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Water Content Water Activity Water Structure On The Stability of Foodstuffs PDF
Water Content Water Activity Water Structure On The Stability of Foodstuffs PDF
www.elsevier.com/locate/foodcont
Abstract
Determination of water content, whatever the accuracy of the analytical method, is not suciently informative in relation to the
stability of the investigated food product. Water activity
aw brings a supplement of information as it accounts for the availability
of water for degradation reactions.
The understanding of why certain products are more stable than others at the same aw needs an elucidation of water structure. Of
particular importance are the interactions (hydrophilic, hydrophobic) between water and the components of the foodstu and the
eect of the soluble molecules of the food on the hydrogen bonding in solvent water.
Studying water in foods should start with an anlytical determination of water content for commercial and legal reasons which are
evident. This has to be completed with the measurement of the thermodynamic activity of water in the food. Such a parameter
aw
should hold an important place in the identi®cation of the food product, especially as regards its shelf life. A further step in unveiling
the behaviour of water in foods consists in determining water molecules in the molecules in the studied food matrix.
The tripartite (analytical, thermodynamical and structural) approach to water in foods will be examined based on examples of
sugars and sugar rich products. Ó 2001 Elsevier Science Ltd. All rights reserved.
0956-7135/01/$ - see front matter Ó 2001 Elsevier Science Ltd. All rights reserved.
PII: S 0 9 5 6 - 7 1 3 5 ( 0 1 ) 0 0 0 3 2 - 9
410 M. Mathlouthi / Food Control 12 (2001) 409±417
types of water vapour sorption isotherms. Soluble foods product. Dierent sources of errors may be found in
like sugar adopt an asymptotic shape of isotherm when oven drying. These are, for example, the incomplete
relative humidity tends towards 100%, and heteroge- removal of water, the loss of other volatiles than water
neous complex foods show a sigmoid curve. Analysis of during the drying period, the formation of a crust at the
the shape of sorption isotherms shows that they may be surface of the product which slows down the escape of
divided into three zones, respectively, corresponding to water, the decomposition of the product and Maillard
monolayer strongly bound water, linear region of less reaction which produces water.
rigidly and capillary adsorbed water and the third region Vacuum-oven drying takes place at lower tempera-
of solvent or free water. tures (70°C) for longer periods (6 h). Such conditions
This description of the three species of water is a may be less destructive for heat sensitive samples.
rough accounting of the dierences in the structure of Nevertheless, duration of drying may not be sucient to
water. Indeed, equilibrium of hydration of a food gen- allow the food to come to steady state. Size of particles
erally described by water activity
aw depends on the in¯uences both oven and vacuum-oven drying.
structure of food components, their eect on the solvent Solvent extraction may be used to extract water form
(water) and other properties like the surface activity of food with an organic solvent prior to its analysis using a
the product. Direct analysis of the structure of water in a chemical titration.
food system is dicult to achieve. Utmost, one can have Distillation which cannot be used for the analysis of
an insight into ``free'' and ``bound'' water from NMR or traces of water has a relatively long duration (P1 h)
other techniques like thermal analysis of frozen systems. (Isengard, 1995).
Amid the water properties that may be used for the
elucidation of its structure, the behaviour as a solvent 2.1.2. Chemical reactions
and the interactions with dierent classes of molecules Dierent quantitative chemical reactions, which pro-
are of great importance. duce gas, exist and can be used to quantify water content
provided that the released gas is accurately analysed.
Amid these reactions, we ®nd
2. Water content
H2 O CaH2 ! CaO H2 O
1
Determination of water content in foodstus can be H2 O CaC2 ! CaO C2 H2
2
performed using either direct or indirect methods. Direct
Reaction (2) is still used in sugar lump workshops to
determinations may be based on some physical separa-
have a rapid (10±20 min) tool of controlling the rate of
tion techniques like distillation, drying or chemical re-
moisturising the sugar prior to pressing and drying the
actions producing gases like H2 or C2 H2 which are
cubes. Volume of C2 H2 , which is directly linked to water
measured using speci®c techniques.
content, might change as a function of temperature in
Indirect determinations rely on the spectroscopic
the workshop.
properties of water molecules. It is the case for NMR,
Karl Fischer titration: Karl Fischer initiated this
infrared and Raman spectroscopy which are non-de-
method in 1935, who introduced a reagent speci®c of
structive techniques, as well as microwave spectroscopy
water which contains pyridine, methanol, sulphur di-
or microwave resonator methods. Some properties of
oxide and iodine. Later, it was shown (Verhoef & Ba-
the foodstus especially sugar solutions and fruit juices
rendrecht, 1976) that pyridine is not necessary and could
allow obtaining of the dry substance (concentration) in
be replaced by other bases with higher basicity whereas
the medium. These properties are either optical (refrac-
methanol which takes part in the reaction cannot be
tometry, polarimetry), gravimetric (density) or electrical
replaced by other alcohols (Isengard, 1995).
