Heliyon

Bentonite-Based Functional Material as Preconcentration System for Determination of

Chromium Species in Water by Flow Injection Analysis Technique
--Manuscript Draft--

Manuscript Number: HELIYON-D-20-00269R2

Article Type: Original Research Article

Section/Category: Chemistry

Keywords: Preconcentration; Trace Analysis; Cr species; modified bentonite; FIA-AAS

Manuscript Classifications: 60.100: Analytical chemistry; 60.160: Physical chemistry; 60.180: Surface Chemistry;
80.160: Agricultural Water Management; 90.160: Environmental Chemistry; 90.200:
Environmental Hazard

Corresponding Author: Handajaya Rusli

Institut Teknologi Bandung
Bandung, West Java INDONESIA

First Author: Muhammad Bachri Amran

Order of Authors: Muhammad Bachri Amran

Sitti Aminah

Handajaya Rusli

Buchari Buchari

Abstract: Chromium species have different level of toxicities. For example, Cr(VI) is 100 times
more toxic than Cr(III). This characteristic makes speciation analysis of chromium
become important. This research will discuss about a development of a Flow Injection
Analysis-Atomic Absorption Spectrometry (FIA-AAS) technique that utilizes bentonite
based functional material as a retention column. The separation, preconcentration and
trace analysis of the Cr species in the water has been successfully performed using a
Bt-MCCs mini-column in the FIA-AAS system. Analytical performance of the developed
method is described as repeatability, linearity, and detection limit. Analytical
performance for Cr(III) are 1.78 %, correlation coefficient 0.9975 for the concentration
range of 50–600 μg.L-1 , and 2.76 μg.L-1 , respectively. Whereas, analytical
performance for Cr(VI) are 0.60 %, correlation coefficient  0.9926 for concentration
range of 50–600 μg.L-1 , and 2.42 μg.L-1 , respectively. This limit detection is better
than the other selective method that has been reported using AAS as detector and the
concentration range can be widened with this limit detection. Evaluation of FIA
performance for both of Cr(III) and Cr(VI) is that it has an enrichment factor of 10 times
higher, it has a concentration efficiency of 12 hour-1 and it has a consumptive index of
12 mL. The analysis that was obtained in Cidurian River, West Java, Indonesia are
38.28 g.L-1 for Cr(III) and 26.73 g.L-1 for Cr(VI), while the accuracy are 98.84 % for
Cr(III), and 100.73 % for Cr(VI).

Suggested Reviewers:

Opposed Reviewers:

Powered by Editorial Manager® and ProduXion Manager® from Aries Systems Corporation
Cover Letter

Chris Russell
Senior Editorial Assistant

Heliyon

May 13th, 2020

Dear Mr. Russell,

I am pleased to resubmit an original research article entitled Bentonite-Based Functional Material as

Preconcentration System for Determination of Chromium Species in Water by Flow Injection
Analysis Technique by Muhammad Bachri Amran, Sitti Aminah, Handajaya Rusli, and Buchari for consideration
for publication in Heliyon. This manuscript is a revision from before manuscript.

In this manuscript, we showed FIA-AAS system with a Bt-MCCs column have good performance for speciation of Cr(III)
and Cr(VI) species and able to determine the concentration each of those species. This technique have been applied
for water sample from Cidurian River, West Java, Indonesia and also give good performance. We believe that, this
manuscript is appropriate for publication by Heliyon because it propose a new, low budget and easy technique to
determine trace concentration of Cr(III) and Cr(VI) in environment, that is still a big problem until now.

This manuscript has not been published and is not under consideration for publication elsewhere. We have no conflicts
of interest to disclose. If you have any question, please tell me. Thank you for your consideration.

Sincerely,

Handajaya Rusli
Analytical Chemistry Research Group
Institut Teknologi Bandung - Indonesia
Response to Reviewers

Editorial office comments:

Please remove “Authors would like to express gratitude to P3MI-ITB Grant and
Ministry of Research, Technology and Higher Education of the Republic of Indonesia
for Doctoral Thesis Research Grant” from your Acknowledgements, as this information
is handled separately.

After discussions with the handling editor, we are not convinced that references
“Separation and preconcentration of trivalent chromium in environmental waters by
using deep eutectic solvent with ultrasound-assisted based dispersive liquid-liquid
microextraction method
Journal of Molecular LiquidsVolume 2911 October 2019Article 111299
Speciation of Cr(III) and Cr(VI) in geological and water samples by ytterbium(III)
hydroxide coprecipitation system and atomic absorption spectrometry
Food and Chemical ToxicologyVolume 49, Issue 7July 2011Pages 1633-1637
3-Ethyl-4-(p-chlorobenzylidenamino-4,5-dihydro-1H-1,2,4-triazol-5-one (EPHBAT) as
precipitant for carrier element free coprecipitation and speciation of
chromium(III) and chromium(VI)
Journal of Hazardous MaterialsVolume 172, Issue 115 December 2009Pages 395-399
Title: Speciation of Cr(III) and Cr(VI) in environmental samples by using
coprecipitation with praseodymium(III) hydroxide and determination by flame atomic
absorption spectrometry
By: Soylak, Mustafa; Aydin, Ayse
Source: JOURNAL OF THE IRANIAN CHEMICAL SOCIETY Volume: 9 Issue: 3 Pages: 263-
267 Published: JUN 2012
Title: Preparation of a chelating resin by immobilizing 1-(2-pyridylazo) 2-naphtol
on amberlite XAD-16 and its application ' of solid phase extraction of Ni(II),
Cd(II), Co(II), Cu(II), Pb(II), and Cr(III) in natural water samples
By: Narin, I; Soylak, M; Kayakirilmaz, K; et al.
Source: ANALYTICAL LETTERS Volume: 36 Issue: 3 Pages: 641-658 Published: 2003
Title: Speciation of Cr(III) and Cr(VI) after column solid phase extraction on
Amberlite XAD-2010
By: Bulut, V. N.; Duran, C.; Tufekci, M.; et al.
Source: JOURNAL OF HAZARDOUS MATERIALS Volume: 143 Issue: 1-2 Pages: 112-117
Published: MAY 8 2007
” are necessary in improving your manuscript. Please reconsider the importance of
these references and remove any that you do not feel are necessary to improve your
manuscript.

Response

We have considered the editor’s input about references and agreed with it. We only included one
reference from Bulut, et al, because its related to our experiment. We have also improved the
acknowledment part according to the input.
Revised manuscript file - highlighting revisions made

Bentonite-Based Functional Material as Preconcentration System for Determination of

