J ournal of Food Engineering 3 (1984) S l-73

The Water-Vapour Sorption Isotherms of

Microcrystalline Cellulose (MCC) and of Purified Potato
Starch. Results of a Collaborative Study
W. Wolf, W.E.L. Spiess, G. Jung
Federal Research Centre for Nutrition, Engesserstrasse 20, 7500 Karlsruhe 1,
West Germany
H. Weisser
University of Karlsruhe, Institute of Food Process Technology, Kaiserstrasse 12.
D-75 Karlsruhe 1, West Germany
H. Bizot
INRA, Laboratory of Food Biophysics, Chemin de la Geraudiere,
F-44 072 Nantes Cedex, France
and
R. B. Duckworth
University of Strathclyde, Department of Food Science and Nutrition,
James P. Todd Building, 131 Albion Street. Glasgow Gl 1 SD, United Kingdom
ABSTRACT
I n a comprehensive collaborative study within the frame-work of COST
(European Cooperation in the Field of Scientific and Technical Research)
_
the COST 90 project - the mean adsorption isotherms of MCC and a
potato starch and the precision data, viz. the repeatability/reproducibility
of the total sorption measurement procedures, were determined.
Recommendations on the practical determination of sorption isotherms
of foods based on this work are made.
INTRODUCTION
Sorption isotherms of well-defined food materials published by
various authors differ, often considerably, from each other.
J ournal of Food Engineering 0160-8774/84/$03.00 - 0 Elsevier Applied Science
Publishers Ltd, England, 1984. Printed in Great Britain
52 W. Wolf et al.
Differences in the sorption properties of such products - like wheat
of a special variety, meat taken from a selected muscle, or milk with a
given fat content - can be attributable to biological variation of the
substrate, differences in equipment design and different handling
procedures.
To study the effect of the last two groups of influences on the
sorption isotherm of a particular material in greater detail, a collabora-
tive research programme was initiated within the COST 90 Project on
physical properties of foodstuffs. The objective was a standard equip-
ment for sorption isotherm measurements on selected materials for
reference purposes.
The programme, in which 3 2 laboratories participated, was carried
out in three stages.
In the first stage, a substance with defined and stable sorption
properties was selected which could be used for the analysis of
methodological problems and which could serve as a reference material
for calibration purposes. In the second stage, sorption equipment was
designed and constructed which allowed well-controlled undisturbed
heat and mass transfer between sorbate source and substrate. In the
third stage, the precision data of the system : reference material-recom-
mended equipment were determined together with the most probable
sorption isotherm of the reference material.
The criteria for the selection of the reference material were:
(1) Stability in sorption behaviour over several adsorption and de-
sorption cycles and to exposure to extreme ambient temperatures
when shipped or stored.
(2) Absence of hysteresis between adsorption and desorption.
(3) Relatively rapid rate of equilibration.
(4) Its sorption isotherm to be sigmoid in shape as are those of many
food products.
(5) Homogeneity of the material in its physical structure and distri-
bution of components.
(6) The determination of its water content to be possible in an un-
equivocal way.
(7) Easy availability and easy handling properties.
Among a variety of homogeneous purified substances, such as starches
and silicic acid, microcrystalline cellulose (Avicel, PH 101 from FMC)
was chosen as the reference material because it fulfilled most of the
above criteria, except that it exhibited hysteresis between adsorption
Sorption isotherms of MCC and purified potato starch 53
and desorption isotherms (Wolf and Spiess, 1980; Wolf et al., 1980).
MCC had also been used in a previous study by Vos and Labuza (1974)
as a reference material when determining the a, of products in the high
water-activity range (0.85-0.98).
When adsorption isotherm measurements are made, three critical
stages are involved:
(1) Desorption in the course of sample preparation to attain zero
water content.
(2) Equilibration at the different water-activity levels.
(3) Transfer of the samples from sorbostat to weighing equipment.
In order to avoid problems connected with these critical stages, it is
necessary to design the equipment and the mode of operation
accordingly.
The equipment components which could influence the sorption
kinetics must be designed in such a way that the resistances to heat and
mass transfer within the equipment are always less than they are within
the product. The product itself should be disposed in such a way that
sorption is complete within a reasonable period of time. Any handling
procedures affecting the sorption process must be carried out in such a
way that the intended processes are achieved; this applies, for example,
to the desorption processes as well as to the adsorption processes.
During manual operations the uptake or loss of water must not be
significant otherwise the time required for individual operations such as
opening the equipment, transferring weighing bottles from the sorption
container to the balance, etc. must be standardized.
