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Low-cost field test kits for arsenic detection in water

a a a b
Joyati Das , Priyabrata Sarkar , Jigisha Panda & Priyabrata Pal
a
Biosensor Laboratory, Department of Polymer Science and Technology , University of
Calcutta , Kolkata , India
b
The Petroleum Institute , Abu Dhabi , U.A.E
Published online: 11 Oct 2013.

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To cite this article: Joyati Das , Priyabrata Sarkar , Jigisha Panda & Priyabrata Pal (2014) Low-cost field test kits for arsenic
detection in water, Journal of Environmental Science and Health, Part A: Toxic/Hazardous Substances and Environmental
Engineering, 49:1, 108-115, DOI: 10.1080/10934529.2013.824764

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 Journal of Environmental Science and Health, Part A (2014) 49, 108–115
Copyright C Taylor & Francis Group, LLC

ISSN: 1093-4529 (Print); 1532-4117 (Online)

DOI: 10.1080/10934529.2013.824764

Low-cost field test kits for arsenic detection in water

JOYATI DAS1, PRIYABRATA SARKAR1, JIGISHA PANDA1 and PRIYABRATA PAL2
1
Biosensor Laboratory, Department of Polymer Science and Technology, University of Calcutta, Kolkata, India
2
The Petroleum Institute, Abu Dhabi, U.A.E

Arsenic, a common contaminant of groundwater, affects human health adversely. According to the World Health Organization
(WHO), the maximum recommended contamination level of arsenic in drinking water is 10 µg/L. The purpose of this research was
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to develop user-friendly kits for detection of arsenic to measure at least up to 10 µg/L in drinking water, so that a preventive measure
could be taken. Two different kits for detection of total arsenic in water are reported here. First, the arsenic in drinking water was
converted to arsine gas by a strong reducing agent. The arsine produced was then detected by paper strips via generation of color
due to reaction with either mercuric bromide (KIT-1) or silver nitrate (KIT-2). These were previously immobilized on the detector
strip. The first one gave a yellow color and the second one grey. Both of these kits could detect arsenic contamination within a range
of 10 µg/L–250 µg/L. The detection time for both the kits was only 7 min. The kits exhibited excellent performance compared to
other kits available in the market with respect to detection time, ease of operation, cost and could be easily handled by a layman. The
field trials with these kits gave very satisfactory results. A study on interference revealed that these kits could be used in the presence
of 24 common ions present in the arsenic contaminated water. Though the kits were meant for qualitative assay, the results with
unknown concentrations of real samples, when compared with atomic absorption spectrophotometer (AAS) were in good agreement
as revealed by the t-test.
Keywords: Arsenic, groundwater, field kit, mercuric bromide, silver nitrate.

