4500-NO2⫺ NITROGEN (NITRITE)*

4500-NO2⫺ A. Introduction

1. Occurrence and Significance

2. Selection of Method
For a discussion of the chemical characteristics, sources, and The colorimetric method (4500-NO2⫺.B) is suitable for
effects of nitrite nitrogen, see Section 4500-N. concentrations of 5 to 1000 ␮g NO2⫺-N/L (See
4500-NO2⫺.B.1a). Nitrite values can be obtained by the au-
tomated method given in Section 4500-NO3⫺.E with the
Cu–Cd reduction step omitted. Additionally, nitrite nitrogen
can be determined by ion chromatography (Section 4110), and
by flow injection analysis (see Sections 4130 and 4500-
* Approved by Standard Methods Committee, 2000. Editorial revisions, 2011. NO3⫺.I).

4500-NO2⫺ B. Colorimetric Method

1. General Discussion b. Filter photometer, providing a light path of 1 cm or longer

and equipped with a green filter having maximum transmittance
a. Principle: Nitrite (NO2⫺) is determined through formation near 540 nm.
of a reddish purple azo dye produced at pH 2.0 to 2.5 by
coupling diazotized sulfanilamide with N-(1-naphthyl)-ethylene- 3. Reagents
diamine dihydrochloride (NED dihydrochloride). The applicable
range of the method for spectrophotometric measurements is 10 a. Nitrite-free water: If it is not known that the distilled or
to 1000 ␮g NO2⫺-N/L. Photometric measurements can be made demineralized water is free from NO2⫺, use either of the fol-
in the range 5 to 50 ␮g N/L if a 5-cm light path and a green color lowing procedures to prepare nitrite-free water:
filter are used. The color system obeys Beer’s law up to 180 ␮g 1) Add to 1 L distilled water one small crystal each of KMnO4
N/L with a 1-cm light path at 543 nm. Higher NO2⫺ concentra- and either Ba(OH)2 or Ca(OH)2. Redistill in an all-borosilicate-
tions can be determined by diluting a sample. glass apparatus and discard the initial 50 mL of distillate. Collect
b. Interferences: Chemical incompatibility makes it unlikely the distillate fraction that is free of permanganate; a red color
that NO2⫺, free chlorine, and nitrogen trichloride (NCl3) will with DPD reagent (Section 4500-Cl.F.2b) indicates the presence
coexist. NCl3 imparts a false red color when color reagent is of permanganate.
added. The following ions interfere because of precipitation 2) Add 1 mL conc H2SO4 and 0.2 mL MnSO4 solution (36.4 g
under test conditions and should be absent: Sb3⫹, Au3⫹, Bi3⫹, MnSO4 䡠 H2O/100 mL distilled water) to each 1 L distilled
Fe3⫹, Pb2⫹, Hg2⫹, Ag⫹, chloroplatinate (PtCl62⫺), and meta- water, and make pink with 1 to 3 mL KMnO4 solution (400 mg
vanadate (VO32⫺). Cupric ion may cause low results by catalyz- KMnO4/L distilled water). Redistill as described in the preceding
ing decomposition of the diazonium salt. Colored ions that alter paragraph.
the color system also should be absent. Remove suspended solids Use nitrite-free water in making all reagents and dilutions.
by filtration. b. Color reagent: To 800 mL water, add 100 mL 85% phos-
c. Storage of sample: Never use acid preservation for samples phoric acid and 10 g sulfanilamide. After dissolving sulfanil-
to be analyzed for NO2⫺. Make the determination promptly on amide completely, add 1 g N-(1-naphthyl)-ethylenediamine
fresh samples to prevent bacterial conversion of NO2⫺ to NO3⫺ dihydrochloride. Mix to dissolve, then dilute to 1 L with water.
or NH3. For short-term preservation for 1 to 2 d, freeze at ⫺20°C Solution is stable for about a month when stored in a dark bottle
or store at 4°C. in refrigerator.
d. Quality control (QC): The QC practices considered to be an c. Sodium oxalate, 0.025M (0.05N): Dissolve 3.350 g
integral part of each method are summarized in Table 4020:I. Na2C2O4, primary standard grade, in water and dilute to
1000 mL.
d. Ferrous ammonium sulfate, 0.05M (0.05N): Dissolve
2. Apparatus 19.607 g Fe(NH4)2 (SO4)2 䡠 6H2O plus 20 mL conc H2SO4 in
water and dilute to 1000 mL. Standardize as in Section
Colorimetric equipment: One of the following is required: 5220B.3d.
a. Spectrophotometer, for use at 543 nm, providing a light e. Stock nitrite solution: Commercial reagent-grade NaNO2
path of 1 cm or longer. assays at less than 99%. Because NO2⫺ is oxidized readily in the

https://doi.org/10.2105/SMWW.2882.088 1
 NITROGEN (NITRITE) (4500-NO2⫺)/Colorimetric Method

