W6 – DETERMINATION OF NITRATE IN WATER BY UV-VISIBLE

SPECTROMETRY, ION CHROMATOGRAPHY

6.1 Introduction

Nitrate and nitrite ions are important indicators of pollution by organic materials as
nitrogen from decomposing organic substances often ends up as nitrate or nitrite ions.
The determination of nitrate is often difficult because of the low levels found , and the
distinct possibility of interfering materials being present.

When determining nitrate it is important to choose a method which suits both the
interferences which are present and the level of analyte in the solution. The UV-
visible method of determining nitrate is a useful method for screening of a solution to
make a rough determination of the level of nitrate. It determines the absorption f the
nitrate ion at 220nm. Good results are obtained if the water contains little or no
organic matter (which also absorbs at 220nm). A correction for organic matter can be
performed by taking absorbance readings at 275nm (where nitrate does not absorb)
and estimating the organic content of the sample.

After screening the sample and determining the amount of nitrate other methods may
be chosen and used to give more accurate estimates of nitrate content of the water
sample. For example the nitrate ion selective electrode is suitable from 0.14 –
1400mg/L, and ion chromatography from 10mg/L upwards.

In this practical you will determine the nitrate ion level of an unknown by first
screening your sample with the UV-visible method, then undertaking analysis by ion
chromatography.

6.2 Procedure

Part 1 – Screening by UV-Visible Spectrophotometry

1. Prepare 100 mL of nitrate calibration standards for 2, 4, 6, 8 and 10mg/L by

serially diluting appropriate amounts of the 100mg/L stock solution, and
making them up to the mark with HCl (1mL) and modulab water.

2. Obtain an unknown nitrate sample and analyse this in parallel with a freshly
obtained creek water sample.

3. Filter a portion of the sample through 0.45µm Millipore paper, then take a 25
mL aliquot and transfer to a 50mL volumetric flask, add HCl (1mL) and make
up to the mark with modulab water.

4. Read the absorbance of the samples and standards at both 220nm and 275nm.
Use modulab water containing HCl as a blank

5. Prepare a calibration graph and determine the concentration of nitrate ion in

the unknown and the creek sample
Part 2 – Ion Chromatographic Method
(Do not use the IC for creek water or high chloride content samples)

1. Prepare calibration standards (1, 2, 5 and 10 mg /L) for the nitrate ion by serial
dilution of the stock solution.

2. Run each of the standards on the ion chromatograph as instructed by the

teacher. The following conditions should be recorded

Column
Flow rate
Eluent solution
Detector type/setting
Sample Loop size

3. Determine the retention time of the nitrate ion from standards and plot a
calibration graph of peak area versus concentration.

4. Filter and de-gas the sample solution through 0.2µm millpore paper.

5. Inject unknown sample into chromatograph and calculate the concentration of

nitrate in your unknown by reference to the calibration graph.

Questions

1. Describe the origins and importance of nitrate as a water pollutant.

2. Why is it important that the levels of nitrate in our water systems are kept to
very low levels?
3. Describe at least one other method for the determination of nitrate in water or
waste water.

References

1. Rump, H.H. & Krist, H., Laboratory Manual for the Examination of
Water, Wastewater & Soil, VCH, 1988. ISBN 0-89573-851-1

2. Rand , M.C., Greenberg, A.E., and Taras, M.J. ed., Standard Methods For
Examination of Water and Wastewater , 18th edition, APHA, AWWA,
WPCF, Chapman & Hall, 1992. ISBN 0-87553-207-1

3. Australian Standard 2383, Standards Association of Australia, 1986.

4. Barker, D., Electroanalytical techniques Theory & Practical notes, Hunter

Institute of Technology Students Union, 1994.

5. Barker, D., Advanced Chromatography Theory & Practical notes, Hunter

Institute of Technology Students Union, 1995