Prez-Olmos et al / Chemistry Journal (2013), Vol. 03, Issue 03, pp.

81-85 ISSN 2049-954X

Research Paper

Sequential Potentiometric Determination of Nitrate and

Chloride in Vegetables
R. Prez-Olmos*, X. Gabiola and J.M. Hurtado

Universidad del Pas Vasco, Departamento Qumica Analtica, E.U.I.T.I., Pichichi n 3, 48.013 Bilbao, Spain
*Tel: 946014428
*E-Mail: qappeolr@lg.ehu.es

Abstract

A rapid and simple sequential potentiometric method is described for the determination of nitrate and chloride ions in
vegetables, by using home-made ion selective electrodes. The extraction of both ions from the vegetable samples was
effected by magnetic stirring of the samples during 30 min in water at room temperature. Direct Potentiometry was used as
analytical technique for measurement. The mean coefficient of variation, the average percentage of spike recovery and the
detection limit for the nitrate determination in 17 samples of different vegetables were found to be 1.7 %, 100.5 % and 0.12
g Kg-1 respectively. These values were found to be 2.3 %, 100.6 % and 0.08 g kg-1 for the chloride ion. The mean nitrate
and chloride ion concentrations determined by application of the proposed potentiometric method were found to be 7.18 g
kg-1 and 5.07 g kg-1, respectively. In addition Comparison of variances (F - test) and linear regression analysis of the results
obtained were carried out to validate the developed sequential potentiometric method.

Keywords: Potentiometry, Nitrate and Chloride Selective Electrodes, Vegetables, Linear Regression

1. Introduction
Nitrate is habitually founded in soils and plants but agron-
omists usually express concern that use of large quantities
of ammoniacal-based fertilizers will results in toxic levels
of nitrate in forage and vegetable crops, mainly in green
leafy vegetables. On the other hand, chloride contained in
soils and irrigation water are readily absorbed by the roots
and accumulated to toxic levels in the leaf margins and
shoot tips. High blood pressure (Weinbey, 1997) and car-
cinogenic nitrosamines production (Glidewell, 1990) are
the main toxicological risks associated with For these rea-
sons, there has been much effort in the development of
analytical methods employing different classical and instr-
umental techniques for the determination of both ions in
waters, vegetables and foodstuff. For these reasons, many
efforts have been devoted to the development of various
classical and instrumental analytical methods for the dete-
rmination of both ions in waters, vegetables and foodstuff.
Mostly Chloride ion is determined by titration using Volh-
ard or Mohr methods and on less extension by potentiom-
etry. Similarly, many methods have been developed for
nitrate determination such as direct or indirect UV/Vis
spectrophotometry, fluorimetry and potentiometry. In add-
ition various liquid chromatographic procedures have also
been proposed because they are less time consuming with
results highly reproducible (Williams, 1979).
In this work a sequential method for the determination of
nitrate and chloride in different types of vegetables using
nitrate and chloride selective electrodes has been develop-
ed (Tables 1-2). In this work a rapid and economical sequ-
ential potentiometric method has been developed for the
determination of nitrate and chloride ions in various vege-
tables using ion selective electrodes. The precisions, accu-
racies and detection limits for both determinations have
been established and the obtained results were compared
with that furnished by reference methods according to the
Spanish legislation (AOAC, 1984 and Ministerio de Sani-
dad y Consumo, 1985). In order to validate the proposed

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 Prez-Olmos et al / Chemistry Journal (2013), Vol. 03, Issue 03, pp. 81-85 ISSN 2049-954X

sequential potentiometric method, linear regression analy- The nitrate and chloride stock solutions 0.1 mol L-1 stock
sis of the obtained data - have also been carried out. solutions of nitrate and chloride ions were prepared from
Table 1. Comparative Study of the Precisions and Accuracies Obtained in the Determination of Chloride in Vegetables by Simultaneous
Application of the Proposed and Reference Methods

Entry Sample Reference Method (Mohr) Proposed Method (ISE)