1 Chromium Species in Water by Flow Injection Analysis Technique
2
3
4
Muhammad Bachri Amran1
5 Sitti Aminah2
6 Handajaya Rusli1*
7 Buchari, Buchari1
8 1Analytical Chemistry Research Group, Faculty of Mathematics and Natural Sciences,
9
10 Institut Teknologi Bandung, Indonesia
11 2Department of Chemistry Education, Tadulako University, Palu-Indonesia
12
13 *corresponding author
14
15
16 ABSTRACT
17
18
19
Chromium species have different level of toxicities. For example, Cr(VI) is 100 times more toxic than
20 Cr(III). This characteristic makes speciation analysis of chromium become important. This research will
21 discuss about a development of a Flow Injection Analysis-Atomic Absorption Spectrometry (FIA-AAS)
22 technique that utilizes bentonite based functional material as a retention column. The separation,
23 preconcentration and trace analysis of the Cr species in the water has been successfully performed
24
using a Bt-MCCs mini-column in the FIA-AAS system. Analytical performance of the developed method
25
26 is described as repeatability, linearity, and detection limit. Analytical performance for Cr(III) are
27 1.78 %, correlation coefficient 0.9975 for the concentration range of 50–600 μg.L-1, and 2.76 μg.L-1,
28 respectively. Whereas, analytical performance for Cr(VI) are 0.60 %, correlation coefficient 0.9926 for
29 concentration range of 50–600 μg.L-1, and 2.42 μg.L-1, respectively. This limit detection is better than
30 the other selective method that has been reported using AAS as detector and the concentration range
31
32
can be widened with this limit detection. Evaluation of FIA performance for both of Cr(III) and Cr(VI)is
33 that it has an enrichment factor of 10 times higher, it has a concentration efficiency of 12 hour-1 and
34 it has a consumptive index of 12 mL. The analysis that was obtained in Cidurian River, West Java,
35 Indonesia are 38.28 g.L-1 for Cr(III) and 26.73 g.L-1 for Cr(VI), while the accuracy are 98.84 % for Cr(III),
36 and 100.73 % for Cr(VI).
37
38
39 Keywords: Preconcentration, trace analysis, Cr species, modified bentonite, FIA-AAS
40
41
42 Introduction
43
44
45 Anthropogenic activities can harm in environment. It uses heavy metal objects as their culture and can
46 result into water pollution when it is being thrown away to the river. The heavy metal ions can give
47 negative impacts on health and the environment due to its high level of toxicity and a risk of spreading
48 its toxic through the food chain of animals. Chromium (Cr) and its compounds are one of the metals
49 that are popular in various industries such as electroplating, tanning, and paint [1–3]. Chromium is
50
51
one of the trace elements that can be very useful for the human being, at the same time pose serious
52 risks depending on their species. Cr(VI) has 100 times toxicity level higher than Cr(III). Cr(III) is essential
53 for glucose, lipid and protein metabolism in mammals. While Cr(VI) is carcinogenic and toxic for
54 biology system [4], so it is important to determine the concentration each of Cr [5,6].
55
56
Many of Cr speciation using column have been developed such as, Solid Phase Extraction-AAS [7–9]
57
58 [7–11], Inductive Coupled Plasma-Mass Spectrometry (ICP-MS) [10,11], Ionic Chromatography-ICP-MS
59 [12], High Performance Liquid Chromatography (HPLC)-ICP-MS [13], and Reverse Phase-HPLC [14].
60 Those methods are difficult to be applied because the instrument is relatively expensive and have
61
62
63
64
65
 complicated procedure analysis. AAS is one of the standard instruments that is used for metal analysis,
1 but its instrument cannot be used for speciation analysis and does not have a good sensitivity for trace
2 analysis. Given the aforementioned explanation, we need separation and pre concentration before
3
the analysis using AAS to deal with the issue.
4
5
6 The alternative method for preconcentration and trace analysis of metal ion is a non-chromatography
7 mini-column technique based on FIA with AAS as detector. The advantages of FIA method compared
8 to other conventional preconcentration method are; low cost, high repeatability, fewer sample
9 needed, relatively short time for analysis, and easy to couple with any detection methods [15,16].
10
11 Many strategies have been developed to find Cr speciation method based on FIA for example using a
12 functional material as sorbent in FIA system. This sorbent can get through modification of a natural
13 material or synthesis of new material. Chelating resin is one of the popular sorbent using in FIA system
14 such as; C-18 column with 1,5-diphenyl carbazide (DPC) as a ligand in FIA-UV-Vis [17], functionalized
15 alumina surfactant with 8-hidroquinoline in FIA-AAS [18], modified XAD in FIA-AAS [19,20][19–21].
16
17
All these FIA systems have a limitation. They can only analyze one of Cr species. So, it is needed to
18 develop an alternative sorbent which has a good performance to determine Cr(III) and Cr(VI) species
19 using FIA system.
20
21 This research managed to develop and evaluate a new FIA-AAS technique utilizing a modified
22
bentonite as a functional material for low cost preconcentration system with good analytical
23
24 performance. Modified bentonite as sorbent has not reported yet to the best of our knowledge. The
25 functional material was made from bentonite which was modified with cetyltrimethylammonium
26 bromide (CTAB) and chitosan. This material could improve the weakness of AAS technique (sensitivity,
27 selectivity and detection limit) for Cr species analysis. The combination with FIA technique can
28 produce a new technique which are able to determine Cr species simultaneously with being selective
29
30 in trace concentration. These abilities expected are expected to give a contribution for determination
31 of Cr(III) and Cr(VI) species in the environment.
32
33
34 Materials and methods
35
36
37 Synthesis of bentonite functional material (Bt-MCCs)
38 The natural bentonite was obtained from Cipatat, West Java, Indonesia and it was used without any
39 further treatment. Cr(NO3)3.9H2O and K2Cr2O7 were used as Cr(III) and Cr(VI) standard solution. CTAB,
40 sodium hydroxide, nitric acid, ammonia solution and acetic acid glacial were taken from Merck-
41 Germany. While chitosan DD 81 % was from Biotech. All chemicals used in this research were pro
42
43 analyst grade except mentioned differently. The Bt-MCCs material has been synthesis with a previous
44 procedure that was reported by Aminah [21].
45
46 Five grams of natural bentonite with particle size +200/-100 mesh dispersed in 250 mL of 0.01 mol.L-1
47 CTAB solution and stirred for 24 hours at room temperature. Then, the product was filtered and
48
49
washed with distilled water until bromide ion became undetectable. Then, dried at 60°C for 24 hours.
50 Afterwards, this aggregate crushed and sieved to obtain particle size +200/-100 mesh. This material is
51 called Bt-MCs. Five grams of Bt-MCs then added to 250 mL of distilled water to get a 2.0 % suspension.
