Food Chemistry 285 (2019) 334–339

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Analytical Methods

A versatile green analytical method for determining chlorine and sulfur in T

cereals and legumes
Filipe S. Rondana, Gilberto S. Coelho Juniora, Rodrigo M. Pereiraa, Alessandra S. Hennb,
Edson I. Mullerb, Marcia F. Meskoa,
⁎

a
Centro de Ciências Químicas, Farmacêuticas e de Alimentos, Universidade Federal de Pelotas, 96160-000 Capão do Leão, RS, Brazil
b
Departamento de Química, Universidade Federal de Santa Maria, 97105-900 Santa Maria, RS, Brazil

ARTICLE INFO ABSTRACT

Keywords: A versatile, rapid and safe green method for chlorine and sulfur determination using ion chromatography in cereals and
Food analysis legumes was developed. Microwave-induced combustion was evaluated for sample preparation. Ultrapure water and
Elemental determination alkaline solutions were assessed for absorption of the analytes. Water was selected because good recoveries (97–109%)
Grains were obtained for both analytes. Low consumption of reagents and small quantities of waste are two important ad-
Microwave-induced combustion
vantages of the proposed method. Accuracy was evaluated by analysis of a standard reference material, which agreed
Ion chromatography
with certified values (91–101%). The results for repeatability (RSDs ≤ 4%) and intermediate precision (RSDs ≤ 7%)
prove the good precision of the proposed method. Limits of quantification were 16 and 17 mg kg−1 for Cl and S,
respectively. Concentrations of Cl and S varied across a wide range (Cl: 35–930 mg kg−1; S: 678–5124 mg kg−1) for 34
samples analyzed, which were, for most of the results, close to the values found in the literature.

1. Introduction Meyers, 2006). Sulfur is involved in protein synthesis process and,

Cereals and legumes are present in most diets globally and are con-
sumed in a variety of forms (e.g. flours in bread, cookies, cakes, ingredients
in soups, etc.) (Arendt & Zannini, 2013; Ruiz-de-Cenzano, Rochina-Marco,
Cervera, & de la Guardia, 2015). These agricultural commodities contain
important nutrients that are essential for human nutrition (Arendt &
Zannini, 2013; Wrigley, Cork, Seetharaman, & Faubion, 2016).
In view of the remarkable increase in the production of cereals and
legumes (FAO, 2018), quality control is extremely important to ensure
food safety. It is important to highlight that this monitoring should not be
restricted to elements that are toxic, but also concentrations of essential
and non-essential dietary compounds. Inadequate intake of some elements
can cause problems in human health (Tarantino et al., 2017). Chlorine (Cl)
and sulfur (S) can be highlighted among essential and non-essential ele-
ments that must be controlled in cereals and legumes since they have
important roles in human metabolism. Concentrations of these elements in
plant foods, including cereals and legumes, derive from soil composition as
well as pesticides and fertilizers (Garratt, Wright, & Leather, 2010).
According to the Institute of Medicine of the National Academies of
Science (United States of America), recommended Cl intake is around
2300 mg day−1. Excess intakes of Cl may cause several problems in
humans, mainly associated with blood pressure (Otten, Hellwig, &
unlike Cl, there is no daily intake recommendation. Nevertheless, some
authors suggest a daily intake of about 850 mg to provide sulfur suffi-
cient for protein synthesis (Antoine, Fung, Grant, Dennis, & Lalor,
2012). Excess intake of S can, in some cases, also cause health pro-
blems, such as ulcerative colitis (Otten et al., 2006).
Techniques commonly used for Cl and S determination in cereals and
legumes are based on instrumental neutron activation analysis (INAA)
and inductively coupled plasma mass spectrometry (ICP-MS) (Al-Dayel &
Al-Kahtani, 2002; Balaji et al., 2000; Barbosa et al., 2013; Laursen,
Hansen, Persson, Schjoerring, & Husted, 2009; Miyamoto, Kajikawa,
Zaidi, Nakanishi, & Sakamoto, 2000; Nyarko, Akaho, Fletcher, Zwicker,
& Chatt, 2006; Singh & Garg, 2006; Soliman & Zikovsky, 1999; Wang &
Li, 2012). Sample preparation step for these methods use concentrated
acids but, when acid solutions are used for further determination of
halogens, target analytes are lost through volatilization (Mesko, Toralles,
et al., 2016). Also, memory effects occur when conventional pneumatic
nebulization is used for sample introduction in ICP-MS (Antes et al.,
2012; Mello et al., 2013). When an ICP-MS equipment with low mass
resolution is used, Cl determination is particularly challenging because of
its low atomic mass (Mesko, Costa, Picoloto, Bizzi, & Mello, 2016). In this
case, polyatomic interferences can occur, requiring the use of reaction
cells (May & Wiedmeyer, 1998). Similarly, S determination using ICP-MS