The equipment finally agreed upon consisted of a rather simple
arrangement: I-litre glass jars with vapour-tight lids as the sorbostat
in the bottom of which the sorbate source was arranged. The substrate
was placed in small weighing bottles standing on trivets directly above
the sorbate source and the jars were submerged in a thermostatically
controlled waterbath almost to the lid. To protect the substrate against
radiative heat exchange with the environment, waterbath and jar were
covered with insulation (Wolf et al. in preparation).
IMPLEMENTATION OF THE COLLABORATIVE STUDY
Participating laboratories
Within the framework of the project, 32 laboratories from 11 countries
took part. Six of the laboratories were industrial research institutions
54 W. Wolf et al.
in which sorption measurements were carried out as routine work
together with other quality-control measures. Of the 26 laboratories
associated with research institutions or universities, 13 groups had
been active in research on water activity for more than 10 years and
had considerable experience in the subject; the remaining teams had
carried through sorption isotherm measurements only occasionally in
the past in the course of other research work such as investigations on
product stability, etc. Their experience in the field of sorption isotherm
measurements could, at the beginning of the study, be regarded as
rather limited. A list of the laboratories which contributed results is
given in Appendix 1.
Materials and methods
The test substrate, Avicel PH 10 1 MCC, was received from the producer
as analytical grade in a loo-kg amount. This quantity was divided into
portions each of 100 g by means of a sample divider. These portions
were packed in polyethylene-coated aluminium pouches, sealed
hermetically and mailed to the participating laboratories together with
recommendations for the design of the standard equipment, the
preparation of the sorbate source and the handling procedures.
The starch samples (purified potato starch) were prepared by one of
the participating groups and distributed in the same way as the MCC.
It was made obligatory that the sorption isotherms were measured on
the standardized equipment. Saturated salt solutions had to be used as
sorbate source. The water activities of the solutions selected were
fairly evenly distributed over the activity range 0.1-0.9. The exact
values are given in Table 1. At each water activity, five replicates were
to be measured in one sorbostat. The main features of the method used
are given in Table 2.
RESULTS
General
Of the 32 laboratories provided with material and instructions, 24
returned pertinent results. Together with the results a completed
questionnaire was received in which the methods and the equipment
used were explained in detail.
Sorption isotherms of MCCand purified potato starch 55
TABLE 1
Salt and Water Quantities to Prepare the Saturated Salt Solutions Required for One
Sorption Container
Salt Relative
humidity
(RHja
(%)
Standard
deviation
Quantity
Salt (gl
Water (ml)
LiCl 11.15
CHsCOOK 22.60
MgClz
32.73
K2C03
43.80
WWM2
52.86
NaBr 57.70
SrC12 70.83
NaCl 75.32
KC1 84.32
BaC12 90.26
0.37
0.5
0.16
0.33
0.23
0.4
0.04
0.12
0.38
150 85
200 65
200 35
200 90
200 30
200 80
200 50
200 60
200 80
250 70
aRH-values and standard deviations (s) of the salts according to Greenspan (I 977).
TABLE 2
Principal Features of the Method used for the Measurement of Sorption Isotherms
of MCC and Starch
Sorbostat 1-litre glass jar
Sorbate source Saturated salt solutions (aqueous)
Measuring method Gravimetric, discontinuous
Mode of sorption Integral
Sample mass 400 mg
Sorption temperature 25 k O.lC
Precision of the balance 0~0001 g
Sample holder Weighing bottle (25 x 25 mm) with ground-in stopper
From this information it became clear that two laboratories had
carried out their experiments in a way which was not in conformity
with the prescribed procedure and so their results were excluded from
subsequent evaluations.
56 W. Wolf et al.
Other deviations from the recommendations reported by the individual
laboratories were as follows:
(1) Instead of a water bath a thermostatically controlled chamber
(incubator) or an air-conditioned room was used for temperature
control. Since the temperature fluctuations were, with one
exception, less than +0.3C, the data from these experiments
were accepted because the influence of temperature on the
sorption isotherms of MCC and of starch is very small (Wolf
et al., 1980).
(2) Instead of the recommended glass equipment (jars and weighing
bottles) equipment showing minor differences in the important
dimensions was used. Since the pathways for the mass transfer
from the sorbate source to the substrate are not affected by these
differences, the data from these experiments were also accepted.
