ANALYTICAL APPLICATIONS OF VOLTAMMETRY

IN TRACE DETERMINATION OF PHOSPHATE

Ρ S h a r m a * , K. G e h l o t a n d S. S o n g a r a

Electroanalytical Laboratory, Department of Chemistry,

J.N.V. University, Jodhpur- 342005, India

ABSTRACT

The suitability of the voltammetric methods has been investigated for the
determination of phosphate anionic species which itself is not reducible at
dropping mercury electrode. The observations have enabled the optimization
of analytical conditions for the micro level determination of phosphate in
environmental samples using differential pulse polarographic (DPP) method.
The limit of determination was achieved 0.05 μ§/ιη1.

Key words: Phosphate, trace determination, DPP, environmental samples.

INTRODUCTION

The determination of trace elements pose challenging problems

particularly when present in the form of an anionic species such as nitrite,
nitrate, sulfide, sulfite, halides, cyanide, fluoride and phosphate. Since most
applications of chemical analysis to environmental samples involve trace
determination at a level of parts per million (ppm) and parts per billion (ppb).
The existing conventional methods for estimation of anionic species (ion
selective electrode, classical colorimetry and spectrophotometry) are not
suitable in terms of sensitivity. High sensitivity of the employed technique
must further be accompanied by sufficient selectivity, precision and accuracy
/1,2/. Voltammetric methods like differential pulse polarography (DPP) and
stripping voltammetry have proved useful in such determinations, possessing
a dynamic linear range of 10 3-10"1(l M. In addition these methods can also be
used under widely differing experimental conditions and the sequential
determination of chemical forms of the concerned ion is possible. It is an

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important aspect in the case of environmental samples such as of soil and

water /3-6/.
The utility of voltammetric methods in analysis of natural waters has been
reported mainly for the determination and speciation of metal ions, even
though many non-metal inorganic species are also electroactive Π/. These
applications have also been limited to determination of halides, sulfides, and
oxyanions of certain elements /8,9/. Thus, it will be of interest to investigate
the suitability of voltammetirc methods for characterising the different
anionic forms in aqueous systems. In the present work attention was focused
on phosphate, which has a definite biological importance, likewise other
anions. For example, a fluoride concentration of 0.8 to 1.0 ppm is required to
prevent dental caries and fluorosis /10/. Iodide is needed for the proper
functioning of thyroxine hormone / l l / . The concentrations of fluoride and
iodide must be maintained within recommended limits. Therefore fluoride
and iodide are added to food supplements. Nitrate is required for the
production of amino acids /12/. Phosphate is an essential nutrient, being
required in biological synthesis and energy transfer processes. However, in
large concentrations phosphoric acid may cause vomiting and diarrhea.
Phosphate when added to city and farm water systems controls hardness,
prevents scale formation and inhibits corrosion. It may occur in surface and
ground waters as a result of leaching from minerals or ores, fertilizers used in
agriculture, and waste water of domestic origin such as human, animal and
plant residues.
Townshend and coworkers /17,18/ have described indirect polarographic
determination of phosphates. The cathodic stripping chronopotentiometric
determination of phosphate at the surface of copper electrode has been given
by Lundquist and Cox /19/. Kanan and Rajagopalan /20/ have estimated
phosphorus with arsenic and silicon by dc polarography using chemical
amplification. Fogg et al. /21-29/ have reported numerous voltammetric
observations for the determination of phosphate in different matrices.
Himeno et al. /30-33/ have also shown voltammetric characterization and
determination of phosphates. Phosphate concentrations in soil extracts by
Potentiometrie flow injection using a cobalt wire electrode has been given by
Chen et al. /34/. Lohithakshan and Aggarwal /35/ have described
Potentiometrie nonaqueous titration method for the determination of
organophosphorous reagents. Haemmerli et al. 136/ have shown
amperometric determination of phosphate by use of a nucleoside
Phosphorylase- xanthine oxidase enzyme sensor. Trace determination of

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Ρ Sharma, Κ. Gehlot and S. Songara Reviews in Analytical Chemistry

dexamethasone sodium phosphate in pharmaceutical formulations by

differential pulse plarography has been given by Jeyaseelan and Joshi /37/.
Astier and Bartlett /38/ have used a disposable micro electrochemical
transistor in the measurement of alkaline phosphatase at nanomolar
concentration.
A microelectrode procedure for determination of silicates and phosphate
in water has been presented by Carpenter and Hodgson /39/. Esson and
Meyerhoff /40/ have reported polyion sensitive membranes electrodes for
detecting polyphosphate rich biological polyanion. Simultaneous
determination of phosphate and pyrophosphate by an amperometric flow
injection analysis system with immobilized enzyme reactors is given by Yao
and Wasa /41/. Brett et al. /42/ have shown voltammetric and impedance
studies of phosphate in its ionised forms. A composite polypyrrole based
Potentiometrie biosensor for phosphate determination in natural water has
recently been reported by Adeloju and Lawal /43Λ Godinez /44/ has
described voltammetric detection of phosphate ion binding to ß-amino-
cyclodextrin. The authors have used the feasibility of selective complexation
of PO4"3 with ammonium molybdate in sodium nitrate containing sulfuric
acid medium for its indirect determination in the form of heteropoly acid
complex to be reduced polarographically at dropping electrode equivalent to
phosphate. The method has successfully been applied for the estimation of
phosphate contents in samples of natural water, soil, and detergents. We have
succeeded in the determination of nitrate- nitrite /45/ bromide / 46/ and iodide
/47/ using DPP procedure.

