International Journal of Food Properties

ISSN: 1094-2912 (Print) 1532-2386 (Online) Journal homepage: https://www.tandfonline.com/loi/ljfp20

Food Stability Beyond Water Activity and Glass

Transtion: Macro-Micro Region Concept in the
State Diagram

Mohammad Shafiur Rahman

To cite this article: Mohammad Shafiur Rahman (2009) Food Stability Beyond Water Activity and
Glass Transtion: Macro-Micro Region Concept in the State Diagram, International Journal of Food
Properties, 12:4, 726-740, DOI: 10.1080/10942910802628107

To link to this article: https://doi.org/10.1080/10942910802628107

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International Journal of Food Properties, 12: 726–740, 2009
Copyright © Taylor & Francis Group, LLC
ISSN: 1094-2912 print / 1532-2386 online
DOI: 10.1080/10942910802628107

FOOD STABILITY BEYOND WATER ACTIVITY

AND GLASS TRANSTION: MACRO-MICRO
REGION CONCEPT IN THE STATE DIAGRAM

Mohammad Shafiur Rahman

Department of Food Science and Nutrition, Sultan Qaboos University, Al Khod,
Oman

The water activity concept proposed that a food product is the most stable at its monolayer
moisture content. Recently, the limitations of the water activity concept were identified and
the glass transition concept was proposed in order to overcome the limitations of water
activity. Based on the glass-transition concept, a food is the most stable at and below its
glass transition point. Recently it has also become evident that the glass transition concept
is not universally valid for stability determination when foods are stored under different
conditions. The glass transition concept was used to develop the state diagram by drawing
another stability map using freezing curve and glass line. Currently, other components
indicating different characteristics are being included in the state diagram. It is being
emphasized in the literature to combine the water activity and glass transition concepts. In
this paper, an attempt is made to combine these two concepts in the state diagram and to
propose a macro-micro region concept for determining the stability of foods.

Keywords: Isotherm, Food stability, Glass transition, BET monolayer, Food spoilage.

INTRODUCTION
Humans in the Atone Age (10,000 years ago) stored nuts and seeds for winter and
discovered that meat and fish could be preserved by drying in the sun. After the discovery
of fire, cooking made food more appetizing and was an aid to preservation. Modern pro-
cessing developed at the end of the 1700s when the Napoleonic wars raged. As Napoleon
pushed forward into Russia, his army was suffering more casualties from scurvy, malnu-
trition, and starvation. The French government offered 12,000 francs to any one who
could develop a method of preserving food. Nicolas Appert took up the challenge. He had
a theory that if fresh foods were put in airtight containers and sufficient heat applied, then
the food would last longer. Appert packed his foods in bottles, corked them, and sub-
merged them in boiling water, thus preserving them without understanding of bacterial
spoilage. After 14 years of experimentation, in 1809 he won the prize and this was given
to him by Napoleon himself. A theoretical understanding of the benefits of canning did not
come until Louis Pasteur observed the relationships between microorganisms and food
spoilage some fifty years later.[1] Eventually canning, drying, and freezing technology

Received 2 October 2008; accepted 10 February 2009.

Address correspondence to Mohammad Shafiur Rahman, Department of Food Science and Nutrition,
Sultan Qaboos University, PO Box 34, Al Khod 123, Oman. E-mail: shafiur@squ.edu.om

726
 MACRO-MICRO REGION CONCEPT IN THE STATE DIAGRAM 727

evolved. Recently new preservation methods have emerged and been adopted by the food
industry. A comprehensive complete coverage of preservation is available in the Hand-
book of Food Preservation and other references.[2–4]
Determining food stability from a scientific basis rather than an empiricism is a
challenge to food scientists and engineers. Microbial death and growth kinetics, deteriora-
tive physical, chemical and biochemical changes during processing and storage depends
on many factors, such as water content, food composition, preservatives, pH, and environ-
mental or processing factors (temperature, pressure, electro-energy, gases or vapors etc).
In the 1950s, the concept of water activity was proposed to determine the stability of
foods, and in the 1980s, significant data on food stability as a function of water activity
was published. In order to avoid the limitations of water activity, the glass transition con-
cept was extensively proposed in the 1980s, although this concept initially appeared in the
literature in the 1960s. These two concepts provide a strong scientific basis of food stabil-
ity during drying and freezing. The stability of foods is of paramount interest to both food
scientists and engineers; and a better understanding of the factors controlling microbial
stability or reaction rates is clearly needed.[2–6] In this paper, the concepts of water activity
and glass transition are discussed along with their applications and limitations. In addition,
an attempt was made to combine these two concepts in the state diagram by proposing a
macro-micro region concept.

