diode array detector was used. Analyzes
were performed using Synergi 4µ Fusion-RP 80Å chromatography column (250 x 4.6 mm) and
Fusion-RP pre-column (4 x 3.0 mm) (Phenomenex, United States). Equipment control and data
acquisition were performed using the YL-Clarity software. The acetonitrile used in the
chromatographic analyzes was HPLC grade, the ultrapure water was purchased from a Milli-Q
system and the methanol was HPLC grade. For these chromatographic conditions it was
obtained the equation of the straight line of y=330.13x + 9.8288, r 2= 0.9996 for propanil and
y=105.86x – 14.545, r2= 0.9955 for 3,4-DCA, both in a linear range of 0.5 to 10 mg L-1. P.
Stratiotes was resistant to concentrations lower than 10 and 7.4 mg L-1 of propanil and 3,4-DCA,
respectively. These concentrations correspond to twice the recommended for the application of
propanil in rice culture, which is 5 mg L-1.3 Evidencing its possible use in water contaminated
with this herbicide and its metabolite. To evaluate the kinetics of degradation P. stratiotes were
treated with solutions of 10 mg L-1 of propanil. The samples were collected on days 0, 7 and 14.
On the 7th day there was only 1.8% of propanil in the control group and it was no longer possible
to quantify it in the treatment group. After 14 days, the concentration of 3,4-DCA had a low
variation in the controls, 26.28% on the 7th day and 24.97% on the 14th day, while in the
treatments the concentration decreased from 16.63 to 5.65%. On the 14th day, it was not
possible to quantify propanil in both control and treatment. P. stratiotes was resistant to twice
the recommended concentration for the application of propanil in agriculture. This work showed
that the culture of P. stratiotes can be an effective and low-cost strategy in the phytoremediation
of waters contaminated by the herbicide propanil and the metabolite 3,4-DCA.
1 Roehrs R, Miguel R, Machado SLO, Zanella R, CLEAN–Soil, Air, Water, 40, 2012, 958.
2 Prajapati SK, Meravi N, Singh S, Proceedings of the International Academy of Ecology and Environmental Sciences. 2,
2012, 136.
3 SOSBAI. Arroz Irrigado: Recomendações Técnicas da Pesquisa para o Sul do Brasil, Porto Alegre, 2014.
[CAPES, CNPq, UNIPAMPA]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

497
CHARACTERIZATION OF BIOCHAR PRODUCED THROUGH SLOW AND FAST
PYROLYSIS OF PINE WOOD AND SUGARCANE STRAW
Lisiane O. Diehl (PG)a, Ana Luiza G. Mendes (PG)b, Samuel R. Waechter (PG)b, Fábio A.
Duarte (R)b Fernanda de Castilhos (R)a, Paola A. Mello (R)b,*
a Federal University of Santa Maria (UFSM), Chemical Engineering Department, Santa Maria, RS, Brazil, 97105-900
b Federal University of Santa Maria (UFSM), Chemistry Department, Santa Maria, RS, Brazil, 97105-900
There is a growing need for alternatives to supply the energy demand. Lignocellulosic biomass is
considered a renewable and abundant resource with the potential to reduce dependence on fossil
fuels.1,2 The use of this biomass requires transformations of the raw material through thermal
processes, such as pyrolysis. Pyrolysis is a thermochemical conversion process that decomposes
biomass in the absence of oxygen, converting it into solid (biochar), liquid (bio-oil) and gaseous
(biogas) products.3 Biochar has been widely studied to apply in adsorption, catalysis, and advanced
carbon materials. Wide varieties of plant residues have been used to produce biochar. 4 The
knowledge of its chemical composition is important since it can impair the application. In the present
study, residues of pine wood (PW) and sugarcane straw (SS) were investigated. Samples were
collected in Brazilian forests and fields and were submitted to slow and fast pyrolysis processes in
the temperature range of 400 to 600 °C to characterize the solid products. The physicochemical and
compositional properties of the biochar were analyzed using different techniques and types of
analysis. Proximate analysis was based on gravimetric methods through calculations of moisture,
ash, volatile matter and fixed carbon contents by using an oven (ETC 45 model, Nova Ética, Brazil)
and a muffle furnace (0913 model, Jung, Brazil). A bomb calorimeter (C2000 model, IKA, Germany)
was used to measure the calorific value of the biochars and the particle size distribution was
calculated using a Malvern Mastersizer 2000 (Malvern Instruments, APA 2000 model, United
Kingdom). An elemental analyzer (Multi EA 5000 model, Analytik Jena, Germany) was used to
determine the amount of N, S, and Cl. The biochars were digested using a single reaction chamber
microwave-assisted wet digestion system (UltraWAVETM model, Milestone, Italy) and the
concentration of metals was determined in the digests using inductively coupled plasma optical
emission spectrometry (ICP-OES, Spectro Ciros model, Spectro Analytical Instruments, Germany).
The functional groups of the biochars were analyzed by Fourrier transform infrared spectroscopy
(FT-IR, IRPrestige-21 model, Shimadzu, Japan). The results showed some differences related to
the change of parameters of the pyrolysis process. Unlike volatile matter content, the contents of
moisture, ash, and fixed carbon increased with the temperature increase. The pyrolysis temperature
changed the elemental composition of the PW and SS biochars. Increasing the temperature from
400 °C to 600 °C the N and Cl content decreased; and there were no significant changes in S content.
FTIR spectra indicated that the investigated biochars were dominated by functional groups typical of
oxygenated hydrocarbons, representing the carbohydrate structures of cellulose, hemicellulose, and
lignin5. The PW and SS biochars showed different particle size distribution under similar grinding
conditions, but both confirm the positive effect of pyrolysis on grindability of biomass. With regard to
the non process elements, Al, Ca and Fe were those that presented higher concentrations. It was
possible to notice that different types of biomasses and process parameters significantly change the
properties of biochars and characterization demonstrated to be a crucial point to understand the
process parameters and its influence on the composition as well as to plan the suitability of the
biochar for further applications.
1 Ghodake SG, Shinde SK, Kadam AA, et al., Journal of Cleaner Production, 297, 2021, 126645.
2 Sonil N, Mohanty P, Pant K, et al., Bioenergy Research 6, 2012, 663.
3 Xiang C, Liu Q, Shi L, et al., Fuel, 277, 2020, 118163
4 Colantoni A, Evic N, Lord R, et al., Renewable and Sustainable Energy Reviews, 64, 2016, 187.
5 Zhang J, Shao J, Jin Q, et al., Fuel, 252, 2019, 101.
[ANP-PRH, CAPES, FAPERGS, CNPq, Bio-Value/BEECOL, UFSM]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

498
STUDYING THE EFFECT OF ARSENIC ON THE METABOLISM OF
UNICELLULAR ALGAE (C. reinhardtii)
Cícero A. Lopes Júnior (R)a,b*, Andrea Raab (R)b, Fernando Fernández-Mendoza (R)c , Martin
Grube (R)c, Jörg Feldmann (R)b
a Federal University of Piauí, Departament of Chemistry, Teresina, Piauí, Brazil, CEP 64049-550
b University of Graz, Institute of Chemistry, Graz, Styria, Austria, CEP 8010
c University of Graz, Institute of Biology, Graz, Styria, Austria, CEP 8010
*e-mail: ciceroalj@ufpi.edu.br
Arsenic (As) is a noxious metalloid universally distributed in both aquatic and terrestrial
ecosystems. However, recent studies with marine macroalgae have revealed that As undergoes
biotransformation and then has a physiological function acting on the antioxidant system 1. So,
As metabolism in algae needs to be better clarified in some aspects. Chlamydomonas reinhardtii
are unicellular green algae widely distributed in soil and fresh water which can uptake and
biotransform arsenic into different classes of metabolites2. The objective of the present study
was to investigate the effect of As on metabolism of C. reinhardtii considering essential elements
(Cu, Co, Fe, Mn, Mo, P e Zn) and polar As species. Algae were mixotrophically cultured in 500
mL Erlenmeyer flasks containing 200 mL of tris-acetate phosphate (TAP, pH 7.0) medium with
500 µg P L-1 in a growth chamber under agitation at 120 rpm at 27 °C and continuous lighting
(2826 lux) for 7 days. The growing conditions were as follows: 0 (control), 2, 10, 50 and 100 µg
As L−1 spiked as sodium arsenate solution (AsHNa2O4·7H2O). Algae growth was checked by
performing optical density measurements at 650 and 730 nm using a UV-VIS. Algae collection
from each regime was performed using centrifugation at 4500 rpm for 10 min (8 °C). The profiles
of the growth curves revealed that the exposure of algae to a concentration from 50 µg As L -1
resulted in inhibition of their growth. The algae biomass obtained in the growth period was 165
± 8 mg, 158 ± 8 mg, 151 ± 6 mg, 129 ± 4 mg and 131 ± 2 mg for the Control, As2, As10, As50
and As100 regimens, respectively. To quantify the total concentration of the elements, the algae
pellets were freezer-dried for 48 h (0.07 mbar) and digested with 4 mL of deionized water and 1
mL of HNO3 in a pressurized UltraClaveIV microwave system with 40 bar of argon. An Agilent
8900 triple quadrupole ICP-MS was used for measurements. The monitored masses were m/z
59 (Co+), m/z 95 (Mo+), m/z 91 (AsO+), m/z 47 (PO+), m/z 56 (Fe+), m/z 65 (Cu+), m/z 55 (Mn+)
and m/z 64 (Zn+). Ir (m/z 93) and Rh (m/z 103) were used as internal standard at 50 µg L-1.
Certified reference materials (CRMs) NIST 1568b, ERM CD-200 and DOLT-2 were used to
confirm the accuracy of the measurements. The total concentration of As found is related to the
algal exposure level in which the As100 regime presented the highest values (20 µg g -1). The
total concentration of P in the algae of the As50 and As100 regimes were ca. 28% and 19%,
respectively, higher than Control. Micronutrient homeostasis (Cu, Fe, Mn, e Zn) was more
sensitive to toxic effects due to algal exposure to arsenic compared to trace elements (Co e Mo).
A sequential extraction was carried out using the solvents hexane, DCM/MeOH (2:1) and water1.
The total arsenic from each step of the extraction protocol was measured for the mass balance.
Recovery values ranged from 76% to 90%. The arsenic polar compounds were the majority in
the algae of the As2 and As100 regimes, 67% and 51%, respectively. The SAX-HPLC-ICP-MS
was used for As speciation in the water fraction for algae of different regimes. Post-column
recovery was in the range of 47% to 105%. The signal of seven arsenic compounds was
detected in water extracts. As(V) is the majority species found in algae, except for the As2
regime, which showed a high concentration of As species at RT 2.1 min. The findings are
relevant to clarify the effects of arsenic on algal metabolism.
1 Pétursdóttir ÁH, Fletcher K, Gunnlaugsdóttir H, Krupp E, Küpper FC, Feldmann, J, Environmental Chemistry, 13, 2016,
21.
2 Mavrakis E, Mavroudakis L, Lydakis-Simantiris N, Pergantis SA, Analytical Chemistry, 91, 2019, 9590.
[CNPq, UFPI, UNI-Graz]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