52 On the other hand, a chitosan solution was prepared by dissolving five grams of chitosan DD 81 % in
53 250 mL of 1.0 % acetic acid solution. The chitosan solution added slowly to suspension and stirred for
54
24 hours at room temperature. The formed materials were washed with distilled water until neutral
55
56 pH and then dried at 60°C for 24 hours. Finally, the aggregate was crushed and sieved to obtain particle
57 size +200/-100 to get a Bt-MCCs materials.
58
59 FIA-AAS system
60
61
62
63
64
65
 In this system, a flow injection was operated based on volume to study eluent volume, eluent
1 concentration, and sample volume effects. Determination of Cr(III) and Cr(VI) was done using AAS. A
2 peristaltic pump, a valve and a loop were used to give elution pressure, elution control, and controlling
3
of sample volume, respectively. For application, 0.2 gram of Bt-MCCs material was packed in mini-
4
5 column. To get the best result, optimization and evaluations of FIA system was done. The evaluation
6 and optimization of sample volume, kind of eluent, eluent concentration, and volume of eluent were
7 applied to get best result. The eluent for Cr(III) and Cr(VI) is nitric acid and ammonia solution,
8 respectively. The carrier in this system was a water of pH 5, which acidified by nitric acid. The
9 concentration of Cr(III) and Cr(VI) that was used in this experiment to study the concentration and
10
11 volume eluent is 250 μg.L-1. The flow of the carrier was 2.0 mL.minute-1. Diagram for FIA-AAS volume-
12 based system illustrated in Figure 1[22].
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35 Figure 1 Diagram of FIA-AAS volume-based system
36
37
(C: Carrier; S: Sample; E = Eluent; P = Peristaltic pump; MC: Bt-MCCs mini-column; FAAS: Flame Atomic
38
39 Absorption Spectrometer)
40
41
42 Results
43
44 Using both CTAB and chitosan together as a modifier shows an increase of retention of Cr(III) and
45
Cr(VI). This phenomenon is caused by the synergistic effect of the active group in chitosan and CTAB
46
47 surfactant. Chitosan have -NH2 and -OH functional groups that has a good affinity for Cr(III) and can
48 interact with anion species such as Cr(VI) depending on the pH of the solution [23]. CTAB as a
49 quaternary ammonium groups can change the charge of bentonite surface from negative to positive,
50 so it will be easy to interact with Cr(VI) [21]. In Figure 2, the synergistic effect show and concludes that
51 Bt-MCCs have better performance for adsorption of Cr(III) and Cr(VI) than one modifier only and
52
53 optimum ratio Bt-MCs and chitosan was 1:1. Effect of another parameter that has an effect on
54 retention capacity in a batch system and physiochemistry characterization have been reported in the
55 previous paper [21].
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25 Figure 2 Synergistic effect between Bt-MCs and chitosan
26
27 Optimization condition
28 The Bt-MCCs mini column showed a good capability to preconcentrate Cr(III) and Cr(VI) ions. Figure 3
29
showed the comparison of the signal with and without preconcentration. A mixture of Cr(III) and Cr(VI)
30
31 with the total concentration 2.0 mg.L-1 that was directly measured using AAS has the same peak height
32 with a 1.0 mL of Cr(III) or Cr(VI) with each concentration of 0.1 mg.L-1 (equal to 0.1 μg Cr) that had
33 preconcentration treatment. Generally, AAS only has the ability to analyze the total Cr in mg.L-1 unit
34 and cannot differentiate Cr species. This fact showed the developed FIA-AAS system is effective to give
35 information about the kind and concentration each of Cr(III) and Cr(VI) species. This results also
36
37 showed possibility to use the Bt-MCCs mini column to preconcentrated Cr(III) and Cr(VI) ions with
38 lower concentration.
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23 Figure 3 Comparison of Cr(III) and Cr(VI) peak height with and without preconcentration
24
25
We found the Bt-MCCs column not only has a good performance for low concentration sample, but
26
27 also good for higher concentration. A mixture standard of Cr(III) and Cr(VI) with each concentration
28 0.5 mg.L-1 as sample and injected 0.5, 1.0 and 1.5 mL of it to get 0.5, 1.0, and 1.5 μg of Cr total. A
29 fiagram in Figure 4 showed the increase of the sample volume with the same Cr species concentration
30 that also increase the peak height of AAS signal. This result indicates that up until the concentration
31 of analyte rises up to 15 times from the initial, the Bt-MCCs mini-column still has a good capability to
32
33 absorb the Cr species. This results can explained from VB capacity of column retention that is 3.13 and
34 2.25 mg.g-1 for Cr(III) and Cr(VI), respectively [21]. For 0.2 g of Bt-MCCs mini column, it can retain up
35 to 626.0 μg Cr(III) and 450.0 μg Cr(VI). That data means the dynamic retention capacity from the Bt-
36 MCCs is still much bigger than the Cr injection from the sample, so both of volume and concentration
37 sample can be enlarged to get bigger preconcentration factor
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Figure 4 Effect of sample volume to fiagram of Cr(III) and Cr(VI)
25
26 Elution is one of importance step after a retention process. A Cr(III) ion, which has a positive charge,
27 can only interact with chitosan. A cation competitor is needed to weaken the interaction between
28 Cr(III) and chitosan. We have used nitric acid to fulfill this condition. On the other hand, a Cr(VI) ion
29
which has a negative charge will interacted with both of quaternary ammonium from CTAB and
30
31 chitosan. With same logic, we have used ammonia to meet the condition. The order of elution will
32 affect the quality of speciation. Half of Cr(VI) was also released when Cr(III) elute first, so we will elute
33 Cr(VI) first. Eluent concentration between 0.1-2.0 mol.L-1 was used to evaluate effect of eluent
34 concentration to quantitative signal of analyte. A 1.0 mL of Cr(III) and Cr(VI) mixture solution with each
35 concentration 0.25 mg.L-1 was used for this analysis. A fiagram in Figure 5 showed that quantitative
36
37 elution (>95%) was reached when minimal concentration 1.0 mol.L-1 for nitric acid to elute Cr(III).
38 Unfortunately, concentration more than 2.0 mol.L-1 made the conditioning time longer and predicted
39 that it will destroy the Bt-MCCs materials. A 0.5 mol.L-1 of ammonia solution already gave a maximum
40 Cr(VI) elution. So for further analysis, nitric acid 2.0 mol.L-1 and ammonia 0.5 mol.L-1 were chosen. A
41 fiagram of Cr(III) and Cr(VI) speciation in optimum concentration of eluent is presented in Figure 6.
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Figure 5 Effect of eluent concentration to fiagram of Cr(III) and Cr(VI)
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
Figure 6 Fiagram of Cr(III) and Cr(VI) separation in optimum eluent concentration
48
49
50 Ideal elution of sample can be reach if the analysis is fast, accurate, precision and consume less
51 eluent[16]. Optimum eluent concentration was used to determine the eluent volume that produce