Introduction hyperkeratosis, pigmentation on skin, etc. There are many

available methods for detection of arsenic in drinking wa-
Arsenic toxicity affects millions of people worldwide. ter e.g., atomic absorption spectroscopy (AAS), cathodic
Among all the heavy metals, inorganic arsenic, a known stripping voltammetry (CSV), inductively coupled plasma-
carcinogen is a common contaminant of potable water mass spectrometry (ICP-MS) and spectrophotometry.[9] A
through out the world. Elevated concentrations of arsenic few decades back, Dal Cortivo et al.[10] developed a spec-
in groundwater in West Bengal and Bangladesh have be- trophotometric method for detection of arsenic by con-
come a major cause for concern in recent years.[1] Arsenic is verting it into arsine and then detecting the arsine gas
a crystal-shape metalloid element, brittle in nature and grey using silver diethyldithiocarbamate dissolved in pyridine
or tin-white in color. Arsenic may be found mainly in two solution. Arsenic in feed additives was also detected by a
forms, arsenate [As (V)] and arsenite [As (III)]. The pres- cathodic stripping voltammetry (CSV) technique using a
ence of arsenic in drinking water is due to either its natural hanging mercury drop electrode.[11] Anderson et al.[12] de-
presence in surface and in groundwaters,[2] or as a result of veloped a novel pneumatic nebulization PN/hydride gener-
human activities such as industrial applications,[3] leather ation (HG) inductively coupled plasma mass spectrometry
and wood treatments,[4] use of pesticides.[5] As per WHO’s (ICP-MS) technique for detection of arsenic.
recommendation, the maximum allowable contamination Although all of these spectrophotometric methods can
level of arsenic in drinking water is 10 µg/L.[6,7] detect arsenic in a very lower detection range but all these
Consumption of arsenic contaminated water may cause instrumental procedures cannot be performed onsite and
many diseases like bladder, lung and skin cancer,[8] skin they are expensive, time consuming and require sufficiently
trained laboratory individuals. In recent years, Sharma
et al.[13] have reported development of one disposable
sensor for quantitative detection of arsenic using MAT-
Address correspondence to Priyabrata Sarkar, Department of LAB software. Although the minimum detection limit of
Polymer Science and Technology, University of Calcutta, 92 this process is 0.018 µg/L, it would be a difficult task to
A.P.C. Road, Kolkata 700009, India; E-mail: sarkarpriya@gmail. analyze the developed color on TLC paper using software
com on-site and moreover it is time consuming. Xia et al.[14]
Received April 12, 2013. developed a colorimetric assay for detection of arsenite
 Low-cost field test kits for arsenic detection 109
using gold nanoparticles and a phyto-chelatin-like peptide. These kits could detect total arsenic (arsenite [As (III)]
This laboratory-based procedure is expensive and its use is + arsenate [As (V)]) present in drinking water. Interference
limited only to laboratory analysis. of 24 ions for both the kits had been tested and in most of
Field kits have been used extensively for qualitative de- the cases it was found that the maximum tolerant level is
termination of arsenic in groundwater.[15] One of the major greater than Merck kit. Because procedure of estimation of
advantages of field test kits is that measurement can be arsenic in a given sample is very simple and also the cost is
conducted on site. Most of the field test kits for detection very low, the users will be able to estimate the concentration
of arsenic in water available in the market utilized Gutzeit of arsenic in drinking water effectively using these kits.
method.[16,17] This involves generation of arsine gas (AsH3 )
by reduction of arsenic using zinc and hydrochloric acid.
The generated arsine can be quantified by trapping it either Materials and methods
in a silver diethyldithiocarbamate solution,[18] or on paper
impregnated with mercuric bromide, [19] both of which act Chemicals
as a detector element. Although field test kits for arsenic Sodium arsenate, heptahydrate (Na2 HAsO4 ·7H2 O)
have an advantage of lower detection time but the major (Merck, India) was used to prepare arsenic (V) stock
drawback of these kits is the use of concentrated hydrochlo- solutions. Sodium arsenite (NaAsO2 ) (Loba Chemie,
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ric acid, which is highly corrosive and thereby deals with Mumbai, India) was used to prepare arsenite (III) stock
handling and transportation problem. solutions. Zinc dust, sodium borohydride, sulfamic acid,
Another major disadvantage of these field kits is that potassium permanganate, potassium peroxodisulfate,
sulfur, selenium compounds may interfere with this assay. silver nitrate, L-ascorbic acid were obtained from Merck.
Many researchers tried to evolve new methods of detec- Mercuric bromide and gluteraldehyde were supplied by
tion. In one such attempt magnesium turnings along with Loba Chemie. Merck kit (Merck Arsenic Test Method:
oxalic acid were used for generation of arsine gas which Colorimetric with test strips, 0.005-0.5 mg/L As3+/5+,
reduced auric chloride to metallic gold for color genera- Prod.: 1.17927.0001, Batch: HC80062, MerckquantR) was
tion.[20] However, the major drawback of this process is the purchased from Merck.
use of auric chloride as the detector element, which is very
costly. The Merck (Mumbai, India) kit (based on Gutzeit
Method; MERCK Arsenic Test Method: Colorimetric with Preparation of arsenic stock solutions
test strips, 0.005–0.5 mg L−1 As3+/5+, Prod.: 1.17927.0001, An arsenic (V) stock solution (4803 µg/L of arsenic) was
Batch: HC80062, MerckquantR) is widely used all over the prepared by dissolving 0.002 g of sodium arsenate, hep-
world for arsenic detection in groundwater and has a de- tahydrate (Na2 HAsO4 ·7H2 O) (Merck) into 100ml of tap
tection capability of 5–500 µg/L of arsenic. Another pro- water. The working standards (10–250 µg/L) were pre-
cedure by Hach (Loveland, CO, USA) uses solid sulfamic pared by proper dilution of the stock solution. Arsenic
acid instead of concentrated hydrochloric acid [20] and it (V) stock solutions were prepared freshly in every 2 to 3
also has a detection capability of 5–500 µg/L. months. An arsenic (III) stock solution (1000 mg/L of
However, both of these kits have a longer detection time arsenic) was prepared by dissolving 0.001 g of sodium
(generally 20–30 mins) and are expensive. Kearns et al.[21] arsenite (NaAsO2 ) (Loba Chemie) into 100 mL tap water.
tried to improve the accuracy and precision of the Hach The working standards (10–250 µg/L) were prepared sim-
EZ test kit for quantifying inorganic arsenic concentration ilarly from this stock solution. Because there was a chance
in drinking water by digital analysis of the color developed of As (III) getting oxidized into As (V), arsenic (III) stock
on the detector strip. The digital analysis would produce solutions were prepared freshly before use.
fewer false positive and false negative results but it would
increase the cost of the test kit and running the test process
Preparation of reagents
for 24 h made the procedure time consuming.
There are demands of low cost field test kits in arsenic A 10% solution of potassium permanganate was prepared
prone areas and to meet this requirement, we have devel- by dissolving the salt in 1% potassium peroxodisulfate so-
oped two user-friendly kits. The detection levels of both lution. This was used for removal of interference (RI), e.g.,
of these kits is 10–250 µg/L of arsenic and the detection sulfide. The reducing agents comprised a mixture of acids
time in both the kits has been reduced to 7 min, which and powdered zinc to produce in-situ nascent hydrogen.
is even lower than the detection time of most well-known However at the time of measurement, the acid mixture and
Merck and Hach arsenic detection kits. In this research, the powdered zinc and borohydride were added separately.
reducing agent was made of a composition of potassium The acid mixture was prepared by mixing sulfamic acid
permanganate, a mixture of sulfamic acid and L-ascorbic and 1% L-ascorbic acid. It has been reported and tested
acid, a mixture of zinc dust and sodium borohydride and that only zinc metal showed slow reactivity towards ar-
the detector materials were mercuric bromide (KIT-1) and sine generation.[15,22] To enhance the reaction, zinc dust
silver nitrate (KIT-2). was mixed with 1% sodium borohydride in both the kits.
 110 Das et al.
It may be noted that all the chemicals, i.e., acid mixtures and shaken for a while. Then the detector strip (either mer-
and reducing agents (measured amount for one test) were curic bromide or silver nitrate) was inserted inside the cap
packed in sealed in pouches in nitrogen atmosphere to avoid of the bottle. The water sample in the bottle was treated
degradation. with 4 g of Zn and borhydride mixture and the cap of the
bottle tightened immediately. Then the bottle was shaken
occasionally for 7 min. Care was taken that the detector
Preparation of mercuric bromide strip for KIT-1 strip was never in contact with water. The cap of the bottle
The detector strip was made of filter paper cut into circular was removed and the detector strip was inspected for visual
discs (1-cm diameter) which were dipped into a 10% solu- observation of the colour. The details of the procedure is
tion of mercuric bromide in methanol and left in dark to depicted in the flow chart (Fig. 1)
dry. A glossy paper was cut in circular shape (3.9-cm diam-
eter), which could fit tightly into the cap of wide-mouthed
reaction bottle. The dried detector strip was pasted at the Results and discussions
central portion of the circular strip of glossy paper using
adhesive. Preparation of arsenic standardization color chart
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Reaction for arsine gas generation. Both the forms of ar-