WATER ACTIVITY CONCEPT

In the 1950s, scientists began to discover the existence of a relationship between the
water contained in a food and its relative tendency to spoil.[7] In the 1980s, Labuza and his
group generated significant data on food stability as a function of water activity. They also
began to realize that the active water could be much more important to the stability of a
food than the total amount of water present. Thus, it is possible to develop generalized
rules or limits for the stability of foods using water activity. For example, there is a critical
water activity below which no microorganisms can grow, this value is about 0.6. Most of
the pathogenic bacteria cannot grow below a water activity of 0.85; whereas most yeasts
and molds are more tolerant to reduced water activity but usually no growth occurs below
a water activity of about 0.62.
Microbial responses to low water activity are shown in Fig. 1. A well-established
response to the temporary loss of turgor pressure after a hyper-osmotic shock (i.e., a
reduction of water activity surrounding the cell) is osmoregulation. Exposure of microorgan-
isms to lower water activity causes an instantaneous loss of water, which is accompanied by
a decrease in the cytoplasmic volume called plasmolysis. This can also cause lysis. Hypo-
osmotic shock generally results in minor changes in cell volume. On the other hand,
hyper-osmotic shock causes considerable shrinkage of the cytoplasmic volume. If the
osmotic shock is not too severe, after an extended lag phase, the cytoplasmic volume
increases as a result of osmotic adjustments made by the cells.[8] After loss of turgor, the
microbial cell raises level of the compatible solutes within the cells.[9] Compatible solutes
including glycerol, sorbitol, cyclohexaneterol, erythritol, arabitol, or mannitol in fungal
cells or amino acids such as proline, aminobutyric acid, and glutamic acid in bacteria.
These compatible solutes assist to prevent dehydration, and thus facilitate metabolic activ-
ities necessary for growth. This results in an increase in internal osmotic pressure and
restores turgor pressure. The type and amount of solute accumulated in cells is not only
influenced by water activity, but also other factors in the environment, including those
728 RAHMAN

CS: Compatible Solutes

Water (if low aw environment)

K+
CS
K+
Turgor Cell K+
CS
Water Low Water Activity CS
Normal (Turgor Cell) accumulation of compatible
solutes to lesser gradient and
reduce water loss

K+
Plasmolysis

K+

Low Water Activity K+

Figure 1 Microbial response at low water activity.

associated with preserving hurdles used to control microbial growth, such as pH and preser-
vatives.[10,11] Besides the accumulation of compatible solutes, changes in the membrane lipid
including structural changes in phospholipids and fatty acid were also observed.[12,13] The
compatible solutes produced internally are highly soluble, pH neutral, and are usually end
product metabolites. They can include sugars from the breakdown of carbohydrates, amino
acids from protein degradation, and cations such as K+. Examples include betaine, trehalose,
glycerol, sucrose, proline, chloline, carnitine, mannitol, glucitol, and ectoine. The cell mem-
brane is selectively permeable to them, allowing the cytoplasmic pool to be determined by
the external osmotic pressure.[14,15] The preferred exogenous bacterial-compatible solutes
are glycine and betaine, which are found in higher plants, and the amino acid proline.[4] In
many foods, however, peptides are more readily available than free amino acids and hence
peptides have become an important source of both nutrients and compatible solutes.[16]
A food product is most stable at its monolayer moisture content, which varies with
the chemical composition, structure and environmental conditions, such as temperature.
However, in many instances the critical limit may also be observed at higher moisture than
monolayer moisture content. Figure 2 shows the monolayer and multilayer water on a
solid surface.[17] This principle was the main reason why food scientists started to empha-
size water activity rather than total water content. Since then, the scientific community has
explored the significance of water activity in determining the physical characteristics, pro-
cesses, shelf life, and sensory properties of foods. It is now used to predict the end point of
drying, in process design and control, in ingredient selection, to predict product stability
and to make packaging selection. One of the earlier food stability maps based on the water
activity concepts considers growth of micro-organisms and different types of bio-chemi-
cal reactions.[18,19] The updated food stability map is presented in Fig. 3. In this present
map, the trends of microbial growth, bio-chemical reactions and mechanical characteris-
tics are presented in the 3 zones of water activity. In general the rule of water activity con-
cept is: Food products are most stable at its monolayer moisture content or monolayer
water activity and unstable above or below monolayer. A water activity map also provides
the following rules: below monolayer (zone I):(i) stability can be decreased with decreasing
 MACRO-MICRO REGION CONCEPT IN THE STATE DIAGRAM 729