(3) Instead of the recommended equilibration times, longer and, in
one case, slightly shorter, equilibration times were used during
pre-drying and adsorption. Because MCC and starch had proved
very stable in their sorption properties against moderate heat
treatments (Wolf et al., 1980) no change in the product and the
results was expected as a result of the longer exposure times in
either stage of the determination.
As the shorter equilibration time used in the one case was
within the tolerance interval encountered when the study was
planned those results were also accepted.
(4) Other minor differences reported by the laboratories were also
considered as not significant.
Statistical treatment and evaluation of precision data for the results
obtained for MCC
The sorption data obtained were, after a first screening test, evaluated
for mean value, standard deviation, repeatability and reproducibility
according to standard test methods based on BS 5497 :Part 1: 1979
issued by the British Standards Institute (1979), DIN-IS0 5735 issued
by the Deutsches Institut fur Normung (1978), and a report on the
planning and statistical evaluation of ring tests issued by the Bundes-
gesundheitsamt, Berlin (Goetsch et al., 1978).
In detail, the procedure comprised the following steps (see Fig. 1. A
Sorption isotherms of MCC and purified potato starch
A
ELlMlNATlON
OF DUlLiLR
GRUBBS-TEST FOR
OlJlLlERS FOR
EAtH LAB FOR
LACIl SALT
ELIPIINATE
THE OUTLIER
CAI CULATE f OR
EACH SALT FOR EAM LAB
MEAN
STANDARD DLVIATION
J
B I
1
EXAMINATION
I
OF SYSTEMATIC
VAR. ANAL, DlXuN TEST
ERRORS
I I
ELIMINATE THE
VALUES OF THIS
LAB WHICH CAUSE
SIGNIFICANT DIFFE
RENCES BETWEEN
D
MEAN VALUES
REPEATABILITY
REPRODUCIBILITY
NO
ci5 _
REPEAT THE CALCULATE
COLLAB. STUDY REPEATABILITY
REPRODUCIBILITY
Fig. 1. Flow diagram of the principal steps in the statistical analysis.
58 W. Wolf et al.
brief description of the statistical tests together with an example of the
results analysed is given in Appendix 3 ):
(A) Identification and elimination of outliers within each individual
experimental data set (Grubbs test).
(B) Comparison of variances (Cochran test j.
(CT) Identification of systematic deviations (Dixon test).
(D) Calculation of mean values, standard deviations, repeatability
and reproducibility on the remaining data.
The two terms which describe variability, viz. repeatability and repro-
ducibility, in most practical cases of routine experiments sufficiently
characterize the precision.
Repeatability refers to tests performed at short intervals in one
laboratory by one operator using the same equipment, while reproduci-
bility refers to tests performed at different laboratories which implies
different operators and different (if not dissimilar) equipment.
The following definitions are used:
( 1) Repeatability, r. The value below which the absolute difference
between two single test results obtained within a short interval of
time by the same method on an identical test material under the
same conditions (same operator, same apparatus, same labora-
tory) may be expected to lie within a specified probability: in the
absence of other indications the probability is 95%;
(3) Reproducibility, R. The value below which the absolute differ-
ence between two single test results obtained with the same
method on an identical test material under different conditions
(different operators, different apparatus, different laboratories
and/or at different times) may be expected to lie within a
specified probability; in the absence of other indications the
probability is 95%.
An analysis of the measured sorption data for the various selected
water activities showed that they are normally distributed, at least on
a 0.1% significance level with the exception of the results for barium
chloride (BaCl,) (a, = 0.9). However, since there is no reason why the
measured values for BaCl, should not be normally distributed, the data
at this water-activity level were still retained for further statistical
treatment.
Sorption isotherms of MCC and purified potato starch 59
Regarding step A, in which the original data were considered, it was
found that outliers could be identified only in a few cases; this con-
firmed at the outset that the experiments had been carried out by all
laboratories with sufficient care; in statistical terminology, the data sets
after the first outlier tests contained, in the majority of all individual
data groups (data for one salt from one laboratory being equal to a
mathematical cell), a similar number of test results. Furthermore it
was found that in no group was the variance of the data zero. Therefore
both the Bartlett and Cochran tests could be applied in step B to the
evaluation of the homogeneity of the variances. The results of either
test are comparable since the tests yielded different results only in a
few cases.
The compilation of the test results shows that variances causing
inhomogeneities are accumulated in certain laboratories. These could be
regarded as laboratories with limited experience, whereas in the labora-
tories with extensive experience practically all variances were homo-
geneous. In accordance with the recommendations of the statistical test
procedures used it was decided that laboratories in which more than
50% of the cell variances were found to cause inhomogeneity were
excluded from further evaluation. From the remaining laboratories
those individual data groups which caused inhomogeneity of the
variances were carefully examined on the basis of the information
obtained from the individual laboratories on the experimental work
and also rejected in all cases.