Preparation of silver nitrate strip for KIT-2 senic [As (V) and As (III)] generated arsine gas by the
reduction with reducing agent (zinc dust and sodium boro-
The same procedure is followed except that 5% silver ni- hydride) under acidic conditions (mixture of sulfamic acid
trate solution was used instead of mercuric bromide. The and L-ascorbic acid). The reactions that could occur inside
preparation of this strip could be done just before the test the bottle are given next.[2] The arsine (AsH3 ) when reacted
since storing of these strips could not be possible due to with mercuric bromide (HgBr2 ) (in case of KIT-1), formed
unstable nature of silver nitrate. yellow-to-brown colored (depending upon arsenic concen-
tration) complex mercuric arsenide [AsH (HgBr)2 (yellow),
As (HgBr)3 (brown)] on the paper strip as per Eq. (4):[2]
Procedure of measurement
As2 O3 + 6 Zn + 12 HCl → 2 AsH3 + 6 ZnCl2 + 3 H2 O
A wide-mouthed plastic bottle (250 mL) was filled with
100 mL of arsenic contaminated water. To the sample two (1)
drops RI solution was added and shaken for a while. The H3 AsO4 + 4 Zn + 8 HCl → AsH3 + 4 ZnCl2 + 4H2 O
water sample was then treated with 2.5 g of acid mixture (2)

