Water and Glycerol as Plasticizers Affect Mechanical and
Water Vapor Barrier Properties of an Edible Wheat
Gluten Film
NATHALIE GONTARD, STEPHANE GUILBERT, and JEAN-LOUIS CUQ
ABSTRACT
Glycerolimprovedfilm extensibilitybut reducedfilm puncturestrength,
elasticity, and water vapor barrier properties.The plasticizingeffect
of waterwas highly temperaturedependent.During hydrationof glu-
ten film, a sharp decreasein puncturestrength,elasticity, and an
increaseof extensibilityand water vapor transmissionrate were ob-
servedat 5, 30 and 5o’C for respectivewater contentsof 30 (0,8 a,),
15 (0,7 a,) and 5% (0,4 a,). This was relatedto disruptivewater-
polymerhydrogenbondingand glass-to-rubber transition.
Key Words:wheat, gluten, film, plasticizers,glycerol
INTRODUCTION
WHEAT GLUTEN is a major functional food ingredient, es-
pecially in baked goods (Wall, 1979; Kasarda et al., 1976),
and an excellent edible film-forming agent (Gontard et al.,
1992; Gennadios and Weller, 1990; Aydt et al., 1989; Anker
et al., 1972; Wall and Beckwith, 1969). Edible films and coat-
ings afford numerous advantagesover conventional nonedible
polymeric packaging. They have been used to protect phar-
maceuticals and improve shelf-life of food products.
Proteins, lipids and polysaccharides can be used as film-
forming agents (Guilbert, 1986; Kester and Fennema, 1986;
Guilbert and Biquet, 1989). In addition to the film-forming
polymer, a major component of edible films is the plasticizer.
The addition of a plasticizing agent to edible films is required
to overcome film brittleness caused by extensive intermolec-
ular forces. Plasticizers reduce these forces and increase the
mobility of polymer chains, thereby improving flexibity and
extensibility of the film (Guilbert and Biquet, 1989; Laguna
et al., 1975; Banker, 1966). This avoids chipping or cracking
of the film during subsequent handling and storage, which
could impair film barrier properties. On the other hand, plas-
ticizers generally increase gas, water vapor and solute perme-
ability of the film (Banker, 1966; Kumins, 1965) and could
decreaseelasticity and cohesion (Delporte, 1981). The plasti-
cizer must be compatible with the polymer and is added at
concentrations ranging from 10 to 60 g/lOOg dry matter de-
pending upon polymer rigidity (Guilbert, 1986). The most
commonly used plasticizers are polyols, mono-, di- or oligo-
saccharides, lipids and derivatives. Because of its ability to
affect the texture of many polymers (Kapsalis, 1975; Katz and
Lubuza, 1981; Boume, 1986, 1987; IX Meste and Colas, 1990),
water is the most ubiquitous and uncontrollable plasticizer dur-
ing storage of food products. Mechanical, water vapor, gas or
solute barrier properties of many edible films could be strongly
affected by environmental conditions such as relative humidity
and temperature. A large increase of water vapor permeability
Authors Gontard and Cuq are with the laboratory of Genie Biol-
ogique et Sciences des Aliments, Universite de Montpellier II,
Sciences et Techniques do Languedoc, place E. Bataillon, 34095
Montpellier, France. Author Guilbert is with the CJRAD-SAR, lab-
oratory of Genie et Technologie Alimentaires, 73 rue Jean-Fran-
Gois Breton, BP 5035, 34032 Montpellier cedex, France. Tel: 67
61 57 59.
2064OURNAL OF FOOD SCIENCE-Volume 58, No. 1, 1993
is frequently observed in high water activity ranges (Landman
et al., 1960; Roth and Mehltretter, 1967; Barrie, 1968; Kam-
per and Fennema, 1984; De Leiris, 1985; Schwartzberg, 1985;
Pascat,1986; Biquet and L.abuza,1988; Ricco-Pefiaand Tortes,
1991).
In a previous investigation (Gontard et al., 1992), an edible
wheat gluten film was developed and the effects of film-form-
ing conditions on film mechanical, optical and water vapor
barrier properties were studied and optimised. The objective
of our current study was to investigate the plasticizing effect
of glycerol and water on wheat gluten film.
MATERIALS & METHODS
Preparation of gl,uten films
Films were preparedfrom a film-forming solution of vital wheat
gluten (TenstarAquitaine, F 33000 Bordeaux)in absoluteethanol/
acetic acid (Aldrich chemicals,Steinhem,Germany)/watersolvent.
The gluten concentration(7.5 g/lOOmLsolution),ethanolconcentra-
tion (45 mL/lOOmLsolution),andpH of the solution(4, adjustedwith
aceticacid)were determinedaccordingto previoustest data(Gontard
et al., 1992).Glycerol(95%, Merck, Darmstadt,Germany)was added
at concentrationsrangingfrom 0 to 33.3 g/lOOgdry film matter.All
components were mixed undermagneticstirring at 40°C.The solution
was then immediatelypouredand spreadonto a level, circular (8.5
cm diam) plexiglassplate. The film-forming solutionwas dried in a
ventilatedoven (12 hr) to evaporatevolatile solvents(ethanol,acetic
acid and water) and a transparentfilm was then formed. For each
experiment,the quantity of solution pouredonto the plate was cal-
culatedso that we obtaineda constantweight of dried matter(gluten
plus glycerol)of 5.9 mgIcm*(e.g. 3.72 mL solutioncontaining16.6g
glycerol/lOOgdry matter).This methodallowedus to producefilms
