Kjendahl: Determination of protein in pork.

Daniel Hernández
Juliana Sánchez

Abstract:
Kjeldahl's method aims to determine the amount of organic nitrogen in a wide range of samples.
Through three fundamental stages: digestion, distillation and titration, this method is commonly
used to estimate the protein content of foods. Kjeldahl's method is based on the digestion of a
sample in concentrated sulphuric acid until the nitrogen in the sample is converted to ammonium
sulphate. By adding an excess of sodium hydroxide solution, which neutralizes the sulfuric acid,
the released ammonia is distilled and collected over a standard HCl solution, of which a fraction is
neutralized by the ammonia. Finally, the unconsumed HCl is titrated by back titration with
standard NaOH and the amount of ammonia collected is found. The Kjeldahl method currently has
a wide variety of applications, from the determination of nitrogen in food and beverages, such as
meat, cereals and fodder. How much nitrogen will be present in a sample of pork?
The main objective of the practice is the determination of the percentage of nitrogen in a given
sample by means of the Kjeldahl method, during which a digestion with sulphuric acid, an
ammonia distillation and a back titration are sought and carried out.

Keywords: titration, protein, nitrogen, percentage, meat

Theoretical frame:
The Kjeldahl method or Kjeldahl digestion, in analytical chemistry, is a chemical analysis process to
determine the nitrogen content of a chemical substance and falls under the category of wet
digestion media. It is commonly used to estimate the protein content of foods. It was developed
by the Danish Johan Kjeldahl in 1883.
Digestion: The goal of the digestion procedure is to break all the nitrogen bonds in the sample and
convert everything organically bound nitrogen in ammonium ions (NH4+). Organic carbon and
hydrogen form carbon dioxide and water. In this process organic matter is charring resulting in the
formation of a black foam. During digestion, the foam decomposes and finally becomes a clear
liquid indicating that the chemical reaction is over. To do this, the simple is mixed with sulphuric
acid at temperatures between 350 and 380 °C. The higher the temperature, the faster will be the
process of digestion. Digestion can also be accelerated with the addition of salts and catalysts. It
adds potassium sulphate to increase the boiling point of the sulphuric acid and catalysts are added
to increase the boiling point of the sulphuric acid and catalysts are added to increase the boiling
point of the sulphuric acid and catalysts are added to increase the boiling point of the sulphuric
acid. increase the speed and efficiency of the digestion process. Oxidizing agents can also be added
to further improve speed. The digestion time depends on the chemical structure of the sample,
the temperature, the amounts of salt sulphate and catalyst.
Kjeldahl Catalysts
Catalysts are made up of more than 97% of a salt that causes a increase in the boiling temperature
of sulphuric acid and 1-3% of a type of sulphuric acid. catalyst or a mixture of catalysts to increase
the speed and efficiency of the catalyst digestion procedure. Typical catalysts are selenium or
metal salts of copper or titanium. The selection of a particular catalyst depends on ecological and
toxic aspects. or for more practical reasons such as reaction time or foaming and the spraying. For
example, the catalyst containing selenium reacts faster but it is toxic, while a catalyst containing
copper is safer for the environment. people and the environment, but gives a slower digestion
process. A The ideal compromise is the mixed catalyst consisting of copper and titanium sulphate.
Distillation
The acid sample is neutralised by means of a concentrated solution of sodium hydroxide. During
the distillation process the ammonium ions (NH4+) are converted into ammonia (NH3) which is
drawn into the receiver vessel by means of a stream of water vapour (steam distillation). The
receiver for the distillate is filled with an absorbent solution to capture the dissolved ammonia gas.
The the most common absorbent solution is boric acid [B(OH)3] in 2-4% aqueous solution. The
following may also be used other acids, accurately dosed, such as sulphuric acid or hydrochloric
acid to capture ammonia in the form of Solvated ammonium ions. Boric acid is the method of
choice because it allows automation.
Valuation
The concentration of the captured ammonium ions can be determined by means of two types of
titration:
- When boric acid is used as an absorbent solution, an acid-base titration is subsequently carried
out using a standardised solution of sulphuric acid or hydrochloric acid. The concentration range of
the solution used varies from 0.01N to 0.5N depending on the amount of ammonium ions present.
Endpoint detection can be performed manually, with colorimetric titration, using a combination of
indicators. The combination of methyl red and methyl blue indicators is frequently used in many
methods. The end point of the titration can also be determined potentiometrically with a pH
electrode. This titration is called direct titration.
- When a titrated solution of sulphuric acid is used as an absorbent solution, the residual sulphuric
acid (i.e. the excess that does not react with NH3) is titrated with a standardised solution of
sodium hydroxide and the residual sulphuric acid (i.e. the excess that does not react with NH3).
the amount of ammonia is calculated by difference. The end point is detected with a color
indicator, the more used is methyl red. This valuation is called indirect valuation or backward
valuation.

Methodology:

Data tables:

Analysis of results:
Image 1. Percentage of meat protein in pork meat

Conclusions:

- Kjeldahl's method has as advantages its applicability to a high range of products, which is
relatively simple, and which is the official method for measuring pure protein content.
- There are several catalysts for use in the Kjeldahl method, among which are mercury oxide
and copper sulfate (which are the most commonly used). However, mercury oxide forms a
stable compound with ammonia that must be broken down with the addition of sodium
thiosulfate and copper sulfate is less effective in recovering nitrogen.
- The realization of the practice allowed to identify different factors that influence the
veracity and accuracy of the results obtained by means of this method.

Bibliography:

- Vinasco, J. (2007). Determination of nitrogen by the micro-Kjeldhal method. Retrieved

October 13, 2013 from website: http://es.scribd.com/doc/504240/informe-microKjendhal
- Romero, N. Methods of Analysis for the Determination of Nitrogen and Nitrogen
Constituents in Foods. Retrieved 13 October 2013 from the website:
http://www.fao.org/docrep/010/ah833s/ah833s17.htm
- Anonymous. Analytical methods for proteins in foods. Retrieved October 13, 2013, from
http://www.fao.org/docrep/006/y5022e/y5022e03.htm#TopOfPage