⁎
Corresponding author.
E-mail address: marcia.mesko@pq.cnpq.br (M.F. Mesko).

https://doi.org/10.1016/j.foodchem.2019.01.169
Received 7 December 2017; Received in revised form 13 January 2019; Accepted 25 January 2019
Available online 01 February 2019
0308-8146/ © 2019 Elsevier Ltd. All rights reserved.
F.S. Rondan, et al.
is hampered by spectral interferences, which form inside the plasma, in
relation to H, O and N (May & Wiedmeyer, 1998). Finally, given the need
for a nuclear reactor to activate samples for INAA, this technique is
unsuitable for routine analysis.
Thus, in this context, ion chromatography (IC) might be an alter-
native for determination of these elements, because it has advantages
over both alternatives, such as the relative low cost of acquisition,
analysis and maintenance (Mello et al., 2013). Ion chromatography is a
multi-elemental technique and has good sensitivity for Cl and S de-
termination. However, as for ICP-MS determination, IC usually requires
complete digestion of the matrix in order to obtain appropriate solu-
tions for analysis. For this reason, an adequate and effective sample
preparation method is required for element determination using IC.
Microwave-induced combustion (MIC) can be useful for the pre-
paration of cereals and legumes, since it is an efficient and robust
method that ensures total elimination of organic matter through oxi-
dation process (Flores et al., 2008). As MIC occurs in closed vessels, it
also minimizes losses through volatilization. Additionally, it is possible
to choose the absorbing solution most suitable for target analytes and
the analytical method (Flores et al., 2008). Thus, the aims of this study
was to develop a versatile, high throughput and green method for de-
termination of total Cl and S in cereals and legumes using IC after prior
digestion by MIC. Sample preparation parameters, such as sample mass
and absorbing solution, were studied. Accuracy was evaluated by
analysis of a standard reference material from rice flour (SRM, NIST
1568a) as well as recovery tests using standard solutions.
2. Materials and methods
2.1. Instrumentation
The samples were ground in a mill (model 226/2, Lucadema
Científica, São José do Rio Preto, Brazil), while the drying process was
performed in an oven (model 400/2ND, DeLeo, Porto Alegre, Brazil).
All mass measurements were carried out using an analytical balance
(model AY220, Shimadzu, Rosario, Philippines), with a resolution of
0.0001 g and a maximum load of 220 g. Ultrapure water (18.3 MΩ cm)
used for all procedures was obtained using a purification system (model
Mega UP, MegaPurity, Seoul, South Korea).
For sample preparation, a microwave oven (model Multiwave 3000,
Anton Paar, Graz, Austria) equipped with eight high pressure quartz
vessels was used (internal volume 80 mL, maximum pressure and
temperature about 8 MPa and 280 °C, respectively). Filter paper and
low-density polyethylene (LDPE) films were decontaminated in an ul-
trasonic bath (40 kHz, 300 W, model USC-2800A, Unique, Indaiatuba,
Brazil). After decontamination, these materials were dried in a class
100 laminar flow hood (model CSLH-12, Veco, Campinas, Brazil).
Chlorine and sulfur were determined using an ion chromatograph
(model 861 Advanced Compact, Metrohm, Herisau, Switzerland) with
suppressed conductivity detection. A Metrosep A Supp 4/5 guard
column and a Metrosep A Supp 5 (150 mm × 4 mm i.d., 5 µm particle
size, anion exchanger functionalized with quaternary ammonium
groups) analytical column were used to separate the analytes. The
mobile phase was composed of 3.2 mmol L−1 sodium carbonate and
1.0 mmol L−1 sodium bicarbonate. The flow rate and sample loop were
set at 0.7 mL min−1 and a 20 µL, respectively. These parameters were
based on previous work and recommendations of the manufacturer
(Coelho Junior et al., 2017; Metrohm; Pereira et al., 2016).
2.2. Reagents and samples
All reagents used in this study were of analytical grade or higher
purity and all solutions used were prepared with ultrapure water. For
decontamination of vessels and quartz holders, 6 mL of nitric acid
(65%, Vetec, Duque de Caxias, Brazil) were used in a microwave