52 optimum peak height. Eluent volume variation was controlled using a loop with volume 0.2, 0.5, and
53 1.0 mL. A fiagram in Figure 7 showed that both of nitric acid and ammonia give optimum elution when
54
55 the volume of eluent 0.5 mL. Volume below 0.5 mL gave lower height peak because the elution is not
56 maximal yet and bigger than 0.5 mL did not gave higher peak because dilution effect. Based on this
57 fact, the next analysis would use a 0.5 mL of eluent.
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24 Figure 7 Effect of eluent volume to fiagram of Cr(III) and Cr(VI)
25
26 Analytical performance
27 To evaluate the analytical performance, we determined linearity, selectivity, repeatability, detection
28 limit, enrichment factor, concentration efficiency and consumptive index. Linearity was determine
29
based on linear regression of a certain range concentration of analyte. In this experiment,
30
31 concentration between 50–500 μg.L-1 has a good relation between peak height (H) and analyte
32 concentration ([Cr]) with dynamic range up to 12-fold. A fiagram profile of various concentration Cr
33 species in optimum condition is presented in Figure 8. A linear regression plotting give correlation
34 coefficient R2 = 0.9905 for Cr(III) and R2 = 0.9926 for Cr(IV). Both R2 value showed good linearity of this
35 FIA-AAS system in the range of analyte concentration.
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60 Figure 8 Fiagram profile of Cr(III) and Cr(VI) on various concentration in optimum condition
61
62
63
64
65
 1 The % variance coefficient (%VC) is used to state quality of the analysis repeatability. Lower %VC
2 means better precision. At optimum condition, fiagram like Figure 9 will be obtained. The %VC value
3
of Cr(III) and Cr(VI) is 1.8 % and 0.6 %, respectively. The %VC value lower than 5.0 % indicates that the
4
5 repeatability of developed analytical methods is very good. The Bt-MCCs mini column have been
6 developed show good stability and performance for routine analysis.
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31 Figure 9 Fiagram profile of Cr(III) and Cr(VI) in optimum condition for five repetitions
32
33 The developed method has a limit detection consecutively 2.8 and 2.4 μg.L-1 for Cr(III) and Cr(VI) for 1
34 mL of sample based on limitation a S/N ratio = 3. The minimal mass detected (MMD) is 2.8 ng for Cr(III)
35 and 2.4 ng for Cr(VI). Saygi et al reports they have a limit detection of 1.94 μg.L-1 as Cr(VI) for 25 mL
36
37 minimal of sample [8]. It means that the MMD for this system is 48.5 ng. So, it can be concluded that
38 this method has a better limit detection than another selective method that has been reported using
39 AAS for ion detection. For too low concentration of Cr, this limit detection can be attained by enlarging
40 the injection volume. But it needs to consider that more volume will make a longer analysis time and
41 an effect to retention-elution process.
42
43
44 An enrichment factor (EF) describe ratio analyte concentration in sample before and after
45 preconcentration. The EF calculated by comparing the slope of sample calibration curve with and
46 without preconcentration. Table 1 showed a linear regression parameter for Cr(III) and Cr(VI) with and
47 without preconcentration in equation
48 H = A + B[Cr], H = peak height; A = constant; B = slope; [Cr] = chromium concentration
49
50 Calculation results showed preconcentration with the developed Bt-MCCs mini-column have EF 10-
51 fold for 1 mL sample. This EF value can still improve by increasing the sample volume. Data in Table 1
52 also can use to calculate sensitivity. Sensitivity can be calculated using equation
53 S = 0.0044/k, S = sensitivity; k = slope of calibration curve
54 Calculation give the sensitivity are 14.7 μg.L-1 for both of Cr species.
55
56
57 Table 1. Regression parameter of Cr(III) and Cr(VI) with and without preconcentration
58 Condition Concentration Species Parameter
-1
59 range (μg.L ) A B R
60 1000–9000 Cr(III) -0.0060 0.00003 0.9984
61
62
63
64
65
 Without Cr(VI) -0.0028 0.00003 0.9964
1 preconcentration
2 With 50–500 Cr(III) -0.0037 0.0003 0.9905
3
preconcentration Cr(VI) -0.0046 0.0003 0.9926
4
5
6 The concentration efficiency (CE) value was determined by counting the analyte peaks. This method
7 can be used in an hour. In this developed method, we can get two analyte peaks (Cr(III) and Cr(VI)) in
8 5 minutes, so CE for this method is 12 hour-1. This value means that the developed method has a short
9 analysis time and worth to use for routine analysis with high number of samples. Beside EF dan CE,
10
11 consumptive index (CI) also can be used to evaluate the FIA performance. CI is a FIA parameter which
12 is related to carrier and eluent volume. A FIA preconcentration supposedly have a small CI value to
13 get an efficiency. In optimum condition, this developed system needs total volume 12.0 mL with detail
14 10 mL of carrier (2.0 mL.minute-1 x 5 minute), 1.0 mL of sample, and 1.0 mL of eluent (2 eluent x 0.5
15 mL) to get 2 signal from different Cr species. The CI value maybe increase if the concentration sample
16
17
very low, but EF value also increases too to compensate it without lose another analytical parameter.
18
19 Application for environmental sample
20 The sample that has been analyzed using this FIA-AAS system came from Cidurian river, West Java,
21 Indonesia and taken in January 2017. The sample was filtered using filter paper to remove an
22
undissolved particle and then the filtrate was analyzed using the optimum condition. To learn the
23
24 matrix effect in analysis, we had used spike method and evaluated the percent recovery. Table 2
25 showed the percent recovery of river water sample for Cr(III) and Cr(VI). A higher percentage of
26 recovery value indicates that the method has high accuracy and sample matrix does not give
27 significant effect to the measurement.
28
29
30 Table 2. Concentration of Cr(III) and Cr(VI) in Cidurian river
31 Sample Cr concentration (μg.L-1) Percent recovery Species
32 Added Found (%)
33 Cidurian 0 38.3 ± 1.8 98.8 Cr(III)
34 river 100.0 136.7 ± 1.8
35
36 water 0 26.7 ± 0.8 100.7 Cr(VI)
37 75.0 101.7 ± 0.8
38
39 Table 2 also showed total concentration of chromium in river sample is 65.0 ± 2.6 μg.L-1. This value is
40
41 too small for FAAS analysis because of inadequate sensitivity. Using the Bt-MCCs mini-column in FIA-
42 AAS system solved that problem. Indonesian government regulation by Ministry of Environment and
43 Forestry decree determine the threshold value for Cr total and Cr(VI) in water are 0.10 mg.L-1 and 0.05
44 mg.L-1 respectively. Sample analysis showed Cr species total is still below this threshold.
45
46
Conclusion
47
48
49 A Bt-MCCs mini-column can be prepared easily because, all of the chemical compounds are relatively
50 easy to get and cheap. The column that has been used in FIA-AAS system shows a good analytical
51 performance and can be applied to trace the analysis of Cr species. The column effectively function as
