INTRODUCTION:
Nitrite is an intermediate oxidation
state of nitrogen both in the oxidation of ammonia to nitrate & in the
reduction of nitrate. Such oxidation & reduction may occur in waste water
treatment, water distribution system & natural water. Nitrite can enter a
supply system through it use as a corrosion inhibitor in industrial process
water.
Nitrite is the actual etiological
agent at methemoglobimia. Nitrous acid which is formed from nitrite in acidic
solution can react with nitro samines , many of which are known to be
carcinogens.
PRINCIPLE:
Nitrite is determined through
formation of reddish purple azo dye produced at pH 2.0 to 2.5 by coupling
diazotize sulphanilamine with N’ (1- napthyl) ethyl diamine dihydrochloride (NED).
The applicable range of the method for photometric measurement is 10-10,000
micro gram NO2/l. Photometric measurement can be made in the range 5 to 50 ยตg
N2/l, if a 5cm light path & a green color filter are used. Higher nitrite
conc. Can be determined by diluting a sample with nitrite free D/W. The
measurement is taken at 543nm using spectrophotometer.
REQUIREMENTS:
- Nitrite frees D/W.
- Water sample.
-
Drinking
water &
-
Waste
water
- Color reagent.
1. 85% phosphotic
acid 100 ml.
2. Sulfonilamide
10 gm
3. Dissolved
property in 800ml D/W.
4. NED
hydrochloride 1 gm.
5. Mix &
final vol. 1000ml with D/W.
6. Solution is
stable for 1 month.
- When stored in dark bottle in refrigerator.
- Stock nitrite solution. (NaNO2 1.232g)
(1ml = 250 ยตg N)
-
D/w
1000 ml
-
Preserve
with 1ml CHCl3.
- Intermediate nitrite solution.
-
Stock
nitrite solution 50ml.
-
D/W 250 ml.
(1ml = 50 ยตg N)
- Standard (working) nitrite solution.
-
Intermediate
nitrite solution 10ml.
-
Make
final vol. 1000ml with D/W
(1ml
= 0.5 ยตg N)
-
Prepare
fresh.
- Test tubes – 11
- Pipettes 1 ml & 10 ml
- Spectrophotometer & quates.
PROCEDURE:
Take different aliquotes of std.
nitrite solution form 0.2, 0.4,…….. 1 ml.
-
In
blank do not add std. solution.
-
Prepare
3 aliquotes of unknown sample.
-
Add
D/W in all tubes to make final vol. 10 ml.
-
Add
0.2 ml color reagent in all tubes.
-
Incubate
all the tubes at room temp. For 10 min.
-
Take
O.D. of all tubes at 543nm using blank as a reference.
-
Prepare
a std. curve by plotting absorbance of std. against nitrite nitrogen (ยต gms) of
std. & find out conc. Of nitrite present in unknown sample.
-
Note
down the result & give your conclusion.
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