heating program (1000 W, 10 min [heating step] and 0 W for 20 min
335
Food Chemistry 285 (2019) 334–339
[cooling step]). The same procedure was repeated using ultrapure
water in order to eliminate acid residues. Oxygen (99.5%, Linde,
Chapecó, Brazil) was used as oxidizing reagent.
Discs of filter paper (17 mm of diameter, Unifil, Hannover,
Germany) and LDPE film (80 mm × 80 mm) were used in the com-
bustion process. These materials were decontaminated previously by
immersion in nitric acid 10% (v/v) and then ethanol 20% (v/v, Synth,
Diadema, Brazil) using an ultrasonic bath for 10 min for each step.
Ammonium hydroxide solutions evaluated as absorbing solution
were prepared by dilution of concentrated solution (27% ammonia,
Synth), whereas the ammonium nitrate solution (6 mol L−1) was pre-
pared by dissolution of the salt (Merck, Darmstadt, Germany).
Calibration curves for Cl and S by IC were prepared using five aqueous
standard solutions, which ranged from 0.1 mg L−1 to 2.0 mg L−1 and
5.0 mg L−1 to 15.0 mg L−1, respectively. These solutions were prepared
by dilution from stock solutions (1000 mg L−1) that were prepared by
dissolving potassium chloride and sodium sulfate salts (Merck) in
water. These solutions were also utilized for recovery tests. The mobile
phase was prepared by dissolution of sodium carbonate and sodium
bicarbonate salts (Merck).
The grains used (rice, maize, wheat, beans, lentils and soybeans)
were purchased from a local market. According the label on the pro-
ducts, the samples were produced in Brazil, Chile, Egypt, Germany,
India, Pakistan, United States of America, and Uruguay. Prior to di-
gestion, about 50 g of each sample were ground in a knife mill to obtain
a fine and, apparently, homogeneous powder. Subsequently, these
samples were dried at 60 ± 5 °C in an oven until they achieved a
constant weight (around 4 h). A standard reference material (SRM)
from the National Institute of Standards and Technology (NIST 1568a,
rice flour) was used to evaluate accuracy; this was prepared as re-
commended by the manufacturer.
2.3. Digestion by microwave-induced combustion
Ground and dried samples of cereals and legumes (100–1000 mg) were
wrapped in LDPE film, which was sealed by heating and the excess re-
moved. Wrappers containing the samples were placed on the base of
quartz holders (specific for the MIC method) already containing a disc of
filter paper moistened with igniter solution (50 µL of 6 mol L−1 ammo-
nium nitrate). These holders were inserted into quartz vessels that were
filled previously with 6 mL of absorbing solution (ultrapure water or 25,
50 or 100 mmol L−1 ammonium hydroxide). Then, the vessels were
closed, fixed on the rotor and pressurized with oxygen at 2 MPa for 1 min.
The microwave irradiation program was 1400 W for 5 min (combus-
tion + reflux step) and 0 W for 20 min (cooling step). At the end of the
process, the pressure was released and the digests transferred to volu-
metric vessels. The digests were made up to 25 mL with water for sub-
sequent Cl and S determination by IC. In each round of digestion, three
samples and one reagent blank were digested to evaluate any potential
contamination from equipment or reagents used during preparation.
Pressurization time, oxygen pressure, concentration and volume of
igniter, volume of absorbing solution and irradiation program were
based on previous studies (Coelho Junior et al., 2017; Costa et al., 2015;
Hartwig et al., 2014; Rondan et al., 2018). All the results were eval-
uated statistically using one-way analysis of variance (ANOVA) fol-
lowed by the Tukey test or Student’s t-test (confidence level of 95%)
with GraphPad InStat version 3.00 (GraphPad, San Diego, USA).
3. Results and discussion
3.1. Evaluation of sample mass and absorbing solution in the MIC method
Microwave induced-combustion has been used for the digestion of
solid matrices previously, e.g., pet food, shrimp, powdered egg, edible
seaweed (Coelho Junior et al., 2017; Corazza et al., 2016; Crizel et al.,
2015; Mesko, Toralles, et al., 2016; Toralles et al., 2017). Although a
F.S. Rondan, et al. Food Chemistry 285 (2019) 334–339