Fig. 1. Flow chart of the experimental procedure (color figure available online).
 Low-cost field test kits for arsenic detection 111
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Fig. 2. Standardization chart (a: KIT-1, mercuric bromide; b: KIT-2, silver nitrate) (color figure available online).

3NaBH4 + 4H3 AsO3 → 4AsH3 + 3H3 BO3 + 3NaOH of 10–15% potassium permanganate and 1% potassium
(3) peroxodisulfate (RI). It could be observed for both the
AsH3 + HgBr2 → AsH(HgBr)2 (yellow)/ kits that detection of arsenic was possible in presence of
10 mg/L of sulfide interference (Table 1).
As(HgBr)3 (brown) (4)
The arsine (AsH3 ) when reacted with silver nitrate (AgNO3 )
[in case of KIT-2], formed a grey-to-black to a silver-black Table 1. Comparison of maximum tolerance level of interfering
colored complex (depending upon arsenic concentration) ions of arsenic test kits (KIT-1, mercuric bromide, KIT-2, silver
nitrate).
silver arsenide (Ag3 As) on the paper strip according to Eq.
(5).[23] Maximum Tolerant
+ + Level
AsH3 + 3 Ag → Ag3 As + 3H (5) Maximum Tolerant
MERCK PST KIT Level PST
Contaminant (mg/L) 1 (mg/L) KIT 2 (mg/L)
Standard arsenic color chart
Al3+ 100 200 150
Depending upon the color variation of the paper strips Ag+ 1 70 70
due to the presence of different concentration of arsenic Ca2+ 1000 1000 1000
in water, the standard color chart for both the kits were Cl− 1000 1500 2000
prepared (Fig. 2). CO2−3 1000 1500 1000
CrO2− 4 1000 1000 1000
Cu2+ 0.5 1 1
Removal of interference F− 500 1000 1000
Fe3+ 1000 1500 1500
In case of field kits, the presence of sulfide in water may Hg2+ 5 15 10
produce H2 S during reduction. This H2 S may interfere in K+ 1000 1500 1500
the change of color of the detector strip. In previously pub- Mg2+ 1000 1500 1500
lished research work, sulfide interference had been removed MnO− 4 500 500 500
by passing the arsine gas stream through a scrubber con- Na+ 1000 1500 1500
taining lead acetate to remove the hydrogen sulfide as lead Ni+ 10 10 10
sulfide. Even zinc and copper scrubbers were also used but NO− 2 100 200 200
they had been found to be less efficient. There are many NO− 3 100 200 200
disadvantages to these methods. Handling and disposal of S2− 0.5 10 10
hazardous lead acetate and lead sulfide, apart from this Sb3− 1 2 2
the rate of gas evolution has to be controlled to allow ad- SeO2−3 1 2 2
equate contact time for the entire sulfide to react with the Sn2+ 100 200 200
scrubber.[2] SO2−
3 1 1 1
The sulphide, if present in water was first oxidized to SO2−
4 1000 1000 1000
sulfate using the oxidizing agent prepared from a mixture Zn2+ 1000 1000 1000
 112 Das et al.
Table 2. Results of detection of arsenic in the field samples using test kits on the spot and comparison of results with Merck test kit.
Test Results
Sample No. Merck Test Kit KIT-1 KIT-2