w w w
w w w
w w w
w w w w
w w w w
Free or Bulk Water (More Active) w w w w w
w w w
p p
w w w
w w w w
w w w p w p

er
w w w w

at
p p

W
w w
w w w

d
w Solid Surface

pe
w w w

r
w w

so
w w

Ad
w w
w w w w Multi-Layers
w w w
w w w
w w w w
w w w w w w w
w w w
w w w
w w w w w w w w
w

er
w w w w w w
p p

at
w w w w
w w w w p p

W
w w w w
w p w p w w

w
w w w p p

Lo
w w w
w p p w
w p p
Solid Surface
w w w Solid Surface
w

Capillary Water BET-Monolayer of Water

p p

er
at
W
p p
P: Polar Site
No

W: Water Molecule p p

Solid Surface

Primary Active Sites

Figure 2 Concepts of BET mono and multi layers water.

Zone I Zone II Zone III e

Adsorbed Water h
c d
i
j n
io
Rate or Growth or Attribute

a ct f
Re
Below Monolayer

l
ica
em
Ch
wth

m
l Gro
Me

Solvent Water
cha

o
obia
nic

b
al P

Micr
rop

a
ert
ies

k l
n q
p
g
0 1
Water Activity

Figure 3 Updated stability diagram based on the water activity concepts.

730 RAHMAN

water activity, for example fat oxidation (line ob); (ii) stability can remain constant (line ab);
and (iii) stability can increase (line nb). In the adsorbed layer (zone II): (i) stability can
increase (line bp); and (ii) stability can decrease (line bc). In multi layer (zone III): stability
can increase, decrease, or remain constant.
Recently, the limitations of water activity concept were identified[7,20–24] as:
(i) Water activity is defined as at equilibrium, whereas foods may not be in a state of equi-
librium, for example low and intermediate moisture foods. (ii) The critical limits of water
activity may also be shifted to higher or lower levels by other factors, such as pH, salt,
anti-microbial agents, heat treatment, electromagnetic radiation, and temperature.
(iii) Nature of the solute used to reduce water activity also plays an important role, for
example some solutes are more inhibiting than others even at the same water activity
(minimum growth of P. fragi is 0.96 using sodium chloride, while for glycerol water
activity is 0.94.[25] (iv) Water activity concept does not indicate the mobility or reactivity
of water and the nature of binding to the substrate. It only provides the information on the
amount of strongly bound and free water without their precise reactivity. (v) Many physical
changes, such as crystallization, caking, stickiness, gelatinization, diffusivity could not be
explained based on the water activity alone. However, these limitations do not completely
invalidate the concept but rather make it difficult to apply universally. In order to find
other alternatives, the glass transition concept was postulated in the literature.