EXPERIMENTAL

Instrumentation

A microprocessor based pulse polarograph (Model CL-362) in

combination with a drop timer, of Elico (India), was used for voltammetric
measurements. Observations were recorded by a HP printer (Model-3820).
The instrumental settings for DPP were as follows: a dropping mercury
electrode was used as the working electrode; pulse amplitude, 50mV; pulse
duration, 57ms; clock time of pulse, 0.5 s and scan rate, 12 mV/s. Saturated
calomel and platinum wire were used as reference and auxiliary electrodes,
respectively.
A Systronics UV -VIS spectrophotometer (Model-108) was also used for

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sample analysis. It has a wavelength range of 190-900nm. Tungsten halogen

deuterium and wide range photo-multiplier were used as the light source and
detector, respectively. The spectral band width of resolution was 2nm.
The pH measurements were made on a digital pH meter (Model -335) of
Systronics, Ahmedabad.

Sample collection and preparation

Glassware and polyethylene containers were soaked in 2M nitric acid for

at least one week and washed with doubly distilled water prior to use. The
samples of industrial waste, lake water, pond water, soil and detergent were
collected for analysis of phosphate contents. Water samples were filtered to
separate particulate matters and also acidified with hydrochloric acid to pH~2
for storage purposes /48/. These (100 ml aliquot) were treated with an
oxidizing mixture of acid to destroy biological materials and heated till the
solution fumed. The contents were transferred to a volumetric flask and made
up to the volume /49/.
In the case of soil, a moisture-free, fine powdered sample was weighted
and leached out overnight with nitric acid. Vigorous mixing and excessive
washing was done and finally the sample was concentrated by evaporation
prior to estimation /50/.
All the experiments were carried out in an air-conditioned laboratory
where the temperature was maintained at 25±1°C. The test solutions were
deaerated by bubbling purified nitrogen for 20 min., prior to voltammetric
recordings. Traces of oxygen present in nitrogen were removed by passing
the gas through ammonium vanadate solution kept in contact with
amalgamated zinc /51/. Chemicals used were of reagent-grade purity.
Solutions were prepared in doubly distilled water. Stock solution of
phosphate was prepared from potassium dihydrogen orthophosphate by Ases
Chemical Works, Jodhpur (Batch No. 203). The test solutions were prepared
each day freshly. Supporting electrolyte used was 1M sodium nitrate
containing 0.25 Μ sulfuric acid.

^t RESULTS AND DISCUSSION

Polarographic Characteristics

Different supporting electrolytes such as citrate buffer, tartrate buffer,

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P. Sharma, Κ. Gehlot and S. Songara Reviews in Analytical Chemistry

sodium Perchlorate and sodium nitrate were studied to observe

electroreduction of molybdenum equivalent of phosphate at dropping
mercury electrode. A combination of 1M N a N 0 3 and 0.25 Μ H 2 S 0 4 was
found a suitable medium where single well- defined polarogaphic wave was
obtained at -0.22V. Relevant data are given in Table 1.

Table 1
Polarographic characteristics of phosphate in different supporting electrolytes

S. No. Supporting electrolyte -Ε1Λ (V) ΐ(μΑ)

I II I II
1 Citrate buffer (pH = 2.5) 0.09 0.53 3.20 4.50
2 IM HCl 0.12 - 0.70 -

3 Tartrate buffer (pH = 2.5) 0.20 0.45 1.50 3.75

4 1 Μ N a N 0 3 + 0.25 Μ H 2 S 0 4 0.22 - 5.47 -

5 0.1 Μ HCl + 0.2 Μ KH 0.28 0.40 2.50 3.80

pthalate
6 IM H 2 S 0 4 0.02 0.35 2.80 4.23
7 Acetate buffer (pH = 3.0) 0.02 0.45 1.75 3.65

The plot between the limiting current (I|) and mercury column height
( Vh) gave a straight line passing through the origin indicating the diffusion-
controlled nature of the wave. The log-plot analysis of the wave illustrated
the reversible nature of the electrode process. A typical DC polarogram of
molybdenum equivalent of phosphate is shown in Fig. 1.

