Accred Qual Assur (2001) 6:317–320

© Springer-Verlag 2001 PRACTITIONER’S REPORT

Judit Budai Experience with wheat flour reference

Judit Fükõ
material

Received: 12 November 1999
Accepted: 16 August 2000
Paper based on a talk given at CERM-2:
2nd Regional European Conference on
Reference Materials, 9–10 September
1999, Prague, Czech Republic
J. Budai (✉) · J. Fükõ
National Office of Measures (OMH),
1124 Budapest, Németvölgyi út 37–39,
Hungary
e-mail: j.budai@omh.hu
Tel.: +36-1-4585800
Fax: +36-1-2143157
Abstract The Chemical Section of
the National Office of Measures,
Hungary (OMH) [1] embarked on
the preparation of a wheat sample
series as a Certified Reference Ma-
terial [2] (CRM) in 1992. The prac-
tical implementation of the wheat
sample series has shown that test-
ing laboratories are in great need
of easy-to-use flour reference sam-
ples to objectively and indepen-
dently qualify wheat. To fill this
need, we have developed a three-
term flour reference sample series.
The certification process was car-
ried out according to ISO recom-
mendations. Investigations of the
long-term stability [3] and the ap-
plication of wheat and flour CRMs
are continuing. Wheat is one of the
most widely grown crops in Hun-
gary and it is one of the major de-
terminative factors in the economy.
Its uniform and objective qualifica-
tion is of outstanding importance.
There are well-equipped laborato-
ries with sufficient experience to
assess wheat flour quality, howev-
er, proficiency testing has shown
that certified samples need to be
used to achieve exact and uniform
measuring results.
Keywords Certified reference
material · Reference material ·
Proficieny testing · Accredited
laboratories · Falling numbers ·
Wet gluten · Water-absorbing
capacity · Ash content · Protein
content · Moisture content

Introduction
Flour testing methods have a long history in Hungary.
Their development dates back to the end of the last
century when the Hungarian milling industry played a
leading role in the world. However, for wheat flour
quality tests are subjective: the results showed great
variability. Even the acceptance price of wheat is deter-
mined by the different parameters of wheat flour. To
make the tests accurate and uniform for the user, it is
expedient to use a simple, uniformly prepared flour
sample series. The need for such samples is justified by
the year to year increase in their demand.
The qualified parameters of certified wheat samples
are: protein, wet gluten, ash and microelement content.
For a longer shelf life, the packed wheat samples are
sterilized by irradiation at 5 kGy.
The qualified parameters of the flour reference sam-
ple are: the falling number, wet-gluten content, water-
absorbing capacity, the quality-value number and the
ash content. Our storage investigations have shown that
irradiation of wheat samples destroys their test quali-
ties. The enzymatic activity of the samples disappears
or diminishes to a ninimum level, so they cannot be
used, or only to a limited extent, for further testing.
Choosing the sample
Reference wheat flour material consist of three sam-
ples. They are produced – in the required quantity –
318

annually, from the new harvest from various growing
regions.
The most important conditions of sample selection
are that i the difference of wet-gluten content and the
value of water absorption must be at least three times
greater than the permissible error measured between
various laboratories.
Different extraction procedures were used for each
and every sample.
Sample preparation
Wheat collected for flour samples was cleaned by the
broken and the other grains and the dust were re-
moved. The homogenization was carried out in a Tur-
bula Shaker. The prepared and milled samples were
dispensed, in the necessary quantity into moisture-
proof triplex foil bags and vacuum-sealed.
Testing for HOMOGENEITY by protein content
measurement results
During sample preparation, sample homogeneity was
based on the protein determination because for homo-
geneity analysis none of the parameters of the flour
testing methods is suitable. Twelve containers to be dis-
tributed to the participants were selected at random
after dividing the bulk sample. The content of each of
the 12 selected containers was homogenized separately
and of them two test portions were taken from each
container. The sampling variance (ss) was estimated
from each of the results of the randomly analysed sec-
ond test portions. The analytical variance (sa) was esti-
mated from all the results by one-way analysis of var-
iance, without exclusion of outliers. There were no sig-
nificant differences between the samples on the basis of
the F-test. So the material was sufficiently homogenous
for the purpose of reference material (RM) preparation.
Certification campaign
The samples were sent to 15 accredited laboratories
which were suitably experienced in cereal testing. They
were to carry out the tests with great care according to
the current standard procedure. Several measurement
methods were specified:
– Wet-gluten determination by manual or mechanical
gluten washing
– Falling number determination by the Hagberg-Pert-
en method
– Determination of water-absorption capacity and
quality-value number by Valorigraf/Farinograf
– Determination of ash content
The organization of the interlaboratory test and the
evaluation of the results were based on ISO Guide 35
[3]. (Certification of reference materials – General and
statistical principles).
Cohran’s test was used for analyzing the deviation in
the laboratory’s replicate measurement results and for
discarding the outlier values. Grubb’s test helps us to
find the outlier results on inter- and intralaboratory ba-
sis, and to discard them from the evaluation. Table 1
and Fig. 1 show the results of protein content measure-