(conductivity) and allow indirect determination of water
Using imidazole (Z) as a base, we have a two-step
content (as a dierence).
reaction at the basis of water titration
A rapid survey of these methods is given together
with the limits and disadvantages they might have. Their CH3 OH SO2 Z ! ZH CH3 OSO2
suitability to the prediction of food product stability is ZH CH3 OSO2 I2 H2 O 2Z
also reported.
! 3ZH CH3 OSO3 2I
2.1. Direct determination The titrating reagent is I2 , which reacts stoichiomet-
rically with water. The ®rst excess of I2 indicates the
2.1.1. Physical separation end-point of reaction. Indication of end-point is ob-
Oven drying for a standardised period and a con- tained through an abrupt drop of voltage due to the
ventional temperature (3 h at 105°C for sugar) is very presence of the redox couple I2 =I at the polarised
often the legal method of determination of water con- platinum electrodes. Karl Fischer titration proves to be
tent. The period of drying is speci®ed for each type of the most accurate method, which can be used for all
M. Mathlouthi / Food Control 12 (2001) 409±417 411
values of water content from traces to high levels. This are used to determine the amount of water content (Is-
method is now computerised and is more and more used engard, 1995). Parameters such as water concentration,
especially as sugars contain only traces of water. density and thickness of the analysed sample may have
an eect on the result. Mobile water can be measured
2.1.3. Chromatography more easily than bound or crystallisation water. A cal-
Gas chromatography has been applied to the deter- ibration is needed. The method may be used for on-line
mination of water content in freeze-dried vaccines measurements provided that the thickness of sample is
(Bervelt, 1975). However, there is a need that water is known.
extracted by organic solvent prior to analysis and that Another application of microwaves to the determi-
the sample is homogeneous. Extraction solvent should nation of water in foods is that of the resonator. If a
have a high anity for water and be protected from the sample containing water is put in the resonator cham-
surrounding atmosphere humidity. Using methanol or ber, the resonance frequency of microwaves shifts under
DMF together with molecular sieves as in Karl Fischer the eect of water as well as the height of the resonance
titration, proves to be an ecient method of extraction. peak. Again a speci®c calibration is needed.
Generally, Porapaq-Q is used to ®ll the column. This
method is relatively rapid providing that the solvent or 2.1.5. Physical properties related to water content
other impurities do not have peaks that obscure the Refractometers are used to determine the percentage
peak of water. of sugar in sugar syrups or fruit juices. By dierence to
100, the water content is deduced. Polarimetry is also
2.1.4. Spectroscopy used for pure sugar solutions or technical sugar solu-
Interaction of the water molecule with electromag- tions after defecation and ®ltration. These optical
netic radiation may be used in the analysis of water methods only determine optically active substances and
content. The fact that hydrogen atoms in water have the water content is only determined by dierence. This
nuclei which possess magnetic properties allowing them might be sucient for a rapid analysis of syrups.
to behave as small bar magnets is exploited in proton Thermal analysis using dierential thermal analysis
NMR for the determination of water in foods (Troller & (DTA) or dierential scanning calorimetry (DSC) may
Christian, 1978). be used during the heating of a frozen sample to deter-
NMR spectroscopy is informative on hydrogen at- mine freezable water, which is approximately the frac-
oms. These are more easily detected in a liquid envi- tion of water, considered as mobile or ``free''. Such
ronment. So that this technique is more adapted to techniques are also informative on the glassy state of
distinguish between free water and bound water (crys- water in foodstus, which might help in the interpreta-
tallisation water for example) than the accurate deter- tion of the behaviour of the product during drying.
mination of water content. Moreover, there is a need of
obtaining precise calibration speci®c of the analysed
3. Water activity and sorption isotherms
product based on a good reference method.