with a controlledconstantthickness(0.050 2 0.003 mm) which was
measuredat severalpositionswith a micrometer(Roth) before ex-
perimentation.
Sorption isotherm measurements
Isothermswere determinedaccordingto the proceduredescribedby
Jowitt et al. (1983) and Spiessand Wolf (1987). Samples(200 mg)
of wheatgluten film were dried in a vacuumdessicatorat 40°C over
P20sfor h 1 wk. For desorptionmeasurements, dried sampleswere
rehydratedby equilibrationvs distilled water vapor. The dried (ad-
sorptionisotherms)andhydrated(desorptionisotherms)sampleswere
equilibratedvs saturatedsalt solutionsof known water activity (a,),
rangingfrom 0.06 to 0.97, at 5, 30 and50°C(SpiessandWolf, 1987;
Multon, 1984;Stampet al., 1984).Equilibratedmoisturecontentwas
calculatedfrom weight gain or loss. Measurements were made in
triplicate. We assumedequilibriumwas reachedwhen the changein
weight (dry basis)did not exceed0.1% for four consecutiveweighings
at 1 day intervals.
Mechanical properties
Puncturetests (Bourne, 1968; Wu and Bates, 1973) were made
using a StevensLFRA TextureAnalyser(modelTA/lOOO)as previ-
ously described(Gontardet al., 1992).Films were cut into 4 cm diam
discs,fixed in an annularring clamp(3 cm diam) and equilibratedvs
appropriatesaturatedsalt solutionsin a small dessicatorat the desired
a, and temperature(5, 30 and 50°C) using the sameprocedurede-
scribedfor adsorptionmeasurements. For each a, and temperature,
the water content of the samples was determined from adsorption
isotherms. A cylindrical probe (0.3 cm diam) was moved perpendic-
ularly at the film surface at a constant speed (1 mm/set) until it passed
through the film. The force-deformation curves were recorded and
strength (N) and deformation (mm) values at the puncture point were
used to determine hardness and deformation capacity of the film.
Viscoelastic properties were evaluated using relaxation curves. The
same procedure was used, but the probe was stopped and maintained
at 3 m m deformation. The film was then allowed to relax. The force-
time relaxation curves were recorded for 1 min following deformation.
The parameterY(l min), a dimensionless ratio, was used to represent
decay of the force and calculated as follows (Peleg, 1979):
Y(lmin) = (F” - F’)/FO
where F” and F1 were forces recorded initially and after 1 min of
relaxation, repectively. A low relaxation coefficient (near 0) indicated
high film elasticity.
Water vapor transmission tests
These tests were conducted using a modified ASTM (1983) pro-
cedure. The film was sealed in a glass permeation cell containing
either silica gel or a saturated salt solution. The glass permeation cells
were 3.4 cm (i.d.) by 5.4 (o.d.) by 3.4 cm deep with an exposed area
of 9.1 cm*. The permeation cells were stored at the desired temper-
ature (5, 30 and 50°C) in small dessicatorseach maintained at a con-
stant relative humidity (RH) with either distilled water or a saturated
salt solution. To maintain a 100% RH gradient across the film, sili-
cage1(in the permeation cell) and distilled water (in the dessicator)
were used. Appropriate saturatedsalt solutions (Spiessand Wolf, 1987;
Multon, 1984; Greenspan, 1977; Stamp et al., 1984) were selected
to maintain a 10 2 1% RH gradient. The RH inside the cell was
always lower than that outside, and water vapor transport was deter-
mined from the weight gain of the cell. Sufficient time (12-48 hr)
was allowed to ensure a stable equilibrium water vapor transmission
rate.After steadystateconditionswerereached,sevenweighingswere
made over a 7-day period. Changes in weight of the permeation cell
were recorded to the nearest O.OOOlg.Slopes (changes in weight of
the cell over time) were calculatedby linear regressionand the cor-
relationcoefficientsfor all reporteddatewere 2 0.99. After the per-
meationtests,film thickeness was measuredwith a micrometer(Roth).
Thesetestswere carriedout with a minimumof 3 replicatesat each
temperatureandRH gradient.Watervapor transmissionrate (WVTR)
and water vapor permeability (WVP) of the film was calculated as
follows (Heiss, 1958; Karel et al., 1959; Labuza and Contreras-Med-
ellin, 1981):
WVTR = (w.x)/A.T = (g).(mm)/(m2).(24 hr)
WVP = (w.x)/A.T. (pZ-p,) = (g).(mm)/(m2).(24hr).(mmHg)
wherew is the weight gain of the cup over 24 hr (T), x is the film
thickness,A is the areaof exposedfilm, p2-p1 is the vapor pressure
differentialacrossthe film.
RESULTS & DISCUSSION
Plasticization effect of glycerol
Unplasticized gluten film could not be measured as it was
too brittle to be handled. In preliminary experimental work
(Gontard, 1991), hydrophilic compounds such as polyols
(glycerol, sorbitol, propanediol) and lactic acid were the only
tested substances to notably plasticize gluten film, thus in-
creasing its fIexibility. Tested amphipolar substances(i.e. gly-
co1 monostearate,acetic ester of monoglyceride, sucrose ester
of stearic acid and diacetyl tartric ester of monoglyceride) had
no substantial plasticizing effect. The hydrophobic substances
investigated, including beeswax and fatty acids (lauric, stearic