general protocol exists for MIC sample digestion, careful evaluation of

parameters, such as sample mass and absorbing solution, should be
carried out for each approach, since incomplete combustion or in-
efficient absorption might lead to inaccurate results.
Initially, six types of cereals and legumes (100–1000 mg) were di-
gested by MIC using water as the absorbing solution and a microwave
irradiation program described previously (Rondan et al., 2018). The
maximum pressure achieved during combustion and characteristics of
the digests were evaluated; the maximum pressure was about 4 MPa,
which is about half the maximum operational pressure of the quartz
vessels (8 MPa). Moreover, it was possible to observe different com-
bustion behaviors for the six types of cereals and legumes used in this
study.
Based on the characteristics of the digests, maximum sample mass Fig. 1. Aspects of the solutions after decomposition of different soybean sample
efficiently digested for each sample was chosen, as presented in Table 1. masses by MIC, using water as absorbing solution. (a) 500 mg (b) 600 mg.
Only clear solutions, without the presence of solid particles or turbidity,
or soot residues in the reaction vessels were considered appropriate.
For masses higher than those in Table 1, the final solution presented
a yellow aspect and/or a significant amount of residue in the quartz
holder and solution, as shown in Fig. 1, where solutions resulting from
digestion of 500 and 600 mg of soybean are shown.
Masses up to 1000 mg of rice were digested satisfactorily, but only
the digestion of 500 mg of soybean resulted in a clear solution. This
behavior is probably related to the composition of food samples. As
shown in Table 1, rice and maize, which have similar compositions, also
presented similar combustion-related behaviors.
In general, the amount of carbohydrates as well as proteins were
proportionally related to the maximum sample mass digested efficiently
for each cereal and legume. Soybean contain the least carbohydrates
and the highest concentration of proteins and lipids compared with
other samples, which probably made it more difficult to digest. Thus,
based on these results, 500 mg was selected for subsequent experi-
ments, since this made it possible to digest all of the samples evaluated.
Subsequently, the most suitable solution for absorption of Cl and S was
also evaluated. For this, the samples were digested by MIC, using water or
ammonium hydroxide solutions (25, 50 or 100 mmol L−1) to absorb the
analytes. The use of these solutions was based on previous studies (Coelho
Junior et al., 2017; Costa et al., 2015; Hartwig et al., 2014; Mesko,
Toralles, et al., 2016). The results obtained are shown in Fig. 2.
According to these results, no statistical differences were observed
between Cl and S concentrations in each cereal and legume when water
or alkaline solutions were used as the absorbing solution (ANOVA/
Tukey test). In view of this, and considering the compatibility with IC,
water was chosen as the absorbing solution. The use of water brought
several benefits to the method, such as reduced reagent consumption
and, consequently, less waste generation as well as greater operational
safety compared with methods that use concentrated acids (Antoine
et al., 2012; Wang & Li, 2012).
Recovery tests were performed using standard solutions for pre-
liminary evaluation of accuracy. For this, 100 µL of standard solutions
Fig. 2. Chlorine and sulfur concentrations obtained for different samples using
(corresponding in mass to about 50% to 100% of Cl and S concentra-
ultrapure water, 25 mmol L−1 NH4OH, 50 mmol L−1 NH4OH or 100
tions present in the samples) were added to each sample before MIC mmol L−1 NH4OH as absorbing solution and 500 mg of sample after decom-
digestion; the results are shown in Fig. 3. position by MIC and determination by IC (n = 3).