Near infrared (NIR) absorption of water occurs at
Most of the methods used for the determination of
dierent wavelengths (1950, 1450 and 977 nm). The ra-
water activity or ERH of foods were originally set for
tios of the intensities of the bands at 1950 and 1450 nm
the measurement of relative humidity in atmospheric air
are used as a measure of water content (Vornhof &
by the meteorologists (Troller & Christian, 1978). When
Thomas, 1970). It is also possible to extract water from
water activity
aw is measured it is generally required to
the food with a solvent (methanol, dimethylformamide)
know if the product has reached the critical zone where
prior to analysis by NIR spectroscopy.
spoilage reactions may occur or not. That is why accu-
Computerised NIR spectrometers are used in dier-
racy within 0.01aw unit is sucient for most food-related
ent food industries for the determination of water con-
applications.
tent and other food constituents. This method requires a
Water activity is de®ned as the ratio of partial pres-
speci®c calibration for the food analysed and the use of
sure of water vapour in the product (p) to that in pres-
statistical methods of exploitation of results which are
ence of pure water
po :
included in the software. The colour, particle size, p
thickness and texture of the product in¯uence results. aw :
po
The re¯ectance technique allows detection of surface
water and might not be representative of the whole If there was no dierence in the interaction between
product if it is not homogeneous. water and water on the one hand and water and solute
Microwave spectroscopy uses the dipolar character of on the other, the determination of water activity would
water molecules. The shift in wavelength and the at- have been easy and its expression
tenuation of the amplitude of the waves when a sample n2
aw Xw
is placed between the emitter and receiver of microwaves n1 n2
412 M. Mathlouthi / Food Control 12 (2001) 409±417
directly obtained from the mole fraction Xw of water though this type of measurement can be rapid and
molecules
n2 to total molecules in the solutions. For precise, it only accounts for surface aw . The surface of
real solutions Aw c Xw , where c is an activity coe- the mirror should be clean and non-contaminated and
cient. The higher the modi®cation of water binding by the amount of condensed water negligible if it is desired
the solute, the more c coecient is dierent from 1. to have the measured relative humidity equal to ERH of
the food.
3.1. Manometry
3.4. Other methods of measurement of relative humidity
As water vapour pressure is given in tables for dif-
ferent temperatures, a direct measurement of water va- Other methods of measurement of relative humidity
pour pressure in the food should give the best direct tool may be used to determine aw . It is the case for the wet
of determination of aw . To achieve this measurement bulb-dry bulb thermometric technique, the hair hy-
there is a need of establishing vacuum and working at grometer, the freezing point depression or the aptitude
temperatures as low as )80°C. Working at zero pressure of certain chemical compounds to change colour at a
in one side of the oil manometer with the freeze trap of given relative humidity. These methods may be used to
moisture and leaving the sample on the other side release give an approximate value of aw during the storage of a
its vapour permits accurate measurement of aw . This food product.
method needs uniform accurate temperature measure-
ment and the device is extremely fragile (Troller & 3.5. Water vapour sorption isotherms
Christian, 1978).
3.5.1. General aspect and meaning
3.2. Electric hygrometer A water vapour sorption isotherm represents the
variation of water content as a function of water activity
The sample is placed in a tight measuring chamber at at a given temperature. The general aspect of a sorption
a controlled stable temperature. During equilibration isotherm usually observed for a food product is sigmoõd
the sample releases humidity and the same relative hu- (Fig. 1).
midity at the level of the electrical sensor and in the
sample is observed. The sensing element may be either a
conducting polymer or an electrolytic element like sat-
urated LiCl solution. Another type of electric hygrom-
eter uses the measurement of electrical conductivity or
capacitance of a hygroscopic substance like LiCl crystal
or the anodised surface of an aluminium rod (Troller &
Christian, 1978). Filters that adsorb contaminants are
often needed. The sensors need frequent calibration with
saturated salt solutions. Except in the extremes of the
water activity range (below 0.15 or above 0.95), the
accuracy of the electric hygrometer is satisfactory.
Among the drawbacks of this type of aw -meter are the
hysteresis observed and the sensitivity of sensor to high
relative humidities (near saturation).
Table 1
LiCIH2 O 0.12
CH3 COOK 0.23
MgCl2 6H2 O 0.33
K2 CO3 0.44
Mg
NO3 2 6H2 O 0.52
NaCl 0.75
NH4 2 SO4 0.79
CdCl2 0.82
Li2 SO4 0.85
K2 CrO4 0.88
KNO3 0.94
K2 SO4 0.97 Fig. 2. Hysteresis of water vapour sorption isotherms.
Na2 HPO4 0.98
Table 2
Comparison of humectancies of cake ingredients with sugar taken as
reference standard (humectancy 1)
Product Humectancy
However, glycerol although a good water activity Luu, Luu, Rull, and Sopron (1982). Such a model may
depressor might not be the ideal ingredient to protect be adopted as a reference for comparative studies of the
against bacterial growth. Water although retained eect of solutes on water structure.
around glycerol molecules is mobile enough to allow
the growth of bacteria (Sperber, 1983). It is also the
4.2. Hydrophilic interactions
case for fructose, which is also known to have much
mobile hydration water around it than sucrose or
Cations of interest in the food industry like
glucose.