2.c

2
c 1.5
zl
f
tj 1.0
!!
f
z
2 0.5
n

0.0
10 20 30 i0 20 30
Glycerol content (g/100 g dry matter) Glycerol content (g/100 g dry matter)

=z 0.0042 -
zalO.
5
a

0.75 I I
i0 20 30 10 20 30
Glycerol content (g/100 g dry matter) Glycerol content (g/100 g dry matter)
Fig, 1 -Effect of glycerol content on mechanical properties at 30°C and 58% RH including puncture strength, puncture deformation,
relaxation coefficient, and water vapor transmission rate of gluten film at 30°C and 100% RH gradient. Variability between replicates
is indicated only when variability exceeded size of the symbol.

Volume 58, No. 1, 1993-JOURNAL OF FOOD SCIENCE-207

EDIBLE WHEAT GLUTEN FILMS. . .
-----o---- Native gluten (adsorption)
c ------IV--
E Native gluten (desorption)
z - Gluten film (adsorption)
E
z-. 40 - ““-.-‘r~-- Gluten film (desorption)
n
5.0 0.2 0.4 0.6 0.8
Water activity
Fig. Z-Water vapor sorption isotherms at 30°C for native wheat
gluten and wheat gluten film.
and oleic acids), had an anti-plasticizing effect on gluten film,
decreasing flexibility.
The importance of glycerol, the most effective plasticizer,
on mechanical and water vapor barrier properties of gluten film
is illustrated in Fig. 1. At concentrations L 15 g/lOOg dry
matter, glycerol decreased the puncture strength of gluten film
(Fig. 1) and increased puncture deformation and the relaxation
coefficient (Fig. 1). Similar results have been reported con-
cerning the plasticizing effect of glycerol on hydrophilic films
such as pectinate (Schultz, 1949), gelatin (Yannas, 1972), starch
and hydroxypropylated starch (Mark et al., 1966) or hydrox-
ypropylmethyl cellulose (Aulton et al., 1980).
The high glutamine content (about 45%) of wheat gluten
proteins is probably responsible for numerous hydrogen bonds
between protein chains. This could contribute to highly co-
operative protein-protein interactions and thus to cohesiveness
and low flexibility of unplasticized gluten film. Glycerol is a
relatively small hydrophilic molecule and could be easily in-
serted between protein chains and establish hydrogen bonds
with amide groups of gluten proteins. When glycerol was in-
corporated in a gluten/protein network, direct interactions and
the proximity between protein chains were reduced. Thus, un-
der stress, movements of protein chains were facilitated on
gluten films plasticized with glycerol.
The water vapor transmission rate through plasticized gluten
film increased with glycerol content (Fig. 1). This could be
related to structural modifications of the protein network which
might become less dense and to the hydrophilic@ of the glyc-
erol molecule which is favorable to adsorption and desorption
of water molecules.
Though flexibility of gluten films was improved by glycerol,
the amount had to be limited due to its adverse effect on film
strength, elasticity and water vapor barrier properties. In the
latter part of our study, we used a minimum glycerol content
of 16,6 g/100 g dry matter for easy handling of the gluten film.
Plasticizing effect of water
Sorption isotherms. Water vapor sorption isotherms of glu-
ten films were compared with sorption isotherms of native
wheat gluten (Fig. 2). The sorption curves of gluten films were
typical of water vapor sensitive polymers such as cellulose