GLASS TRANSITION CONCEPT

Glassy materials have been known for centuries but the glass transition concept was
first applied to foods with scientific understanding in the 1980s.[26] A low glass transition
means that at room or mouth temperature, the food is soft and relatively plastic, and at
higher temperatures it may even flow. In contrast, a food with a high glass transition
temperature is hard and brittle at ambient temperature. Early attempts to describe glassy
phenomena concluded that glass is a liquid that has lost its ability to flow, thus instead of
taking the shape of its container, glass itself can serve as the container for liquids. Food
materials are in an amorphous or non-crystalline state below the glass transition tempera-
ture and are rigid and brittle. Glasses are not crystalline with a regular structure, but retain
the disorder of the liquid state. Physically it is a solid but resembles closely a thermody-
namic liquid. Molecular mobility increases 100-fold above glass transition. In kinetic
terms, Angell [27] described a glass as any liquid or super-cooled liquid whose viscosity is
between 1012 and 1013 Pa s, thus effectively behaving like a solid, which is able to support
its own weight against flow due to gravity. To put this viscosity into context, a super-
cooled liquid with a viscosity of 1014 Pa s would flow 10−14 m/s in the glassy state, in con-
trast the flow rate of a typical liquid is in the order of 10 m/s. In other words, a glass is a
liquid that flows about 30 μm in a century.[28] This is evidenced by the fact that ancient
stained glass windows are thicker at their base due to flow under gravity.[29]
The early papers related to glass transition in food and biological systems appeared
in the literature in the 1960s.[30–32] White and Cakebread[31] first highlighted the impor-
tance of the glassy state of foods in determining its structural stability. They were perhaps
the first food scientists to discuss the importance of the glassy and rubbery states in rela-
tion to the collapse of a number of high solid systems. The significant applications of the
glass transition concept emerged in food processing in the 1980s, when Levine and Slade
[33]
and Slade and Levine[34] identified its major merits and wide applications. In the
1990s, Roos, Karel, and other groups generated significant data on the glass transition and
 MACRO-MICRO REGION CONCEPT IN THE STATE DIAGRAM 731

A Free
4
zing Tgs
Curv
e
Rubber

Temperature
Ice 3
Tg′
c
Glass
2 F

ine
Tgw Gla
ss L 1
G

0 Xs′ 1
Solute Mass Fraction

Figure 4 State diagram proposed by Levine and Slade showing 5 macro-regions.

components (i.e., characteristic curves) of state diagrams for a number of food ingredients.
In 1990s, Chirife, Buera, Bell, Karel, Roos, Labuza, and others started to present data on
food stability based on the glass transition and water activity concepts. It has been men-
tioned in the literature that foods can be considered very stable at the glassy state, since
below glass temperature compounds involved in the deterioration reactions take many
months or even years to diffuse over molecular distances and approach each other to
react.[35] A hypothesis has recently been stated that glass transition greatly influences food
stability, as the water in the concentrated phase becomes kinetically immobilized and
therefore does not support or participate in reactions. Formation of a glassy state results in
a significant arrest of translational molecular motion, and chemical reactions become very
slow[36]. The rules of glass-transition concept are: (i) Foods are most stable at and below
glass transition; and (ii) the higher the T-Tg or T/Tg (i.e above glass transition), the higher the
deterioration or reaction rates. These conditions identify two macro regions (i.e., 3 regions
above and one region below) in the state diagram (Fig. 4). Similarly, mechanical and
transport properties could also be related with glass transition. It is very interesting to see
that this concept has been so widely tested in foods as evident from the literature. In many
instances, glass transition concept does not work alone, thus it is now being recommended
to use both the water activity and glass transition concepts in assessing process-ability,
deterioration, food stability, and shelf-life predictions.[37]

GLASS TRANSITION AND GLASSY STATE

Phase transitions in foods can be divided into two groups: first-order and second-
order. At the first-order transition temperature, the physical state of a material changes
isothermally from one state to another (e.g., solid to liquid, liquid to gas) by release or
absorption of latent heat (e.g., melting, crystallization, condensation, evaporation).
Second-order transition occurs (e.g., amorphous state to glassy state) without release or
absorption of latent heat.[38] Glass transition is a second-order, time-temperature depen-
dent transition, which is characterized by a discontinuity or change in slope of physical,
732 RAHMAN

onset and end onset (Tgi)

Tgi= Tge
A B
Heat Flow (W/g)

Heat Flow (W/g)

peak (Tgp)

end (Tge)

Temperature (°C) Temperature (°C)

Figure 5 Second order transition in foods identified by DSC thermogram. A: ideal second order transition,
B: non-ideal second order transition.