Na ; Ca2 or Mg2 are known to immobilise several
layers of hydration water. Generally such ions are
known to have three layers of water around them. A
4. Water structure ®rst layer strongly bound to the ion, a second layer of
perturbed water because it is submitted to the attrac-
Most of the reactions occurring during the storage tion due to electrical ®eld of the ion on the one hand
of a foodstu, like lipid oxidation, enzyme degrada- and to the ``cooperativity'' of H-bonds with water
tion, Maillard reaction may ®nd an interpretation re- molecules of the third layer where water is less in¯u-
lated to the structure of water. Likewise the speci®c enced by the ion.
interactions of the dry substance with water, namely Beside this type of attraction of water molecules
hydrophilic or hydrophobic interactions can be helpful around a hydrophilic solute, the hydration of small
to understand the behaviour of the product towards non-electrolytes and biopolymers is generally consid-
water vapour during its preservation. A brief recall of ered as hydrophilic. Small hydrophilic molecules like
the structure of pure liquid water will be reported be- sugars have a sphere of hydration characterised by a
fore the description of solute±solvent interactions in number of hydration (®ve for sucrose) and a ``structure
foods and their consequences on the stability of food maker'' eect as they have a long-range eect of ori-
products. entation of bulk water molecules. The hydration of
food polymers is sensitive to their conformations and
this can be measured by dierential scanning calori-
4.1. Liquid water structure
metry (Silvonen, Lindberg, Seppala, Evasti, & Hauta-
hoo, 1982). It is also important to take into account
The chemical structure of H2 O, where an atom of
the eect of the phase (gel, emulsion,) in which the
oxygen occupies the centre of a regular tetrahedron,
biomolecules are embedded, which may change their
two vertices of which are the H atoms and the two
conformation. In particular globular proteins may
others the lone pairs of electrons of the oxygen atom, is
change conformation as concentration is increased and
at the origin of its 3D structure. Indeed, such a con-
temperature raised to expose their hydrophobic back-
®guration allows each water molecule to be associated
bone to water and establish intramolecular H-bonds.
to four molecules by H-bonds. Moreover, hydrogen
Interactions of water with the hydrophobic surface of
bonds are sensitive to charge transfer from donors to
folded proteins corresponds to the so-called hydro-
acceptors in a sequence which produces an eect called
phobic hydration.
``cooperativity'' (Frank, 1974). Other liquids like alco-
hols form chains of H-bonds because they have only
one hydroxyl group whereas water forms tetrahedral 4.3. Hydrophobic interactions
clusters.
Water structure was one of the most studied subjects Hydrophobic interactions are due to the hydration of
these last years using either computer models or exper- apolar groups like methyl groups. Interaction between
imental spectroscopic techniques. However, no theory such a group and water is weaker than water±water
or conclusions drawn from experimental results can hydrogen bonding, so that, when several water mole-
account for its particular physical properties. Computer cules surround an apolar group, they do not have a
models for water structure generally describe this liquid direct contact with the group and hence reinforce their
as a continuous network of H-bonds randomly ar- water±water bonds. This layer of water is much less
ranged. These models have two disadvantages: the lack mobile than bulk water. Its enthalpy is decreased and its
of details and a static character. Experimental tech- entropy is increased. Hydrophobicity of proteins may be
niques such as neutron scattering, FTIR and Raman at the origin of the stability of emulsions. Processed
spectroscopy were used to investigate structure and dy- meat products may be considered as emulsions where
namics of the hydrogen bonding in liquid water. De- hydrophobic hydration of the proteins together with the
composition of the experimental Raman band in four hydrophilicity of salt and glycerol tends to immobilise a
components assigned to four species of clusters with H- maximum of water and to increase the shelf-life (Lacroix
bonds diering in strength and number was proposed by & Castaigne, 1984).
416 M. Mathlouthi / Food Control 12 (2001) 409±417
Verhoef, J. C., & Barendrecht, E. (1976). Mechanism and reaction rate Vornhof, D. W., & Thomas, J. H. (1970). Determination of moisture
of the Karl Fischer titration reaction ± Part I: Potentiometric in starch hydrolysates by near-infrared and infrared spectropho-
measurements. Journal of Electroanalytical Chemistry, 71, 305. tometry. Analytical Chemistry, 42, 1230.