52 a preconcentrator for low concentration Cr(III) and Cr(VI) in real sample matrix without losing the
53
54 speciation performance. It is possible to decrease the concentration range of chromium species with
55 increasing the volume sample injected to column. Increasing the sample volume of course effects the
56 time to flow all the sample through the mini-column.
57
58 Speciation that was done using 0.5 mol.L-1 ammonia solution and 2.0 mol.L-1 nitric acid solution as
59
60
specific eluent for each of chromium species. This eluent combination did not damage the column
61
62
63
64
65
 because the peak heigth height has a good consistency after using it in many cycles. We suggest this
1 technique for Cr(III) and Cr(VI) routine analysis in environmental sample because this technique have
2 advantages like accurate, precise, and robust methods, modest equipment, short time for analysis,
3
easily automated, low sample volume, easily to tandem with another equipment and low cost. An
4
5 analytical performance summary is showed at Table 3.
6
7 Table 3. Summary of analytical parameter
8 Parameter Value
9 Cr(III) Cr(VI)
10
11 Concentration range (μg.L-1) 50–500 50–500
12 Correlation coefficient 0.9926 0.9975
13 Sensitivity (μg.L-1) 14.7 14.7
14 Repeatability (%) 1.8 0.6
15 Limit detection (μg.L-1) 2.8 2.4
16
17
EF 10 10
18 CE (hour-1) 12 12
19 CI (mL) 12 12
20 Accuracy (%) 98.8 100.7
21
22
Acknowledgment
23
24
25 Authors would like to express gratitude to to P3MI-ITB Grant and Ministry of Research, Technology
26 and Higher Education of the Republic of Indonesia for Doctoral Thesis Research Grant and M. A. Majid
27 Al Kindi for proofreading this manuscript.
28
29
30 References
31
32 [1] M.K. Hasan, A. Shahriar, K.U. Jim, Water pollution in Bangladesh and its impact on public
33 health, Heliyon. 5 (2019) e02145. doi:10.1016/j.heliyon.2019.e02145.
34 [2] R.R. Elmorsi, K.S. Abou-El-Sherbini, G.A.-H. Mostafa, M.A. Hamed, Distribution of essential
35
heavy metals in the aquatic ecosystem of Lake, Heliyon. 5 (2019).
36
37 doi:10.1016/j.heliyon.2019.e02276.
38 [3] L.N. Nthunya, M.L. Masheane, S.P. Malinga, E.N. Nxumalo, B.B. Mamba, S.D. Mhlanga,
39 Determination of toxic metals in drinking water sources in the Chief Albert Luthuli local
40 municipality in Mpumalanga, South Africa, Phys. Chem. Earth, Parts A/B/C. 100 (2017) 94–
41 100. doi:10.1016/j.pce.2017.04.006.
42
43 [4] B. Kiran, N. Rani, A. Kaushik, Environmental toxicity: Exposure and impact of chromium on
44 cyanobacterial species, J. Environ. Chem. Eng. 4 (2016) 4137–4142.
45 doi:10.1016/j.jece.2016.09.021.
46 [5] J. Kotaś, Z. Stasicka, Chromium occurrence in the environment and methods of its speciation,
47 Environ. Pollut. 107 (2000) 263–283. doi:10.1016/S0269-7491(99)00168-2.
48
49
[6] V. Gómez, M.P. Callao, Chromium determination and speciation since 2000, Trends Anal.
50 Chem. 25 (2006) 1006–1015. doi:10.1016/j.trac.2006.06.010.
51 [7] V.N. Bulut, C. Duran, M. Tufekci, L. Elci, M. Soylak, Speciation of Cr(III) and Cr(VI) after column
52 solid phase extraction on Amberlite XAD-2010, J. Hazard. Mater. 143 (2007) 112–117.
53 doi:10.1016/j.jhazmat.2006.08.074.
54
[8] K.O. Saygi, M. Tuzen, M. Soylak, L. Elci, Chromium speciation by solid phase extraction on
55
56 Dowex M 4195 chelating resin and determination by atomic absorption spectrometry, J.
57 Hazard. Mater. 153 (2008) 1009–1014. doi:10.1016/j.jhazmat.2007.09.051.
58 [9] B. Saha, R.J. Gill, D.G. Bailey, N. Kabay, M. Arda, Sorption of Cr(VI) from aqueous solution by
59 Amberlite XAD-7 resin impregnated with Aliquat 336, React. Funct. Polym. 60 (2004) 223–
60
61
62
63
64
65
 244. doi:10.1016/j.reactfunctpolym.2004.03.003.
1 [10] Y.C. Sun, C.Y. Lin, S.F. Wu, Y.T. Chung, Evaluation of on-line desalter-inductively coupled
2 plasma-mass spectrometry system for determination of Cr(III), Cr(VI), and total chromium
3
concentrations in natural water and urine samples, Spectrochim. Acta Part B. 61 (2006) 230–
4
5 234. doi:10.1016/j.sab.2006.01.007.
6 [11] B. Wen, X. Shan, J. Lian, Separation of Cr(III) and Cr(VI) in river and reservoir water with 8-
7 hydroxyquinoline immobilized polyacrylonitrile fiber for determination by inductively coupled
8 plasma mass spectrometry, Talanta. 56 (2002) 681–687.
9 [12] G. Choppala, M.M. Rahman, Z. Chen, N. Bolan, M. Mallavarapu, Simultaneous determination
10
11 of chromium species by ion chromatography coupled with inductively coupled plasma mass
12 spectrometry, in: 19th World Congr. Soil Sci. Soil Solut. a Chang. World, International Union
13 of Soil Sciences, 2010: pp. 114–116.
14 [13] Y. Martinez-Bravo, A.F. Roig-Navarrro, F.J. Lopez, F. Hernandez, Multielemental
15 determination of arsenic, selenium and chromium(VI) species in water by high-performance
16
17
liquid chromatography – inductively coupled plasma mass spectrometry, J. Chromatogr. A.
18 926 (2001) 265–274.
19 [14] M.A. Hossain, M. Kumita, Y. Michigami, T.S. Islam, S. Mori, Rapid speciation analysis of Cr(VI)
20 and Cr(III) by reversed-phase high-performance liquid chromatography with UV detection, J.
21 Chromatogr. Sci. 43 (2005) 98–103. doi:10.1093/chromsci/43.2.98.
22
[15] D. Harvey, Modern Analytical Chemistry, McGraw-Hill, 2000.
23
24 [16] F. Zhaloun, Flow injection separation and preconcentration, Wiley-VCH, 1993.
25 [17] K. Grudpan, N. Worakijchar oenchai, O. Tue-Ngeun, P. Sooksamiti, J. Jakmunee, Flow injection
26 spectrophotometry for speciation of Cr(VI) and Cr(III) using diphenylcarbazide and solid phase
27 extraction with C18 in-valve mini-column, Sci. Asia. 25 (1999) 99–106.
28 [18] S.H. Ahmadi, A.M. Haji Shabani, S. Dadfarnia, M. Taei, On-line preconcentration and
29
30 speciation of chromium by an 8-hydroxyquinoline microcolumn immobilized on surfactant-
31 coated alumina and flow injection atomic absorption spectrometry, Turkish J. Chem. 31
32 (2007) 191–199.
33 [19] A. Ahmad, J.A. Siddique, M.A. Laskar, R. Kumar, S.H. Mohd-setapar, A. Khatoon, R.A. Shiekh,
34 New generation Amberlite XAD resin for the removal of metal ions: A review, J. Environ. Sci.
35
31 (2015) 104–123. doi:10.1016/j.jes.2014.12.008.
36
37 [20] M. Wulandari, Resin Pengkhelat Amberlit XAD-16-1,5-difenilkarbazid untuk prakonsentrasi
38 dan analisis selektif Cr(VI), Institut Teknologi Bandung, 2010.
39 [21] S. Aminah, Buchari Buchari, Muhammad Bachri Amran, Bentonite based functional material
40 for chromium species adsorption, Int. J. Appl. Sci. Technol. 7 (2017) 77–86.
41 [22] M.B. Amran, A.S. Panggabean, A. Sulaeman, M. Rusnadi, Preparation of a chelating resin and
42
43 its application as a preconcentration system for determination of cadmium in river water by
44 flow injection analysis, Internastional J. Environ. Res. 5 (2011) 531–536.
45 doi:10.22059/IJER.2011.337.
46 [23] E. Guibal, Interactions of metal ions with chitosan-based sorbents: A review, Sep. Purif.
47 Technol. 38 (2004) 43–74. doi:10.1016/j.seppur.2003.10.004.
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
Manuscript File Click here to view linked References