The remaining data sets, however, qualified for the Dixon test, step
C; according to the test procedure, this test was applied to identify
mean values which differed significantly from the comparable data
base. Only four individual data groups (cells) had to be excluded after
this test procedure.
From the remaining data for each salt (on the average 80 measure-
ments) the mean value, the standard deviation, the repeatability
standard deviation and the reproducibility standard deviation were
calculated (step D); from the latter information, the repeatability and
the reproducibility values were derived (see Table 3).
The standard deviations of the equilibrium water contents vary from
SmiIl
= 0.19 to s,,, = O-3 1 g H,O/lOO g DM (dry matter). Since there is
no clear correlation of standard deviation and other precision data with
water activity, the mean standard deviation and the means of the pre-
60 W. Wolj.et al.
TABLE 3
Adsorption Isotherm Data for MCC at 25C; Repeatability and Reproducibility
Values of a Precision Study with 24 Laboratories (Mean Standard Deviation 0.25;
Mean Repeatability 0.17; Mean Reproducibility 0.72; all in g HzO/l 00 g DM)
0,
Mean water
content (X)
Standard Repeatability Reproducibility
deviation (s)
(r) (R)
0.1 1 2.02 0. 25 0.13 0.72
0.23 3.19 0.21 0.11 0.63
0.33 4.06 0.75 0.11 0.73
0.44 5.04 0.19 0.14 0.54
0.53 5.82 0.27 0.23 0.79
0.58 6.48 0.21 0.07 0.61
0.7 1 8.21 0.20 0.22 0.57
0.75 8.83 0.31 0.21 0.89
0.84 10.95 0.29 0.19 0.85
0.90 12.96 0.30 0.26 0.88
cision data can be calculated; the mean standard deviation within the
range covered by the experiments is s, = 0.25 g H,O/lOO g DM, the
mean repeatability is r, = 0.17 g H20/ 100 g DM, and the mean repro-
ducibility is R, = 0.72g H,O/lOOg DM.
From sorption measurements carried out at higher temperatures by
the same method as recommended, similar precision data were obtained
(Wolf et al., 1980). It can therefore be concluded that the values of
mean repeatability r and mean reproducibility R derived from the
results of the collaborative study are also valid for experiments with
MCC in the standardized equipment in the temperature range 1.535C.
The calculated mean values of the equilibrium water contents for the
10 water activities were used to construct the most probable sorption
isotherm for MCC as the mean sorption isotherm.
For the computer plot of the sorption data the G.A.B. Model
(Guggenheim-Anderson-De Boer according to Van den Berg (1981))
was used to interpolate sorption isotherms:
X Cka,
-=
xln
(1 -ka,)(l -_a, +Cka,)
Sorption isotherms of MCCand purified potato starch
61
where
a,
= Water activity,
C = Guggenheim constant = C exp [(H, - H,)/RTl,
HI = Total heat of sorption of the first layer in primary sites,
H, = Heat of condensation of pure water vapour,
H, =Total heat of sorption of the multilayer which differs from
the heat of condensation of pure water,
k = Factor correcting properties of the multilayer molecules
relative to the bulk liquid (k = k' ( H, - HJ RT) ,
X =Water content on dry basis,
f
c z 0.4 0
/
I
.t 5 0.8 1.0
Water activity a,
Fig. 2. Adsorption isotherm for MCC at 25C; result of a collaborative study with
24 laboratories. (G.A.B. constants: X, = 4.064; C = 8.776; k = 0.772.)
62 I V. Wolf et al.
X, = Water content on dry basis corresponding to saturation of all
primary adsorption sites by one water molecule (formerly
called monolayer in BET theory).
As the most prominent result of the entire study, Fig. 2 shows the mean
MCC isotherm with standard deviations at the measuring points. Since
each of these points represents 75-85 individual values (depending on
the salt in question) the confidence intervals of the mean values (at the
95% probability level) have a width of 0.11 g H,O/lOO g DM (for the
calculated mean standard deviations).
Sorption and precision data for potato starch
The results of the sorption isotherm measurements on potato starch
were evaluated in the same way as described above for MCC.
The results are presented in Table 4 and Fig. 3.