499
A NEW APPROACH TO SCREENING ANALYSIS USING NIR SPECTROSCOPY
AND CHEMOMETRICS APPLIED TO BULK SWEETENERS
Jamille C. Souza (PG)a, Celio Pasquini (R)b, Maria C. Hespanhol (R)a*, Fábio R. P. Rocha (R)c
a Federal
University of Viçosa, Chemistry Departament, Viçosa, MG, Brazil, 36570-900
b State
University of Campinas, Chemistry Institute, Campinas, SP, Brazil, 13083-862
c Centre for Nuclear Energy in Agriculture – CENA-USP, Piracicaba, SP, Brazil, 13416-000
The consumption of natural sweeteners has been increasing worldwide to meet dietary restrictions
imposed by diabetes or low calories diets. Several sweeteners, such as erythritol, maltitol, xylitol,
maltodextrin and sorbitol are purchased in bulk. Due to their similar appearance to the naked eye
(white powder), they are prone to adulteration. The most usual technique to analyze these
sweeteners is high-performance liquid chromatography (HPLC), which is an expensive and laborious
technique.1 Thus, this work proposes a new analytical strategy based on NIR spectroscopy and
information provided by multivariate techniques and simple spectrum graphical evaluation, using a
minimum number of reference samples. The sample set were constituted of 103 bulk sweeteners,
purchased from Brazilian suppliers, contemplating most of the sources of variability (granulometry,
batch and physical appearance). Reference samples were purchased from Sigma-Aldrich with 98-
99% purity. A compact NeoSpectra micro-FT-NIR spectrophotometer (Si-Ware Systems, Egypt),
operating in the 1350 – 2550 nm range, was employed. The compact NIR spectrophotometer was
attached to a sample probing device, which simultaneously rotates and moves the sample vial
linearly over the measurement window, to obtain a representative NIR spectrum of the samples.2
The spectral data set, the PCA, the correlation analysis, MCR-ALS, and PLS models were processed
using the Unscrambler 11.0 software (Aspen Technologies). Original spectra have been smoothed
(Savitzky-Golay, 11 points, 2nd-degree polynomial) and transformed by standard normal variate
(SNV) to minimize scattering effects. Data were centered for PLS regression. MCR-ALS employed
only non-negative concentrations as a constraint and the authentic commercial sweeteners (pure)
spectra for initial algorithm guess. Figure 1 shows the PCA scores plot for the first (PC1) and second
(PC2) principal components and PC1 and PC3. Together the three PCs capture 94% of the variance
of the data set. Bulk sweeteners are grouped and promptly identified.
4
3 A B
A4
2 A4
A8 2
1 A7
PC-2 (12 %)
PC-3 (9 %)
A6
0 0
A5 A8
-1 A7
A1
-2 A2 -2 A6
A3 A5
-3
A1-3
-4
-8 -6 -4 -2 0 2 4 -8 -6 -4 -2 0 2 4
PC-1 (73 %) PC-1 (73 %)
Figure 1. PCA scores distribution obtained for the PCA of the whole sample spectra including the reference sweeteners.
(A), PC1 x PC2; (B), PC1 x PC3. A1-A8, possible counterfeit commercial samples. fructose, reference fructose,
sorbitol, reference sorbitol, sucrose, reference sucrose, stevia, sucralose, maltodextrin, reference
maltodextrin, maltitol, reference maltitol, erythritol, reference erythritol, xylitol, and reference xylitol.
Qualitative and quantitative characterization of the samples was effectively achieved. The cost of
method implementation is substantially reduced using a compact, NIR spectrophotometer, few
references samples, and avoiding expensive analytical techniques commonly employed to construct
classification and regression multivariate models. Several non-conformities of commercial bulk
sweeteners (samples A1-A8 in Fig. 1) were identified in the present study using the proposed
method.
1 Saraiva A, Carrascosa C, Raheem D, Ramos F, Raposo A, International Journal of Environmental Research and Public
Health, 17, 2020, 5227.
2 Pasquini C, Hespanhol MC, Microchemical Journal, 170, 2021, 106747.
[INCTAA, CNPq, FAPESP, FAPEMIG, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

500
NANOFIBERS PVDF/REDUCED GRAPHENE OXIDE FILMS OBTAINED BY
SOLUTION BLOW SPINNING FOR REMOVAL OF METHYLENE BLUE.
Luiz F. Malmonge (R)a*, Gabriel C. Dias (R)a, Mirian C. Santos (R)b
a Department of Physics and Chemistry (DFQ), São Paulo State University (UNESP), Ilha Solteira, São Paulo, Brazil,
15385-000
b Department of Analytical, Physical-Chemistry and Inorganic Chemistry (DQAFQI), Institute of Chemistry, São Paulo
State University (UNESP), Araraquara, São Paulo, Brazil. 14800-900

*e-mail: luiz.malmonge@unesp.br
Annually tons of toxic materials harmful to living beings are emitted by chimneys and sewage from
industries, agricultural activity, mining have reached the hydrosphere polluting water bodies around
the world. The technique used to obtain nanofibers presented in this work together with the
adsorption methodologies is promising and robust for water purification at low cost in addition to
presenting high efficiency. In this work Poly(Vinylidene Fluoride) (PVDF) nanofibers with Reduced
Graphene Oxide (RGO) were obtained by Solution Blow Spinning (SBS) from a commercial airbrush
in different incorporations for testing with removal of organic pollutants, such as the dye Methyl Blue
(MB).The concentration of methylene blue contaminants not adsorbed by the nanocomposite was
determined by UV-visible. The concentration of dye in the medium was determined using equation
1. The amount of material adsorbed per gram of composite, as well as the percentage of adsorbed
concentration of material were calculated using equations 2 and 3. The results showed the
predominance of the β phase of PVDF. The incorporation of RGO particles in the nanofibers from
the applied methodology does not change the conformation and configuration of the matrix, in
addition to good thermal stability. As for the adsorption tests, the material is potentially effective for
removing organic dyes (Table 1). The kinetic model was governed by the pseudo-second order
model, in addition to the Langmuir-Freundlich (L-F) isotherm kinetic model (Figure 1) being the most
appropriate to describe the kinetic and adsorption isotherm data, respectively, in aqueous medium,
which validates its application as an adsorbent material.
Table 1 - Kinetic results of PVDF/RGO nanofibers in aqueous medium, Figure 1 - Fit to the Freundlich-
for concentration, adsorbed mass per gram and percentage of Langmuir isotherm model. PVDF
adsorption, both as a function of time. Nanofibers with Reduced Graphene
Oxide
contact Methylene Blue(MB) 2,2
5mg RGO - MD - F -L
Sample time Concentration qeq Adsorption 2,0
(h) (ppm) (mgg-1) (%) 1,8
1,6
Pure PVDF 120 2,790 0,500 44,97
1,4
eq
PVDF/RGO 120 3,075 0,518 39,65

C1/n/q
eq
1,2
1mg 1,0
PVDF/RGO 120 3,190 0,483 40,42 0,8
3mg 0,6
PVDF/RGO 120 0,968 1,161 83,04 0,4
5mg 0,8 1,0 1,2 1,4 1,6
C1/n
eq
𝐴𝑏𝑠±𝑦 (𝐶0 −𝐶𝑡)𝑉 𝑞𝑡

𝑐𝑡 = (1); 𝑞𝑡 = (2) ; 𝑎𝑑𝑠(%) = × 100 (3)
𝑎 𝑚 𝑞𝑚𝑎𝑥
1 Chowdhury S, Balasubramanian R, Advances in Colloid and Interface Science, 204, 2014, 35.
2 Konios D, Stylianakis MM, Stratakis E, Kymakis E, Journal of Colloid and Interface Science, 430, 2014,108.
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

501
AGAROSE GEL BASED DGT DEVICES FOR SIMULTANEOUS DETERMINATION
OF CATIONIC AND ANIONIC TRACE METALS IN AQUATIC SYSTEMS
Carlos E. Eismann (PG)ª*, Edson G. O. Junior (UG)ª, Lucas P. Elias (PG)ª, José L. M. Viana
(PG)b, Melina B. T. Zanatta (R)ª, Laurent Barbiero (R)c, Anne H. Fostier (R)b, Amauri A.
Menegario (R)ª
a São Paulo State University, Environmental Studies Center, Rio Claro, SP, Brazil, 13506900
b University of Campinas, Institute of Chemistry, Campinas, SP, Brazil, 130083970
c Université P. Sabatier, Géoscience Environment Toulouse, Toulouse, France
*e-mail: kadueismann@gmail.com
DGT technique uses passive samplers that accumulate analytes in a binding phase after their
diffusion through a hydrogel. DGT devices are capable of pre-concentrate the analytes, giving time
integrated information of the labile concentration of trace elements in situ. Conventional DGT devices
use a polyacrylamide hydrogel as the diffusive layer and the Chelex 100 resin immobilized in
polyacrylamide gel as the binding phase1. Alternative binding agents are used to perform chemical
speciation (e.g. Cr, Mn), or in situations where the performance of the Chelex 100 is not satisfactory
(e.g. low pH or to determine As, Hg, Se, U)2. Some mixed binding layers were also proposed in order
to simultaneously determine cationic and anionic species3. Even tough a great number of binding
layers have been proposed, the majority of works use the polyacrylamide to immobilize the binding
agents. The present work uses a self-made diffusive layer of agarose and a mixed binding layer of
Zr oxide and Chelex 100 immobilized in agarose hydrogel to simultaneously determine the labile
concentration of cationic and anionic species of trace elements in aquatic systems. The use of self-
made binding and diffusive layers using agarose instead the carcinogenic polyacrylamide as the
hydrogel significantly reduces the costs and simplify the manufacturing process. The proposed
devices were evaluated through recovery tests and deployment curves performed in bulk solutions
containing As, B, Cd, Co, Cu, Mn, Mo, Ni, Sb, Se, V and Zn in order to establish the elution factors
and diffusion coefficients to be used in the calculations required by DGT. The performance of the
devices was evaluated in conditions of pH (5,0 – 12,0) and Ionic strength (0,005; 0,05; 0,25 and 0,5
mol L-1 NaNO3). The proposed DGT devices and conventional Chelex and Zr oxide DGT devices
were deployed in waters collected from the Ribeirão Claro River to evaluate the efficiency of the
proposed device in natural waters. The concentration of the trace elements was determined by
Inductive Coupled Plasma Mass Spectrometry (ICP-MS). The elution factors of As, B, Cd, Co, Mn,
Mo, Ni, Se, V and Zn obtained in the recovery testes ranged from 73 to 87% and were considered
satisfactory, but the recoveries for Cu (121 ± 39%) were not reproductive and for Sb (16 ± 1%) were
very low. The relationship between the accumulated mass and the time of all the studied elements
was linear for up to 24h (r² > 0,9). The diffusion coefficients obtained were consistent with the
literature4 and ranged from 2,59 to 9,16 cm² s-1. The variation in the accumulation of the As and Se
was not significant in all the pH range studied. However, a decrease in the accumulation of B, V, Mn,
Co and Ni in pH above 9 was verified, while for Se and Mo the decrease occurs above pH 10,0. The
trace metals concentrations in the experiments using 0,005 mol L-1 NaNO3 were underestimated for
the majority of the studied elements, indicating that charges that are present in the agarose gel could
influence the mass accumulation in low ionic strengths.
1 Zhang H, Davison W, Analytycal Chemistry, 67, 1995, 3391.
2 Menegario AA, Yabuki LNM, Luko KS, Williams PN, Blackburn DM, Analytica Chimica Acta, 983, 2017, 54.
3 Wang Y, Ding S, Shi L, Gong M, Xu S, Zhang C, Analytica Chimica Acta, 972, 2017, 1.
4 Wang Y, Ding S, Gong M, Xu S, Xu W, Zhang C, Analytica Chimica Acta, 945, 2016, 47.
[FAPESP]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

502
DEVELOPMENT OF A NEW PROCESS USING ULTRASOUND ENERGY FOR THE
TREATMENT OF H2S: INITIAL RESULTS
Ana Luísa S. Machado (UG)a, Milena A. Mambrin (UG)a, Karen Giacobe (PG)a
Cezar A. Bizzi (R)a, Rodrigo C. Bolzan (R)a*
a Universidade Federal de Santa Maria, Departamento de Química, Santa Maria, RS, Brasil, 97105-900
*e-mail: rodrigo.bolzan@ufsm.br
Hydrogen sulfide (H2S) is one of the main sulfur compounds present in petroleum. This compound
is a colorless gas with characteristic smell, extremely toxic and corrosive, which presents adverse
effects on human health, and on the environment. In oil, sulfur compounds as H2S are responsible
for increasing the stability of emulsions, and for contamination of catalysts.1 The processes currently
used by the oil industry to remove H2S cause a high increase in the cost of production of this material
and mainly include the use of scavenger, such as bases, oxidizing agents, metal compounds and
others. The process for the H2S treatment is problematic because usually produce harmful
compounds that damages the distillation towers and generates insoluble precipitates. 1 In addition,
ultrasound (US) can be an important tool in the development of new processes when chemical
reactions are necessary.2 Thus, this work aims to evaluate the effect of sonication on the formation
of acids from H2S.These acids could be easily separated and eliminated. In this work, H2S generated
from thioacetamide decomposition (10 mL of 1 mol L-1 thioacetamide, in an acidic medium), is
bubbled into a special designed reactor containing 20 mL water in the presence/absence of O3 (also
bubbled into the reactor - from a O3 generator). The reactor was sonicated using several US systems,
including the indirect (baths and cup horn), and the direct (probes) sonication of the reaction medium,
with different frequencies (20 to 130 kHz) and nominal powers (100 to 750 W), during 15 minutes at
40 oC. The formation of sulfur containing acids was monitored in the end of process. The acid
formation was monitored using volumetric titration with NaOH solution and the forming sulfur
containing species were evaluated by ICP-OES and ion chromatography. The sonication using baths
in the frequency of 25 kHz increased the acid formation leading to a consumption of 14.55 ± 0.05
mg NaOH mL-1 (mean ± standard deviation) for neutralization (the blanks consumed only 0.04 mg
NaOH mL-1). In this condition the individual bubbling of H2S was sufficient for acid formation. It is
important to point that the addition of O3 into the medium did not increase the acid formation. The
sonication using cup horn systems (20 kHz/130 W and 20 kHz/750 W) and probes (20 kHz/130 W
and 20 kHz/750 W) did not significantly increased the acid formation. The sulfur contents in the
reaction medium, determined using ICP-OES, were higher when only H2S was bubbled, and US was
applied using the baths (6.5 mg L-1 at 45 kHz) and the cup horn (20 kHz/750 W) (7.6 mg L-1) systems
(S content if no sonication was applied = 2.7 mg L-1). If O3 was also bubbled in the medium the total
amount of sulfur decreases. In addition, the results obtained after analysis using ion chromatography
showed that the bubbling of H2S + O3 led to a major SO42- formation when all the sonication systems
were applied (for example, SO42- concentration was 4.2 ± 0.8 mg L-1 / probe 20 kHz/130 W – if no
sonication was applied SO42 concentration was 2.4 ± 0.9 mg L-1). These results allow to conclude
that the sonication can be an important component in the development of a new process dedicated
to the H2S conversion treatment.
1 Thomas JE, Fundamentos de Engenharia de Petróleo, Interciência, 2ª edição, 2004.
2 Mason TJ, Sonochemistry: The uses of ultrasound in chemistry, Royal Society of Chemistry, 1990.
[ANP, PRH 52.1, UFSM]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