Table 1 Certification campaign. Measurement Results and Evaluation of Protein Content in Flour Sample

Labora- Results of replicate Aver- St dev Cochran’s Grubb’s test Grubb’s test
tory measurements % m−m age test C within laboratory between laboratories
number
1 2 3 4 5 % m−m % m−m Gmax Gmin Gmin Gmax

1. 14.11 14.15 14.17 14.23 14.27 14.19 0.064 0.048 1.32 1.19 0.15
2. 13.93 14.02 14.07 14.07 14.10 14.04 0.067 0.053 0.93 1.62 0.29
3. 14.14 14.16 14.18 14.19 14.24 14.18 0.038 0.017 1.54 1.11 0.13
4. 14.64 14.88 14.90 14.96 15.03 14.88 0.147 0.258 0.53 1.64 2.18
5. 13.86 13.94 13.95 13.98 13.99 13.94 0.051 0.031 0.90 1.64 0.56
6. 14.30 14.30 14.30 14.40 14.40 14.34 0.055 0.036 1.10 0.73 0.60
7. 14.01 14.01 14.10 14.10 14.18 14.08 0.072 0.061 1.39 0.98 0.16
8. 14.05 14.06 14.10 14.14 14.15 14.10 0.045 0.024 1.10 1.10 0.10
9. 13.05 13.19 13.25 13.32 13.33 13.23 0.115 0.156 0.89 1.55 2.65
10. 13.84 13.84 13.86 13.89 13.89 13.86 0.025 0.007 1.04 0.96 0.79
11. 14.10 14.20 14.20 14.20 14.30 14.20 0.071 0.059 1.41 1.41 0.19
12. 14.10 14.11 14.11 14.14 14.16 14.12 0.025 0.007 1.43 0.96 0.03
13. 14.16 14.24 14.34 14.25 0.090 0.096 1.03 0.96 0.32
14. 14.29 14.34 14.39 14.42 14.46 14.38 0.067 0.053 1.20 1.35 0.71
15. 14.12 14.24 14.24 14.25 14.37 14.24 0.088 0.093 1.42 1.40 0.32
From all results After exclusion the outliers
Mean: 14.14 Mean: 14.14
St dev: 0.342 St dev: 0.291
 319

Fig. 1 Results of moisture

content in the flour sample
from the certification cam-
paign

Table 2 Certified values of wheat flour reference materials (RMs) in 1999

Certified Sample 1 Sample 2 Sample 3

values
X X U xa X U xa X U xa

Protein content % (m/m) (f=5.7) 13.73 ±0.1 13.45 ±0.1 12.49 ±0.2
Falling numbers 352 ±8 297 ±4 219 ±5
Wet-gluten content % (m/m) 28.3 ±0.3 26.7 ±0.3 24.9 ±0.3
Water absorbing capacity % (m/m) 61.9 ±0.6 60.4 ±0.5 60.9 ±0.5
Quality-value number 67.0 ±2.1 57.0 ±1.7 46.6 ±2.2
Ash content % (m/m) dry matter 0.75 ±0.01 0.51 ±0.01 0.60 ±0.01
a Ux(k=2) = uncertainty of certified value