In contrast to the precision data for MCC in the case of starch r and
R are related to the water activity of the sample. The repeatability r
and the reproducibility R of the water activity are represented by the
TABLE 4
Adsorption Isotherm Data for Potato Starch at 25C; Repeatability and Reproduci-
bility Values (in g H,O/lOO g DM) of a Precision Study with 23 Laboratories
a,
Mean water Standard Repeatability Reproducibility
content(X) deviation (s)
(r) (RI
0.11 4.69 O-38 0.13 1.12
0.23 7.54 o-45 0.32 1.31
0.33 9.74 0.49 o-19 1.42
0.44 12.10 0.43 o-14 l-25
0.53 13.92 0.82 0.25 2.39
0.58 15.34 0.52 0.40 1.50
0.71 19.28 0.55 0.30 1.59
0.75 20.93 0.66 0.36 1.91
0.84 24.95 0.97 0.42 2.80
0.90 28.83 I .64 0.60 4.76
Sorption isotherms of MCC and purified potato starch 63
Water activity a,
Fig. 3. Adsorption isotherm of potato starch at 25C; result of a collaborative
study with 23 laboratories. (G.A.B. constants: X,,, = 10.312; C= 7.976; k = O-734.)
following linear regression equations which are valid for the range
0.1-0.9:
I- = 0.424~~~ + 0.08
(2)
R = 3.035a, + 0.36
(3)
In general at lower water activities measurements were carried out with
greater precision than at higher activities. The reason for this is not
completely clear; it could, however, be that at higher activities the time
needed to incorporate water molecules into the starch varies and is in
some cases extremely long so that equilibrium is not reached even after
7 days.
64 W. Wolf et al.
CONCLUSIONS AND RECOMMENDATIONS
It is evident that the determination of sorption isotherms can be rather
difficult and cumbersome; if, however, sufficient care is taken they may
be achieved with precision. The reference and precision data obtained
may help to improve inadequate methods.
For practical applications, the method used in this study. namely
to evaluate first the sorption behaviour of a relatively simple uniform
product before more complicated systems are introduced, seems to be
a useful approach to sorption measurements.
When sorption measurements are approached in such a way, errors
caused by insufficient treatment of the substrate can be detected and
distinguished from errors caused by equipment deficiencies more easily
than when complicated substrates are used from the outset.
For practical sorption measurements the following procedure is
recommended.
( 1 ) Measure the adsorption isotherm of MCC (Avicel) according to
the standardized method (five replicates).
(3) Compare the results obtained with the mean value and precision
data of the collaborative study.
(3) If good agreement is obtained, proceed with the material to be
investigated. (a) Evaluate the necessary desorption (pre-drying)
conditions for achieving zero water content. (b) Evaluate the
necessary exposure time to reach equilibrium conditions.
(4) if the agreement is poor. search for and eliminate errors and
repeat the measurements on MCC until good agreement is reached.
The question as to whether the agreement obtained is good or poor has
to be decided with the aid of the precision data (reference value,
repeatability. reproducibility ):
(1) If II determinations performed by one laboratory under repeat-
ability conditions produce a mean value R that is to be compared
to a reference value X,, then good agreement, at the 95% prob-
ability level, is obtained if the difference lx--X0 I is equal to or
smaller than a so-called critical difference D,,, the value of which
is defined by the term:
(4)
(2)
Sorption isotherms of MCCand purified potato starch
For the precision data obtained in the collaborative study:
r=Ol7gH,O/lOOgDM
R = 0.72 g HzO/ 100 g DM
65
n = five replicates in the laboratory under scrutiny
the critical difference has the value
D,, = 0.498
In a random case, for example; with X0= 5.6 1 g H,O/lOO g DM
(equilibrium water content of MCC for a water activity a, = 0.5,
according to the sorption isotherm developed in the study) mean
values of X which are within the range 5.1 l-6.1 1 are then in
good agreement with the recommended sorption isotherm. Mean
values falling outside the range are in poor agreement and should
be considered as questionable. In most cases in which the differ-
ence I X-X,, I is larger than the critical difference Dcr, systematic
errors in the experimental procedure are very likely.
Where the mean value X of five measurements differs from the
reference value X0 by not more than the critical difference, the
standard deviation s obtained has to be compared to the repeat-
ability standard deviation s, as established in the collaborative
study for the experimental procedure in question, The repeat-
ability standard deviation is defined as:
r
s =283
r (5)
where the factor 2.83 corresponds to the 95% probability level.
(If another probability level is sought, this factor becomes 2.32
at the 90% and 3.65 at the 99% probability levels.)