208~JOURNAL OF FOOD SCIENCE-Volume 58, No. 1, 1993
0.2 0.4 0.6 0.8
Water activity
Fig. 3- Water vapor adsorption isotherms of wheat gluten film
at 5 and 50°C.
films (De Leiris, 1985). The curves showed a relatively slight
slope at low a,,,,with an exponential increase at high a, (above
0.7). Native wheat gluten showed lower sorption capacity
throughout the high a,.,range, thus at 0.90 a, the gluten film
contained 39 g water/lOOg dry matter while the native wheat
gluten contained 26g. These differences in sorption capacity
may have been due to modifications of the protein conforma-
tion during film formation, i.e. unfolding and exposure of po-
lar residues which become accessible to water molecules, and
to the high hydrophilic@ of the plasticizer. As with most water-
polymer systems, a comparison of adsorption and desorption
isotherms (Fig. 2) showed an apparent hysteresis between 0.2
and 0.8 a,,,. Hysteresis could result from slow swelling due to
conformational changes involving structural relaxation (Bryan,
1987; Slade et al., 1989). Hysteresis was greater for native
gluten than for the gluten film. Based on the behavior of var-
ious polymers, hysteresis is often tie dependent (Barrie, 1968;
Neogi, 1983; Van den berg, 1981) and the relaxation rate in
the swelling/deswelling process of the polymer matrix could
be higher for gluten film than for native wheat gluten.
The effect of sorption temperature (Fig. 3) showed that the
isotherm at 5°C was slightly above 50°C as expected. Accord-
ing to Clausius-Clapeyron equation, the amount of adsorbed
water at each a, should decreasewith increasing temperatures.
Our results showed that the temperature had the expected effect
on the sorption isotherm but differences between the sorption
measurement at 5,30 and 50°C (Fig. 2 and 3) were quite small.
Mechanical properties. The effect of a, and temperature
on puncture strength of gluten film is shown in Fig. 4. The
shape of the curves was similar despite temperature variations,
i.e. puncture strength increased at low a,., range (below 0.5)
but decreased with further hydration. During hydration to a,+,
0.5, puncture strength increased identically at 5, 30 and 50°C.
However, the decrease occurred at a temperature dependant
a/zw. Puncture strength declined at water content of 30, 15
and 5 g/lOOg dry matter at 5, 30 and 50°C respectively (Fig.
5).
Puncture deformation was also strongly affected by a, and
temperature (Fig. 6). The effect of gluten film hydration on
puncture deformation could be divided into two stages. At low
a the curve had a similar slope at all temperatures. At a
tckperature dependant L, there was an abrupt increase in
puncture deformation. At a,,, limits 0.5 puncture deformation
increased sharply.
 ______ 0 ----. fj”C
......-. *” ......

m i

0.2 0.4 0.6 0?8

Water activity
Fig. 4-Puncture strength
of wheat gluten film as a function of
water activity at 5, 30 and 50°C. Variability between replicates 0.0 0.2 0.4 0.6 0.8 1 .o
is indicated only when variability exceeded size of the symbol.
Water activity
Fig. B-Puncture deformation of wheat gluten film as a function
of water activity at 5, 30 and 50°C. Variability between replicates
is indicated only when variability exceeded size of the symbol.