presence of moisture, use a fresh bottle of reagent for preparing the let temperature fall below 85°C. If necessary, warm beaker
stock solution and keep bottles tightly stoppered against the free contents during titration; 100 mg will consume about 6 mL
access of air when not in use. To determine NaNO2 content, add a solution. Run a blank on distilled water and H2SO4.
known excess of standard 0.05N KMnO4 solution (see ¶ h below),
g Na2C2O4
discharge permanganate color with a known quantity of standard Normality of KMnO4 ⫽
reductant, such as 0.025M Na2C2O4 or 0.05M Fe(NH4)2(SO4)2, and (A ⫺ B) ⫻ 0.067
back-titrate with standard permanganate solution. where:
1) Preparation of stock solution—Dissolve 1.232 g NaNO2 in
water and dilute to 1000 mL; 1.00 mL ⫽ 250 ␮g N. Preserve A ⫽ mL titrant for sample and
with 1 mL CHCl3. B ⫽ mL titrant for blank.
2) Standardization of stock nitrite solution—Pipet, in order,
Average the results of several titrations.
50.00 mL standard 0.05N KMnO4, 5 mL conc H2SO4, and
50.00 mL stock NO2⫺ solution into a glass-stoppered flask or
4. Procedure
bottle. Submerge pipet tip well below surface of permanganate-
acid solution while adding stock NO2⫺ solution. Shake gently a. Removal of suspended solids: If sample contains suspended
and warm to 70 to 80°C on a hot plate. Discharge permanganate solids, filter through a 0.45-␮m-pore-diam membrane filter.
color by adding sufficient 10-mL portions of standard 0.025M b. Color development: If sample pH is not between 5 and 9,
Na2C2O4. Titrate excess Na2C2O4 with 0.05N KMnO4 to the adjust to that range with 1N HCl or NH4OH as required. To
faint pink endpoint. Carry a water blank through the entire 50.0 mL sample, or to a portion diluted to 50.0 mL, add 2 mL
procedure and make the necessary corrections in the final cal- color reagent and mix.
culation as shown in the equation below. c. Photometric measurement: Between 10 min and 2 h after
If standard 0.05M ferrous ammonium sulfate solution is substi- adding color reagent to samples and standards, measure absor-
tuted for Na2C2O4, omit heating and extend reaction period between bance at 543 nm. As a guide use the following light paths for the
KMnO4 and Fe2⫹ to 5 min before making final KMnO4 titration. indicated NO2⫺-N concentrations:
Calculate NO2⫺-N content of stock solution by the following
equation:
Light Path Length NO2⫺-N
[(B ⫻ C) ⫺ (D ⫻ E)] ⫻ 7 cm ␮g/L
A⫽
F
1 2–25
5 2–6
where: 10 ⬍2
A ⫽ mg NO2⫺-N/mL in stock NaNO2 solution,
B ⫽ total mL standard KMnO4 used, 5. Calculation
C ⫽ normality of standard KMnO4,
D ⫽ total mL standard reductant added, Prepare a standard curve by plotting absorbance of standards
E ⫽ normality of standard reductant, and
against NO2⫺-N concentration. Compute sample concentration
F ⫽ mL stock NaNO2 solution taken for titration.
directly from curve.
Each 1.00 mL 0.05N KMnO4 consumed by the NaNO2 solu-
tion corresponds to 1725 ␮g NaNO2 or 350 ␮g NO2⫺-N. 6. Precision and Bias
f. Intermediate nitrite solution: Calculate the volume, G, of
In a single laboratory using wastewater samples at concentrations
stock NO2⫺ solution required for the intermediate NO2⫺ solution
of 0.04, 0.24, 0.55, and 1.04 mg NO⫺3 ⫹ NO2⫺-N/L, the standard
from G ⫽ 12.5/A. Dilute the volume G (approximately 50 mL)
deviations were ⫾0.005, ⫾ 0.004, ⫾ 0.005, and ⫾ 0.01, respec-
to 250 mL with water; 1.00 mL ⫽ 50.0 ␮g N. Prepare daily.
tively. In a single laboratory using wastewater samples at concen-
g. Standard nitrite solution: Dilute 10.00 mL intermediate
trations of 0.24, 0.55, and 1.05 mg NO3⫺ ⫹ NO2⫺-N/L, the
NO2⫺ solution to 1000 mL with water; 1.00 mL ⫽ 0.500 ␮g N.
recoveries were 100%, 102%, and 100%, respectively.1
Prepare daily.
h. Standard potassium permanganate titrant, 0.05N: Dissolve
7. Reference
1.6 g KMnO4 in 1 L distilled water. Keep in a brown glass-
stoppered bottle and age for at least 1 week. Carefully decant or
1. U.S. ENVIRONMENTAL PROTECTION AGENCY. 1979. Methods for Chem-
pipet supernate without stirring up any sediment. Standardize ical Analysis of Water and Wastes; Method 353.3. Washington, D.C.
this solution frequently by the following procedure:
Weigh to the nearest 0.1 mg several 100- to 200-mg samples
of anhydrous Na2C2O4 into 400-mL beakers. To each beaker, in 8. Bibliography
turn, add 100 mL distilled water and stir to dissolve. Add 10 mL BOLTZ, D.F., ed. 1958. Colorimetric Determination of Nonmetals. Inter-
1 ⫹ 1 H2SO4 and heat rapidly to 90 to 95°C. Titrate rapidly with science Publishers, New York, N.Y.
permanganate solution to be standardized, while stirring, to a NYDAHL, F. 1976. On the optimum conditions for the reduction of nitrate
slight pink endpoint color that persists for at least 1 min. Do not by cadmium. Talanta 23:349.

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