01 (BARASAT Akrampur Pilot Plant Latitude: Above 250 µg/L∗ Above 250 µg/L∗ Above 250 µg/L∗
22.2300◦ N Longitude: 88.4500◦ E)
02 (BARASAT Nivedita Palli, Latitude: ∼300 µg/L∗ ∼300 µg/L∗ ∼300 µg/L∗
22.2300◦ N Longitude: 88.4500◦ E)
03 (BARASAT R.K.Mission Bamunmura, ∼185 µg/L–190 µg/L∗ ∼185 µg/L–190 µg/L∗ ∼185 µg/L–190 µg/L∗
Latitude: 22.2300◦ N Longitude: 88.4500◦ E)
∗
Higher ranges detected via dilution of samples.

Removal of interference of other ions and comparison with Detection limit and detection time of the kits
Merck kit (Prod.: 1.17927.0001, Batch: HC80062,
The lower visual detection limit of both the kits is 10 µg/L
MerckquantR)
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of arsenic and the maximum visual detection limit of both

A typical groundwater may contain many minerals in the the kits is 250 µg/L of arsenic. Both the kits could detect
form of anions and cations e.g., sulfide, iron, nitrate, mag- arsenic in water in only 7 min where as in case of Merck
nesium, sodium, calcium etc. We studied interference of kit the detection time is 20 min and in the case of Hach kit
most of these ions (24 in number) and compared the results the same is 30 min. The comparison of the performance the
with those of Merck Kit. The entire interference tests had reported kits vis-à-vis some test kits available in the market
been performed with of 100 µg/L arsenic solution. It had are given in Table 3.
been found that the maximum tolerant level of interfering
ions, in both the kits, was either higher or similar to the
Merck kit. The results are represented in Table 1. Validation of arsenic test kit by laboratory instrument
Arsenic concentrations determined by field kits were com-
pared with measurements by atomic absorbsion spec-
Field study and precision of the kits trophotometry (AAS). We had collected samples of arsenic
contaminated water from Baruipur, 24 Parganas (South),
To study the reproducibility and precision of the two kits, West Bengal, Chakdah, Nadia, South 24 Praganas, West
we had conducted field experiments in three different estab- Bengal, Ashokenagar, Kalyanghar, North 24 Praganas,
lishments of CGCRI (Central Glass and Ceramic Research West Bengal. We performed AAS with those samples. The
Institute, Kolkata) at Barasat, 24 Parganas (North), West concentration of arsenic present in those samples was also
Bengal, where arsenic contaminated groundwater was pu- detected by our two test kits [KIT-1 (mercuric bromide),
rified and arsenic was removed by the CGCRI technology. KIT-2 (silver nitrate)] (Table 4).
We measured the concentrations of arsenic in contami- The correlation coefficient between arsenic concentra-
nated water using our two kits and also by the Merck kit. tions measured by KIT-1 and AAS was found to be 0.9952
We found that in all the cases our results matched with the (Fig. 3). Similar results were found in case of KIT-2, which
Merck kit results (Table 2). This proved reproducibility and suggested that the arsenic test kits had a good agreement
precision of both the kits. with results obtained using AAS. It might be noted that

Table 3. Comparison of performance with other test kits available in market.

Sulfide Interference Cost Detection Incubation
Kits (maximum tolerable) Shelf life ($ per test) limit in µg/L time in Min

KIT-1 (Mercuric bromide) 10 mg/L One year 0.6 10–250 07

KIT-2 (silver nitrate) 10 mg/L Not applicable (since strips 0.6 10–250 07
made on the spot)
MERCK 2 mg/L 2 years 1.84 0–500 20
HACH COMPANY, USA Not available Not available 1.42 0–500 30
Wagtech WTD, Palintest Not available Not available 2.05 2–100 Less than 20
Ltd., United Kingdom
MACHEREY-NAGEL Not available Not available 1.01 0–500 10
GmbH & Co. KG, Düren,
Germany
 Low-cost field test kits for arsenic detection 113
Table 4. Comparison of the results of proposed kits with that obtained by AAS.
Place from where Arsenic Concentration Arsenic Concentration Arsenic
Arsenic contaminated detected by Kit-1 detected by Kit-2 Concentration
Sample No. sample collected (Mercuric Bromide) (Silver Nitrate) detected by AAS