0.9 1

0.5 -

I I I I
0.3 I . 1. I .I .I .
10 20 30 40 0.3 0.4 0.5 0.6 0.7 0.8 (

Water content (g/100 g dry matter) Water activity

Fig, 5-Puncture strength of wheat gluten film as a function of Fig. 7-Relaxation coefficient of wheat gluten film as a function
water content at 5, 30 and 50°C. Variability between replicates of water activity at 5, 30 and 50°C. Variability between replicates
is indicated only when variability exceeded size of the symbol. is indicated only when variability exceeded size of the symbol.

The effect of a,.,and temperature on elasticity was deduced
from the relaxation measurements(Fig. 7). Our procedure re-
quired minimal extensibility thus the relaxation coefficient could
not be measured at a, level < 0.55. Variations in relaxation
coefficient during hydration of film was highly temperature
dependent.
At 3O”C, hydration had a positive effect on mechanical prop-
erties of the gluten film, which became more resistant, elastic
and slightly more extensible, until a,,,reached 0.7. During hy-
dration of biopolymers from the dry state, hydration of charged
groups occurred first. Then side chain polar groups and finally
polar residues involved in peptide bonds and secondary struc-
ture were hydrated (Poole and Finney, 1983). In a protein
network, water molecules may develop interactions between
polar groups (e.g. -CO and -NH) which are too far apart form
direct hydrogen bonds (Blond and Le Meste, 1988). At 3O”C,
during hydration of gluten film, the first water fraction to be
sorbed (from 0 to 5 g/lOOg dry matter) improved film elasticity
and puncture resistance, probably due to the formation of sup-
plementary hydrogen bonds between protein chains. The later-
sorbed water fraction (beyond 15 g/lOOg d.m.) reduced strength
required to break the protein chain bonds, and the behaviour