If s is equal to or smaller than s,, the experimental procedure
tested can be considered as good; if s is larger than s,, the
variability in the results is too high; in this case a more careful
repetition of the measurements is necessary.
REFERENCES
British Standards Institute. (1979). Precision of test methods. Part 1. Guide for the
determination of repeatability and reproducibility for a standard test method.
BS 5497 : Part 1.
66 W. Wolf et al.
Deutsches lnstitut fiir Normung. (1978). Bestimmung von Wiederhoibarkeit und
Vergleichbarkeit. Entwurf DIN IS0 5725.
Goetsch, P. H., Krijnert, W., Olschimke, O., Otto, U. and VierkGtter, S. (1978).
Planung und statistische Auswertung von Ringversuchen. Teil des Berichtes des
Max von Pettenkofer-Institutes des Bundesgesundheitsamtes l/78, Berlin,
Dietrich Reimer Verlag, Berlin.
Greenspan, L. (1977). Humidity fixed points of binary saturated aqueous solutions.
J. Research of the National Bureau of Standards. A. Physics and chemistry,
81A(l), 89.
Van den Berg, C. (1981). Vapour sorption equilibria and other water-starch inter-
actions; a physico-chemical approach. PhD Thesis, Agricultural University
Wageningen .
Vos, P. T. and Labuza, T. P. (1974). Technique for measurement of water activity
in the high a, range. J. Agric. Fd. Chem., 22 (2), 326.
Wolf, W. and Spiess, W. E. L. (1980). Use of reference materials for the measure-
ment of water-vapor sorption isotherms of foodstuffs. fioceedings of the I nter-
national Symposium Production and use of reference materials. Bundesanstalt
fiir Materialpriifung, Berlin, p. 263.
Wolf, W., Spiess, W. E. L., Weisser, H., Gril, S. and Bizot, H. (1980). Mikrokristalline
Zellulose als Referenzmaterial zum Bestimmen des Wasserdampf-Sorptions-
verhaltens von Lebensmitteln. ZFL, 31 (3), 148.
Wolf, W., Spiess, W. E. L., Weisser, H., Bizot, H., Duckworth, R. B. and Jung, G.
(to be published). Standardization of water sorption isotherms: description of
equipment and measuring procedure.
APPENDIX 1: PARTICIPANTS AND THEIR ORGANIZATIONS
Bauder, U.
Bizot, H.
Casey, J. C.
Casolari, A.
Delmer, M.
Duckworth, R. B.
Eichner, K.
Evans, E. W.
Gil, S.
Knorr, Nihrmittel GmbH, Thayngen.
INRA, Nantes.
Meat Research Institute, Langford, Bristol.
Statione Sperimentale per 1Industria delle Conserve
Alimentari, Parma.
CTU, Nogent sur Marne.
Department of Food Science and Nutrition, Uni-
versity of Strathclyde, Glasgow.
Institut ftir Lebensmitteltechnologie und Ver-
packung, Fraunhofer-Gesellschaft, Munich.
National Institute for Research in Dairying, Shin-
field, Reading.
Organische Chemische Institut, Universittit Bern,
Bern.
Sorption isotherms of MCC and purified potato starch 61
Hallstrom, B.
Higgs, D.
Jason, A. C.
Kleeberg, H.
Kluge, G.
Linko, P.
Lintas, C.
McKenna, B.
Multon, J. L.
Pixton, S. W.
Poulsen, K. P.
Rtiegg, M.
Schutz, M.
Simatos, D.
Tanner, S. F.
Tobback, P.
Weisser, H.
Wolf, w.
Division of Food Engineering, University of Lund,
Alnarp.
Brooke Bond Liebig, Croydon.
Torry Research Station, Aberdeen.
Institut fur Physikalische Chemie, Universitlt
Marburg, Marburg.
Pfanni Werke GmbH, Munich.
Department of Chemistry, University of Technology,
Helsinki.
Institute Nazionale della Nutrizione, Rome.
Department of Agricultural Engineering, University
College, Dublin.
INRA, Nantes.
Ministry of Agriculture, Fisheries and Food, Slough.
Food Technology Laboratory, Danmarks Tekniske
Hojskole, Lyngby.
Eidgen. Forschungsanstalt fur Milchwirtschaft,
Liebefeld.
Ecole National Superieur des Industrie Textiles,
Mulhouse.
Laboratoire de Biologie Physico-Chimique, Dijon.
Food Research Institute, Norwich.
Laboratory of Food Preservation, Catholic Univer-
sity of Leuven, Leuven.