503
INFLUENCE OF SILICON IN MITIGATION OF STRESS BY Cd2+ IN TRANSGENIC
SOYBEAN LEAVES: A METABOLOMIC APPROACH BY NMR
Amanda L. Quintela (PG)a,b*, Daniel C. Freitas (PG)b, Maria F. C. Santos (R)a, Cláudio F.
Tormena (R)a, Marco A. Z. Arruda (R)b
a Laboratory of Physical Organic Chemistry, Institute of Chemistry, University of Campinas, Campinas, SP, Brazil, CEP
13083-970
b Spectrometry, Sample Preparation and Mechanization Group, Institute of Chemistry, University of Campinas,

*e-mail: a234902@dac.unicamp.br
Brazil is the world's leader of soy (Glycine max) producer with a production of 124 million tons
2021/2022, which ranks it as one of the main commodities in the world1. In this context, the levels of
environmental contaminants have increased and impacted soil fertility and grain yield. Cd 2+ is an
element that presents high toxicity to organisms even at low concentrations, being considered an
environmental contaminant that negatively affects plant growth, interruption of physiological
processes and causes changes in plant metabolism. On the other hand, silicon (Si) is an element
known to mitigate several types of abiotic and biotic stresses2. The goal of this work was to
investigate the mitigation responses of Si on the effects of Cd2+ on transgenic soybean leaves
applying 1H NMR-based metabolomics approach. For this, two varieties of transgenic soybean seeds
(Roundup Ready® and Intacta) obtained through a process of genetic modification by inserting the
cp 4 EPSPS gene were cultivated with four different treatments: TC (control, without addition of Si
or Cd2+), T1 (with addition of Si), T2 (with addition of Cd2+) and T3 (with addition of Si and Cd2+). The
metabolite extraction procedure was adapted from the method developed by Kim et al. (2010) 3, in
which 40 mg of soybean leaves were frozen and sprayed with N2(l) and subjected to microextraction
in 1 mL of CD3OH (0.05% TMS). The extracts were homogenized for 1 min by vortexing. Then, the
samples were centrifuged for 5 min at 12000 rpm. A 600 μL aliquot of the supernatant was
transferred to the 5 mm NMR tube. The 1H and 2D NMR spectra were acquired on a Bruker AVANCE
III NMR spectrometer, operating at 14.1 T, where 1H is observed at 600.13 MHz and 13C at 150 MHz
equipped with a TBI probe. The 1H NMR experiments were performed at 298 K with the pulse
sequence (zgpr) using the following parameters: spectral window (SW) of 12019,230 Hz (~ 20 ppm),
number of points (TD) 64k, delay time between the pulses (D1) 1s and 32 scans. Metabolites were
identified in comparison with the literature and with direct 1H-13C (HSQC) and long-distance 1H-13C
(HMBC) correlation 2D NMR experiments. From the analysis of the 1H NMR spectral profiles of
soybean leaves, it was possible to observe that both genotypes (RR and Intacta) presented
qualitatively similar profiles, being identified 32 chemical compounds, highlighting the following
classes: amino acids, fatty acids, organic acids, sugars and polyphenols. In this way, a comparative
study of the chemical profiles of the two transgenic soybean varieties will be carried out from leaves
to roots and stems to investigate the influence of Cd2+ stress on soybean metabolic pathways.
1 CONAB - Companhia Nacional de Abastecimento. Acompanhamento da Safra Brasileira de Grãos, Brasília, DF, v. 9,
safra 2021/22, n. 10 décimo levantamento, julho 2022. Disponível: https://www.conab.gov.br/info-
agro/safras/graos/boletim-da-safra-de-grãos. Acesso em 07/07/2022.
2 Carneiro JMT, Chacón-Madrid K, Galazzi, RM, Campos BK, Arruda SCC, Azevedo RA, Arruda MAZ, Journal of Trace
Elements in Medicine and Biology, 44, 2017, 50.
3 Kim HK, Choi YH, Verpoorte R, Nature protocols, 5, 2010, 536.
[CAPES, FAPESP, INCTBio]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

504
COMPARATIVE STUDY OF ASPHALT AGING BASED ON THE MOLECULAR
FLUORESCENCE EMISSION OF MALTENIC AND ASPHALTENIC FRACTIONS
Kezia de Souza (UG)a, Patrícia Schmidt (PG)a, Leandro Carvalho (R)a
a Federal University of Santa Maria (UFSM), Department of Chemistry, Santa Maria, RS, Brazil, 97105-900
*e-mail: keziadesouza@hotmail.com
It is well-known that asphalt can be separated into two major fractions known as asphaltenes
and maltenes. The asphaltene fraction is the most complex fraction of the asphalt, with the
highest polarity and molecular weight, composed mainly of aromatic chains and molecular
aggregates with heteroatomic functional groups1. On the other hand, the maltene fraction is
soluble in light alkanes and, compared to asphaltenes, presents the lower molecular weight
hydrocarbons, which can be additionally separated into resins, saturated and aromatic
fractions2. The asphalt analysis using fluorescence techniques is used for sample
characterization and identification, since the fluorescence of the samples derives from the
aromatic hydrocarbon fraction, and the fluorescence emission is strongly influenced by the
chemical composition and physical characteristics. Keep it in mind, fluorescence spectroscopic
studies of asphalts and its fractions can provide a solid base for the identification of these
structures. In this work, the maltenic and asphaltenic fractions were analyzed by UV-
fluorescence using a series of solvents with different polarities for evaluating firstly the
differences in solubility and fluorescence characteristics. According to Figure 1, it can be seen
that the fraction of maltenes presented the highest signal intensity for all solvents analyzed,
followed by the whole asphalt and finally the fraction of asphaltenes. The decrease in the
fluorescence of the whole asphalt sample compared to the maltenic fraction suggests that the
presence of asphaltenes can decrease the fluorescence of the maltenic fraction3. Besides that,
it is possible to notice a shift in the emission peak between the asphalthenic and maltenic
fractions. While for the asphalt and maltenic fraction the emission maximum is located at around
400 nm, for the asphaltic fraction it shifts to higher wavelengths. The spectra shift to higher
wavelengths (>400 nm) can be related to the presence of longer chain hydrocarbons 4.
Furthermore, spectra shifts to lower wavelengths (~400 nm) can be related to a decrease in the
proportion of saturates to aromatics and an increase in the proportion of resins and
asphaltenes5. The maximal emission of both asphaltenic and maltenic fractions were used to
establish a new aging index for asphalts based on the decrease of fluorescence emission of
asphaltenes and maltenes. The aging index was calculated after performing the thermal- and
pressure-based aging processes of 4 asphalt samples using standardized methods (RTFOT
and RTFOT/PAV methods).
Figure 1. Fluorescence spectra of total asphalt, asphaltene and maltene fractions in 14 different solvents analyzed.
1 Subramanian S, Simon S, Sjöblom J, Journal of Dispersion Science and Technology, 37, 2016, 7.
2 Ashoori S, Sharifi M, Masoumi M, Salehi MM, Egyptian Journal of Petroleum, 26, 2017, 209.
3 Wang F, Xiao Y, Cui P, Lin J, Li M, Chen Z, Construction and Building Materials, 250, 2020, 118824.
4 Ryder AG, Reviews in Fluorescence, 2007.
5 Stasiuk LD, Gentzis T, Rahimi P, Fuel, 79, 2000, 7.
[CAPES, CNPq, Petrobras]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

505
SOLID-PHASE EXTRACTION FOR SPECTROPHOTOMETRIC
DETERMINATION OF MERCURY IN WATERS AND URINE USING A
DITHIZONE-LOADED POLY(SMA-CO-EDMA) MONOLITH
Fernando H. do Nascimento (R)a, Jorge C. Masini (R)a
a Universidade de São Paulo, Instituto de Química, São Paulo, SP, Brazil, 05508-000
*e-mail: fernandoslifer@gmail.com
Mercury and mercurial compounds are highly toxic species that bioaccumulate in the food chain,
causing several health problems for living beings, including humans, even at low concentrations.
Several methods employ spectroscopic techniques1 for determining Hg but involve high
instrumentation and operation costs. UV-vis measurements are simple and robust, relying on
portable instrumentation widely available in research and industrial laboratories2. Molecular
absorption spectrophotometry reaches satisfactory sensitivities for Hg(II) if preconcentration steps
involving liquid-liquid (LLE) or solid-phase extractions (SPE) are undertaken. Conventional LLE
procedures use toxic solvents, are laborious and time-consuming, not being compatible with field
analyses. Polymeric monoliths are well-established porous materials used in SPE procedures3. This
work proposes the fast preparation of a dithizone (DTZ) modified C18 polymeric monolithic column
synthesized inside polypropylene ink-pen tubes by photo copolymerizing stearyl methacrylate (SMA)
with ethylene glycol dimethacrylate (EDMA) in the presence of n-propanol and 1,4 butanediol
(porogens). The resulting poly(SMA-co-EDMA) monolith retains DTZ, which concentrates mercury
from aqueous media, followed by spectrophotometric detection of the mercury-dithizone complex at
460 nm after elution with ethanol. Limits of detection (LOD) and quantification (LOQ) were 7.9 and
26 µg L-1, respectively. The adsorption capacity for a 40 × 2 mm poly(SMA-co-EDMA) column was
7.8 µg g-1 (94 µg m-1), whereas a 25 mm column performed at least ten cycles of SPE with
quantitative recoveries of about 100%. The enrichment factor was about 10 (83% recovery), which
was estimated by comparison between absorbance measurements for a solution with (150 µg L-1)
and without preconcentration (1,8 mg L-1). EDTA circumvented potential interferences of Cu2+, Cd2+,
Pb2+, Zn2+ and Fe3+, and the variation observed for the measurement in the absence and presence
of these metals was less than 6.8%. Furthermore, chloride, humic and fulvic acids caused variations
of -3.3%, +2.5% and -7.5% in the Hg(II) measurements, respectively, suggesting a greater affinity
between Hg(II) and dithizone than the other complexing/chelates species. Spiking and recovery
studies evaluated the method's accuracy by using external calibration in water and urine samples
with and without enrichment. Samples enriched with 30, 60 and 90 µg L-1 Hg(II) exhibited recoveries
greater than 90%, whereas samples analyzed without enrichment had Hg(II) concentrations lower
than the LOQ. The poly(SMA-co-EDMA) column was of rapid preparation (20 min) and exhibited
permeability (2 x 10-13 m2), enabling the sample loading (12 mL) at 1.0 mL min-1 assisted by a syringe
pump. The proposed methods have the potential for field analyses, given the procedures simplicity
and the instrumentation portability. It attends to the demand for cleaner analytical methods using
reduced amounts of dithizone (6.8 nmol per column) and ethanol for elutions, thus avoiding toxic
solvents such as the CCl4 classically used in ancillary dithizone-based spectrophotometric methods
for Hg(II).
1 Kozaki D, Mori M, Hamasaki S, Doi T, Tanihata S, Yamamoto A, Takahashi T, Sakamotoc K, Funado S, Analytical
Methods, 13, 2021, 1106.
2 Hatiboruah D, Das T, Chamuah N, Rabha D, Talukdar B, Bora U, Ahamad KU, Nath P, Measurement, 154, 2020, 107507.
3 Namera A, Nakamoto A, Saito T, Miyazaki S, Journal of Separation Science, 34, 2011, 901.
[CAPES, CNPq]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

