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Kjeldahl Method for Total Nitrogen Analysis

The document outlines the Kjeldhal method for determining Total Nitrogen and Crude Protein in food samples. It describes the principle of converting nitrogen to ammonium sulfate, followed by steam distillation and titration to quantify nitrogen content. The procedure includes sample preparation, digestion, and distillation, with specific calculations for nitrogen and crude protein percentages.

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74 views3 pages

Kjeldahl Method for Total Nitrogen Analysis

The document outlines the Kjeldhal method for determining Total Nitrogen and Crude Protein in food samples. It describes the principle of converting nitrogen to ammonium sulfate, followed by steam distillation and titration to quantify nitrogen content. The procedure includes sample preparation, digestion, and distillation, with specific calculations for nitrogen and crude protein percentages.

Uploaded by

Be Frank
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

DETERMINATION OF TOTAL NITROGEN /CRUDE PROTEIN USING

THE KJELDHAL METHOD

AIM:

To determine the Total Nitrogen / Crude Protein for the given Food sample using Kjeldhal
Method.

PRINCIPLE:

The Nitrogen in protein or any other organic material is converted to ammonium sulfate by
Sulfuric Acid during digestion. This salt on steam distillation liberates ammonia which is
collected in boric acid solution and titrates against standard acid. 1 ml of 0.1N acid is equivalent
to 1.401mgN, calculation is made to arrive the Nitrogen content of the sample.

Materials Required

 Sample.
 Boric acid.
 Conc. Sulfuric Acid.
 Potassium sulfate.
 Copper sulfate.
 40% NaOH.

STEPS:

 Weighing sample.
 Adding catalyst, salt and Sulfuric acid.
 Digestion.
 Distillation.
 Titration.
 Nitrogen is determined and crude protein is calculated.

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CALCULATION:

(ml HCl sample - ml HClblank) x Con HCl x 14.01


Calculation of % of Nitrogen = --------------------------------------------------------- x 100
1000 x Weight of sample (g)
Calculation of % of crude Protein = % of nitrogen x conversion factor for crude protein

The factor 6.25 is used for almost all feed. The conversion factor varies for different samples.

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PROCEDURE:

1. Sample is weighed (0.2g to 0.5 g) into 250ml glass tubes

2. Kjeltabs containing K2SO4 and CuSO4 are added into each glass tube (* CuSO4 is used as a
catalyst, * K2SO4 is used to increase the boiling point)

3. 15ml of concentrated H2SO4 is added and the sample is mixed well.

Digestion Process:

1. Heating block should be warmed to 420 C. The rack with the glass tubes is placed on the
heating block.

2. The heating block is boiled for 75 min.

3. After digestion is complete, we get a clear green color which indicates a complete
chemical digestion.

Digestion equation:

Protein + H2SO4  2NH4+ + SO4- + CO2 + SO2 + H2O

Distillation Process:

1. 25ml of 4% Boric acid is added to a conical flask and then the glass tube is attached to the
rubber holder and then the safety door is closed.

2. The distillation cycle is started.

3. 60ml of 40% NaOH is pumped into the glass tube steam is created in the steam generator
and the distillation will start.

4. Distillation time is 5 min. The ammonia distillate drops into the boric acid in the conical
flask.

5. Distillation cycle is finished.

Distillation equations:

Liberation of ammonia

2NH4+ + SO4- + 2NaOH  Na2SO4 + 2H2O + 2 NH3

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