DPP Conditions for the Determination of Phosphate

A sharp DP pPeak (Ep= -0.22V) was also obtained for molybdenum

equivalent of phosphate in presence of 1M N a N 0 3 + 0.25 Μ H 2 S 0 4 (Fig 2).
It was found that the peak current increased linearly with the concentration of
phosphate up to 3.0 ppm as presented in Table 2. The calibration
characteristics (Fig. 3) were as follows:
Slope, 0.515; intercepts, 0.061 and coefficient of correlation (r), 0.996

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Fig. 1: DC polarogram of phosphate in 1M N a N 0 3 + 0 . 2 5 M H 2 S 0 4

(a) P 0 4 " 3 = 1.05 χ 10"5 Μ
(b) Blank solution of supporting electrolyte.

Fig. 2: DP curve of phosphate in 1M N a N 0 3 + 0 . 2 5 M H 2 S 0 4

P 0 4 " 3 = 1.00 ppm
Modulation amplitude = 25mV;
Drop time = 0.5s and scan rate = 12 mV/s

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C o n c e n t r a t i o n in p p m

Fig. 3: The plot of calibration curve, concentration (ppm) vs. peak current
(μΑ)

Table 2
Relationship between phosphate concentration and peak current

S.No. P 0 4 3 Cone, (ppm) Peak current (μΑ) Ip/C

1. 0.1 0.066 0.660
2. 0.2 0.143 0.715
3. 0.4 0.299 0.749
4. 0.8 0.539 0.673
5. 1.0 0.616 0.616
6. 1.2 0.660 0.550
7. 1.4 0.748 0.534
8. 1.8 0.980 0.544
9. 2.0 1.090 0.545
10. 2.4 1.310 0.545
11. 3.0 1.600 0.533

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Interference

The possible associated anionic forms of nitrate and fluoride gave no DP

peak in I M N a N O j + 0 . 2 5 M H 2 S 0 4 , and thus did not interfere. Similarly
antimony, arsenic and germanium are reduced at more negative potentials
and produced distinguishable DP peaks. However, silicon caused significant
interference during phosphate determination because it also forms
molybdosilisic acid complex. It was eliminated by selective extraction of
phosphate with diethyl ether.

Limit of Determination

The limit of determination achieved was 0.05 μg/ ml using DPP

technique under these experimental conditions.

Analytical Applications

Phosphate has been determined indirectly by estimation of

phosphomolybdate complex. Phosphate forms a heteropoly acid complex
with acidic ammonium molybdate solution. T h e complex is selectively
separated by solvent extraction and heteropoly acid is decomposed by
treatment with an alkaline buffer solution to liberate the molybdenum, which
has finally been determined equivalent of phosphate by DPP.

Determination of Phosphate in Aqueous Matrices

50 ml aliquot of the prepared sample was taken into the polarographic

medium. DP polarograms were recorded in the potential range of 0.0 to -1.2
V and peak currents were measured at -0.22 V after making blank
corrections.
Quantitation in all observations was made by the standard addition
method 1551 T h e results of determination of phosphate in industrial waste,
lake water, pond water, soil samples and detergents are summarized in Table
3.
The U V - V I S spectrophotometric method was used to compare the results
obtained by DPP. The observations are included in Table 4.

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P. Sharma, Κ. Gehlot and S. Songara Reviews in Analytical Chemistry

Table 3
DPP determination of Phosphate

Sample PO 4 - Cone, (ppm) SD cv

Min. Max. Avg. (±) (%)
Soil 0.158 0.162 0.159 0.002 1.304
Detergent 0.128 0.140 0.133 0.006 4.583
Industrial waste 0.249 0.252 0.250 0.001 0.610
House hold waste 0.438 0.440 0.439 0.001 0.228
Pond water 0.311 0.321 0.315 0.005 1.679
Lake water 0.394 0.412 0.401 0.009 2.313

Table 4
Comparison of results obtained by DPP and UV- VIS Spectrophotometry

S.No. Sample P 0 4 3 Conc.(ppm)*

DPP UV-VIS
1. Common effluent 0.538 0.589
2. Industrial waste 0.458 0.478
3. Lake water 0.591 0.611
4. Pond water 0.408 0.403
5. Soil samples 0.181 0.187
6. Detergent 0.638 0.653
* Average of three determinations

CONCLUSION

The reported DPP method for the determination of microgram level

phosphate in environmental samples is more selective, rapid and of simpler
approach in comparison to the commonly employed spectrophotometric
method, due to lack of required sensitivity /52-54/. Further it suffers
interferences from a number of intermediate products during molybdate
reaction and sample preparation. The results obtained by DPP are in good
agreement with that of UV-VIS spectrophotometry suggesting the validity of
measurements. The proposed method is also quantitative in terms of
sensitivity (detection limit = 0.05 μg/ ml).

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ACKNOWLEDGEMENT

CSIR is thankfully acknowledged for awarding Junior Research

Fellowship (NET-JRF) to one author (S. Songara).

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