Table 1 As shown in Fig. 3, good recoveries for all the samples analyzed
Maximum sample mass efficiently digested by MIC, and major composition of were obtained (97–105% for Cl, 98–109% for S). Accuracy of the pro-
cereals and legumes obtained from the manufacturer’s label (%, w/w). posed method was also evaluated via the digestion of SRM NIST 1568a,
Sample Maximum sample mass digested Carbohydrates Proteins Fiber using water as the absorbing solution. This SRM was chosen because it
(mg) has a composition similar to the majority of the samples evaluated.
Agreement with the reference values for Cl and S were 101 ± 2% and
Rice 1000 78 7 5
Maize 900 74 6 5
91 ± 1% (n = 5), respectively. In turn, precision was assessed at two
Wheat 600 57 10 10 levels, repeatability and intermediate precision, according to EURAC-
Bean 600 53 23 27 HEM guidelines (Eurachem, 2014). The results obtained for repeat-
Lentil 600 60 23 4 ability (RSD ≤ 4%) and intermediate precision (RSD ≤ 7%) demon-
Soybean 500 22 40 8
strated good applicability of the proposed method. Values for Cl and S

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F.S. Rondan, et al. Food Chemistry 285 (2019) 334–339

Table 2
Concentrations of Cl and S in several types of cereals and legumes from dif-
ferent countries, determined by IC after MIC digestion (x¯ ± sd , n = 5).
Sample (origin) Concentration (mg kg−1)