mechanical, electrical, thermal, and other properties of a material, when plotted as a function
of temperature.[38] This transition is most commonly identified as a shift in the thermogram
line measured by Differential Scanning Calorimetry (DSC). The process is considered to be
a second order thermodynamic transition in which the material undergoes a change in
state but not in phase. It is more meaningfully defined as analogue as nature to second-
order, since each measurement technique is based on monitoring change in a specific
property, and since change or break in properties are achieved within a certain temperature
range rather than at a specific temperature.[22] A perfect second-order transition occurs at a
specific temperature. During heating, glass transition of food materials does not occur at a
fixed point with the change of specific heat. Instead networks soften or transform over
quite a large temperature range.[29] However, glycerol produces a step change in heat
capacity as a function of temperature at 190 K.[39] Figure 5 shows the ideal second order
transition (A) (i.e., shift at a specific temperature) and non-ideal second order transition
(i.e., shift over a temperature range); (B) as evident by DSC thermogram.

STATE DIAGRAM AND ITS COMPONENTS

State Diagram
A state diagram is another stability map of different states of a food as a function of
water or solids content and temperature.[40] Levine and Slade[33] presented a state diagram
of providone N-vinyl pyrrolidone (PVP) illustrating glass line, freezing curve, and inter-
section of these lines as Tg′ (in this paper, the author is noted as Tg′′). The intersection was
identified by extrapolation of extending the freezing curve by maintaining a similar curva-
ture. This is most probably the first state diagram in the food science literature. Many of
his publications later presented[34] the state diagram of starch by quoting the source refer-
ence of van den Berg.[41] Later he presented another state diagram showing glass line,
freezing curve, melting line, eutectic point, Tg′ and vapor line.[42] Starting from the 1990s,
Roos, Karel, Kokini, and others presented state diagrams of a number of food components
and food products. The main advantages of drawing a map are to help understanding the
complex changes when the water content and temperature of foods are changed.[37,43–49] It
also assists in identifying stability of foods during storage as well as selecting a suitable
condition of temperature and moisture content for processing.[21,50–51] The regions of
drying and freezing can be easily visualized in the diagram, and product stability can be
 MACRO-MICRO REGION CONCEPT IN THE STATE DIAGRAM 733

U (Tds)

Liquid
Highest Molecular Mobility L (Tms)

Water vapor 13 Solid

D
M
Tbw Crystal 10
E (Tgs)
A 12 Collapse 11
Temperature

Solution Stickiness
Rubber
9 Softening zone
G ice + solution (solute+free water) B Tgiv
P Reaction zone
Tu Entangle flow
H 8 ice + solution (solute crystal+free water)
Tm′ C N 1
I 7 ice + rubber (solid matrix + un-freezable water) R
Tg″′

BET-Monolayer
ice + glass at Tg″′ Q
J 6
Tg″
5 ice + glass Tg″ Glass
K
Tg′
4 ice + glass Tg′
S 2
Tgw
F
3 glass

0 Xs′ Xs″′ Xs″ O 1

Solute Mass Fraction

Figure 6 State diagram showing different macro-micro regions (updated from Rahman [22]).

assessed based on moisture content and temperature. Figure 6 shows a state diagram indi-
cating the different states as a function of temperature and solute mass fraction (updated
from Rahman [22]).

Components of State Diagram

Earlier state diagrams were constructed with only the freezing curve and glass tran-
sition line. Recently, attempts have been made to add other structural changes along with
the glass line, such as solubility line. Numbers of micro-regions and new terminologies
are being included in constructing the state diagram. The presented state diagram shown in
Fig. 6 is updated from Rahman.[22,40] In Fig. 6, the freezing line (ABC) and solubility line
(BDL) are shown in relation to the glass transition line (EQFS). Point F (Xs′ and Tg′) which
is lower than Tm′ (point C) is a characteristic transition (maximal-freeze-concentration
condition) in the state diagram, defined as the intersection of the vertical line from (Tm′)a
to the glass line EQFS.[22] At this maximal-freeze-concentration, all possible freezable
water is transformed into ice. The water content at point F or C is considered as the
un-freezable water (i.e., 1- Xs′). The un-freezable water mass fraction is the amount of
water remaining unfrozen even at very low temperature. It includes both un-crystallized
free water and bound water attached to the solids matrix. Point Q is defined as Tg′′ and Xs′′
as the intersection of the freezing curve to the glass line by maintaining the similar curvature
of the freezing curve ABC.
734 RAHMAN

A B

Temperature (°C)
(Tg′′′)a

Heat Flow (W/g)

(Tm′)a
(Tm′)a
Tm′
Tg′′′
(Tg′′′)a Tg′ Tg′′

Temperature (°C) Solute Mass Fraction

Figure 7 Identification of (Tm′)a, (Tg′′′)a, Tm′, Tg′′′, Tg′′, or Tg′ in the state diagram.