Xa C.Vb Rc Xa C.V.b Rc REd F

1 Celery 6.530.12 1.8 101.8 6.680.07 1.0 101.1 +2.3 2.9
2 Lettuce 3.390.06 1.8 100.9 3.500.03 0.9 101.2 +3.2 4.0
3 Spinach 15.390.06 0.4 101.1 15.190.10 0.7 101.0 -1.3 2.8
4 Cucumber 0.650.03 4.6 101.1 0.620.02 3.2 101.0 -4.6 2.3
5 Parsley 0.970.03 3.1 106.7 1.010.02 2.0 101.1 -4.1 2.3
6 Radish 7.830.12 1.5 100.4 7.510.07 0.9 101.2 -4.1 2.9
7 Carrot 2.300.07 3.0 102.3 2.290.05 2.2 101.3 -0.4 2.0
8 Cabbage 2.480.04 1.6 101.6 2.530.03 1.2 98.7 -2.0 1.8
9 Green beans 6.370.10 1.6 100.6 6.210.07 1.1 98.7 -2.5 2.0
10 Endive 4.200.10 2.4 98.0 4.290.09 2.1 100.3 +2.1 1.2
11 Brode beans 6.420.11 1.7 100.5 6.710.09 1.3 101.1 +4.5 1.5
12 Escarole 5.250.17 3.2 99.3 5.140.13 2.5 101.3 -1.8 1.7
13 Artichoke 7.460.16 2.1 101.5 7.630.14 1.8 100.9 +2.3 1.3
14 Borage 8.120.10 1.2 98.9 7.810.07 0.9 99.0 -3.9 2.0
15 Cauliflower 4.300.18 4.2 102.8 4.430.12 2.7 102.0 +3.0 2.2
16 Chicory 1.150.02 1.8 101.2 1.190.03 2.5 99.7 +3.4 2.6
17 Watercress 3.520,11 3.1 99.8 3.460,09 2.6 100.8 -1.7 1.5
a
Mean Concentration and Standard Deviation (g kg-1), bCoefficient of Variation (%),cMeanspike Recovery (%), dRelative Error of the Potentiometric
Method Versus The Reference Method (%),The Critical F-value, Considering a 95% Confidence level and 16 Degrees of Freedom, for a Two-Tailed
Test is 2.77

2. Experimental Section
2.1. Apparatus and Materials
Nitrate and chloride selective electrodes were constructed,
without inner reference solution, by direct applying of the
respective PVC and homogeneous crystalline membranes
onto a conductive epoxy resin support, as previously desc-
ribed (Lima & Machado, 1986 and Lima & Rocha, 1990).
The potentials were measured with a Crison 2002 digital
potentiometer ( 0.1 mV sensitivity) coupled to a home-
made electrodes switcher (5 channels) (Barros et al,
1983). An Orion 90-02-00 double junction reference elect-
rode was used with a 0.033 mol L-1 potassium sulphate
(K2SO4) solution as an outer reference solution. A Perkin-
Elmer Lamda 3B UV-Vis spectrophotometer was used for
the nitrate determination (brucine reference method). A
Selecta agitamatic-N magnetic stirrer and a Selecta 981
drying oven were also used.
2.2. Reagents and Solutions
NaCl and potassium nitrate KNO3 dried at 110 C for 24
h. The working standards were prepared by subsequent
dilution of the stock solutions. 0.33 mol L-1 K2SO4 soluti-
on was used as ionic strength adjuster solution (ISA). The
outer filling solution of the reference electrode was prepa-
red by rigorous dilution (1/10) of the ISA solution and the
inner chamber was filled with the same solution as shipp-
ed with the electrode.
Analytical grade reagents were purchased and used witho-
ut further purification. Deionised water was used through-
out the experimental work, with conductivity less than 0.1
S cm-1
2.3. General Analytical Procedure
Approximately 5.0 g of previously homogenised and dri-
ed (overnight at 60 C) vegetables were extracted in 400
mL of deionised water by magnetic stirring for 30 minut-
es, at room temperature (Prez-Olmos et al, 2001). The
extract was filtered and diluted up to 500 mL using water.