Bentonite-Based Functional Material as Preconcentration System for Determination of

1 Chromium Species in Water by Flow Injection Analysis Technique
2
3
4
Muhammad Bachri Amran1
5 Sitti Aminah2
6 Handajaya Rusli1*
7 Buchari, Buchari1
8 1Analytical Chemistry Research Group, Faculty of Mathematics and Natural Sciences,
9
10 Institut Teknologi Bandung, Indonesia
11 2Department of Chemistry Education, Tadulako University, Palu-Indonesia
12
13 *corresponding author
14
15
16 ABSTRACT
17
18
19
Chromium species have different level of toxicities. For example, Cr(VI) is 100 times more toxic than
20 Cr(III). This characteristic makes speciation analysis of chromium become important. This research will
21 discuss about a development of a Flow Injection Analysis-Atomic Absorption Spectrometry (FIA-AAS)
22 technique that utilizes bentonite based functional material as a retention column. The separation,
23 preconcentration and trace analysis of the Cr species in the water has been successfully performed
24
using a Bt-MCCs mini-column in the FIA-AAS system. Analytical performance of the developed method
25
26 is described as repeatability, linearity, and detection limit. Analytical performance for Cr(III) are
27 1.78 %, correlation coefficient 0.9975 for the concentration range of 50–600 μg.L-1, and 2.76 μg.L-1,
28 respectively. Whereas, analytical performance for Cr(VI) are 0.60 %, correlation coefficient 0.9926 for
29 concentration range of 50–600 μg.L-1, and 2.42 μg.L-1, respectively. This limit detection is better than
30 the other selective method that has been reported using AAS as detector and the concentration range
31
32
can be widened with this limit detection. Evaluation of FIA performance for both of Cr(III) and Cr(VI)is
33 that it has an enrichment factor of 10 times higher, it has a concentration efficiency of 12 hour-1 and
34 it has a consumptive index of 12 mL. The analysis that was obtained in Cidurian River, West Java,
35 Indonesia are 38.28 g.L-1 for Cr(III) and 26.73 g.L-1 for Cr(VI), while the accuracy are 98.84 % for Cr(III),
36 and 100.73 % for Cr(VI).
37
38
39 Keywords: Preconcentration, trace analysis, Cr species, modified bentonite, FIA-AAS
40
41
42 Introduction
43
44
45 Anthropogenic activities can harm in environment. It uses heavy metal objects as their culture and can
46 result into water pollution when it is being thrown away to the river. The heavy metal ions can give
47 negative impacts on health and the environment due to its high level of toxicity and a risk of spreading
48 its toxic through the food chain of animals. Chromium (Cr) and its compounds are one of the metals
49 that are popular in various industries such as electroplating, tanning, and paint [1–3]. Chromium is
50
51
one of the trace elements that can be very useful for the human being, at the same time pose serious
52 risks depending on their species. Cr(VI) has 100 times toxicity level higher than Cr(III). Cr(III) is essential
53 for glucose, lipid and protein metabolism in mammals. While Cr(VI) is carcinogenic and toxic for
54 biology system [4], so it is important to determine the concentration each of Cr [5,6].
55
56
Many of Cr speciation using column have been developed such as, Solid Phase Extraction-AAS [7–9],
57
58 Inductive Coupled Plasma-Mass Spectrometry (ICP-MS) [10,11], Ionic Chromatography-ICP-MS [12],
59 High Performance Liquid Chromatography (HPLC)-ICP-MS [13], and Reverse Phase-HPLC [14]. Those
60 methods are difficult to be applied because the instrument is relatively expensive and have
61
62
63
64
65
 complicated procedure analysis. AAS is one of the standard instruments that is used for metal analysis,
1 but its instrument cannot be used for speciation analysis and does not have a good sensitivity for trace
2 analysis. Given the aforementioned explanation, we need separation and pre concentration before
3
the analysis using AAS to deal with the issue.
4
5
6 The alternative method for preconcentration and trace analysis of metal ion is a non-chromatography
7 mini-column technique based on FIA with AAS as detector. The advantages of FIA method compared
8 to other conventional preconcentration method are; low cost, high repeatability, fewer sample
9 needed, relatively short time for analysis, and easy to couple with any detection methods [15,16].
10
11 Many strategies have been developed to find Cr speciation method based on FIA for example using a
12 functional material as sorbent in FIA system. This sorbent can get through modification of a natural
13 material or synthesis of new material. Chelating resin is one of the popular sorbent using in FIA system
14 such as; C-18 column with 1,5-diphenyl carbazide (DPC) as a ligand in FIA-UV-Vis [17], functionalized
15 alumina surfactant with 8-hidroquinoline in FIA-AAS [18], modified XAD in FIA-AAS [19,20]. All these
16
17
FIA systems have a limitation. They can only analyze one of Cr species. So, it is needed to develop an
18 alternative sorbent which has a good performance to determine Cr(III) and Cr(VI) species using FIA
19 system.
20
21 This research managed to develop and evaluate a new FIA-AAS technique utilizing a modified
22
bentonite as a functional material for low cost preconcentration system with good analytical
23
24 performance. Modified bentonite as sorbent has not reported yet to the best of our knowledge. The
25 functional material was made from bentonite which was modified with cetyltrimethylammonium
26 bromide (CTAB) and chitosan. This material could improve the weakness of AAS technique (sensitivity,
27 selectivity and detection limit) for Cr species analysis. The combination with FIA technique can
28 produce a new technique which are able to determine Cr species simultaneously with being selective
29
30 in trace concentration. These abilities expected are expected to give a contribution for determination
31 of Cr(III) and Cr(VI) species in the environment.
32
33
34 Materials and methods
35
36
37 Synthesis of bentonite functional material (Bt-MCCs)
38 The natural bentonite was obtained from Cipatat, West Java, Indonesia and it was used without any
39 further treatment. Cr(NO3)3.9H2O and K2Cr2O7 were used as Cr(III) and Cr(VI) standard solution. CTAB,
40 sodium hydroxide, nitric acid, ammonia solution and acetic acid glacial were taken from Merck-
41 Germany. While chitosan DD 81 % was from Biotech. All chemicals used in this research were pro
42
43 analyst grade except mentioned differently. The Bt-MCCs material has been synthesis with a previous
44 procedure that was reported by Aminah [21].
45
46 Five grams of natural bentonite with particle size +200/-100 mesh dispersed in 250 mL of 0.01 mol.L-1
47 CTAB solution and stirred for 24 hours at room temperature. Then, the product was filtered and
48
49
washed with distilled water until bromide ion became undetectable. Then, dried at 60°C for 24 hours.
50 Afterwards, this aggregate crushed and sieved to obtain particle size +200/-100 mesh. This material is
51 called Bt-MCs. Five grams of Bt-MCs then added to 250 mL of distilled water to get a 2.0 % suspension.
52 On the other hand, a chitosan solution was prepared by dissolving five grams of chitosan DD 81 % in
53 250 mL of 1.0 % acetic acid solution. The chitosan solution added slowly to suspension and stirred for
54
24 hours at room temperature. The formed materials were washed with distilled water until neutral
55
56 pH and then dried at 60°C for 24 hours. Finally, the aggregate was crushed and sieved to obtain particle
57 size +200/-100 to get a Bt-MCCs materials.
58
59 FIA-AAS system
60
61
62
63
64
65
 In this system, a flow injection was operated based on volume to study eluent volume, eluent
1 concentration, and sample volume effects. Determination of Cr(III) and Cr(VI) was done using AAS. A
2 peristaltic pump, a valve and a loop were used to give elution pressure, elution control, and controlling
3
of sample volume, respectively. For application, 0.2 gram of Bt-MCCs material was packed in mini-
4
5 column. To get the best result, optimization and evaluations of FIA system was done. The evaluation
6 and optimization of sample volume, kind of eluent, eluent concentration, and volume of eluent were
7 applied to get best result. The eluent for Cr(III) and Cr(VI) is nitric acid and ammonia solution,
8 respectively. The carrier in this system was a water of pH 5, which acidified by nitric acid. The
9 concentration of Cr(III) and Cr(VI) that was used in this experiment to study the concentration and
10
11 volume eluent is 250 μg.L-1. The flow of the carrier was 2.0 mL.minute-1. Diagram for FIA-AAS volume-
12 based system illustrated in Figure 1[22].
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35 Figure 1 Diagram of FIA-AAS volume-based system
36
37
(C: Carrier; S: Sample; E = Eluent; P = Peristaltic pump; MC: Bt-MCCs mini-column; FAAS: Flame Atomic
38
39 Absorption Spectrometer)
40
41
42 Results
43
44 Using both CTAB and chitosan together as a modifier shows an increase of retention of Cr(III) and
45
Cr(VI). This phenomenon is caused by the synergistic effect of the active group in chitosan and CTAB
46
47 surfactant. Chitosan have -NH2 and -OH functional groups that has a good affinity for Cr(III) and can
48 interact with anion species such as Cr(VI) depending on the pH of the solution [23]. CTAB as a