506
ANALYSIS OF PAH AND DERIVATIVES IN SURFACE WATER BY GCxGC/Q-
TOFMS/MS USING A NEW PASSIVE SAMPLER
Nathan S. Coelho (PG)a*, Nayara S. Fraga (PG)a, Helvécio C. Menezes (R)a, Zenilda L.
Cardeal (R)a
a Universidade Federal de Minas Gerais, Department of Chemistry /ICEx, Belo Horizonte, M.G., Brazil, 31270-901
*e-mail: nathancoelho@ufmg.br
The assessment of water resources contamination is usually done through conventional sampling,
which commonly consumes time, financial resources and does not allow the detection of
contamination episodes over a long period1. Valenzuela et al1, proposed a simple device for
pesticides passive sampling using the hollow fiber liquid phase microextraction (HF-LPME)
technique. Besides, Dos Santos used solid phase microextraction with polydimethylsiloxane, PDMS,
for the analysis of PAHs and derivatives2. Thus, the objective of this work was to develop new PDMS
fibers for HF-LPME passive sampling analysis of PAHs and its oxygenated and nitrated derivatives
in surface water with determinations by GCxGC/Q-TOFMS/MS.The comprehensive two-dimensional
gas chromatography system comprises a GCxGC Agilent system model 7890 coupled to an Agilent
7250 Q-TOF-MS/MS, equipped with a split/splitless injector, an GC autosampler 80 and a Zoex ZX2
thermal modulator. Separation was performed using HP-5MS UI column in 1D and DB-17HT column
in 2D. All work was based on 16 PAHs, 4 Oxi-PAHs and 4 Nitro- PAHs.
Figure 1. Process of production, cleaning and

extraction of the analytes with the HF-LPME.
Figure 2. Color diagram of a standard solution of 16 PAHs and 8
derivatives: 1_Naphtalene, 2_Acenaphthleny, 3_Acenaphthene,
4_Fluorene, 5_9-Fluorenone, 6_Phenanthrene, 7_Anthracene, 8_9.10-
Anthraquinone, 9_Fluoranthene, 10_2-Methylanthra-quinone, 11_2-
Nitrofluorene, 12_Pyrene, 13_9-Nitroantracene,
14_Benzo[a]anthracene,15_Chrysene, 16_3-Nitrofluoranthene, 17_1-
Nitropyrene, 18_5,12-Nafthacenequinone, 19_Benzo[b]-fluoranthene,
20_Benzo[k]f-luoranthene, 21_Benzo[a]pyrene, 22_Indeno[123-
c,d]pyrene, 23_Dibenzo[a,h]anthracene, 24_Ben-zo[g,h,i]perylene.
A calibration study for the passive sampling of analytes using the new sampler was also performed
and the parameters of method validation were tested following Eurachem Guide.
The new HF-LPME fiber showed good results for passive sampling of PAHs and derivatives in
surface waters. The developed GCxGC/Q-TOFMS method showed good performance for
environmental determination.
1 Valenzuela EF, Menezes HC. Cardeal ZL Environmental Chemistry Letters, Springer, 2020.
2 Santos RR, Leal LDV, Cardeal ZL, Menezes HC, Journal of Chromatography A, 1584, 2019, 64.
[CEMIG, CNPq]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

507
COLORIMETRIC METHOD BASED ON DIGITAL IMAGES FOR THE
DETERMINATION OF THE DITHIOCARBAMATES USING SMARTPHONE
Fernanda C. O. L. Martins (R)a* and Wanessa R. Melchert (R)b
a Center for Nuclear Energy in Agriculture, University of São Paulo, Piracicaba, SP, Brazil.
b College of Agriculture “Luiz de Queiroz”, University of São Paulo, Piracicaba, SP, Brazil.
*e-mail: fernandacolm@hotmail.com
The development of the analytical techniques for the control of pesticides, mainly dithiocarbamates,
in the food samples is very important. They were prohibited by the European Union, the United
States, and Codex Alimentarius because of their adverse effects (neurotoxic and endocrine effects,
Parkink’s diseases, and cancers, among others) on human health with the high ingestion them 1. In
this context, the purpose of the work was the development of the methodology for the determination
of the dithiocarbamates using digital image analysis by colorimetric reaction of dithiocarbamates and
copper (Cu2+) for the quantification of it in food samples. The analytical responses of the digital image
analysis and spectrophotometer were very similar, permitting the determination of the
dithiocarbamates by canal B. Preliminary tests evaluated the preconcentration using a dodecanol
and cloud point (7% (m/v) Triton X-114), demonstrating that the use of cloud point pre-concentrated
the reduced dithiocarbamate on the surfactant rich-phase, which was analyzed directly. The cloud
point preconcentration optimization was made 27-3 factorial designs (screenings) to the concentration
of solution EDTA, Cu2+, and Triton X-114, Triton X-114 volume, bath time, centrifugation speed, and
time, where was demonstrated that only the concentration of solution EDTA and Triton X-114 volume
had significant effects. The concentration of Cu2+ and Triton X-114, centrifugation speed and time,
and bath time were 80 mg L-1; 6.0% (m/v); 2500 rpm; 10 min, and 10 min, respectively, to be quick
and require a small quantity. Posteriorly, the 22 factorial designs with five central and axial points
were made of mode randomized order. The conditions were concentration of solution EDTA (x1) from
0.1 to 2.5 mmol L-1 and Triton X-114 volume (x2) from 192 to 1057.5 µL, only the coefficients x0 and
x22 were significant. The proposed model was expressed by the equation (Analytical signal = 10.25
– 2.00x222), and obtained the response surface, as presented in Figure 1. The proposed model was
used to calculate the concentration of solution EDTA and Triton X-114 volume of 1.30 mmol L-1 and
625 µL, respectively. The analytical curve had a
linear range from 1.00 to 30.00 µmol L-1
dithiocarbamate (Analytical signal = 1.13C (µmol
L-1) + 122.65) (R = 0.9954), and a detection limit
of 0.33 µmol L-1. It was calculated by the ratio
between the slope of the analytical curve in the
extracting phase and the initial slope of the
analytical curve the enrichment factor of 10.75,
which was very similar to the expected due to
1000 µL preconcentration to approximately 100
µL. Thus, with the methodology proposed was
possible determine the dithiocarbamates using Figure 1 - Response surface and level curves for
optimization of the solution EDTA concentration and
digital image analysis by colorimetric reaction of Triton volume of the preconcentration for the
dithiocarbamates and copper, showing that the dithiocarbamates determination using digital image
method is simple, quick, and follows the guidelines analysis.
of the Green Analytical Chemistry.
1 Martins FCOL, Batista AD, Melchert WR, Comprehensive Reviews in Food Science and Food Safety, 20, 2021.
[CAPES, FAPESP, CENA]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

508
SELENIUM INFLUENCE AGAINST MERCURY UPTAKE IN EDIBLE
MUSHROOMS: BIOACCUMULATION AND Hg SPECIATION
Aline P. de Oliveira (PG)a,b*, Juliana Naozuka (R)b, Julio A. Landero-Figueroa (R)a
a University of Cincinnati, Department of Chemistry, Cincinnati, OH, USA, 45221
b Universidade Federal de Sao Paulo, Department of Chemistry, Diadema, SP, Brazil, 09972-270
*e-mail: apoliveira@unifesp.br
Mushrooms are bioaccumulators and have been used as bioindicators as well as bioremediators of
environmental pollution, while nutritional proposals seek to produce mainly Se-enriched edible
mushrooms.1,2 Since mushrooms can bioaccumulate both potentially toxic and essential elements,
cultivation strategies development must be proposed in terms of food safety, aiming to reduce toxic
elements bioaccumulation and/or favor toxic species interconversions to less harmful species.3
Studies have shown that Se plays effective roles against Hg uptake, bioaccumulation, and toxicity
in several organisms due to Se-induced interconversion of toxic species as well as inhibition of Hg
translocation.3,4 Therefore, the aim of this study was to evaluate the protective role of different Se
species (Se(IV): SeO32- and Se(VI): SeO42-) against Hg in white oyster (Pleurotus ostreatus) and pink
oyster (Pleurotus djamor) mushrooms in order to provide a better understanding of Hg-Se
interactions in edible mushrooms. For this, the effects of supplementing the growth medium with
different levels of Se(IV) or Se(VI) (12.8 and 51.2 µg g-1) and/or Hg (2.4 and 9.6 µg g-1) were
evaluated in terms of fruiting bodies morphology, Hg and Se bioaccumulation, protein-bound Hg and
Se distribution as well as Hg(II) and MeHg distribution. Microwave-assisted micro-scaled acid
digestion was carried out and Se and Hg were determined by ICP-MS. Afterwards, proteins and Hg
species extractions were performed followed by SEC-UV-ICP-MS and HPLC-ICP-MS analysis,
respectively. Then, it was possible to evaluate Se- and Hg-proteins distribution and also to perform
speciation analysis of Hg. Pink oyster mushrooms showed greater potential for Hg uptake and
bioaccumulation as well as greater ability to methylate Hg(II) from growth substrate, in which MeHg
corresponded from 2.0 to 25.3% of the total Hg. As far as the antagonism between Se and Hg was
concerned, Se(IV)-induced protective effect was found to be more effective than Se(VI) in terms of
Hg incorporation for both mushroom species, decreasing up to 96% the total Hg concentration. In
this way, addition of Se(IV) (51.2 µg g-1) to Hg-contaminated growth medium (2.4 µg g-1) produced
mushrooms with Hg concentration as low as 0.46±0.03 µg g-1 and Se concentration as high as 193±1
µg g-1 in the edible part of the fungus. Furthermore, Se-induced inhibitory effects on Hg methylation
were also observed, MeHg species content decreased up to 100% in mushrooms exposed to Se(IV)
(51.2 µg g-1). Additionally, it was also found that supplementation, mainly with Se(IV) reduced the
fraction of Hg bound to medium molecular weight compounds (17 - 44 kDa) up to 80%. On the other
hand, the results do not suggest Hg-Se-protein complex formation in these mushroom fruiting
bodies. In conclusion, the addition mainly of Se(IV) to potentially Hg-contaminated growth substrates
proved to be promising and deserves to be highlighted in terms of food safety by reducing Hg in the
food chain in addition to favoring the production of Se-enriched edible mushrooms, simultaneously.
1 Kapahi M, Sachdeva S, Bioresources and Bioprocessing, 4, 2017, 1.
2 Oliveira AP, Naozuka J, Landero-Figueroa JA, Food Chemistry, 383, 2022, 132582.
3 Wang X, Wang WX, Environmental Pollution, 231, 2017, 1543.
4 Tang W, Dang F, Evans D, Zhong H, Xiao L, Chemosphere, 169, 2017, 369.
[FAPESP, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

509
PHYSICAL-CHEMICAL ANALYSIS AND SPECTROPHOTOMETRIC
PROFILE OF PITAYA (HYLOCEREUS POLYRHIZUS)
Lendeson B. S. Sousa (UG)a,b, Jonas O. Vinhal (R)*a
a Instituto
Federal de Educação, Ciência e Tecnologia do Maranhão, Campus Açailândia, Avenida Projetada, s/n, Vila
Progresso II, Açailândia, Maranhão, Brazil, 65930-000
b Universidade Federal do Maranhão, Campus Imperatriz, Pós-Graduação em Ciência dos Materiais, Rua Urbano
Santos, s/n, Centro, Imperatriz, Maranhão, Brazil, 65900-410

*e-mail: jonas.vinhal@ifma.edu.br
The red Pitaya (Hylocereus polyrhizus) is an exotic and little-known fruit. It is known by different
names, such as dragon fruit, and is named according to some of its observed physical
characteristics. The fruit has been gaining prominence around the world due to its potential as a
source of vitamins and minerals1. From this perspective, this work had two main objectives. The first
consisted of performing a physical-chemical analysis of Pitaya trough the determination of moisture
and ash contents, water activity (WA), total soluble solids (TSS) and titratable acidity. The second
was to study the spectrophotometry profile of extracts obtained from Pitaya pulp in different solvents.
The Pitaya fruits were obtained in local market in the city of Açailândia – MA. After acquisition of the
samples, it were carried to the Chemistry Laboratory of IFMA for cleaning and adequate storage at
time of 0 (T0), 7 (T1), 14 (T2), 30 (T3) and 60 days (T4) to -19 ºC for further analysis. The physical-
chemical analyses followed the standards of the Adolf Lutz Institute2 and were performed at different
storage times. The spectrophotometric profile analyzes were performed by experiments with pulp
extracts obtained by solubilisation in water, ethanol and acetone solvents, and later analyzed in a
UV-Vis spectrophotometer in a range of 380-800 nm. The obtained results revealed that there were
no differences between the storage periods for the physical-chemical characterization, such shown
in Table 1. The highest average value found for moisture was 87.43%. For the ash content, variations
from 0.53 to 0.77% were noted. The water activity revealed similar values for all storage times,
between 0.931 and 0.934. As for the highest and lowest averages found for the TSS content, a total
of 17 ºBrix was obtained to T0 (fresh fruit) and 9.70 ºBrix to T4 (60 days of freezing), respectively.
With regard to acidity results, the highest average obtained was 0.35 mg of citric acid/100 g of
sample. Regarding the spectrophotometric profile, Pitaya showed a better affinity for water and
ethanol solvents, since there is a high content of compounds with polar groups in betacyanins,
present in large amounts in Pitaya, as well as higher water content. The wavelength of maximum
absorption in these solvents was the same under different dilution conditions, with value around 532
nm. Acetone proved to be unfeasible for the procedure, since it did not completely solubilised the
fruit pulp, making impossible an adequate analysis by spectrophotometric.
Table 1. Physical-chemical analysis of Pitaya (Hylocereus polyrhizus).
Freezing storage
Parameters*
T0 T1 T2 T3 T4
85.83 ± 87.27 ± 87.37 ± 85.54 ±
Moisture (g/100g (%)) 87.43 ± 2.444
0.003 0.003 0.002 0.005
Ash (g/100g (%)) 0.53 ± 0.001 0.56 ± 0.000 0.77 ± 0.000 0.70 ± 0.000 0.70 ± 0.000
0.934 ± 0.931 ± 0.932 ± 0.934 ±
WA 0.931 ± 0.001
0.001 0.000 0.001 0.001
TSS (ºBrix) 17 ± 0.000 10 ± 0.006 10 ± 0.006 10 ± 0.006 9.70 ± 0.006
Acidity
0.27 ± 0.000 0.31 ± 0.000 0.35 ± 0.000 0.32 ± 0.000 0.31 ± 0.000
(mg of citric acid/100 g)
Note: T0 (fresh fruit); T1 (7 freezing days); T2 (14 freezing days); T3 (30 freezing days); T4 (60 freezing days).
*Average content ± standard deviation.
1 Nunes EN, Sousa ASB, Lucena CM, Silva SM, Lucena RFP, Alves RE, Gaia Scientia, 8, 2014, 90.
2 Métodos químicos e físicos para análise de alimentos, 4ª edição, IAL editora, 2008.
[IFMA, FAPEMA, CAPES, CNPq]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

