TEST Document No.

UT/Lab/TM/W/021
METHOD Issue Date 01-04-2009
1. TITLE DETERMINATION OF TOTAL KJELDAHL NITROGEN.
2. SCOPE This procedure covers instructions for determination of determination
of total kjeldahl's nitrogen by distillation and titrimetric method.
3. REFERENCE APHA 19th Edition1995, 4500 NOrg B.Macro Kjeldahl Method.
4. AUTHORITY & This procedure is approved by Managing Director. It is issued by
RESPONSIBILITY Management Representative. This procedure is implemented by
Laboratory Department.

5. PROCEDURE STEPS

5.1PRINCIPLE Total Kjeldahl Nitrogen is defined as the sum of free ammonium and
organic nitrogen compounds which are converted to ammonium
sulphate (NH4)2SO4, under the condition of digestion. This method is
applicable for samples containing either low or high concentration of
organic nitrogen In presence of sulphuric acid, potassium sulphate and
mercuric sulphate catalyst organically bound nitrogen gets converted
into ammonium sulphate. Potassium sulphate is added to raise the
boiling point of sulphuric acid from 345oC to 370oC. The digested is
diluted, made alkaline with NaOH and distilled. The liberated ammonia
is absorbed in boric acid. The absorbed ammonia is determined either
by titration or direct nesslerization.

5.2 APPARATUS 1. Kjeldahl Digestion assembly

2. Distillation apparatus
3. Spectrophotometer for use at 400 – 425 nm with a light path 1cm or
more.

5.2REAGENTS • TKN Digestion Reagent: Dissolve 134 g K2SO4 and 7.3 g CuSO4
in about 800 ml water. Add 134 ml Conc. H2SO4.Dilute to 1 lit.
• Sodium Hydroxide-Sodium thiosulfate reagent: Dissolve 500 gm
NaOH and 25 gm Na2S2O3.5H2O in ammonia free distilled water
and dilute to 1000 ml.
• Borate buffer solution: Add 88 ml 0.1 N NaOH to 500 ml
app.0.025 M Sodium tetraborate and dilute to 1 L.
• Sodium Hydroxide,6 N
• Standard Sulphuric Acid Titrant (0.02 N): 0.1N H2SO4: Prepare
stock solution of 0.1N sulphuric acid by diluting 3.0 ml conc.
H2SO4 to 1000 ml with distilled water.

• Standardisation: Standardize against 10 ml 0.02N Na2CO3

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 TEST Document No. UT/Lab/TM/W/021
METHOD Issue Date 01-04-2009
solution, in a beaker by titrating to pH of about and titrate with
0.02N H2SO4 acid using phenolphthalein and methyl orange
indicator.
10 x 0.02 (N of Na2CO3)
N of H2SO4 = ---------------------------------
B. R. (ml of H2SO4)

Standard Sulphuric acid, 0.02N: Dilute 200 ml of the 0.1N standard

acid to 1000 ml with distilled water. Standardize as above by taking
10 ml of Na2CO3. 1ml = 0.28 mg N.
• Mixed indicator: Dissolve 200 mg Methyl red and 100 mg
methylene blue in 100 ml 95% ethyl or iso-propyl alcohol.
• Indicating Boric acid solution (2%): Dissolve 20g of H3BO3 in
1000 ml of distilled water. Add 10 ml of mixed indicator and dilute
to 1000 ml.
• NaOH/EDTA solution: Dissolve 400 gm NaOH in 800 ml water.
Add 45.2 g EDTA and stir to dissolve and dilute to 1000 ml.
• Stock NH4Cl solution: Dissolve 3.819 g NH4Cl (dried) in water and
dilute to 1000 ml.
(1 ml = 1mg N=1.22 mg NH3)

5.3PROCEDURE A. Digestion
1. Take suitable volume (25 ml) of well mixed sample in Kjeldahl
flask.
2. Add 50 ml digestion reagent mix the contents in flask.
Add few glass beads.
3. Continue digestion for another 30 min. after the sample appears
clear to ensure complete decomposition of organic matter.
4. Cool and dilute with D/W. to 150 or 300 ml depending on capacity
of the flask.
5. Add 50 ml Sodium hydroxide reagent to form an alkaline layer.
B. Distillation
1. Place the flask in its proper position in distillation apparatus and
Turn on heat. Distill and collect 200 ml distillate, use 50 ml
indicating boric acid as absorbent solution.
2. Extend the tip of the condenser well below the level of boric acid
solution. Do not let temp in condenser rise above 29oC.
3. When distillation complete remove the flask containing distillate
first and then put off heat to avoid back suction.
4. Titrate distillate with 0.02 N H2SO4 till the indicator turns a pale
lavender color.

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 TEST Document No. UT/Lab/TM/W/021
METHOD Issue Date 01-04-2009

5.4CALCULATION
1. Titrimetric :
(A-B) x 280
Total Kjeldahl Nitrogen mg/L = ------------------
ml of sample

If normality of the H2SO4 is not 0.02 N,

(A - B) x N x 1000 x 14
Total Kjeldahl Nitrogen mg/L = ---------------------------------
ml of sample
where,
A = ml of standard H2SO4 solution used in titrating sample.
B = ml of standard H2SO4 solution used in titrating blank.
N = Normality of H2SO4 (0.02 N)
14 = Milliequivalent weight of Nitrogen (14 mg).

Action by/ Responsibility : Chemist