INTERNATIONAL
STANDARD
INTERNATIONAL ORGANIZATION FOR STANDARDIZATIONWlEIKJjYHAPO,QHAR OPrAHM3AL&lR fl0 CTAH~APTM3AL&WWORGANISATION INTERNATIONALE DE NORMALISATION
Starches and derived products - Determination of nitrogen
content by the Kjeldahl method - Titrimetric method
Amidons, fdcules et produits d&iv& - Dosage de l azote selon Ia methode de Kjeldahl - Mt!thode titrimetrique
First edition - 1978-08-15
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ISO 3188:1978
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372388bc295e/iso-3188-1978
Ei
- UDC 664.2 : 546.17 : 543.24 Ref. No. ISO 3188-1978 (E)
Descriptors : starches, Chemical analysis, determination of content, nitrogen, volumetric analysis, Kjeldahl method.
Price based on 3 pages
FOREWORB
ISO (the International Organization for Standardization) is a worldwide federation
of national Standards institutes (ISO member bodies). The work of deweloping
International Standards is carried out through ISO technical committees. Ewery
member body interested in a subject for which a technical committee has been set
up has the right to be represented on that committee. International organizations,
governmental and non-governmental, in liaison with ISO, also take part in the work.
Draft International Standards adopted by the technical committees are circulated
to the member bodies for approval before their acceptance as International
Standards by the ISO Council.
International Standard iTeh STANDARD PREVIEW
ISO 3188 was developed by Technicai Committee
ISO/TC 93, Starch (including derivatives and by-products), and was circulated
to the member bodies in August 1975. (standards.iteh.ai)
lt has been approved by the member bodies of the following
ISOcountries
3188:1978:
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Australia 1ran Thailand
372388bc295e/iso-3188-1978
Czechoslovakia Netherlands Turkey
France Poland United Kingdom
Germany Romania Yugoslavia
Hungary South Africa, Rep. of
The member body of the following country expressed d isapprova I of the docume nt
on technical grounds :
U.S.A.
@ International Organisation for Standard ization, 9 978 l
Printed in Switzerland
INTERNATIONAL STANDARD ISO 31884978 (E)
Starches and derived products - Determination of nitrogen
content by the Kjeldahl method - Titrimetric method
1 SCOPE AND FIELD OF APPLICATION 5 REAGENTS
This International Standard specifies a titrimetric method The reagents shall be of recognized analytical quality.
for the determination, by the Kjeldahl method, of the Ammonia-free distilled water or water of at least equivalent
nitrogen content of starch and its derived products whose purity shall be used.
presumed nitrogen content is greater than 0,Ol % (mlm)? )
5.1 Sulphuric acid, concentrated, p20 1,84 g/ml [96 %
NOTE - In starches and derived products to which nitrogenous
materials have not been added, the nitrogen is present essentially (mhd].
in the form of Protein and/or amino acids.
5.2 Sodium hydroxide, solution 30 % (mlm), p20
1,33 g/ml.
2 REFERENCES
NOTE - This Solution may be more concentrated.
ISO 1227fAdd. 2, Starch, including derivatives and by-
products - Vocabulary. iTeh STANDARD 5.3PREVIEW Boric acid, 20 g/ solution.3)
ISO 1871, Agricultural food products - (standards.iteh.ai)
General directions
for the determination of nitrogen by the Kjeldahlmethod. 5.4 Compound catalyst4), consisting of, for example :
ISO 5378, Starches and derived products - DeterminationISO 3188:1978 - potassium sulphate : 97 g;
of nitrogen content by https://standards.iteh.ai/catalog/standards/sist/e0bfc8aa-f1b5-43ba-b584-
the Kjeldahl method - Spectro-
pho tome tric me thod, - copper( II) sulphate, anhydrous : 3 g.
372388bc295e/iso-3188-1978
5.5 Sulphuric acid, approximately 0,02 N or- 0,l N
3 DEFINITION Standard volumetric Solution,
nitrogen content : The value found using the procedure
specified. lt includes the nitrogen content of free amino 5.6 Colorimetric indicator, prepared by mixing 2 Parts
acids, of compounds producing amino acids on hydrolysis by volume of a cold saturated Solution of neutral methyl
and of ammonium compounds. lt does not include the red in 50 % (WV) ethanol with 1 part by volume of a
nitrogen of nitrate and nitrite radicals, the nitrogen 0,25 g/l Solution of methylene blue in 50 % (VW) ethanol.
attached directly to another nitrogen atom or the nitrogen Store the indicator in a brown glass bottle.
attached to an Oxygen atom.
6 APPARATUS
4 PRINCIPLE
Usual laboratory equipment, and in particular
Destruction of organic matter by sulphuric acid in the
presence of a compound catalyst*), alkalization of the 6.1 Kjeldahl flask, of suitable capacity, usually 500 to
reaction products, distillation of the Iiberated ammonia 800 ml, preferably with a ground glass joint, and provided
and collection in a boric acid Solution, followed by with a pear-shaped glass bulb fitting loosely in the top of
titration with a Standard volumetric sulphuric acid Solution. the neck of the flask.
1) For products whose presumed nitrogen content is less than 0,025 % (m/m), see ISO 5378.
2) See ISO 1871.
3) For anticipated low nitrogen contents, a less concentrated Solution may be needed in Order to obtain greater accuracy.