510
3D PRINTED MEASURING DEVICES FOR QuEChERS METHOD SORBENTS
Tainá M. Saúgo (PG)a, Marlon C. M. Guimarães (UG)a, Lenise G. de Oliveira (R)a, Márcia H.
Kurz (R)a , Fábio F. Gonçalves (R)a, Manoel L. Martins (R)a*
a Federal University of Rio Grande – Campus Santo Antônio da Patrulha – R. Barão do Cahy, 125 - Cidade Alta, Escola
de Química e Alimentos, Rio Grande do Sul, Brazil, 95500-000
*e-mail: mlmartins@furg.br
In modifications of the QuEChERS method, sorbents such as C18, PSA, active carbon, Z-sep and
others are used in varying amounts according to the matrix under study. The type and amounts of
sorbents are usually optimized for efficient extraction and removal of interferents1. Routine use of a
procedure based on the QuEChERS method requires to exactly weigh the adsorbents in varying
amounts, according to the type of sample and optimized method conditions demanding labor and
time to weigh, with greater manipulation of the sorbents (particle size of 40 µm)3 and use of analytical
balance. In order two minimize these drawbacks, the aim of this work is to develop simple and
customizable devices to accurately measure the amounts of sorbents. The prototypes were designed
with FreeCAD ver. 0.18 (The FreeCAD Team) and 3D printed (Model CL-1 Black Edition Printer,
Cliever, Porto Alegre, Brazil) using polylactic acid (PLA) filament with a diameter of 1.75 mm. The
apparent densities were determined for C18 (Supelco, Lot 574608, 50 μm particle size): 480 ± 7 mg
cm-1, PSA (Supelco, Lot 5739201, 50 μm p. s.): 503 ± 12 mg cm-1 e Zsep (Supelco, 50 μm p. s.):
575 ± 6 mg cm-1 (n=3) by weighing the quantity measured on an analytical balance (Shimadzu,
ATX224). A spatula (measuring device) sized to obtain 100 mg of PSA (Figure 1) resulted in an
average mass of 104 ± 4 mg, n=6.
A
Figure 1. (A) 3D CAD model and (B) 3D printed spatula.

B
Spatulas for 230 mg of C18, 184 mg of PSA, and 184 mg of Zsep, quantities needed to determine
pyrethroid residues in fish by QuEChERS modified method2, give averages masses ± standard
deviations of 237 ± 9,5 mg for C18, 184 ± 1,9 mg for PSA, and 184 ± 3,4 mg for Zsep (n=20), with
relative average errors of 3,0%, 0,7% and 0,3%, respectively. For comparison, an analytical balance
read at mg units (fourth decimal = uncertainty plate) gives 0,43%, 0,54% and 0,54% for the same
masses, respectively. During a validation of the analytical method2, parallel series of spiked Trahira
samples, at 5, 10, and 75 μg kg-1 were prepared with sorbents measured with these spatulas. After
GC-MS determination, the trueness was evaluated through recoveries for accuracy and RSD for
precision. Recoveries results ranged from 75 to 112% with RSD ranging from 3.7 to 22,8%, when
sorbents masses were weighed with an analytical balance and ranged from 77 to 116% with RSD
ranging from 1.6 to 19.9%, when sorbents amount were measure with 3D printed spatulas. In both
cases the recoveries results are considered satisfactory and equivalent, within the acceptable range
of 70 to 120%. As the spatulas are easily customizable through software resizing and 3D printing for
each case, the results indicate the possibility of its practical use for variable amounts of C18, PSA
and Zsep in the application of optimized methods in laboratory routine, with less labor, time and
chemical materials handling.
1 Zanella R, Prestes OD, Adaime MB, Martins, ML, QuEChERS In: Borges KB, Figueiredo EC, Queiroz MEC, Preparo de
amostras para análise de compostos orgânicos, LTC, 2015.
2 Oliveira LG, Kurz MHS, Guimarães MCM, Martins ML, Prestes OD, Zanella R, Ribeiro JNS, Gonçalves FF, Food
Chemistry, 297, 2019, 124944.
3 Cabrera LC, Martins ML, Primel EG, Prestes OD, Adaime MB, Zanella R, Scientia Chromatographica, 4, 2012, 227.
[CAPES, PPGQTA, PRAE-FURG-SAP]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

511
5-HYDROXYMETHYLFURFURAL DEGRADATION BY OZONATION IN APPLE
JUICE
Alessandra C. Romero (R)a*, Pedro E.D. Augusto (R)b, Wanessa R. Melchert (R)a
a College of Agriculture “Luiz de Queiroz”, University of São Paulo, Piracicaba, SP, Brazil.
b Université Paris-Saclay, CentraleSupélec, Laboratoire de Génie des Procédés et Matériaux, Centre Européen de
Biotechnologie et de Bioéconomie (CEBB), Pomacle, France.
*e-mail: acromero@usp.br
Ozonation is a technique widely used for food and feed decontamination and for the increasing shelf-
life of food products.1 5-Hydroxymethylfurfural (5-HMF) is a toxic compound formed after the thermal
processing of foods that contain reducing sugars such as fructose.2 Thus, thermally processed fruit
juices can present 5-HMF at varying levels, depending on the intensity of thermal treatment. 2 This
study aims to evaluate the effectiveness of ozonation in eliminating 5-HMF in apple juices. The
ozonation time (0 – 45 min) was performed by adding a high concentration of 5-HMF standard (2.975
± 63 µg mL-1) in apple juice and in pure water (0 up 30 min). To assess whether the profile of the
chromatogram eluted peaks of apple juice samples would show variations over the ozonation time
and also as the effect of the 5-HMF concentration would change the efficiency of ozonation, a sample
of apple juice containing a low native concentration of 5-HMF (1 µg mL-1) was ozonated (0 up 30
min). Similarly, an apple juice sample was submitted to O2 flux to verify its effect on 5-HMF. Ozone
processing was performed using a glass reactor under magnetic stirring coupled to an ozone
generating unit (Ozone & Life, O&G Model RM 3.0, São José dos Campos, Brazil), and a monitor to
measure ozone concentrations (2B Technologies Inc., 106-H). The gas flow was 1.0 L min-1 and the
ozone concentration was 52 mg L-1. Aliquots (1 mL) of apple juice were filtered (45 µm pore) before
the liquid chromatography development (Agilent 1100, UV 280 nm, ODS-Zorbax 250 x 4.5 x 5 id).
Figure 1. Effect of ozonation on 5-HMF reduction (left side); peaks profile (right side): apple juice (green line), apple juice
after ozonation (blue line, 45 min of treatment), and 5-HMF peak before ozonation (highlighted in blue, 0 min).
The O2 treatment showed no effect on the 5-HMF concentration. Ozonation was effective in
degrading 100% of 5-HMF in apple juice. The native concentration of 5-HMF in apple juice (1 µg mL-
1
) was faster degraded by ozonation (20 min) than the high concentration added to the sample (25
min) (Figure 1, left side). In general, the chromatograms of apple juice showed a reduction of eluted
peaks at long ozonation time regarding before the process (Figure 1, right side). The ozonation
process proved to be efficient for the degradation of 5-HMF due to the peak reduction in 6.29 min.
1 Pandiselvam R, Subhashini S, Banuu Priya EP, Kothakota A, Ramesh SV, Shair S, Ozone: Science & Engineering, 41,
2019, 17.
2 Czerwonka M, Opilka J, Tokarz A, European Food Research Technology, 244, 2018, 11.
[CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

512
Mn-FIBERS AND Mn-CARTRIDGES: COMPARISION OF 224Ra RETENTION
EFFICIENCY
Alessandra G. da Costa(PG)a*, Mariele L. de Paiva(R)a, Taís B. de Oliveira(UG)a, Renata R. de
Moura (R)a, Carlos F. F. de Andrade (R)a
a Universidade Federal do Rio Grande - Campus Carreiros – Avenida Itália, Km 08, Instituto de Oceanografia, Rio
Grande, RS, Brasil, CEP 96201-900
*e-mail: ale_gcosta@hotmail.com
Natural radium isotopes, due to their diversity in half-life times (223Ra = 11.4 days; 224Ra = 3.6 days;
226
Ra = 1600 years; 228Ra = 5.7 years) are indispensable for understanding processes environmental
factors (e.g. submarine groundwater discharge in coastal zones)1. Making necessary for
radioanalitical analises as Radium Delayed Coincidence Counting (RaDeCC) the development fibers
and cartridges chemically modified by the reduction of KMnO4 to MnO2, called reaction impregnation,
for pre-concentration the analyte from the interaction and electrostatic frames on the surface of the
adsorbent2. Therefore, the objective of the present work was to apply the impregnation method3 for
cellulose cartridges and polyacrylonitrile fibers and compare the 224Ra isotope retention efficiencies
between polymers and the influence on reaction times. The polymers, previously washed with
ultrapure H2O (Milli-Q® resistivity < 18.2 MΩ cm-1), were impregnated with KMnO4 (0.5 mol L-1; 20°C)
for reaction times of 24, 48 and 68 hours. Afterwards, the polymers were removed from the solution,
leaving them to dry on absorbent paper (20°C; 12 hours). Subsequently, washing was carried out
with ultrapure H2O (pH 8.5) and then were lyophilized to constant weight. For retention efficiency,
samples with natural radiological activities were used. Aliquots of 35 L were percolated in the Mn-
fibers, in series, by gravity (flow ≤ 1L min-1) and in the Mn-cartridges in series, by pumping (flow < 8
L min-1). Then, the polymers were washed with ultrapure H2O (pH 8.5) and the humidity adjusted
(50% mMn-polymers/mwater) by gravimetry. 224Ra activities were performed from 0 – 4 days, after pre-
concentration, via the RaDeCC System3,4. In figure 1, the results are presented. For retention
efficiency for each set of polymers, no significant difference was observed, even for polymers with a
greater amount of MnO2 adsorption sites. Despide that, for Mn-fibers, values above 99.8% were
found, while for Mn-cartridges, percentages from 93.1 – 94%. Therefore, these polymers showed a
significant difference in Ra retention, in which the fibers practically adsorbed the total fraction of Ra
but the cartridges made it possible to pre-concentrate 2.5 times faster.
Figure 1. Comparision Mn-fibers and Mn-cartridges (A) Retention efficiency (n=3) (B) Impregnation of MnO2 as a function
of reaction time (n=6).
*Equal letters - no
significant difference
(p<0.05) between the means by the Tukey test
It was concluded that the cartridge impregnation method can be applied to fibers. Furthermore, even
with the variation of adsorption sites, the shortest reaction time can be applied to polymers without
losing their adsorptive properties.
1 Szabo Z, Depaul VT, Fischer JM, Kraemer TF, Jacobsen E, Applied Geochemistry, 27, 2012, 729.
2 Thakur P, Ward AL, González-Delgado AM, Journal of Environmental Radioactivity, 228, 2021, 106522.
3 Henderson PB, Morris PJ, Moore WS, Charette MA, Journal of Radioanalytical and Nuclear Chemistry, 296, 2012, 357.
4 Scholten JC, Pham MK, Blinova O, Charette MA, Dulaiova H, Eriksson M, Marine Chemistry 121, 2010, 206.
[CAPES, CNPq, PPGQTA, FURG]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––
513
DETECTION AND QUANTIFICATION OF TITANIUM DIOXIDE
NANOPARTICLES (TiO2-NPs) BY spICP-MS
Santos A. V. Neto (R)a*, Cristiane Barata-Silva (R)a, Fabio S. Bazilio (R)a, Silvana C. Jacob
(R)a, Josino C. Moreira (R)a, Lisia M. G. Santos (R)a
a Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (Fiocruz), Departamento de
Química/ Setor de Eelementos Inorgânicos, Rio de Janeiro, Rio de Janeiro, Brazil, CEP: 21.040-900
*e-mail: cristiane.barata@fiocruz.br
The growing interest in nanotechnology has led to an increase in the production and application of nanoparticles
worldwide1. Due to the unique functional properties of nanoparticles, these materials are being used by many industries,
including the agricultural and food sectors 2. Among the commercially available nanomaterials, it is possible to highlight
those produced with titanium dioxide nanoparticles3. One of the most promising techniques for the analysis of metallic
nanoparticles is the Inductively coupled plasma mass spectrometry (ICP-MS) performed in single particle mode (spICP-
MS)4. However, the use of the technique in the detection and measurement of nanoparticles requires that the equipment
be operated differently than when used to analyse dissolved metal solutions 5. The aim of the present study was to validate
methodology single particle inductively couple plasma mass spectrometry (spICP-MS) (Nexion 300D, Perkin Elmer)
according to the parameters as describe in ISO/IEC 17025, because is the unique technique which allows ability to provide
information on nanoparticle size, size distribution, element composition, and number concentration in single, rapid analysis
of dissolved analyte in the sample. The limit of detection for nanoparticles diameter and for number concentration were
determined with ultrapure water and dwell time of 60 µs and integration time 100 seconds. A LOD size of 69 nm, equivalent
to a titanium mass of 775 ag, and a LOD number distribution of 4.9 x 10 4 L-1, were achieved. The LOD(I) 0.5 µg L -1 for
dissolved titanium was obtained in standard mode conditions, calculated as three times the standard deviation of the blank
divided by sensitivity. Suspensions containing titanium dioxide nanoparticles at number concentrations below 10 5 L-1 and/or
dissolved Ti(I) were introduced into the ICP-MS by Meinhard nebulization for determining the recovery and precision. The
recovery study for Ti(I) 98% and TiO2NP 109% and the relative standard deviation was < 5%. The method proved to be
adequate for the purpose, indicated the possibility of using the technique to determine the size and concentration of titanium
dioxide nanoparticles in products of sanitary interest.
1 Kulthong K, Srisung S, Boonpavanitchakul K, Kangwansupamonkon W, Maniratanachote R, Particle and Fibre

