PAGE 1 OF 4

APPLICATION VAPODEST
RAW PROTEIN IN GRAIN AND GRAIN PRODUCTS

1. Principle
The organic nitrogen of the sample is digested with concentrated sulphuric acid, potassium sulphate and a
catalyst. The nitrogen thus is broken down to ammonium sulfate. By adding caustic soda in excess, ammonia
is released by water steam distillation and trapped in a solution of boric acid. This solution is titrated with a
standard acid.
2. Source
2.1. Method
Based on
Cooperative Wheat Research Group, Standard-Methods for wheat, flour and bread, Determination of the
protein in wheat and wheat products, ICC-Standard No. 105
AOAC Official Method 979.09. Protein in Grain
2.2. Product Table see item 7.
th
Food Composition and Nutrition Tables; S. W. Souci, W. Fachmann, H. Kraut; 5 revised and completed
edition, medpharm Scientific Publishers, Stuttgart 1994, CRC PRESS, Tokyo
3. Chemicals
Quality: p.a.
Water: demineralised or distilled
3.1. Sulphuric acid 98 % min.
3.2. Catalyst tablets Kjeltabs CX, cat. no. 12-0328
3.3. Caustic soda 32 %
3.4. Boric acid solution 2 %
3.5. Indicator solution M5 (Merck) or similar
3.6. Standard solution: hydrochloric acid c = 0.1 mol/l, alternatively sulphuric acid c = 0.05 mol/l
4. Instruments
4.1. Bruised grain mill or star mixer
4.2. Analytical Balance (0.0001 g)
4.3. Kjeldahl digestion block KJELDATHERM, TURBOTHERM or flask heater for Kjeldahl flask with wide
neck opening
4.4. TURBOSOG scrubber, cat. no. 12-0057
4.5. VAPODEST distillation system
4.6. Burette, 50 ml nominal capacity, with a scale on 0.05 ml, or titration system
(not necessary using VAP 50s) or pH meter with combined electrode
5. Analysis
5.1. Sample Preparation
There should be a representative sample of at least 40 g. Coarse-grained samples have to be crushed so
that at least 90 % of the substance can pass through a test sieve of 1.0 mm mesh size.
The prepared samples are weighed into the digestion tube using weighing paper, chemicals are added and
sulphuric acid is used to wash down any sample residue remaining at the glass wall.
Chemicals
Sulphuric acid (3.1.)
Kjeltabs (3.2.)

20 ml
2

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APPLICATION VAPODEST
RAW PROTEIN IN GRAIN AND GRAIN PRODUCTS

5.2. Digestion with KJELDATHERM

When using a KB system with 250 ml digestion tubes, the following parameters are recommended:
Time min
30

Temperature C
400

30

400

Comments
The digestion tubes are put into the preheated block;
time until sample turns translucent

5.3. Digestion with TURBOTHERM

When using a TURBOTHERM system with 12 x 250 ml digestion tubes, the following parameters are
recommended:
Time min
20
40

Power %
100
70 to 80

Comments
Time until the digestion solution boils
After 20 to 30 minutes the digestion solution should have
turned translucent

5.4 . Digestion with Flask Heater

When using a flask heater with 500 or 750 ml flasks with wide neck opening, the following parameters are
recommended:
Time min
20
40

Power Level
3
1.5

Comments
Time until the digestion solution boils and white fume forms
After 20 to 30 minutes the digestion solution should have
turned translucent. Any sample residue remaining at the
glass walls is washed down into the flask by condensing
sulphuric acid.

5.5. Suction of the Digestion Gases

During the entire digestion period the scrubber should be working. About 1200 ml of an approx. 15 % caustic
soda solution is recommended for the washing bottle; this amount is sufficient to neutralize digestion gases of
about 60 digestions. The cooling down period after lifting the insert rack or after turning off the heating is
about 30 minutes. During this time the scrubber should continue working.
5.6. Distillation
After the digested sample has cooled down, water steam distillation is effected according to the following
program:
Program parameter
H2O Addition
NaOH Addition
Reaction Time
Distillation Time
Steam Power
H3BO3 Addition
Suction Sample
Suction Receiver
Titration
Calculation
1)
2)

VAP 10/s
1)
~90 ml
~80 ml
240 s
100 %
1)
70 - 80 ml
manual
manual
manual
manual

VAP 20s
1)
~90 ml
~80 ml
0s
240 s
100 %
1)
70 - 80 ml
manual
manual
manual
manual