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 Prez-Olmos et al / Chemistry Journal (2013), Vol. 03, Issue 03, pp. 81-85 ISSN 2049-954X

Table 2. Comparative Study of the Precisions and Accuracies Obtained in the Determination of Nitrate in Vegetables by Simultaneous
Application of the Proposed and Reference Methods

Reference Method (UV-VIS) Proposed Method (ISE)

Entry Samples
a b c a
X C.V R X C.Vb Rc REd F
1 Celery 12.990.17 1.3 101.0 12.820.25 2.0 101.2 -1.3 3.8
2 Lettuce 7.520.09 1.2 101.6 7.360.06 0.8 101.4 -2.1 2.2
3 Spinach 10.610.12 1.1 100.6 10.750.10 0.9 101.2 +1.3 1.4
4 Cucumber 4.010.04 1.0 100.7 3.930.03 0.8 100.5 -2.0 1.8
5 Parsley 8.520.15 1.8 99.9 8.800.11 1.2 101.3 +3.2 1.9
6 Radish 10.040.10 1.0 99.3 9.750.08 0.9 101.2 -2.9 1.6
7 Carrot 3.520.004 0.1 101.2 3.500.004 0.1 100.9 -0.6 1.0
8 Cabbage 4.520.06 1.3 101.6 4.610.07 1.5 98.2 +2.0 1.4
9 Green beans 2.500.05 2.0 100.6 2.490.040 0.8 99.4 -0.4 1.6
10 Endive 2.010.05 2.3 98.0 1.920.04 1.8 100.6 -4.4 1.6
11 Brode beans 18.640.04 0.2 98.2 18.900.02 0.5 100.5 +1.4 4.0
12 Escarole 7.140.11 1.5 101.8 7.170.13 1.8 101.6 +0.4 1.4
1 Artichoke 10.500.12 1.1 101.6 10.650.17 1.6 101.1 +1.4 2.0
14 Borage 3.870.10 2.6 98.4 3.760.07 1.9 99.7 -2.8 2.0
15 Cauliflower 2.020.07 3.5 99.0 1.960.05 2.5 98.9 -3.0 2.0
16 Chicory 8.750.07 0.8 100.6 8.680.10 1.2 101.1 -0.8 0.5
17 4.960.12 Watercress 2.4 99.5 5.020,08 1.6 100.3 +1.2 0.4
a
Mean Concentration and Standard Deviation (g kg-1), bCoefficient of Variation (%),cMeanspike Recovery (%),d Relative Error of The Potentiometric
Method Versus the Reference Method (%),The Critical F-value, Considering a 95% Confidence Level and 16 Degrees of Freedom, for a Two-Tailed
Test is 2.7

An aliquot of 50 mL was pippetted from prepared solution
directly into a 100 mL beaker, 5 mL of the ISA solution
were added and the reference, nitrate-selective and chlori-
de-selective electrodes were immersed. The nitrate and
chloride ions concentrations were determined by direct
potentiometry.
3. Results and Discussion
Taking into account the nitrate and chloride (Tables 1-2)
contained in different types of vegetables, determined by
application of the reference methods, and the value of the
potentiometric selectivity coefficient of the nitrate selecti-
ve electrode (based on tetraoctylammonium bromide as
sensor) (Alegret et al, 1987) for the chloride ion, only the
addition of an ISA solution was necessary to carried out
during the potentiometric procedure to avoid the chloride
interference. If any sample could contain abnormal high
chloride concentration, the use of some interference supp-
ressor-solution, usually silver (Milham et al, 1970) or lead
(Alexiades & Mitrakas, 1990) salts, is recommended. Unf-
ortunately, in that case the analytical procedure must be
carried out in two different steps being necessary the prec-
ipitation of chloride after its determination, and the filtra-
tion of the precipitate obtained, previous the nitrate deter-
mination. Since nitrate solutions are a culture medium for
bacteria and algae, boric acid could be added, to prevent
biological growth, if extracted samples need to be stored.
Due to the fact that the extracted samples of some veget-
ables are coloured, an error in the spectrophotometric ref-
erence method used in the nitrate determination can arise.
This problem can be circumvented by adding active carb-
on to the sample extracts before the analysis.
Table 1 summarizes the result obtained for, 17 samples of
various vegetable simultaneously analysed by the sequent-
ial potentiometric proposed method and the reference
techniques. The nitrate content was found to be in range
of 1.92 to 12.82 g kg-1 with an average value of 7.18 g
kg-1. The chloride content ranged from 0.62 to 15.19 g kg-
1
with a average value of 5.07 g kg-1. In each case, the
precision was evaluated by their application to 11 samples
of the same product and was expressed in terms of their
standard deviations and coefficients of variation. The
obtained data confirmed that the potentiometric proposed
method showed adequate precisions since the mean variat-
ion coefficients for the nitrate and chloride determinations
were 1.7% and 2.3%, respectively. To test whether the