Cl S

White rice 1 (BRA) 199 ± 8 919 ± 30

White rice 2 (URU) 222 ± 9 900 ± 45
White rice 3 (CHL) 242 ± 11 813 ± 48
White rice 4 (PAK) 247 ± 10 1200 ± 69
White rice 5 (GER) 130 ± 8 837 ± 49
White rice 6 (IND) 236 ± 12 1320 ± 44
White rice 7 (BRA) 219 ± 10 688 ± 23
White rice 8 (USA) 209 ± 13 755 ± 39
White rice 9 (EGY) 270 ± 11 1234 ± 41
Parboiled rice 1 (BRA) 192 ± 7 930 ± 28
Parboiled rice 2 (GER) 103 ± 5 5124 ± 322
Parboiled rice 3 (GER) 106 ± 7 678 ± 46
Parboiled rice 4 (URU) 233 ± 15 1248 ± 73
Brown rice 1 (BRA) 190 ± 12 1017 ± 41
Fig. 3. Recoveries obtained for chlorine and sulfur in cereals and legumes Brown rice 2 (BRA) 149 ± 7 1220 ± 72
obtained using 500 mg of sample and ultrapure water as absorbing solution in Brown rice 3 (GER) 141 ± 9 991 ± 60
MIC and determination by IC (n = 3). Brown rice 4 (USA) 208 ± 8 981 ± 34
Brown rice 5 (USA) 329 ± 14 935 ± 59
Brown rice 6 (USA) 364 ± 23 1328 ± 87
in reagent blanks were appropriate, considering analyte concentrations Wild rice (BRA) 358 ± 22 1724 ± 117
(< 0.13 mg L−1 and < 0.3 mg L−1, respectively). Moreover, the pro- Black rice 1 (BRA) 166 ± 8 1004 ± 40
posed method had good limits of detection (LOD) and quantification Black rice 2 (USA) 648 ± 43 1721 ± 108
Yellow maize canjica (BRA) 455 ± 17 759 ± 24
(LOQ) for Cl and S, which were 10 and 9 mg kg−1, and 16 and
Popcorn (BRA) 470 ± 19 838 ± 18
17 mg kg−1, respectively. LOD and LOQ were calculated from the White maize canjica (BRA) 812 ± 55 1226 ± 61
average values of blanks and the standard deviations of 10 replicates Wheat (BRA) 394 ± 23 1291 ± 53
(INMETRO, 2018), considering the final volume of digests (25 mL) and Wheat canjica (BRA) 381 ± 26 1347 ± 83
Black bean (BRA) 196 ± 12 1902 ± 79
sample mass (500 mg). Depending on sample composition, better LOD
Pinto bean (BRA) 147 ± 7 2077 ± 58
and LOQ can be obtained with greater sample masses, as observed for White bean (BRA) 71 ± 4 1982 ± 92
rice (1000 mg) and maize (900 mg). These results demonstrate that the Lentil 1 (BRA) 930 ± 31 1874 ± 91
proposed method is versatile, accurate and precise for Cl and S de- Lentil 2 (BRA) 635 ± 28 1955 ± 80
termination in cereal and legumes. Soybean 1 (BRA) 35 ± 2 2861 ± 145
Soybean 2 (BRA) 52 ± 3 2899 ± 121

3.2. Chlorine and sulfur determination in cereals and legumes
The proposed method (using 500 mg of sample and water as ab-
sorbing solution) was applied to food samples and the results obtained
are shown in Table 2.
As can be seen in Table 2, S concentrations were higher (1.6–85
times) than Cl concentrations in all samples. This is in agreement with
the results observed by Soliman and Zikovsky (1999). Additionally,
there was a wide variation in Cl and S concentrations (Cl:
35–930 mg kg−1; S: 678–5124 mg kg−1).
More specifically, for Cl, concentrations in rice samples did not vary
widely and analysis did not reveal any patterns in the concentrations for
the foods evaluated. Despite the small variations, it was possible to verify
that, among the white rice samples, the highest Cl concentrations were
observed in samples from Egypt (White rice 9), followed by Pakistan
(White rice 4) while, for parboiled rice, higher Cl concentrations were
observed in samples from Uruguay (Parboiled rice 4) and Brazil
(Parboiled rice 1). For brown rice, two samples from USA (Brown rice 5
and Brown rice 6) had higher concentrations of Cl than samples of the
same type. Moreover, USA black rice (Black rice 2) has Cl concentrations
about four times higher than black rice from Brazil (Black rice 1).
There were no statistical differences (ANOVA/ Tukey test,
P = 0.3074) when samples of the same brand but of different types
were compared (White rice 1, Parboiled rice 1 and Brown rice 1). For
Parboiled rice 2 and Parboiled rice 3 samples from Germany (Student’s
t-test, P = 0.4579), which belong to the same brand and type, i.e. no
differences were observed. For White rice 2 and Parboiled rice 4 sam-
ples, both from Uruguay, Cl concentrations were not statistically dif-
ferent (Student’s t-test, P = 0.1973) and, although they were not the
same brand, they were produced by the same company. Despite Brown
rice 2 and Black rice 1 being the same brand, there was a statistical
Brazil (BRA), Chile (CHL), Egypt (EGY), Germany (GER), India (IND), Pakistan
(PAK), United States of America (USA), Uruguay (URU).
difference (Student’s t-test, P = 0.0002), i.e. between type. Thus, based
on these results, it can be inferred that Cl concentrations in rice are
related to genetic factors as well cultivation, and not related processing,
such as polishing and parboiling.
There was no statistical difference among the three maize samples
evaluated (Student’s t-test, P = 0.2248), but White maize canjica con-
tained the most Cl. The differences in Cl concentrations among maize of
different colors may also be related to genetic factors rather than lo-
cation or processing methods. There were no statistical differences
between the two wheat samples analyzed (Student’s t-test, P = 0.4267).
In legume samples, there were more significant differences among
the three types of sample (i.e. beans, lentils and soybeans) regarding Cl
concentrations. Among the bean samples, White beans had the lowest
Cl concentrations, while Black beans had the highest. There were sta-
tistical difference within lentil (Student’s t-test, P < 0.0001) and soy-
bean samples (Student’s t-test, P < 0.0001), which belonged to dif-
ferent brands.
Concerning the S concentration in rice, as observed for Cl con-
centrations, there were no obvious trends as related to type. Even
samples belonging to the same brand were different. For example,
Parboiled rice 2 and Parboiled rice 3 samples, which belonged to the
same brand from Germany, contained the highest and the lowest S
concentrations, respectively. However, this difference was associated
with protein content, as stated on the manufacturer’s labels.
Among the maize samples, White maize contained the most S and
trends that were observed followed those seen in Cl concentrations in
the same samples. Yellow maize had the lowest Cl concentration, and