VAP 30s
~90 ml
~80 ml
0s
240 s
100 %
1)
70 - 80 ml
30 s
manual
manual
manual

VAP 45s
~90 ml
~80 ml
0s
240 s
100 %
70 - 80 ml
30 s
30 s
2)
automatic
manual

VAP50s
~90 ml
~80 ml
0s
240 s
100 %
70 - 80 ml
30 s
30 s
automatic
automatic

All activities or additions that have to be done manually are written in italic print.
external titrator

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APPLICATION VAPODEST
RAW PROTEIN IN GRAIN AND GRAIN PRODUCTS

5.7. Titration (automatically with Vap 50s)

About 3-4 drops of the mixed indicator (3.5.) are added to the receiving solution which is then titrated with the
standard solution (3.6.) until the color changes from green to grey/violet.
If a pH meter or a titration system is used for the end-point determination, no mixed indicator is necessary.
5.8. Blank Value
For the determination of the blank value the analysis (digestion and distillation) is run without any sample
material but using the above listed chemicals. The amount of consumed standard solution is to be considered
when the calculation is done.
5.9. Quality Check
The reliability check of the method is done using a reference substance which protein content was
determined in a ring test. In addition to the protein content, the repeatability and the comparability have to be
indicated.
6. Interpretation of Results
6.1. Calculation
1.4007

(V - Vb)

%N=
E
+

c - H ion concentration of the standard acid solution: hydrochloric acid c = 0.1 mol/l.
alternatively sulphuric acid c = 0.05 mol/l
V - consumption of standard acid solution of sample ml
Vb - consumption of standard acid solution of blank sample ml
E - initial sample weight g
% Raw Protein = % N

Protein factor

Protein factor:
6.25
5.7 for wheat and wheat products
6.2. Repeatability
The maximum deviation in a double analysis or between repetitions must not exceed 0.2 % protein.

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APPLICATION VAPODEST
RAW PROTEIN IN GRAIN AND GRAIN PRODUCTS

7. Product Table
Sample Type
Amaranth, Seeds
Buckwheat, peeled
Buckwheat groats*
Buckwheat whole grain flour*
Barley, no peel, whole grain
Peeled barley
Barley groats
Spelt flour, whole grain flour
Spelt
Spelt flour
Oats, no peel, whole grain
Oat meal
Oat groats
Oat flour
Corn, whole grain
Corn Flakes
Corn meal
Quinoa
Rice, unpolished
Rice, polished
Rice, polished, boiled, drained
Rice flour
Rye, whole grain
Rye flour, Type 815
Rye flour, Type 997
Rye flour, Type 1150
Rye flour, Type 1370
Rye flour, Type 1800
Rye germs
Sorghum
Triticale
Wheat, whole grain
Seminola*
Wheat flour, Type 405*
Wheat flour, Type 550*
Wheat flour, Type 630*
Wheat flour , Type 812*
Wheat flour, Type 1050*
Wheat flour, Type 1700*
Wheat germs*
Wheat bran, bran*

Amount of Sample
g +/- 10 %
1.5000
1.5000
1.5000
1.2000
1.5000
1.5000
1.5000
1.2000
1.5000
1.2000
1.5000
1.5000
1.5000
1.2000
1.5000
1.2000
1.5000
1.2000
1.5000
1.5000
2.0000
1.2000
1.5000
1.2000
1.2000
1.2000
1.2000
1.2000
1.0000
1.5000
1.5000
1.5000
1.5000
1.2000
1.2000
1.2000
1.2000
1.2000
1.2000
1.0000
1.0000

Theoretical content
% Protein
15.58 15.93
8.12 10.03
4.56 9.31
10.21 10.85
9.70 11.30
7.80 12.00
6.70 10.50
14.36
11.59
8.90 11.90
11.90 13.20
12.00 14.40
11.40 16.30
14.20 15.91
7.72 10.60
6.90 8.10
6.72 10.00
13.11 14.88
7.56 7.88
7.04 7.88
2.10
6.70 7.50
8.08 11.10
6.94
6.90 7.94
6.66 10.80
7.14 11.70
8.91 11.80
39.30 44.70
7.30 18.90
13.13 14.35
10.02 12.98
8.43 10.03
9.40 9.95
9.12 10.31
10.03 11.49
10.95 12.23
10.21 11.85
10.40 12.13
26.14
12.68 16.69

Products with protein factor 5.7 are written in italics and marked with *.
Products with protein factor 6.25 are written in regular font.

A.1.1.5. Raw Protein in Grain and Grain Products.doc

10.01.2011 mkr/ak-rbr