Toxicology, 7, 2010, 8.
2 Capelezzo AP, Mohr LC, Dalcanton F, Barreta CRDM, Martins MAPM, Fiori MA, De Mello JMM, Quimica Nova, 2018,
41, 367.
3 Grasso A, Ferrante M, Zuccarello P, Filippini T, Arena G, Fiore M, Cristaldi A, Conti GO, Copat C, International Journal
of Environmental Research and Public Health, 2020, 17, 9547.
4 Stephan C, Hineman A, Iron Nanoparticles by SP-ICP-MS: Overcoming Spectral Interferences Using Universal Cell
Technology, Walthan, 2014.
5 Peters RJB, Rivera ZH, Van Bemmel G, Marvin HJP, Weigel S, Bouwmeester H, Analytical and Bioanalytical Chemistry,
2014, 406, 3875.
6 Internacional Standard Organization; General Requirements for the Competence of Testing and Calibration Laboratories,
ISO/IEC 17025, 2017.
[FAPERJ, CAPES]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

514
ANTIOXIDANT ACTIVITY OF HONEYS FROM THE WESTERN REGION OF
PARANÁ
Juliana Ferreira (PG)a*, Cristian A. Godoy (UG)a, Daniela Q. Pedroso (UG)a, Luiz Roberto
Faria Junior (R)a, Marcela Boroski (R)a
a Federal University of Latin American Integration (UNILA), ILACVN, Foz do Iguaçu, PR, Brazil, 85867-970
*e-mail: j.ferreira.2020@aluno.unila.edu.br
Honey produced by Apis mellifera, bees from plant nectar and/or from secretions of living plants or excretion of plant-
sucking insects is a complex natural sweet product. The composition of honey is quite variable and depends mainly on the
floral source; however, certain external factors also play a role, such as seasonal and environmental factors and
processing. Honey is a supersaturated solution of sugars, of which fructose (38%) and glucose (31%) are the main
contributors. This natural food is produced in almost all countries and widely consumed as food. The popularity of this
complex natural sweet product has increased in recent years due to claims of health benefits, being considered a desirable
ingredient in food products1. Honey contains numerous compounds, such as organic acids, proteins, amino acids,
minerals, flavonoids, polyphenols, vitamins (ascorbic acid) and aromatic compounds2. Foods rich in antioxidants play an
increasingly important role in promoting health, as a result of the presence of of bioactive compounds, found naturally in
foods and that have aroused great interest due to their beneficial effects, highlighting their action in terms of disease
prevention3.This study aimed to evaluate the antioxidant activity of 15 samples of honey from municipalities located in
western region of Paraná, comprising the cities of Santa Terezinha, São Miguel do Iguaçu, Matelândia, São Pedro do
Iguaçu, Alvorada do Oeste, Diamante do Oeste and Foz do Iguaçu (comprising the region of the Iguaçu National Park).
These are regions with different floral availability, which permeate natural landscapes and agricultural areas. To evaluate
the quality of the honey, analyzes of pH, electrical conductivity, free acidity, moisture and antioxidants (total phenolics,
flavonoids, ferrocyanide, DPPH and ABTS) were performed. Acidity values ranged from 19 to 33 mEq/kg. The pH values
of the samples were slightly acidic, 3.58 to 4.36, as is characteristic of honeys. The phenolic compounds ranged from 66
to 111 mg EAG/kg. Considering the reduction power by the ferrocyanide assay, there was a variation from 29 to 48 mg
EAG/kg. There was a correlation between lower pH values and the highest concentration of total phenolic compounds,
and this result was obtained in samples collected in the surroundings of the Park Nacional do Iguaçu, probably due to the
greater diversity of flora for visitation. t was possible to observe that the humidity obtained in the samples maintained the
range of 20%, contributing to avoid the process of deterioration of the honey. It was observed the differentiation of the
samples in relation to the IC 50 values obtained from the DPPH analysis, referring to the flowering of soybean and corn,
showing an influence on the occurrence of bioactive compounds in honey. It was possible to establish a correlation between
the antioxidant activity, phenolic compounds and flavonoids of honey and the study of the hive location landscape.
1 Tang FHM, Lenzen M, McBratney A, Maggi F, Nature Geoscience 14, 2021, 206.
2 Schäfer RB, Liess M, Altenburger R, Filser J, Hollert H, Roß‑Nickoll M, Schäfer A, Scheringer M, Environmental Sciences
Europe, 31, 2019, 21.
3 Castilhos D, Dombroski JLD, Bergamo GC, Gramacho KP, Gonçalves LS, Apidologie, 50, 2019, 657.
[PRPPG/UNILA, Fundação Araucária, CAPES, SETI]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

515
DETECTION OF Fe(III)-D2EHPA COMPLEXES BY UV-VIS SPECTROMETRY
Marcel Felipe Alves de Souza (R)a*, Ludmila Querem Teixeira (R)a, Silvéria Neves de Paula e
Souza (R)a, Fernando Mota de Oliveira (R)a
aInstituto Federal de Educação, Ciência e Tecnologia de Minas Gerais (IFMG), Betim, Minas Gerais, Brasil, 32677-562
*e-mail: marcel.souza@ifmg.edu.br
Iron is an important micronutrient for many species. In this sense, such element can be found in
drugs and supplemental foods. Moreover, in society, there are different metal alloys contain iron
which are applied in the equipment or building construction. However, high levels of iron cations in
water are toxic to human health. Then, the iron analysis becomes a necessity for quality control of
drugs, foods and metal alloys as well as for water characterization. The compound di-2-ethylhexyl
phosphoric acid (D2EHPA) has been widely used in metallurgy industry for separation and
recuperation of several cations.1 The D2EHPA molecules extract cations from aqueous phase
exchanging them with protons from its structures and forming complexes.2 Therefore, this study
goals to evaluate the detection of Fe(III) extracted by D2EHPA by UV-vis Spectrometry. Thereby,
the extraction of Fe(III) was performed using a two-phase system: the aqueous phase contained
Fe(III) with known initial concentration; the organic phase contained D2EHPA in heptane (0.01 mol
L−1). Initial analyses were accomplished in a beaker with 50 mL of each phase stirring them with a
magnetic bar. The stirring was allowed for 3 min at room temperature and, after the phases
separation, the organic phase was analyzed by UV-vis spectrometry. In the spectrum obtained, it
was possible to observe an absorbance band in the ultraviolet region due the presence of Fe(III) in
aqueous phase before the extraction. Such statement can be confirmed by increasing the initial
concentration of Fe(III) in aqueous phase. So, probably D2EHPA forms charge-transfer complexes
with Fe(III). The absorbance signal, at 266 nm, was a linear function of the initial concentration of
Fe(III) in aqueous phase in range 2.0 to 8.0 × 10−6 mol L−1. As soon, fixing the volume of the phases
and the D2EHPA initial concentration in organic phase, at room temperature, the quantity of cations
extracted presented the same proportion for different initial concentrations of Fe(III) in aqueous
phase. No pH control was necessary for the solutions used. That can be explained by the fact that
Fe(III) extraction is around 100 % if the pH value of aqueous phase is equal or higher than 2. 1 In
order to make the analysis simpler and with less cost, the extractions were also accomplished using
a covered glass bottle (15 mL) for which 6.0 mL of each phase were transferred. In this case, the
stirring was manual. Similar results are achieved evidencing that the analyses can be performed with
little quantity of phases. It is worth mentioning that the baseline of the spectra needed to be corrected.
For this goal, a simple approach was applied: an absorbance signal at a wavelength in which no
absorbance is expected (400 nm was used) was subtracted from spectra. In Figure 1 the spectra
are shown as well as the absorbance of the organic phase after the extraction vs. Fe(III) initial
concentration in aqueous phase ([Fe(III)]aq). Studies regarding interfering ions, repeatability and
reproducibility will be carried out. After, the methodology developed will be tested in different samples
containing Fe.
Figure 1: Absorbance of organic phase after extraction of Fe(III), spectrum (a) and analytical curve (b).
1 Virolainen S, Salmimies S, Hasan M, Häkkinen A, Sainio T, Hydrometallurgy, 140, 2013, 181.

2 Wilson AM, Bailey PJ, Tasker PA, Turkington JR, Grant RA, Love JB, Chemical Society Reviews, 43, 2014, 123.
[IFMG]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