01 (tap water) Kolkata, West Bengal 0 µg/L 0 µg/L 0 µg/L

Latitude: 22◦ 34 10.92N
Longitude: 88◦ 22 10.92E
02 Baruipur, South 24 10 µg/L 10 µg/L 11 µg/L
Praganas West Bengal.
Latitude: 22.57485◦ N
Longitude: 88.3983◦ E
03 Chakdah, Nadia, 20 µg/L 20 µg/L 21 µg/L
South 24 Praganas,
West Bengal.
Latitude: 23.07605◦ N
Longitude: 88.52546◦ E
04 Ashokenagar, Kalyanghar, 50 µg/L 50 µg/L 45 µg/L
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North 24 Praganas, West Bengal.

Latitude: 22.833◦ N
Longitude: 88.633◦ E
05 Baruipur, South 24 100 µg/L 100 µg/L 92 µg/L
Praganas West Bengal.
Latitude: 22.57485◦ N
Longitude: 88.3983◦ E
06 Ashokenagar, Kalyanghar, 200 µg/L 200 µg/L 210 µg/L
North 24 Praganas,West Bengal.
Latitude: 22.833◦ N
Longitude: 88.633◦ E
07 Chakdah, Nadia, 250 µg/L 250 µg/L 273 µg/L
South 24 Praganas,
West Bengal.
Latitude: 23.07605◦ N
Longitude: 88.52546◦ E
08 Baruipur, South 24 ∼ 400 µg/L ∼ 400 µg/L 393 µg/L
Praganas West Bengal.
Latitude: 22.57485◦ N
Longitude: 88.3983◦ E

the kits were capable of only estimating four concentra- The seven samples for field study (Table 4) were again
tions namely 10, 50, 100, 250 µg/L. However, any sample tested 4 times each with the KIT-2 and statistical t-test was
could be diluted suitably and later on multiplied by dilution conducted with test results from AAS considering P-value
factor to get as close as the actual concentration. 0.05 (i.e., 95% confidence limit) and the results are tab-
ulated in Table 5. In all the cases the calculated t-values
did not exceed the t-value from table with number of sam-
ples 8 (4 for kit and 4 for AAS) and degrees of freedom 6
at 95% confidence level. This indicated that there was no
significant difference between the results of the proposed
method and the spectrophotometric method. This proved
the authenticity, high sensitivity and reliability of our two
test kits.

Cost estimation and shelf life of the arsenic test kits

We had developed two cost-effective kits, which could be
handled by any layman. The test kits would only require
an estimated $0.6 per test (sale price) and the cost is much
lower compared to a laboratory analysis.
Fig. 3. Comparison of arsenic detection with KIT-1 and analysis The shelf life of KIT-1(mercuric bromide) was approx-
by AAS (color figure available online) imately one year and shelf life of KIT-2(silver nitrate) is
 114 Das et al.
Table 5. ‘T’ test results with t-values from t-table considering 95% work could be our society and common people who would
confidence limit. be able to estimate the concentration of arsenic in drink-
∗
Calculated ∗
Tabulated t- value
ing water effectively using our two kits at a very low cost
Sample name t-value (P = 0.05) for (n = 4) (estimated to be 0.6$ per measurement).

Sample 2 0.146 2.45

Sample 3 0.0561 2.45
Sample 4 0.0193 2.45 Acknowledgments
Sample 5 0.110 2.45
Sample 6 0.146 2.45 The authors are indebted to DST (WTI) for a project grant
Sample 7 0.039 2.45 and grateful to DST-CSIR Sensor Hub at CGCRI, Kolkata
Sample 8 0.045 2.45 for providing infrastructural facilities.
∗
In all the cases the calculated t-values did not exceed tabulated t-value
at 95% confidence level.
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