ment. The outlier result and the laboratory number are
marked by a shadowed background in the Table. Based
on the results the consensus value is certified and its
uncertainty is defined.
Table 2 contains the certified values of the 1999
RMs [3]. Reference samples – prepared with the meth-
od above – were kept at room temperature. During
their storage they were tested three times, i.e. every 6
months at 18 laboratories regarding their certified and
other parameters. The results show that the enzymatic
processes can’t be stabilized, so the applicability of the
parameters is limited to half a year and the useability to
1 year. These results prove that the packaging and stor-
age we applied were suitable for flour RMs. The deter-
mination of the reference values was again carried out
by the previously mentioned method.
ratories. The organization of test and the evaluation of
the data were based on the ISO Guide 43–1 (Proficien-
cy Testing by interlaboratory comparison-Part 1: De-
velopment and operation of proficiency testing sche-
mes) [4]. The laboratories were classified based on the
resulting “z-score” values (Fig. 2). The results of sev-
eral laboratories were significantly different from each
other and from the average of the measurement, they
exceed the allowed target value for standard deviation
(σ). The calculated z-score of laboratory measurement
clearly shows the direction and the extent of the devia-
tion of laboratory result from the consensus value (Ta-
ble 3). If the z-score value is >±3 of a measurement,
the laboratory has to carry out improvements using our
samples.

Using the flour samples Conclusion

Because the samples for the certification campaign
were sufficiently homogenous we also used them for
proficiency testing.
In the test more then 20 laboratories took part, all of
them were independent of the above-mentioned labo-
The test described above, as well as the need for labo-
ratories, producers of instruments, and services for our
samples prove that our goal has been achieved and we
have succeeded in developing a practical, usable RM
series.
320

Fig. 2 z-Score values of wet-

gluten content determination
in the wheat flour samples

Table 3 Proficiency test. Measurement results and z-score for moisture content in flour sample

Labora- Results of replicate Average After exclusion

tory measurements the outliers
number
% m−m % m−m Average and z-score value
1 2 3 4 5 % m−m z

16. 13.06 13 12.95 13.01 12.94 12.99 12.99 0.48

17. 12.9 12.85 12.85 12.88 12.9 12.88 12.88 −0.29
18. 12.98 13.01 12.92 12.94 12.95 12.96 12.96 0.27
19. 12.52 12.44 12.5 12.6 12.55 12.52 12.52 −2.65
20. 12.76 12.9 12.98 12.85 12.94 12.89 12.89 −0.23
21. 12.68 12.71 12.72 12.6 12.61 12.66 12.66 −1.71
22. 12.18 12.1 12.19 12.16 −5.09
23. 13.1 13.2 13.1 13.1 13 13.10 13.10 1.20
24. 13.05 13 13.04 12.97 12.95 13.00 13.00 0.55
25. 13.24 13.24 13.23 13.23 13.24 13.24 2.10
26. 12.87 12.95 12.95 12.89 12.92 12.92 12.92 −0.03
27. 13.05 13.16 13.2 13.23 13.11 13.15 13.15 1.53
28. 13.06 13 13.05 13.03 13.04 13.04 13.04 0.77
From all results After exclusion the outliers
Mean: 12.88 Mean: 12.94
St dev: 0.29 St dev: 0.20
σ*: 0.15 σ*: 0.15
a σ: target value for standard deviation is the measurement repeatability given by the Hungarian Standards

The proposed applications of the RM series are: The results of our examinations prove that our
– Assessment of different measurements wheat flour samples are suitable for checking and de-
– Adjustment of the calibrated measuring instrument tection of mistakes and for unifying measurement re-
– Validation of different measurements. sults.

References
1. Hungarian Law on Legal Metrology
No XLV/1991
2. OMH (1996) Measures and Servies of
the National Office of Measures in
1996. OMH, Budapest, Hungary
3. ISO Guide 35 (1989) Certification of
reference materials – General and sta-
tistical principles. ISO, Geneva Swit-
zerland
4. ISO Guide 43–1 (1997) Proficiency
testing by interlaboratory comparison
– Part 1: Development and operation
of proficiency testing schemes. ISO,
Geneva, Switzerland