49 quaternary ammonium groups can change the charge of bentonite surface from negative to positive,
50 so it will be easy to interact with Cr(VI) [21]. In Figure 2, the synergistic effect show and concludes that
51 Bt-MCCs have better performance for adsorption of Cr(III) and Cr(VI) than one modifier only and
52
53 optimum ratio Bt-MCs and chitosan was 1:1. Effect of another parameter that has an effect on
54 retention capacity in a batch system and physiochemistry characterization have been reported in the
55 previous paper [21].
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25 Figure 2 Synergistic effect between Bt-MCs and chitosan
26
27 Optimization condition
28 The Bt-MCCs mini column showed a good capability to preconcentrate Cr(III) and Cr(VI) ions. Figure 3
29
showed the comparison of the signal with and without preconcentration. A mixture of Cr(III) and Cr(VI)
30
31 with the total concentration 2.0 mg.L-1 that was directly measured using AAS has the same peak height
32 with a 1.0 mL of Cr(III) or Cr(VI) with each concentration of 0.1 mg.L-1 (equal to 0.1 μg Cr) that had
33 preconcentration treatment. Generally, AAS only has the ability to analyze the total Cr in mg.L-1 unit
34 and cannot differentiate Cr species. This fact showed the developed FIA-AAS system is effective to give
35 information about the kind and concentration each of Cr(III) and Cr(VI) species. This results also
36
37 showed possibility to use the Bt-MCCs mini column to preconcentrated Cr(III) and Cr(VI) ions with
38 lower concentration.
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23 Figure 3 Comparison of Cr(III) and Cr(VI) peak height with and without preconcentration
24
25
We found the Bt-MCCs column not only has a good performance for low concentration sample, but
26
27 also good for higher concentration. A mixture standard of Cr(III) and Cr(VI) with each concentration
28 0.5 mg.L-1 as sample and injected 0.5, 1.0 and 1.5 mL of it to get 0.5, 1.0, and 1.5 μg of Cr total. A
29 fiagram in Figure 4 showed the increase of the sample volume with the same Cr species concentration
30 that also increase the peak height of AAS signal. This result indicates that up until the concentration
31 of analyte rises up to 15 times from the initial, the Bt-MCCs mini-column still has a good capability to
32
33 absorb the Cr species. This results can explained from VB capacity of column retention that is 3.13 and
34 2.25 mg.g-1 for Cr(III) and Cr(VI), respectively [21]. For 0.2 g of Bt-MCCs mini column, it can retain up
35 to 626.0 μg Cr(III) and 450.0 μg Cr(VI). That data means the dynamic retention capacity from the Bt-
36 MCCs is still much bigger than the Cr injection from the sample, so both of volume and concentration
37 sample can be enlarged to get bigger preconcentration factor
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Figure 4 Effect of sample volume to fiagram of Cr(III) and Cr(VI)
25
26 Elution is one of importance step after a retention process. A Cr(III) ion, which has a positive charge,
27 can only interact with chitosan. A cation competitor is needed to weaken the interaction between
28 Cr(III) and chitosan. We have used nitric acid to fulfill this condition. On the other hand, a Cr(VI) ion
29
which has a negative charge will interacted with both of quaternary ammonium from CTAB and
30
31 chitosan. With same logic, we have used ammonia to meet the condition. The order of elution will
32 affect the quality of speciation. Half of Cr(VI) was also released when Cr(III) elute first, so we will elute
33 Cr(VI) first. Eluent concentration between 0.1-2.0 mol.L-1 was used to evaluate effect of eluent
34 concentration to quantitative signal of analyte. A 1.0 mL of Cr(III) and Cr(VI) mixture solution with each
35 concentration 0.25 mg.L-1 was used for this analysis. A fiagram in Figure 5 showed that quantitative
36
37 elution (>95%) was reached when minimal concentration 1.0 mol.L-1 for nitric acid to elute Cr(III).
38 Unfortunately, concentration more than 2.0 mol.L-1 made the conditioning time longer and predicted
39 that it will destroy the Bt-MCCs materials. A 0.5 mol.L-1 of ammonia solution already gave a maximum
40 Cr(VI) elution. So for further analysis, nitric acid 2.0 mol.L-1 and ammonia 0.5 mol.L-1 were chosen. A
41 fiagram of Cr(III) and Cr(VI) speciation in optimum concentration of eluent is presented in Figure 6.
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
Figure 5 Effect of eluent concentration to fiagram of Cr(III) and Cr(VI)
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
Figure 6 Fiagram of Cr(III) and Cr(VI) separation in optimum eluent concentration
48
49
50 Ideal elution of sample can be reach if the analysis is fast, accurate, precision and consume less
51 eluent[16]. Optimum eluent concentration was used to determine the eluent volume that produce
52 optimum peak height. Eluent volume variation was controlled using a loop with volume 0.2, 0.5, and
53 1.0 mL. A fiagram in Figure 7 showed that both of nitric acid and ammonia give optimum elution when
54
55 the volume of eluent 0.5 mL. Volume below 0.5 mL gave lower height peak because the elution is not
56 maximal yet and bigger than 0.5 mL did not gave higher peak because dilution effect. Based on this
57 fact, the next analysis would use a 0.5 mL of eluent.
58
59
60
61
62
63
64
65
 1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24 Figure 7 Effect of eluent volume to fiagram of Cr(III) and Cr(VI)
25
26 Analytical performance
27 To evaluate the analytical performance, we determined linearity, selectivity, repeatability, detection
28 limit, enrichment factor, concentration efficiency and consumptive index. Linearity was determine
29
based on linear regression of a certain range concentration of analyte. In this experiment,
30
31 concentration between 50–500 μg.L-1 has a good relation between peak height (H) and analyte
32 concentration ([Cr]) with dynamic range up to 12-fold. A fiagram profile of various concentration Cr
33 species in optimum condition is presented in Figure 8. A linear regression plotting give correlation
34 coefficient R2 = 0.9905 for Cr(III) and R2 = 0.9926 for Cr(IV). Both R2 value showed good linearity of this
35 FIA-AAS system in the range of analyte concentration.
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60 Figure 8 Fiagram profile of Cr(III) and Cr(VI) on various concentration in optimum condition
61
62
63
64
65
 1 The % variance coefficient (%VC) is used to state quality of the analysis repeatability. Lower %VC
2 means better precision. At optimum condition, fiagram like Figure 9 will be obtained. The %VC value
3
of Cr(III) and Cr(VI) is 1.8 % and 0.6 %, respectively. The %VC value lower than 5.0 % indicates that the
4
5 repeatability of developed analytical methods is very good. The Bt-MCCs mini column have been
6 developed show good stability and performance for routine analysis.
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31 Figure 9 Fiagram profile of Cr(III) and Cr(VI) in optimum condition for five repetitions
32
33 The developed method has a limit detection consecutively 2.8 and 2.4 μg.L-1 for Cr(III) and Cr(VI) for 1
34 mL of sample based on limitation a S/N ratio = 3. The minimal mass detected (MMD) is 2.8 ng for Cr(III)
35 and 2.4 ng for Cr(VI). Saygi et al reports they have a limit detection of 1.94 μg.L-1 as Cr(VI) for 25 mL
36
37 minimal of sample [8]. It means that the MMD for this system is 48.5 ng. So, it can be concluded that
38 this method has a better limit detection than another selective method that has been reported using
39 AAS for ion detection. For too low concentration of Cr, this limit detection can be attained by enlarging
40 the injection volume. But it needs to consider that more volume will make a longer analysis time and
41 an effect to retention-elution process.
42
43
44 An enrichment factor (EF) describe ratio analyte concentration in sample before and after
45 preconcentration. The EF calculated by comparing the slope of sample calibration curve with and
46 without preconcentration. Table 1 showed a linear regression parameter for Cr(III) and Cr(VI) with and
47 without preconcentration in equation
48 H = A + B[Cr], H = peak height; A = constant; B = slope; [Cr] = chromium concentration
49
50 Calculation results showed preconcentration with the developed Bt-MCCs mini-column have EF 10-
51 fold for 1 mL sample. This EF value can still improve by increasing the sample volume. Data in Table 1
52 also can use to calculate sensitivity. Sensitivity can be calculated using equation
53 S = 0.0044/k, S = sensitivity; k = slope of calibration curve
54 Calculation give the sensitivity are 14.7 μg.L-1 for both of Cr species.
55
56
57 Table 1. Regression parameter of Cr(III) and Cr(VI) with and without preconcentration
58 Condition Concentration Species Parameter
-1
59 range (μg.L ) A B R
60 1000–9000 Cr(III) -0.0060 0.00003 0.9984
61
62
63
64
65
 Without Cr(VI) -0.0028 0.00003 0.9964
1 preconcentration
2 With 50–500 Cr(III) -0.0037 0.0003 0.9905
3
preconcentration Cr(VI) -0.0046 0.0003 0.9926
4
5
6 The concentration efficiency (CE) value was determined by counting the analyte peaks. This method
7 can be used in an hour. In this developed method, we can get two analyte peaks (Cr(III) and Cr(VI)) in
8 5 minutes, so CE for this method is 12 hour-1. This value means that the developed method has a short