Volume 58, No. 1, 1993-JOURNAL OF FOOD SCIENCE-209

EDIBLE WHEAT GLUTEN FILMS. . .
0.00~
I in the gluten film from the glassy to the rubbery state.
i Water vapor transmission rate. WVTR of gluten films
1
0.000 0.0 0.2 0.4 0:6 0.8 1.0
Water activity
Fig. 8-Water vapor transmission rate through wheat gluten film
exposed to a constant relative humidity gradient (10 c 1%) as
a function of average water activity of the ftim at 5, 30 and 50°C.
of the gluten film changed from elastic to viscous. In the high
hydration range, water-polymer interactions probably devel-
oped to the detriment of polymer-polymer bonds (Barrie, 1968).
As a consequence of the rupture of the interchain bonds, the
protein chains became more mobile and the gluten protein net-
work lost cohesiveness and elasticity.
However, it was difficult to interpret the temperature de-
pendance of the effect of hydration on mechanical properties
of gluten film in terms of disruptive water-polymer hydrogen
bonding in a polymer hydrogen-bonded network. The critical
role of water as plasticizer of gluten film appeared to be highly
temperature dependant. According to Levine and Slade (1987)
and Slade et al. (1989), the structure/property relationships of
hydrated proteins, and particularly of gluten, could be better
understood through the theories of glass transition used in pol-
ymer science, in terms of critical variables of time, temperature
and moisture content. Glass transition is typically described as
a transition from a brittle glass to a highly viscous or rubbery
solid. It is well established that plasticization by water affects
the glass transition temperature of amorphous or partially crys-
talline proteins such as gluten (Hoseney et al., 1986), gelatin
(Yannas, 1972; Marshall and Petrie, 1980), collagen (Batzer
and Kreibich, 1981) and elastin (Kakivaya and Hoeve, 1975;
Scandola et al., 1981), thus resulting in a drop in the glass
transition temperature. Water, which is a low molecular weight
component, increases free volume allowing increased back-
bone chain segmental mobility. Gluten is a highly amorphous
multipolymer system, which is water plasticizable but not water
soluble (Hoseney, 1986; Slade, 1984; Slade et al., 1989). Ac-
cording to the graph of glass transition temperature vs water
content of gluten presented by Hoseney at al. (1986), the glass
transition temperature was lowered to about lo”C/l% of water
due to the plasticizing effect of water. For 1, 6 and 16% water
content, glass transition temperatures were 160, 66 and 20°C
respectively. Gluten was thus capable of undergoing glass-to-
rubber transition at the temperatures and moisture contents in-
vestigated in our experiments. Thus during hydration of gluten
film, the glass transition temperature might be lowered until it
equalled our experiment temperatures. Ferry (1980) reported
that dependance of the ultimate mechanical properties (rupture
stress and strain) on temperature and rate of deformation was
210--JOURNAL OF FOOD SCIENCE-Volume 58, No. 1, 1993
similar to that of viscoelastic properties and therefore reflected
the configurational rearrangement rates. The sharp decrease of
film puncture strength and elasticity, and the increase of film
extensibility at 5, 30 and 50°C for respective water contents
of 30, 15 and 5 g/100 dry matter, could be attributed to changes
was strongly dependant on a, and temperature (Fig. 8). As
expected in a hydrophilic and/or edible film (Barrie, 1968; De
Leiris, 1985; Pascat, 1986; Schwartzberg, 1985; Crank, 1975;
Kamper and Fennema, 1984; Biquet and Labuza, 1988), in-
creasing a,,,induced an increase in WVTR. For the same dif-
ferential relative pressure across the film (DHR = lo? 1%) at
3o”C, increasing the mean a, of the film from 10 to 90% led
to a fifty-fold increase in WVTR. The high plasticizing effect
of water on gluten film has been demonstrated with sorption
and mechanical measurements. At high a,.,, extensive swelling
of the protein network with water probably enhances water
molecule diffusion and such films would clearly not be effi-
cient water vapor barriers. The effect of a, or water content
on WVTR in gluten film was more complex when temperature
was taken into account (Fig. 8). At 5°C increasing the a, had
a moderate effect on WVTR, with the slope relatively constant
and low. At 3O”C, the curve had two stages. For a, 0 to 0.8,
it had a relatively low slope and at a, > 0.8, a steep slope.
At 5O”C, there was a significant increase in WVTR at relatively
low a,,,and at a,,,0.6, the WVTR remained constant and very
high. We calculated permeability from the WVTR resulted
from the fact that the differential pressureacrossthe film changed
with temperature fluctuations. At 3o”C, permeability was 1.1
at 0.50 a, and 8.2 g.mm/m*.mmHg.24 hr at 0.93 G. We were
unable to evaluate the diffusion coefficient from the standard
water vapor permeability test and sorption results. Water sorp-
tion isotherms were highly non-linear and in the high a,,,range,
water content dropped very rapidly with comparatively small
decrease in a,,, (Fig. 1). Schwartzberg (1985) observed that
mass transfer resistance in adjacent gas layers was not sepa-
rated from that of the film itself. Thus the estimated water
content of upstream and downstream surfaces of the film from
sorption measurementsshould be respectively higher and lower
than the actual surface water contents. Thus, calculated dif-
fusion coefficients might be substantially underated.
As for the mechanical properties, glass transition could ex-
plain the discontinuous behaviour of the gluten film WVTR in
response to changes in moisture content and/or temperature.
The anomalous diffusion behaviour of glassy polymers may be
directly related to the influence of the changing polymer struc-
ture on solubility of the penetrant and diffusional mobility of
the penetrant (Crank, 1975).
Our study clearly demonstrated that a, and temperature were
crucial parameters in gluten films and water plasticized films
in general. The a,,,when structure/property modifications oc-
curred in the gluten film, moved upward in the RH spectrum
as temperature decreased. Based on mechanical and water va-
por transmission measurements, this important modification
occurred at 30°C and 0.7-0.8 a, at 50°C and 0.5-0.6 a,,,and
at 5°C when a, was higher than 0.8. Below these a,-temper-
ature dependant values, the wheat gluten film retained rela-
tively good mechanical and water vapor barrier properties.
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MS received 12/18/91; revised 6/6/92; accepted g/19/92.
This researchwas supportsdby s gent from the Mini&m de la Rechercheet de la
Technologie,Paris and the BSN company, Paris. Thanks to A. Verel and 0. Moniod
for technicalassistance.
OF FOOD SCIENCE-21 1