Standard Methods for the Examination of Water and Wastewater

1.04 mg NO–3 + NO2–-N/L, the standard deviations were ±0.005, ± 0.004, ± 0.005, and ± 0.01,
respectively. In a single laboratory using wastewater samples at concentrations of 0.24, 0.55, and
1.05 mg NO3– + NO2–-N/L, the recoveries were 100%, 102%, and 100%, respectively.1

7. Reference
1. U.S. ENVIRONMENTAL PROTECTION AGENCY. 1979. Methods for Chemical Analysis of
Water and Wastes. Method 353. 3. U.S. Environmental Protection Agency,
Washington, D.C.

8. Bibliography
BOLTZ, D.F., ed. 1958. Colorimetric Determination of Nonmetals. Interscience Publishers, New
York, N.Y.
NYDAHL, F. 1976. On the optimum conditions for the reduction of nitrate by cadmium. Talanta
23:349.

4500-NO2– C. (Reserved)

4500-NO3– NITROGEN (NITRATE)*#(60)

4500-NO3– A. Introduction

1. Selection of Method
Determination of nitrate (NO3–) is difficult because of the relatively complex procedures
required, the high probability that interfering constituents will be present, and the limited
concentration ranges of the various techniques.
An ultraviolet (UV) technique (Method B) that measures the absorbance of NO3– at 220 nm
is suitable for screening uncontaminated water (low in organic matter).
Screen a sample; if necessary, then select a method suitable for its concentration range and
probable interferences. Nitrate may be determined by ion chromatography (Section 4110) or
capillary ion electrophoresis (Section 4140). Applicable ranges for other methods are: nitrate
electrode method (D), 0.14 to 1400 mg NO3–-N/L; cadmium reduction method (E), 0.01 to 1.0
mg NO3–-N/L; automated cadmium reduction methods (F and I), 0.001 to 10 mg NO3–-N/L. For
higher NO3–-N concentrations, dilute into the range of the selected method.

© Copyright 1999 by American Public Health Association, American Water Works Association, Water Environment Federation
 Standard Methods for the Examination of Water and Wastewater

Colorimetric methods (B, E) require an optically clear sample. Filter turbid sample through
0.45-µm-pore-diam membrane filter. Test filters for nitrate contamination.

2. Storage of Samples
Start NO3– determinations promptly after sampling. If storage is necessary, store for up to 2
d at 4°C; disinfected samples are stable much longer without acid preservation. For longer
storage of unchlorinated samples, preserve with 2 mL conc H2SO4/L and store at 4°C. NOTE:
When sample is preserved with acid, NO3– and NO2– cannot be determined as individual
species.

4500-NO3– B. Ultraviolet Spectrophotometric Screening Method

1. General Discussion
a. Principle: Use this technique only for screening samples that have low organic matter
contents, i.e., uncontaminated natural waters and potable water supplies. The NO3– calibration
curve follows Beer’s law up to 11 mg N/L.
Measurement of UV absorption at 220 nm enables rapid determination of NO3–. Because
dissolved organic matter also may absorb at 220 nm and NO3– does not absorb at 275 nm, a
second measurement made at 275 nm may be used to correct the NO3– value. The extent of this
empirical correction is related to the nature and concentration of organic matter and may vary
from one water to another. Consequently, this method is not recommended if a significant
correction for organic matter absorbance is required, although it may be useful in monitoring
NO3– levels within a water body with a constant type of organic matter. Correction factors for
organic matter absorbance can be established by the method of additions in combination with
analysis of the original NO3– content by another method. Sample filtration is intended to remove
possible interference from suspended particles. Acidification with 1N HCl is designed to prevent
interference from hydroxide or carbonate concentrations up to 1000 mg CaCO3/L. Chloride has
no effect on the determination.
b. Interference: Dissolved organic matter, surfactants, NO2–, and Cr6+ interfere. Various
inorganic ions not normally found in natural water, such as chlorite and chlorate, may interfere.
Inorganic substances can be compensated for by independent analysis of their concentrations and
preparation of individual correction curves. For turbid samples, see ¶ A.1.

2. Apparatus
Spectrophotometer, for use at 220 nm and 275 nm with matched silica cells of 1-cm or
longer light path.
© Copyright 1999 by American Public Health Association, American Water Works Association, Water Environment Federation
 Standard Methods for the Examination of Water and Wastewater

3. Reagents
a. Nitrate-free water: Use redistilled or distilled, deionized water of highest purity to
prepare all solutions and dilutions.
b. Stock nitrate solution: Dry potassium nitrate (KNO3) in an oven at 105°C for 24 h.
Dissolve 0.7218 g in water and dilute to 1000 mL; 1.00 mL = 100 µg NO3–-N. Preserve with 2
mL CHCl3/L. This solution is stable for at least 6 months.
c. Intermediate nitrate solution: Dilute 100 mL stock nitrate solution to 1000 mL with
water; 1.00 mL = 10.0 µg NO3–-N. Preserve with 2 mL CHCl3/L. This solution is stable for 6
months.
d. Hydrochloric acid solution, HCl, 1N.

4. Procedure
a. Treatment of sample: To 50 mL clear sample, filtered if necessary, add 1 mL HCl solution
and mix thoroughly.
b. Preparation of standard curve: Prepare NO3– calibration standards in the range 0 to 7 mg
NO3–-N/L by diluting to 50 mL the following volumes of intermediate nitrate solution: 0, 1.00,
2.00, 4.00, 7.00 . . . 35.0 mL. Treat NO3– standards in same manner as samples.
c. Spectrophotometric measurement: Read absorbance or transmittance against redistilled
water set at zero absorbance or 100% transmittance. Use a wavelength of 220 nm to obtain NO3–
reading and a wavelength of 275 nm to determine interference due to dissolved organic matter.

5. Calculation
For samples and standards, subtract two times the absorbance reading at 275 nm from the
reading at 220 nm to obtain absorbance due to NO3–. Construct a standard curve by plotting
absorbance due to NO3– against NO3–-N concentration of standard. Using corrected sample
absorbances, obtain sample concentrations directly from standard curve. NOTE: If correction
value is more than 10% of the reading at 220 nm, do not use this method.

6. Bibliography
HOATHER, R.C. & R.F. RACKMAN. 1959. Oxidized nitrogen and sewage effluents observed by
ultraviolet spectrophotometry. Analyst 84:549. GOLDMAN, E. & R. JACOBS. 1961.
Determination of nitrates by ultraviolet absorption. J. Amer. Water Works Assoc. 53:187.
ARMSTRONG, F.A.J. 1963. Determination of nitrate in water by ultraviolet spectrophotometry.
Anal. Chem. 35:1292.
NAVONE, R. 1964. Proposed method for nitrate in potable waters. J. Amer. Water Works Assoc.
56:781.
© Copyright 1999 by American Public Health Association, American Water Works Association, Water Environment Federation