Experiment No.

Name of the Practical : Determination of Nitrate-nitrogen (NO3-N), Ammonical

Nitrogen (NH4-N) & TKN of given wastewater sample.

Theory: Nitrogen is an essential nutrient for the growth of photosynthetic organisms, thus
it is important to monitor and control the discharge into the environment. Four forms of
Nitrogen namely; Nitrate Nitrogen (NO3-N), Nitrite Nitrogen (NO2-N), Ammonical
Nitrogen (NH4-N) and organic nitrogen may be present in the water and wastewater. The
Kjeldahl method is commonly used to determine the nitrogen content of the samples.
 The Kjeldahl procedure involves three major steps:

1) Digestion - Organic nitrogen is converted into NH4+

2) Distillation - NH3 is distilled and retained in a receiver vessel

3) Titration - Nitrogen is determined

The aim of the digestion procedure is to break all nitrogen bonds in the sample
and convert all of the organically bonded nitrogen into ammonium ions (NH4 +).
 Digestion Procedure

• The sample is mixed with sulfuric acid at temperatures between 350-380 ºC

• Potassium sulphate is added in order to increase the boiling point of sulfuric
acid.
• After digestion is completed the sample is allowed to cool to room
temperature, then diluted with water and transferred to the distillation unit.
Estimation of Nitrogen : Kjeldahl Method
 Reagents Required
A) Total Kjeldahl Nitrogen:

N.B.: For 0.02N, 1.00 mL = 280 µg N

B) Ammonical Nitrogen:
C) Nitrate Nitrogen:

Concentrated HCl
Procedure for TKN determination:
Procedure for Ammonical Nitrogen determination:
Procedure for Nitrate Nitrogen determination:

275

N.B.: 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
Plot calibration curve by taking absorbance at different known concentration
(prepared by stock solution)
Observations:

Note down the value of A & B:

A - Volume of Standard 0.02N H2SO4 solution used for sample titration.

B - Volume of Standard 0.02N H2SO4 solution used for blank
Calculations
Environmental Significance:
Video Links for Demonstration:

https://www.youtube.com/watch?v=KjNOFBc6_t8

https://www.youtube.com/watch?v=6iB9XlRIbck

https://www.youtube.com/watch?v=h84cOeJNSC0&t=112s