516
CHARACTERIZATION OF ALCOHOL CONTENT, COPPER AND LEAD IONS
CONCENTRATION BY F AAS IN SAMPLES OF CACHAÇA FROM NORTHERN
RIO GRANDE DO SUL, BRAZIL
Mistura, Clóvia Marozzin (R)a*, Guilherme Mattia Guarnieri (UG)a, Marcos Luis Salvador (R)a,
Alana Neto Zoch (R)a
a Universidade de Passo Fundo, ICEG, Passo Fundo, RS, Brazil, 99074000
*e-mail: clovia@upf.br
“Cachaça” is the typical and exclusive hard liquor made from distilled sugarcane produced in Brazil,
with an alcohol content varying from 38% to 48% by volume at 20 °C. There are over 30,000 different
producers in the country, most of them small independent craftsperson, with an estimated total yield
of about 1.3 billion L/year (2016)1,5. Exports in 2016 amounted to US$ 18 million going to 66 different
countries1. Given the importance of this product for small artisanal producers, a study regarding the
standardization and quality of this spirit was carried out. Twelve samples of alembic cachaça from
different sources were collected in the northern region of Rio Grande do Sul (RS), Brazil, region
ranked as the sixth state in Brazil in number of producers according to the Ministry of Agriculture2.
The distillation equipment used by small producers, as a rule of thumb, is made of metallic copper
that is known to contaminate its final product by dissolving ions3 in the liquor, mostly when using
tin/lead solders in its seams4. The objective of this study was to determine the ethanol content as
well as the concentration of copper and lead ions in cachaças through Flame Atomizer Atomic
Absorption Spectrophotometry (F AAS) with standard addition and matrix effect of the composition
in ethyl alcohol. For quality assurance of this technique, the method performed met the recovery
criteria, coefficient of variation and known concentration standard which proved suitable for the given
samples. The 12 samples were obtained from off the shelf products or directly from the still’s owners
in the region. From standard solutions of 100 mg L-1 Cu2+ and Pb2+, analytical curves were evaluated
from 0.005 to 1.0 mg L-1 Pb2+ and from 0.025 to 25 mg L-1 Cu2+. Two standard curves were
conducted, one with ultrapure water and one with matrix effect made from ultrapure water/ethyl
alcohol (50:50% v/v), Alcohol content was measured by alcoholometer density (0 to 100 °GL/%
scale). The analytical quality was verified through recovery tests, with the addition of a standard
solution of metal ions in aqueous/alcoholic medium with 5% nitric acid: 0.05 mg L-1, 0.30 mg L-1 and
0.75 mg L-1. The results are shown in Table 1.
Table 1: Alcohol content, concentration of Cu and Pb ions mg L-1 (mean and standard deviation SD) along with its
necessary dilutions for the results produced
Concentration of Concentrarion of Concentration of Concentrarion of
Alcohol Alcohol
Sample Pb2+ ions Cu2+ ions* Sample Pb2+ ions Cu2+ ions*
Content** Content**
n=3 (SD) n=3 (SD) n=3 (SD) n=3 (SD)
A 43,8 0,05 (±0,12) 14,469 (±0,002) G 45,3 0,14 (±0,06) 0,619 (±0,004)
B 41,2 0,06 (±0,11) 6,050 (±0,011) H 45,5 0,15 (±0,05) 0,049 (±0,063)
C 46,3 0,05 (±0,02) 8,330 (±0,005) I 45,4 0,16 (±0,02) 2,140 (±0,002)
D 46,2 0,03 (±0,01) 32,870 (±0,001) J 43,1 0,18 (±0,03) 2,340 (±0,072)
E 41,2 0,07 (±0,04) 9,429 (±0,050) K 41,0 0,14 (±0,02) 0,029 (±0,004)
F 43,7 0,16 (±0,02) 1,600 (±0,003) L 47,4 0,13 (±0,07) 0,690 (±0,046)
*Dilution required just to determine Cu ions. Samples A, B, C, E and H required 10x dilution while sample D required 100x dilution and
samples F, G, I, J, K and L did not required dilution. **in ethilic alcohol % v/v6.
The concentration of Cu2+ ions generated from samples A to E varied from 6 to 32 mg L-1, which is
above the limits set by the Brazilian regulations, that is ≤5 mg L-1, while its Pb2+ ions turned out below
such limits, that is set at ≤0.2 mg L-1.5,7 Samples F to L have shown to be within the legal limits.
1 IBRAC, Instituto Brasileiro da cachaça, 2022.
2 MAPA, Ministério da Agricultura, Pecuária e Abastecimento, 2021.
3 Catarino S, Pinto D, Curvelo-Garcia AS, Ciência e Técnica Vitivinícola, 18, 2003, 3.
4 INMETRO/SEBRAE, Certificação da cachaça de alambique, 2022.
5 BRASIL. Portaria 339, 2021. Lei 6.871. 2009. dispõe de registro, padronização, classificação, inspeção e a fiscalização
da produção de bebidas.
6 Garbin R, Bogusz Jr S, Montano MA, Ciência Rural, 35, 2005, 1436.
7 Marinho AV, Rodrigues JPM, Siqueira MI, Estudos Vida e Saúde, 36, 2009, 785.
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

517
EVALUATION OF THE CONTINUOUS USE OF PROTON PUMP INHIBITORS ON
THE BIOAVAILABILITY OF Fe, Ca, Zn, Mg, Cu AND K
Andréa Santana de Brito (PG)a, Rafaela Vidigal (UG)b, Alexandre Minami Fioroto (PG)c,
Giuliana Petri (R)b, Fernando Luiz Affonso Fonseca (R)b*, Angerson Nogueira (R)a**
a Federal University of São Paulo, Department of Exact and Earth Sciences, Diadema, São Paulo, Brazil, 09972-270
b FMABC University Center, Santo André, São Paulo, Brazil, 09060-870
c University of São Paulo, Institute of Chemistry, São Paulo, São Paulo, Brazil, 05508-010
*e-mail: affonso.fonseca@unifesp.br
**e-mail: angerson.nogueira@unifesp.br
Many studies have demonstrated a relationship between the prolonged use of proton pump inhibitors
(PPIs) and the reduced bioavailability of vitamins and minerals, bringing risks to patients who use
these drugs. However, certain relationships between PPIs and minerals are not well established1,2,3,4.
Therefore, this study aimed to analyze the bioavailability of Fe, Ca, Zn, Mg, Cu, and K in rats treated
with omeprazole, a PPI very used in Brazil5,6. Thirty-six Wistar rats weighing 200 and 300g were
divided into two groups (control and treated) and different treatment periods (10, 30, and 60 days).
The animals were euthanized at the end of each treatment, and the blood was collected. Thus, the
sample was immediately analyzed to get the biochemical and hematological parameters. The organs
(liver, spleen, and stomach) were freeze-dried and digested in a microwave oven and the elemental
analysis was performed by ICP-MS (Inductively Coupled Plasma Mass Spectrometry). The results
obtained from treated groups showed changes in the following parameters: hematological series,
hemoglobin, and hematimetric indices. In addition, there were variations in the leukocytes and
lymphocytes. These hematological alterations may suggest an inflammatory process and anemias
in the rats. The determination of analytes using ICP-MS in biological samples indicates a reduction
of Cu in the liver, stomach, and spleen samples and an increase in the concentration of Fe, Zn, K,
Mg, and Ca. Therefore, using omeprazole can interfere with the reserve of these minerals in the
body, resulting in osteoporosis and anemia.
1 Wiesner A, Zwolinska-Wcislo M, Pasko P, International Journal of Environmental Research and Public Health, 18, 2021,
3527.
2 Kaczmarczyk O, Przybylska-Feluś M, Piątek-Guziewicz A, Wcisło K, Krośniak M, Kryczyk-Kozioł J, Kleszcz K, Zagrodzki
K, Cibor D, Mach T, Zwolińska-Wcisło M, Polish Archives internal Medicine, 130, 2020, 179.
3 Morschel CF, Mafra D, Eduardo JCC, Brazilian Journal of Nephrology, 40, 2018, 301.
4 Yanagihara GR, Paiva AG, Neto MP, Torres LH, Shimano AC, Louzada MJQ, Annoni R, Penoni ACO, Revista Brasileira
de Ortopedia, 50, 2015, 232.
5 Mota DB, Borja A, Riscos relacionados ao uso prolongado de inibidores da bomba de prótons (omeprazol): importância
da atenção farmacêutica. Revista Oswaldo Cruz, 23, 2018, 6.
6 Salgado AL, Palma ALR, Ramos LP, Miranda PE, Oliveira FG, Cortelli AFD, Fernades WS, Lapena SAB, Brazilian
Journal of Health Review, 2, 2019, 5883.
[FINEP, FAPESP]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

518
SENSORY AND CHEMICAL EVALUATION BY SPECTROSCOPY OF DIFFERENT
METHODS OF COFFEE EXTRACTION
Cleidiana Vieira Guimarães (R)a, Marinalva Maria Bratz Simmer (R)a, Christiane Fileti Altoé
(UG)a, Danieli Grancieri Debona (PG)b, Taís Rizzo Moreira (R)c, Emanuele Catarina da Silva
Oliveira (R)a, Lucas Louzada Pereira (R)a*
a Federal Institute of Espírito Santo, Department of Coffee Research Analysis Laboratory - LAPC, Avenida Elizabeth
Minete Perim, S/N, Bairro São Rafael, CEP: 29375- 000, Venda Nova do Imigrante, Espírito Santo, Brazil.
b Federal University of Espírito Santo/UFES, Department of Chemistry, Campus Goiabeiras, CEP 29075-910, Vitória,
Espírito Santo, Brazil.

c Federal University of Espirito Santo/UFES, Department of forest and wood sciences, Center of Agrarian Sciences and
Engineering, Av. Governador Lindemberg, 316, CEP: 29550-000, Jerônimo Monteiro, Espirito Santo, Brazil .
*e-mail: lucas.pereira@ifes.edu.br
Sensory analysis techniques are constantly used in the food sector as tools to assess consumer
acceptance or preference for products, processes, and methods. Coffee becomes an important
target of sensorial analyses that reflect its valorization and acceptance in the market for presenting
a complex chemical matrix and highly influenced by factors that determine quality. However,
instrumental techniques are used to complement the sensory analysis, providing a large amount of
information and a complete view of the product. Thus, this study evaluated the acceptance and
preference of specialty coffees in relation to different preparation methods, with untrained
consumers, correlating sensory analysis with chemical aspects. For this, Coffea arabica obtained by
the Hario V60, Chemex, French Press and Espresso extraction methods were evaluated by affective
sensory tests, and their respective extraction residues were characterised by the infrared spectrum
in the range of 4000 to 630 cm-1. In the sensory panel, the Chemex and Hario V60 methods were
preferred, followed by Espresso and French Press, respectively. This result can be explained by the
type of granulometry used in each method, as well as the changes in the microstructural properties
of the coffee, due to the percolation rate, caffeine content, solids content, pH, and acidity. The
principal component analysis (PCA) allowed us to observe four distinct groups, suggesting that
different methods can also influence the chemical composition of coffees. The methods that most
differed in relation to chemical classes were French Press and Hario V60. The main regions
responsible for the separation of the groups were from 3000-3100 cm-1, which shows OH stretches
present in carboxylic acids and alcohols, which can characterise the sugars. In addition to this
stretch, this region characterises the primary amines through the NH stretches. Absorptions in the
region from 2921 to 2852 cm-1 are present aldehydes, 1740 cm-1 C=O of carboxylic acid and
carboxylic acid ester, the latter being found in triesters, present in lipid carbonyls. Thus, the mid-
infrared associated with the analysis of principal components demonstrated efficiency in the
separation of the classes of compounds between the extraction methods, French Press and Hario
V60, confirming that the technique can be used as an adjunct in quality control.
[Sicoob, CAPES, IFES, CNPq]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

519
EVALUATION OF IMPREGNATION AND RETENTION EFFICIENCY OF 224Ra
PAN FIBERS
Bruna B. Silva (UG)a*, Alessandra G. da Costaa, Mariele L. de Paiva(R)a, Gabriel K. de Souzaa
(R), Renata R. de Moura (R)a, Carlos F. F. de Andrade (R)a
aUniversidade Federal do Rio Grande - Campus Carreiros – Avenida Itália, Km 08, Instituto de Oceanografia, Rio
Grande, RS, Brasil, CEP 96201-900
* e-mail: botelhobrunab@gmail.com
Radium (Ra) is a chemical element that has 4 naturally occurring isotopes with different half-lives (223Ra = 11.4 days; 224Ra
= 3.6 days; 226Ra = 1600 years; 228Ra = 5.7 years) enabling its application in the understanding of oceanographic processes
(e.g. water residence time). However, for quantification by Radium Delayed Coincidence Counting (RaDeCC) is necessary
the pre-concentration of the analyte by electrostatic affinity in chemically modified polymers by the impregnation reaction
that enriches the polymer with nanoparticles of MnO2 by the reduction of KMnO4, such as Mn-fibers1. Therefore, the
objective in this work was to study the impregnation method2 in polyacrylonitrile fibers (PAN fibers) evaluating the influence
of reaction time and its relationship with the retention efficiency of 224Ra. The PAN fibers were washed with ultrapure H2O
(MILLI-Q® resistivity < 18.2 MΩ cm-1) and impregnated for reaction times of 24, 48 and 68 hours with KMnO4 (0.5 mol L-1)
at 20°C. Afterwards, were removed from the solution, leaving them to dry on absorbent paper at 20°C for 12 hours. Next,
washing was carried out with ultrapure H2O (pH 8.5) and then were lyophilized to constant weight and the percentage of
MnO2 was determined by gravimetric. For retention efficiency, the Mn-fibers were disposed in series and used samples
with natural radiological activities were used. Aliquots of 35 L were percolated by gravity (flow ≤ 1L min-1). Finally, the
polymers were washed with ultrapure H2O (pH 8.5) and the humidity adjusted (50% m Mn-fiber/mwater) by gravimetric. 224Ra
activities were performed from 0 – 4 days, after pre-concentration, via the RaDeCC System2,3. Table 1 shows the results
of better impregnation of the adsorption sites of Ra (MnO2) with increasing reaction time. However, it was found that there
is no significant difference in retention efficiency with increasing adsorption sites. In addition, the RE equal to or greater
than that reported in the literature4.
Table 1. Percentage of MnO2 (n=6) and retention efficiency (RE) (n=3) with variation of reaction time (t).
t (hours) MnO2 (%) RSD RE* (%) RSD

24 1.97 0.07 99.94a -
48 5.26 0.46 100a -
68 7.71 0,18 99.77a -
*Equal letters - no significant difference (p<0.05) between the means by the Tukey test
It was concluded that the impregnation method can be applied to fibers. In addition, the shorter
reaction time can be adopted, without it losing its adsorptive properties for Ra.
1 Garcia-Orellana J, Rodellas V, Tamborski J, Diego-Feliu M, Van Beek P, Weinstein Y, Charette M, Alorda-Kleinglass A,