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F.S. Rondan, et al. Food Chemistry 285 (2019) 334–339

the two wheat samples were statistically identical for S (Student’s t-test,
P = 0.2393).
Disregarding Parboiled rice 2, S concentrations in legumes were
higher than in cereal. This might be due to higher protein contents in
legumes, as shown in Table 1. There were no statistical difference be-
tween Black and White bean samples (Student’s t-test, P = 0.1784) with
respect to S concentrations, while Pinto bean had higher S concentra-
tions than either of the other types. There were no statistical differences
between lentil (Student’s t-test, P = 0.1733) or soybean samples (Stu-
dent’s t-test, P = 0.6647).
3.3. Comparison of obtained results with other studies
The results obtained in this study were compared with others re-
ported in the literature (Table 3).
According to the literature, generally, in white rice samples, Cl
concentrations were higher in samples from China and Japan (Singh &
Garg, 2006; Wang & Li, 2012), while S concentrations varied in a
narrow range. With respect to brown rice, Soliman and Zikovsky (1999)
found the highest S concentration in a sample from Canada, which also
had a Cl concentration similar those evaluated here (from Brazil, Ger-
many and USA). For maize samples, Nyarko et al. (2006) found the
highest Cl concentration in a sample from Ghana, while Cl concentra-
tions found by Singh and Garg (2006) were within the range found in
the present study (from Brazil).
With regard to wheat samples, Balaji et al. (2000) and Singh and
Garg (2006) found the highest Cl concentrations in samples from India,
while the highest S concentration was observed by Soliman and
Zikovsky (1999) in a sample from Canada; this was about 2.3-times
higher than the Brazilian sample in the present study. Lastly, soybean
samples, which are the only legumes shown in Table 3, had the lowest
Cl concentrations.
Thus, based on the results obtained, it is possible to affirm that the
proposed method, which consists of sample preparation using MIC and
IC determination, is suitable for total Cl and S determination in cereals
and legumes. Using the proposed method, only water was required as
the absorbing solution, which reduced the consumption of reagents and
the generation of waste, and provides greater operational safety when
compared to existing methods. In addition, the proposed method is
versatile, allowing digestion and analysis of a wide variety of cereals
and legumes. Good accuracy and precision were observed as well as
appropriate LODs and LOQs. In this sense, the proposed method is an
interesting alternative to routine analysis, since it also presents poten-
tial for high throughput analysis (sample preparation: 16 samples h−1;
determination: 4 samples h−1).
The proposed method demonstrate several advantages over other
existing methods, which use concentrated acid for sample preparation
and expensive determination techniques, such as ICP-MS and INAA.
Antoine et al. (2012) and Wang and Li (2012), for example, used con-
centrated acids for sample preparation. In both studies, prior to diges-
tion, samples remained overnight in acid. Importantly, however, none
of these methods allowed simultaneous determination of Cl and S.
Furthermore, complete analysis (irradiation, decay and measurement)
using INAA vary from 5 min to 3 months, depending on the radio-
nuclide used, which reduces considerably the sample throughput. In
comparison, the present study used fewer reagents and reduced waste
generation as well as enabling multi-elemental analysis. Finally, in
previous studies, such as Barbosa et al. (2013), the authors have used
dynamic reaction cells with ammonia, as the reaction gas, to minimize
interference as well as an alkaline solution to absorb the analyte. In the
present study, only water was used to absorb Cl and S for further de-
termination by IC.
4. Conclusion
The proposed method is suitable for digestion of different cereals
and legumes and, subsequently, total Cl and S determination. The main
advantage of the method developed here is its versatility, since it is
possible to digest several types of grains with different compositions.
Low LODs and LOQs were obtained, when compared to other studies
reported in the literature. Additionally, the proposed method meets
some of the principles of green chemistry, since it requires only water,
reduces the consumption of reagents and, consequently, also the gen-
eration of waste. Furthermore, the proposed method uses microwave
irradiation, which increases energy efficiency, since only 5 min irra-
diation is sufficient for complete digestion. Our results demonstrated
that the proposed method is accurate and precise for both elements and
is, potentially, an alternative for routine analysis. Finally, regarding the
elements, variations observed in the food samples might be related to
different factors, such as soil composition as well as genetic factors,
depending on the crop of interest. The use of fertilizers and/or pesti-
cides might also be related to the variations observed. Although