Institut fur Lebensmittelverfahrenstechnik, Univer-
sitat Karlsruhe, Karlsruhe.
Bundesforschungsanstalt fur Ernahrung, Karlsruhe.
APPENDIX 2
In the following, the selected method of statistical evaluation will be
explained by means of an example of the results obtained (see Table
A. 1). In Table A. 1 each line indicates first the laboratory code number,
followed by the five measured values (water content/dry matter)
obtained by the individual laboratories, calculated means and the
standard deviations calculated from these five measurements.
Considering now step A in Fig. 1 in which the original data were
considered, first an outlier test according to Grubbs was made for the
values submitted by each individual laboratory.
68 W. Wolf et al.
TABLE A. 1
Adsorption Data for MCC, Salt 5 (Mg(NO&), a, = 0.529. (All Values in g H20/
100 g DM.)
Labora- XI
tory
x2
1
7
;
6
7
11
13
13
14
15
16
17
19
20
7,
_I
25
26
28
30
31
32
6.32 6.18
6.67 6.65
5.14 5.18
6.04 60.5
5.88 5.98
5.96 6.00
6.04 6.00
5.61 5.63
5.83 5.81
5.97 5.96
6.03 5.94
5.70 5.55
5.94 5.91
6.3 I 6.42
5.93 5.85
5.65 5.59
4.81a 5.81
5.79 6.15
5.38 5.55
5.65 5.71
6.23 6.28
x3 x4
6.36 6.19
6.66 6.67
5.17 5.16
6-08 6.08
5.97 5.95
6.01 6.84
5.89 6.46
5.88 5.57
5.81 5.79
6.05 5.83
6.14 628
5.91 5.80
5.90 5.83
6.44 5.94
5.66 5.94
5.69 5.65
5.84 5.69
5.88 5.78
5.47 5.58
5.64 5.66
6.24 5.96
6.87
6,79a
5.13
6.08
5.90
5.99
5.94
5.75
5.82
6.08
6.04
561
5.85
5.99
5.77
5.70
5.73
5.97
d
_
5,73
5-91
6.224 0.117
6.662 b 0.010
5.1 56b 0.02 1
6.066 0.019
5.936 0.044
6.000 0.029
6.066 0.228
5.672 0.098
5.812 0.015
6.018 0.052
6.086 0.130
5.714 0.145
5.886 0.045
6.220 o.239c
5.830 0.117
5.656 0.043
5.767 0.069
5.888 0.157
5.475 0.07 1
5.678 0.040
6.124 o.174c
x5 Mean Standard
deviation
fs)
a Outlier within individual laboratory. eliminated (Grubbs test, 9970).
b Mean value outlier, eliminated (Dixon test, 95%).
Standard deviation causing inhomogeneity of the variances, eliminated (Cochran
test. 95%).
d - No measured value.
If the nulnber of participating laboratories is m then the laboratories
are characterized by the index i:
i = I . 7.. . . m (A.11
Sorption isotherms of MCCand purified potato starch
69
Within each individual laboratory, the measured values are characterized
by the index k; the number of measured values in laboratory i is n(i):
k = 1, 2,. . . , n(i) (A.2)
Then, the measured value number k in laboratory i is denoted by Xik,
the total number of all measurements in all laboratories is:
N = f n(i)
i= 1
The arithmetic mean in laboratory i, denoted xi, is:
xi = 1 y -&
n (9 k=l
and the standard deviation in laboratory i, denoted as si, is
TABLE A.2
Critical Values of the Grubbs Test
According to Goetsch et al. (1978)
n fil
Significance level
5% 1%
3 1.153 1.155
4 1.463 1.492
5 1.672 1.749
6 1.822 1.944
7 1.938 2.097
8 2.032 2.22 1
9 2.110 2.323
10 2.176 2.410
11 2.234 2.485
12 2.285 2.550
(A.3)
(A.4)
(A.5)
70 W. Wol f et al .
According to the Grubbs test, a measured value is identified as an out-
lier if its absolute deviation from the mean value 1 Xi, -Xi I divided by
the standard deviation si is larger than a corresponding value given in
Table A.?.
Any measured value with a superscript a in Table A.1 has been
identified, by means of the outlier test, as an outlier and is not included
in further calculations. The remaining values of the individual labora-
tories were then used to calculate the corresponding mean values and
standard deviations.
In step B of Fig. 1 the homogeneity of the variances was then
checked by means of the Cochran test. According to the Cochran test
the variances sf for all laboratories (eqn (AS)), and hence the largest
variance Sk,, , have to be determined.