Michael HA, Stieglitz T, Earth-Science Reviews, 220, 2021, 103681.
2 Henderson PB, Morris PJ, Moore WS, Charette MA, Journal of Radioanalytical and Nuclear Chemistry, 296, 2012, 357.
3 Scholten JC, Pham MK, Blinova O, Charette MA, Dulaiova H, Eriksson M, Marine Chemistry, 121, 2010, 206.
4 Lopez CV, Murgulet D, Santos IR, Journal of Hydrology, 590, 2020, 125395.
[CAPES, CNPq, FURG]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

520
TEACHING CHEMISTRY TO COMPUTERS: EXPLORING THE CHEMICAL SPACE
THROUGH TEXT REPRESENTATION
Lucas B. Ayres (PG)a*, Armele Varillas (R)b, Jeb R. Linton (R)a, Carlos D. Garcia (R)a
a Department of Chemistry, Clemson University, Clemson SC 29634, USA
b SC Governo’s School for srience & Mathematics
*e-mail: ldayres@clemson.edu
The advances of Artificial Intelligence (AI) in the past two decades enabled algorithms to perform
human-like daily tasks such as driving cars, playing complex games, composing classical music and
even generating realistic images only by using text as input parameter. These achievements can be
linked to the implementation of Deep Neural Network architectures in combination of the use of large
databases as well as the increase in computing power. This strategy has also been showing promise
in several sub fields of natural sciences such as chemistry, biology, and physics through speech
recognition, data analysis, and computer vision. While deep learning has been used to predict
properties of molecules and predict a number of chemical reactions, these advances have always
been linked to large databases containing millions of entries associated to a single molecule, for
example. Considering that such databases are not always available and performing those
experiments is simply an unreasonable task in a human time scale, we propose to use the Simplified
Molecular Input Line Entry System (SMILES)1 and the use of Natural Language Processing (NPL)
tools such as transformers aiming to teach chemistry to computers. The central idea of our strategy
is based on the fact that those AI tools are capable of extracting useful information from the chemical
space only by means of text. In this sense, our work has demonstrated that it is possible to train a
general chemistry model in less than 12 hours, reaching a constant learning rate (training loss <3.5)
after the epoch number 20. Then, the last layer of neurons of the resulted model was re-trained with
a relatively small database containing less 1000 entries aiming to leverage their knowledge in
specific fields of chemistry. We demonstrated that typical overfitting issues commonly present in this
step can be overcame by rearranging the input array of data and substituting any integer number
related to stoichiometric by text. As a result, the neural network model reached a maximum in
performance (e.g., phi coefficient > 0.70 and F1-score > 0.90) at around 15 to 20 rearrangements
per combination, also displaying a performance 3 times higher compared to stoichiometric numbers
represented in integers. In this sense, the strategy herein proposed is promising as an efficient and
alternative way to teach chemistry to computers (AI models) where the available database limits the
expansion in chemical knowledge of those models.
1 Weininger D, The Journal for Chemical Information and Computer Scientists, 28, 1988, 31.
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

521
EVALUATION OF THE MULTIPLE PROBES OF QD GSH-CdTe IN
MULTIPLEXED ANALYSIS EMPLOYING HG HS-SDME SYSTEMS
Leila S.V. Barbosa (R)a*, Maria das Graças A. Korn (R)a, Rodolfo M.M Santana (R)a
a Federal University of Bahia, Analytical Departament, Chemical Institute, Salvador, Bahia, Brazil,
*e-mail: leilasvbarbosa@gmail.com
The hydride generation with headspace single drop microextraction (HG HS-DME) systems were
used for individual determination of volatile species employing quantum dots (QD) as extractant
phase and photoluminescent probes.1 Therefore, the present study aimed to evaluate these systems
for simultaneous determination of two or more volatile species by multiplex analysis using quantum
dot multiple probes. Hydride was generated by derivatization with 4.8% NaHB4 (m v-1, in NaOH 0.25
mol L-1) in acidic medium (HCl 2.0 mol L-1). For extraction in headspace different CdTe mixture were
exploited as extractant droplet. All fluorescent assays were carried out by molecular fluorescence
(FL), synchronous fluorescence (SFL). First derivative fluorescence (DFL) was evaluated for data
treatment. To ensure this system a better analytical response against the target species, physical
and chemical parameters of the medium were evaluated, such as: pH (5.5 to 10.5), size and height
of the microdrop (4.0 µL to 8.0 µL; 1.5 cm to 4.0 cm, respectively) and time of hydride generation (1
to 10 min). Then, to evaluate the behavior of the mixtures of analytes, concentration ranges (10 µg
L-1 to 200 µg L-1) were evaluated both for the individual species and for mixture. Initially, the
separated QDs emission spectra and the binary mixtures were recorded in order to observe these
bands overlap degree. Therefore, sensitization assays against target species (Hg, Cd, Se, Sb, As,
Bi, Pb) individual solutions were performed, using the GSH-CdTe QDs probes (2.9 nm and 3.5 nm).
Considering the evaluated species only As(III) and Cd2+ caused an enhancement in the QD emission
signals of 3.5 nm and 2.9 nm, respectively. In the mixture, the same phenomenon was observed for
the As(III) and a quenching was observed for Cd2+ in front of the binary probe. Thus, it was observed
through FL that As(III) and Cd2+ presented linear behavior in the concentration range from 50 to 200
µg L-1. Under optimal conditions (pH 8.5; 8.0 µL; 1.5 cm and 3 min), analytical performance
parameters were obtained as a linear working range from 50.0 to 200.0 µg L-1, for both analytes,
LOD of 0.194 µg L-1 and 0.159 µg L-1 and LOQ of 0.589 µg L-1 and 0.481 µg L-1, with good linearity
(R=0.994 and R=0.996 for As(III) and Cd2+, respectively) precision (RSD < 10%; C=125.0 µg L-1,
n=6). Analyte addition assays were performed on tap water samples with recoveries between 86%
and 112% for As(III) (C=125 µg L-1 and 175 µg L-1 ), and 80% and 96% for Cd2+( C=125 µg L-1) in a
binary mixture of the hydrides. From this evaluation, it was possible to observe that these multiple
QD probes have great potential in the application of multiplexed analyses, that can be used to
simultaneously identify, and determine two volatile species.
1 Costas-Mora I, Romero V, Pena-Pereira F, Lavilla I, Bendicho C, Analytical Chemistry, 86, 2011, 2388.
[CAPES, CNPq, FAPESB, UFBA]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

522
QUANTIFICATION OF ESTOLIDES SAMPLES OBTAINED FROM CASTOR OIL
OLIGOMERIZATION USING 13C NMR SPECTROSCOPY
Lucas N. de Melo (PG)a, Simoni M. P. Meneghetti (R)a, Mario R. Meneghetti (R)a, Janaína H.
Bortoluzzi (R)a*
a Universidade Federal de Alagoas, Instituto de Química e Biotecnologia, Maceió, AL, Brazil, 57072-900
*e-mail: janaina.bortoluzzi@iqb.ufal.br
Estolides are oligomers composed of fatty acid monomers linked by secondary ester bonds in the
middle of its alkyl chains.1 These molecules can be synthetized from triacylglycerols (TAGs),
monoesters or free fatty acids and their size and properties depend on their degree of oligomerization
and on the presence of other functionalizations. The extension of the oligomerization is usually
measured as estolide number EN (EN = n + 1), which represents the average number of fatty acids
added to the base fatty acid moeity.1 Gel permeation chromatography (GPC) can be used to quantify
different types of oligomers and polymers based on their size, but this technique is time consuming
and requires large volumes of solvent.2 When TAG sources, such as fats and oils, are used as raw
material, the estolides produced can be found not just as TAGs, but also as di and monoacylglycerols
(DAGs, MAGs), which make EN determinations with 1H NMR spectroscopy difficult due to signal
overlapping. Alternatively, these glycerol-derived compounds can be quantified without previous
separation using 13C NMR spectroscopy as long as proton decoupled spectra are obtained only
during acquisition and waiting time between pulses is five times the longitudinal relaxation time. 3 In
addition, when equivalent resonances are compared, long relaxation delays may not be required to
obtain satisfactory product ratios using standard broadband decoupled experiments.4 The 13C NMR
spectra of 12 estolide samples, produced from TAGs of castor oil at different reaction conditions, 5
were obtained using a Bruker Avance 400 NMR Ultra Shield spectrophotometer (9,4 Tesla),
operating at 151 MHz, with a 0.9044 s acquisition time, spectral width of 36.2 KHz, 3072 scans,
using a 5 mm probe at 295.15 K. The J-modulating spin-echo sequence (J-mod) was used, providing
spectra in which the methine and methyl resonances have opposite phase to those of quaternary
and methylene signals. Approximately 40 mg of the samples were dissolved in 0.6 mL of CDCl3. The
EN of all samples was determined by the ratio between the integration of the C-O-C (73.7 ppm) and
C-O-H (71.5 ppm) signals (Fig. 1).2 EN values from 0.00 to 3.99 were obtained for all samples
studied. As it can be seen in Fig. 2, the EN measurements are in good agreement with kinematic
viscosity at 40 °C determinations (ASTM D445), since the viscosity tends to increase with the
average molecular weight and EN of the sample.
Figure 1. 13C NMR spectra of estolide sample with Figure 2. Correlation between EN and
characteristic C-O-C (73.7 ppm) and C-O-H (71.5 ppm) signals. kinematic viscosity of estolide samples.
1 Chen Y, Biresaw G, Cermak SC, Isbell TA, Ngo HL, Chen L, Durham
AL, Journal of the American Oil Chemists' Society, 97, 2020, 231.
2 Greco-Duarte J, Collaço ACA, Costa AMM, Silva LO, Da Silva JAC, Torres AG, Fernandez-Lafuente R, Freire, DMG
Fuel, 245, 2019, 286.
3 Casas A, Ramos MJ, Pérez A, Simón A, Lucas-Torres C, Moreno A, Fuel, 92, 2012, 180.
4 Otte DAL, Borchmann DE, Lin C, Weck M, Woerpel KA, Organic Letters, 16, 2014, 1566.
5 Melo LN, Santana LA, Rocha PF, Meneghetti SMP, Meneghetti MR, Bortoluzzi JH, Energy and Fuels, 34, 2020, 5948.
[CNPq, CAPES, FINEP, FAPEAL, INCT-Catalysis]
––––––––––––––––––––– Miscellaneous –––––––––––––––––––––

523

Book of Abstracts

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Book of Abstracts

Uploaded by

Copyright:

Available Formats

BOOK OF

Welcome to 20th ENQA/8th CIAQA.

Érico Marlon de Moraes Flores

Federal, Brazil, 70610-200

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

1,8 (a) (b) FIT

log Rredox ()

1 Olean-Oliveira A et al., ACS Sensors, 4, 2019, 118.

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

2 4 6 8 10 0.0 0.5 1.0 1.5 2.0 2.5 3.0

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

Current Density ( cm

Potential (E vs ECS) Potential (E vs ECS)

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

Electrochemical sensors based on electrochemical impedance spectroscopy have gained

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

0.2 0.4 0.6 0.8 1.0

1 Espinosa BM et al., J. Pharm Biomed Anal, 42, 2006, 291

[PIBIC/UFT, FAPT, CNPq, CAPES, FAPESP]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

[FAPESP, Brazilian Synchrotron Light Laboratory]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

Thermoplastic filaments containing conductive carbon materials have contributed tremendously to

[CAPES, CNPq, FAPEMIG, FAPESP, INCTBio]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

Department of Chemistry, Institute of Chemistry, Campinas, Sao Paulo, Brazil, 13083-970

[CAPES, CNPq, INCTBio, UEL, IFPR]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

The analytical performance of electrochemical biosensors is significantly reduced when subjected to

[CNPEM, UNICAMP, UFABC, CAPES, FAPESP, CNPq]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

[FAPESP, CAPES, CNPq]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

1 Huang XJ., O’Mahony AM, Compton RG, Small, 5, 2009, 776

[UFU, CNPq, FAPEMA, FAPEMIG, CAPES]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

Electroanalytical is an inexpensive and high-performance analytical technique. However, their lack

Miniaturized device 600 - 10000 0.99866 30.0 100.0 0.007

Miniaturized device integrated to gas

[UFU, CAPES, FAPEMIG, CNPq, FAPESP]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

[CAPES, CNPq, FAPESP]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

3D-Printing or additive manufacturing is an emerging technology with several fields of application,

[FAPEMIG, CAPES, CNPq, InovaLeite e Instituto Sua Ciência]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

Mathematics, Sorocaba, São Paulo, Brazil, CEP 18052-780

[CNPq, FAPESP, CAPES]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

[FURG, EQA, CNPq]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

[CNPq, FAPESP CAPES]

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

Departament of Chemistry, Institute of Chemistry, Campinas, Sao Paulo, Brazil, 13083-970

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––

1 Werion A, et al., Kidney Int. 98, 2020, 1296.

––––––––––––––––––––– Eletrochemistry and Eletroanalytical Chemistry –––––––––––––––––––––