9 analysis time and worth to use for routine analysis with high number of samples. Beside EF dan CE,
10
11 consumptive index (CI) also can be used to evaluate the FIA performance. CI is a FIA parameter which
12 is related to carrier and eluent volume. A FIA preconcentration supposedly have a small CI value to
13 get an efficiency. In optimum condition, this developed system needs total volume 12.0 mL with detail
14 10 mL of carrier (2.0 mL.minute-1 x 5 minute), 1.0 mL of sample, and 1.0 mL of eluent (2 eluent x 0.5
15 mL) to get 2 signal from different Cr species. The CI value maybe increase if the concentration sample
16
17
very low, but EF value also increases too to compensate it without lose another analytical parameter.
18
19 Application for environmental sample
20 The sample that has been analyzed using this FIA-AAS system came from Cidurian river, West Java,
21 Indonesia and taken in January 2017. The sample was filtered using filter paper to remove an
22
undissolved particle and then the filtrate was analyzed using the optimum condition. To learn the
23
24 matrix effect in analysis, we had used spike method and evaluated the percent recovery. Table 2
25 showed the percent recovery of river water sample for Cr(III) and Cr(VI). A higher percentage of
26 recovery value indicates that the method has high accuracy and sample matrix does not give
27 significant effect to the measurement.
28
29
30 Table 2. Concentration of Cr(III) and Cr(VI) in Cidurian river
31 Sample Cr concentration (μg.L-1) Percent recovery Species
32 Added Found (%)
33 Cidurian 0 38.3 ± 1.8 98.8 Cr(III)
34 river 100.0 136.7 ± 1.8
35
36 water 0 26.7 ± 0.8 100.7 Cr(VI)
37 75.0 101.7 ± 0.8
38
39 Table 2 also showed total concentration of chromium in river sample is 65.0 ± 2.6 μg.L-1. This value is
40
41 too small for FAAS analysis because of inadequate sensitivity. Using the Bt-MCCs mini-column in FIA-
42 AAS system solved that problem. Indonesian government regulation by Ministry of Environment and
43 Forestry decree determine the threshold value for Cr total and Cr(VI) in water are 0.10 mg.L-1 and 0.05
44 mg.L-1 respectively. Sample analysis showed Cr species total is still below this threshold.
45
46
Conclusion
47
48
49 A Bt-MCCs mini-column can be prepared easily because, all of the chemical compounds are relatively
50 easy to get and cheap. The column that has been used in FIA-AAS system shows a good analytical
51 performance and can be applied to trace the analysis of Cr species. The column effectively function as
52 a preconcentrator for low concentration Cr(III) and Cr(VI) in real sample matrix without losing the
53
54 speciation performance. It is possible to decrease the concentration range of chromium species with
55 increasing the volume sample injected to column. Increasing the sample volume of course effects the
56 time to flow all the sample through the mini-column.
57
58 Speciation that was done using 0.5 mol.L-1 ammonia solution and 2.0 mol.L-1 nitric acid solution as
59
60
specific eluent for each of chromium species. This eluent combination did not damage the column
61
62
63
64
65
 because the peak height has a good consistency after using it in many cycles. We suggest this
1 technique for Cr(III) and Cr(VI) routine analysis in environmental sample because this technique have
2 advantages like accurate, precise, and robust methods, modest equipment, short time for analysis,
3
easily automated, low sample volume, easily to tandem with another equipment and low cost. An
4
5 analytical performance summary is showed at Table 3.
6
7 Table 3. Summary of analytical parameter
8 Parameter Value
9 Cr(III) Cr(VI)
10
11 Concentration range (μg.L-1) 50–500 50–500
12 Correlation coefficient 0.9926 0.9975
13 Sensitivity (μg.L-1) 14.7 14.7
14 Repeatability (%) 1.8 0.6
15 Limit detection (μg.L-1) 2.8 2.4
16
17
EF 10 10
18 CE (hour-1) 12 12
19 CI (mL) 12 12
20 Accuracy (%) 98.8 100.7
21
22
Acknowledgment
23
24
25 Authors would like to express gratitude to M. A. Majid Al Kindi for proofreading this manuscript.
26
27 References
28
29
30 [1] M.K. Hasan, A. Shahriar, K.U. Jim, Water pollution in Bangladesh and its impact on public
31 health, Heliyon. 5 (2019) e02145. doi:10.1016/j.heliyon.2019.e02145.
32 [2] R.R. Elmorsi, K.S. Abou-El-Sherbini, G.A.-H. Mostafa, M.A. Hamed, Distribution of essential
33 heavy metals in the aquatic ecosystem of Lake, Heliyon. 5 (2019).
34 doi:10.1016/j.heliyon.2019.e02276.
35
[3] L.N. Nthunya, M.L. Masheane, S.P. Malinga, E.N. Nxumalo, B.B. Mamba, S.D. Mhlanga,
36
37 Determination of toxic metals in drinking water sources in the Chief Albert Luthuli local
38 municipality in Mpumalanga, South Africa, Phys. Chem. Earth, Parts A/B/C. 100 (2017) 94–
39 100. doi:10.1016/j.pce.2017.04.006.
40 [4] B. Kiran, N. Rani, A. Kaushik, Environmental toxicity: Exposure and impact of chromium on
41 cyanobacterial species, J. Environ. Chem. Eng. 4 (2016) 4137–4142.
42
43 doi:10.1016/j.jece.2016.09.021.
44 [5] J. Kotaś, Z. Stasicka, Chromium occurrence in the environment and methods of its speciation,
45 Environ. Pollut. 107 (2000) 263–283. doi:10.1016/S0269-7491(99)00168-2.
46 [6] V. Gómez, M.P. Callao, Chromium determination and speciation since 2000, Trends Anal.
47 Chem. 25 (2006) 1006–1015. doi:10.1016/j.trac.2006.06.010.
48
49
[7] V.N. Bulut, C. Duran, M. Tufekci, L. Elci, M. Soylak, Speciation of Cr(III) and Cr(VI) after column
50 solid phase extraction on Amberlite XAD-2010, J. Hazard. Mater. 143 (2007) 112–117.
51 doi:10.1016/j.jhazmat.2006.08.074.
52 [8] K.O. Saygi, M. Tuzen, M. Soylak, L. Elci, Chromium speciation by solid phase extraction on
53 Dowex M 4195 chelating resin and determination by atomic absorption spectrometry, J.
54
Hazard. Mater. 153 (2008) 1009–1014. doi:10.1016/j.jhazmat.2007.09.051.
55
56 [9] B. Saha, R.J. Gill, D.G. Bailey, N. Kabay, M. Arda, Sorption of Cr(VI) from aqueous solution by
57 Amberlite XAD-7 resin impregnated with Aliquat 336, React. Funct. Polym. 60 (2004) 223–
58 244. doi:10.1016/j.reactfunctpolym.2004.03.003.
59 [10] Y.C. Sun, C.Y. Lin, S.F. Wu, Y.T. Chung, Evaluation of on-line desalter-inductively coupled
60
61
62
63
64
65
 plasma-mass spectrometry system for determination of Cr(III), Cr(VI), and total chromium
1 concentrations in natural water and urine samples, Spectrochim. Acta Part B. 61 (2006) 230–
2 234. doi:10.1016/j.sab.2006.01.007.
3
[11] B. Wen, X. Shan, J. Lian, Separation of Cr(III) and Cr(VI) in river and reservoir water with 8-
4
5 hydroxyquinoline immobilized polyacrylonitrile fiber for determination by inductively coupled
6 plasma mass spectrometry, Talanta. 56 (2002) 681–687.
7 [12] G. Choppala, M.M. Rahman, Z. Chen, N. Bolan, M. Mallavarapu, Simultaneous determination
8 of chromium species by ion chromatography coupled with inductively coupled plasma mass
9 spectrometry, in: 19th World Congr. Soil Sci. Soil Solut. a Chang. World, International Union
10
11 of Soil Sciences, 2010: pp. 114–116.
12 [13] Y. Martinez-Bravo, A.F. Roig-Navarrro, F.J. Lopez, F. Hernandez, Multielemental
13 determination of arsenic, selenium and chromium(VI) species in water by high-performance
14 liquid chromatography – inductively coupled plasma mass spectrometry, J. Chromatogr. A.
15 926 (2001) 265–274.
16
17
[14] M.A. Hossain, M. Kumita, Y. Michigami, T.S. Islam, S. Mori, Rapid speciation analysis of Cr(VI)
18 and Cr(III) by reversed-phase high-performance liquid chromatography with UV detection, J.
19 Chromatogr. Sci. 43 (2005) 98–103. doi:10.1093/chromsci/43.2.98.
20 [15] D. Harvey, Modern Analytical Chemistry, McGraw-Hill, 2000.
21 [16] F. Zhaloun, Flow injection separation and preconcentration, Wiley-VCH, 1993.
22
[17] K. Grudpan, N. Worakijchar oenchai, O. Tue-Ngeun, P. Sooksamiti, J. Jakmunee, Flow injection
23
24 spectrophotometry for speciation of Cr(VI) and Cr(III) using diphenylcarbazide and solid phase
25 extraction with C18 in-valve mini-column, Sci. Asia. 25 (1999) 99–106.
26 [18] S.H. Ahmadi, A.M. Haji Shabani, S. Dadfarnia, M. Taei, On-line preconcentration and
27 speciation of chromium by an 8-hydroxyquinoline microcolumn immobilized on surfactant-
28 coated alumina and flow injection atomic absorption spectrometry, Turkish J. Chem. 31
29
30 (2007) 191–199.
31 [19] A. Ahmad, J.A. Siddique, M.A. Laskar, R. Kumar, S.H. Mohd-setapar, A. Khatoon, R.A. Shiekh,
32 New generation Amberlite XAD resin for the removal of metal ions: A review, J. Environ. Sci.
33 31 (2015) 104–123. doi:10.1016/j.jes.2014.12.008.
34 [20] M. Wulandari, Resin Pengkhelat Amberlit XAD-16-1,5-difenilkarbazid untuk prakonsentrasi
35
dan analisis selektif Cr(VI), Institut Teknologi Bandung, 2010.
36
37 [21] S. Aminah, Buchari Buchari, Muhammad Bachri Amran, Bentonite based functional material
38 for chromium species adsorption, Int. J. Appl. Sci. Technol. 7 (2017) 77–86.
39 [22] M.B. Amran, A.S. Panggabean, A. Sulaeman, M. Rusnadi, Preparation of a chelating resin and
40 its application as a preconcentration system for determination of cadmium in river water by
41 flow injection analysis, Internastional J. Environ. Res. 5 (2011) 531–536.
42
43 doi:10.22059/IJER.2011.337.
44 [23] E. Guibal, Interactions of metal ions with chitosan-based sorbents: A review, Sep. Purif.
45 Technol. 38 (2004) 43–74. doi:10.1016/j.seppur.2003.10.004.
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65