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 Prez-Olmos et al / Chemistry Journal (2013), Vol. 03, Issue 03, pp. 81-85
proposed method and reference methods applied to the
nitrate and chloride determination differ in their precision,
a significance F-test (two-tailed test) was carried out. All
the F-values, with the practical exception of one sample
for the nitrate determinations, were less than the tabulated
F-value (2.77), so there are no significant differences bet-
ween the standard deviations at the 95% confidence level
(Miller & Miller, 1984).
After several additions of small volumes of nitrate and
chloride standard solutions to the extracted vegetable sam-
ples, the concentration of both ions were determined using
the same analytical methods and the mean percentages of
spike recovery were used to evaluate the accuracies. From
the data obtained, it is possible to affirm that the accuracy-
es of the sequential potentiometric proposed method were
good since the mean percentages of spike recovery obtain-
ed were 100.5% and 100.6% for the nitrate and chloride
determinations respectively. On the other hand, the perce-
ntages of difference obtained between the proposed and
references methods ranged from -4.4% to +3.2% with an
average value of -0.6% for the nitrate, and from -4.6% to
+4.5% with an average value of -0.3% for the chloride
determination. From this data it is possible to state that
exist a very slight tendency to get lower values with the
potentiometric proposed method than with the reference
methods.
The linear regression analysis of the results obtained by
the simultaneous application of the proposed potentiomet-
ric and reference methods to all samples showed the follo-
wing equations:
For nitrate determination
CUV - Vis = (0.988 0.017) CISE + (0.087 0.145)..1
r = 0.999 t = 86.537
For chloride determination
Ctitr. = (1.016 0.023) CISE + (0.073 0.151)...2 Analysis. 14th ed. Williams, S. ed. Arlington, Association
r = 0.998 t = 61.145
The confidence limits of the slopes, intercepts of the
regression lines and the t values for the correlation coeffi-
cients at 95 % confidence level and n2 degrees of freed-
om were also determined. From these data, it is possible
to affirm that the calculated slopes and intercepts do not
significative differ from the ideal values of 1 and 0 respec-
tively. Thus, there is no evidence of systematic differences
between the proposed and reference methods. From the
correlation coefficients, the calculated t values were great-
er than the tabulated t value (t = 2.13 for n = 15), hence
showing that significant correlations exist between both
methods (Miller & Miller, 1984).
Available online at www.scientific-journals.co.uk
ISSN 2049-954X
The detection limits for nitrate and chloride ions determi-
nations were obtained as recommended by the Analytical
Method Committee (Analytical Method Committee, 1987)
(+3 blank) and they resulted to be 0.12 g kg-1 and 0.08 g
kg-1, respectively.
4. Conclusions
The current study demonstrates the possibility of the sequ-
ential determination of nitrate and chloride in various veg-
etables by using potentiometric selective electrodes to each
ion. The developed analytical procedure resulted to be
rapid, economic, and easy to operate with adequate values
of precisions and accuracies. A statistical concordance of
the results for the developed and reference methods to 17
extracts of different vegetables were obtained by judging
by the linear regression analysis.
Acknowledgement
We thank to Instrumentacin Analtica S.A. for its finance-
al support.
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