Table 3
Comparison between results for Cl and S in cereals and legumes reported in the literature and from this study.
Sample Origin Concentration (mg kg−1) Reference

Cl S

White rice BRA/CHL/EGY/GER/IND/PAK/URU/USA 130–270 688–1320 Present study

CAN 243–251 1116–1410 Soliman and Zikovsky (1999)
IND 520 – Singh and Garg (2006)
CHN/JAP 705–1299 – Wang and Li (2012)
JAM – 912–1328 Antoine et al. (2012)
Brown rice BRA/GER/USA 141–364 935–1328 Present study
CAN 222 1865 Soliman and Zikovsky (1999)
JAM – 1118–1671 Antoine et al. (2012)
Maize BRA 455–812 759–1226 Present study
GHA 1000 < 160* Nyarko et al., 2006
IND 570 – Singh and Garg (2006)
Wheat BRA 381–394 1291–1347 Present study
CAN 719 3040 Soliman and Zikovsky (1999)
IND 1700 – Balaji et al. (2000)
IND 1420 – Singh and Garg (2006)
Soybean BRA 35–52 2861–2899 Present study
CHN/JAP/USA 47–87 – Miyamoto et al. (2000)
BRA 22 – Barbosa et al. (2013)

Brazil (BRA), Canada (CAN), Chile (CHL), China (CHN), Egypt (EGY), Germany (GER), Ghana (GHA), India (IND), Jamaica (JAM), Japan (JAP), Pakistan (PAK)
United States of America (USA), Uruguay (URU).
* LOD.

338
F.S. Rondan, et al.
concentrations of elements in the plant foods select were not very dif-
ferent from those reported elsewhere in the literature, the method is
more user-friendly and the results suitable for making recommenda-
tions regarding consumption.
Conflict of interest
The authors declared that there is no conflict of interest.
Acknowledgments
This study was financed in part by the Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior - Brazil (CAPES) -
Finance Code 001. The authors are also grateful to Conselho Nacional
de Desenvolvimento Científico e Tecnológico - Brazil (CNPq) - Code
409357/2016-2 and Fundação de Amparo à Pesquisa do Estado do Rio
Grande do Sul - Brazil (FAPERGS) – Code 16/2551-0000-516-8, for
supporting this study.
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