If the value of the expression:
SZW
C=--..--
(A.@
Jsi
is larger than the corresponding value (with due regard to m and n)
from Table A.3, it may be concluded that the variance of this labora-
tory is larger than the variances of the remaining laboratories and
causes inhomogeneity. A superscript c in Table A.1 identifies the
standard deviations causing inhomogeneity of the determined variances.
The data sets of the laboratories concerned were not taken into account
for any further calculations. The Dixon test was subsequently applied
to the data sets screened in this way, to find out whether significant
differences exist between the mean values.
According to the Dixon test the given data set has first to be arranged
in order of magnitude:
X(h), h = 1, 2,3,. . . , H
(A.7)
Then (in the case of 3 <H < 7) the larger of the two expressions:
X(2) -X(l)
X(H) -X(l)
and X(H)--XH-- 1)
X(H)--(l)
(A-8)
has to be compared to the critical values in Table A.4.
A superscript b in Table A.1 marks all means which differ signifi-
cantly from the joint mean value: the data sets of these laboratories
TABLE A.3
Critical Values of the Cochran Test (Significance Level 5%) According to Goetsch
ef al. (1978).
m n=2 n=3 n=4 n=5 n=6
2
-
0.975
3 0.967 0.87 1
4 0.906 0.768
0.939 0.906 O-877
0.798 0.746 0.707
O-684 0.629 0.590
5 0.841 O-684 0.598
6 0.781 0.616 0.532
7 0.727 0.561 0.480
8 0.680 0.516 0.438
9 O-638 0.478 0.403
0.544 0.506
0.480 o-445
0.43 1 o-397
0.391 0.360
O-358 0.329
10 0.602 o-445 0.373 0.331 0.303
11 0.570 0.417 0.348 0.308 0.281
12 0.541 0.392 0.326 0.288 0.262
13 o-515 0.371 0.307 0.27 1 0.246
14 0.492 0.352 O-29 1 0.255 0.232
15 0.47 1 o-335 0.276 0.242 0.220
16 0.452 0.319 0.262 0.230 0.208
17 0.434 o-305 0.250 0.219 0.198
18 0.418 0.293 0.240 0.209 0.189
19 0.403 0.281 0.230 0.200 0.181
20 O-389 0.270 0.220 0.192 0.174
21 0.377 0.261 o-212 O-185 0.167
22 0.365 0.252 0.204 0.178 0.160
23 0.354 0.243 0.197 O-1 72 0,155
24 0.343 0.235 o-191 0.166 o-149
2s 0.334 0.228 0.185 0.160 0.144
26 0.325 o-22 1 0.179 0.155 0.140
27 0.316 0.215 0.173 0.150 0.135
28 0.308 0.209 0.168 0.146 0.131
29 0.300 0.203 0.164 0.142 0.127
30 0.293 0.198 0.159 0.138 0.124
31 0.286 0.193 0.155 o-134 0.120
32 0.280 O-188 0.151 0.131 0.117
33 0.273 0.184 0.147 0.127 0.114
34 0.267 0.179 0.144 0.124 0.111
35 0.262 o-175 o-140 0.121 0.108
36 0.256 0.172 0.137 0.111 0.106
37 0.25 1 0.168 0.134 0.116 0.103
38 0.246 0.164 0.131 0.113 0.101
39 0.242 0.161 0.129 0.111 0.099
40 O-237 0.158 0.126 0.108 0.097
72 W. Wolf et al.
TABLE A.4
Critical Values of the Dixon Test
According to Goetsch et aE. ( 1978)
n fil
Significance level
8
9
10
11
12
5% 1%
___
0.970 0.994
0.819 0.926
0.710 0.82 1
0.628 0.740
0.569 0.680
0.608 0.717
0,564 0.672.
0.530 0.635
0.501 0.605
0.479 0.579
had to be eliminated as well (however, before elimination, a background
analysis should be made in each case).
Using the remaining data sets the repeatability Y and the reproduci-
bility R are calculated using eqns [A.B) to (A. 15):
I = ' 7. 83~~
CA. 91
R =2. 83~~
where the factor 3.83 represents the 95% probability level.
s,, the repeatability standard deviation. is defined as:
(A.lO)
and sR. the reproducibility standard deviation, is defined as:
I<
(A.1 1)
CA.12)
Sorption isotherms of MCC and purified potato starch 73
where
m n(i)2
a=
&+-I~
(
i=l
1
(A.13)
(A. 14)
(A.15)