4) See ISO 1871, sub-clause 5.2,
1
ISO 31884978 (E)
6.2 Digestion stand, on which the Kjeldahl flask (6.1) Mix the contents of the flask by swirling the flask gently
tan be heated in an inclined Position in such a way that until the mixture is free from lumps and the test Portion
heat is applied only to that part of the flask wall which is completely wetted. In Order to avoid super-heating,
is below the liquid level at all stages. add a boiling aid (for example glass beads). Insert the
pear-shaped glass bulb (see 6.1) in the neck of the flask
6.3 Distillation or steam distillation apparatus, with a and place it in an inclined Position on the digestion stand
200 ml graduated dropping funnel and an efficient splash (6.2).
head, the latter connecting the Kjeldahl flask (6.1) to the Heat with care until the liquid in the flask boils gently.
condenser. Continue to heat for 1 h after the liquid becomes clear.
Any apparatus that satisfies the control tests given in In the case of digestion apparatus heated by gas, ensure
ISO 1871 is permitted. that the flame does not extend beyond the part of the
flask filled with liquid, in Order to avoid loss of nitrogen.
6.4 Burette, 25 ml with 0,05 ml graduations, or 10 ml
with 0,Ol ml graduations.
7.4 Distillation and titration
6.5 Mechanical grinder or mortar.
Allow the contents of the flask to cool and rinse the
pear-shaped glass bu!b and the inner neck of the flask
6.6 Sieve, with nominal mesh openings of 0,6 mm,
with a few millilitres of water, allowing the rinsings to
complying with the requirements of ISO 565.
run into the flask. Add, with care, between 50 and 200 ml
of water (according to the apparatus used), whilst swirling
6.7 Analytical balance. the contents of the flask. Connect the flask to the
distillation or steam distillation apparatus (6.3), previously
7 PROCEDURE freed from ammonia by steaming.
7.1 Preparation of the test Sample
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Adjust the Iower end of the condenser so that it just
touches the bottom of a 500 ml conical flask containing
Mix the Sample thoroughly
(standards.iteh.ai)
and rapidly by shaking or
a known volume (varying between 25 and 50 ml) of the
boric acid Solution (5.3) and 3 to 5 drops of the titration
stirring with a spatula in the Sample container? If the
indicator (5.6). Render the digestion liquid alkaline by
Sample Container is too small for this purpose, transferISO 3188:1978
slowly adding, through the graduated separating funnel
the entire Sample to another https://standards.iteh.ai/catalog/standards/sist/e0bfc8aa-f1b5-43ba-b584-
predried Container of a
(See 6.3) placed in the neck of the flask, between 150 and
suitable size to facilitate mixing. 372388bc295e/iso-3188-1978
200 ml of the sodium hydroxide Solution (5.2), ensuring
lt may be necessary to grind the Sample, in which case that the stem of the funnel does not become empty. Mix
it shall all pass through the sieve (6.6) without leaving weil, then turn on the condenser water and Start heating;
any residue. the ammonia then begins to be carried over.
The indicator contained in the flask turns immediately
7.2 Test Portion to its alkaline colour.
Weigh, to the nearest 0,001 g, up to IO g (mass m) of the During distillation, ensure that steam generation is kept
test Sample (7.1), according to the presumed nitrogen constant. Distillation is complete when 200 ml of liquid
content, and transfer to the predried Kjeldahl flask (6.1), have been collected in 20 to 30 min.
taking care that none of the product adheres to the inner
wall of the neck of the flask. Turn off the heat and lower the conical flask. Allow the
condenser to drip for a few minutes into the flask and
In the case of a viscous liquid or a product in Paste form, rinse the tip of the condenser with water, collecting the
the test Portion may be weighed in a small glass Container rinsings in the conical flask.
or on a sheet of aluminium, Paper or plastics which does
not yield nitrogen, or whose nitrogen content is known, The liquid contained in the flask should be green.
and which is left in the flask. In the case of a Container
which yields nitrogen, this should be taken into account Titrate the contents of the flask with either 0,02 N or 0,l N
sulphuric acid Solution (5.5), using a IO ml or 25 ml burette
in the blank test (7.6).
(6.4) as appropriate, until the colour of the contents
turns reddish violet.
7.3 Destruction of organic matter -.
Add 10 g of the compound catalyst (5.4) and, using a
suitable measuring cylinder, add- the appropriate. volume, 7.5 Number of determinations
in millilitres, of the concentrated sulphuric acid (5.1),
calculated by the formula 20 + 4 m, in such a way that Carry out two determinations on the same test
the acid rinses the inner wall of the neck of the flask. Sample (7.1).
1) In the case of glucose syrup, remove the surface layer (about 5 mm) before mixing.
2
ISO 3188-1978 (E)
7.6 Blank test V, is the volume, in millilitres, of the sulphuric acid
Solution used in the blank test (7.6);
Carry out a blank test on the reagents only. If the test por-
tion has been weighed in a Container which yields nitrogen V, is the volume, in millilitres, of the sulphuric acid
(see 7.2), carry out the blank test using an identical Container. Solution used in the determination (7.4);
m is the mass, in grams, of the test Portion (7.2).
7.7 Check tests
Express the result as the arithmetic mean of the two
Carry out the check tests specified in ISO 1871. determinations if the volumes of the sulphuric acid Solution
used do not differ by more than 0,l ml. Otherwise, repeat
8 EXPRESSION OF RESULTS the determinations on the same test Sample.
The nitrogen content is given, as a percentage by mass, by
the formula
100 9 TEST REPORT
0,01401 xTx(V, -V,) x-
m
The test report shall indicate the method used and the
1,401 T(V, - V,) results obtained. lt shall also mention all operating
= conditions that are not specified in this International
m
Standard, or are regarded as optional, as weil as any
where circumstances that may have influenced the results.
T is the normality sf the sulphuric acid Solution (5.5) The test report shall include all details required for
used in the two titrations; complete identification of the Sample.
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