The apparent glass transition (Tg′′′)a of the solids matrix in the frozen sample is usu-
ally determined by Differential Scanning Calorimetry (DSC) below (Tm′)a (Fig. 7A). This
is due to the formation of a similar solid matrix associated with un-freezable water and
transformation of all free water into ice, although the sample contains different levels of
total water before the start of DSC scanning.[52] The values of apparent (Tg′′′)a and (Tm′)a
decreased with increasing solids content and reached to a nearly constant value. The inter-
sections of the lines in Fig. 7B show the Tg′′′ (point R in Fig. 6) and Tm′ (point C in Fig. 6).
If the sample containing moisture close to the points R and C, these apparent values
become similar to Tg′′′ and Tm′ as marked in Fig. 7.
In the region AGB as shown in Fig. 6, the phases present are ice and solution. Below
point B, the first crystallization of solute occurs, transforming the GBCH region to three
states: ice, solution and solute crystal. There is no free water (i.e able to form ice) to the
right side of point C (Tm′, end point of freezing with maximal freeze-concentration-condition)
and below the very concentrated solution is transformed to the rubber state. The maximal-
freeze-concentration condition can be achieved using optimum conditions by slow cooling
and/or annealing of the samples so that all freezable water can be transformed into ice.
The region HCRI contains ice, rubber, and solute crystal. Point F is the Tg′, below this
point all portion of the rubber state is transformed to the glass state, thus the region KFS
contains glass, ice, and solute crystal. The rate of cooling can shift the points B, C, Q and
F. More detailed effects of cooling on the shift are discussed by Rahman.[40] The glass line
should follow the line EQFS if maximum ideal plasticization occurs in the solid matrix
with the addition of water. The line EQF could be shifted upward if water causes different
degrees of plasticization (as shown by line EP) in the sample containing un-freezable
water. The vertical line from F intersects at point P and the temperature is defined as Tgiv.
In this situation, fitting the glass transition data to the Gordon-Taylor equation, consider-
ing the two ends as Tgs and Tgw, may not be valid. Based on the author’s hypothesis of dif-
ferent characteristic temperatures Tgiv, Tm′, Tg′′′, Tg′, and Tg′′, the modified equation could
be proposed as:

X sTgs + kc X wTc
Tg = (1)
X s + kc X w

where Xw and Xs are the mass fraction of water and solids; Tc and Tgs are the characteristic
temperature and glass transition of solids; and kc is the characteristic of the material, respec-
tively. In the case of the Gordon-Taylor equation, the characteristic temperature is the
glass transition of water instead of Tc (i.e., Tgiv, Tm′, Tg′′′, Tg′, or Tg′′).
 MACRO-MICRO REGION CONCEPT IN THE STATE DIAGRAM 735

The region BQEL is important in food processing and preservation, many character-
istics, such as crystallization, stickiness, and collapse are phenomena that are observed in
this region.[37,53] In case of cereal proteins, Kokini et al.[48] determined an entangled poly-
mer flow region and a reaction zone based on the mechanical characteristics. The line
BDL is the melting line which is important when products are exposed to high tempera-
ture during processing, for examples frying, baking, roasting, and extrusion cooking. In
the case of a multi-component mixture such as food a clear melting is difficult to observe
at high temperature due the reactions or interactions between the components. In this case,
Rahman.[40] and others have defined melting as the decomposition temperature. Line
MDL is the boiling/evaporation line for water from the liquid phase (line MD) and solid
matrix with a degree of saturation with water (line DL). It is possible for the melting and
evaporation lines to intersect since water evaporation could happen in a saturated matrix
before melting of the solid matrix mixed with water.
It was identified that further work is required on the relationships between glass
transition, water activity and food stability.[3] It appears that the interrelationships can be
very complex, depending on the complexity of the food system and on the type of stability
being studied. Recently many papers have presented data on both water activity and glass
transition as a function of water content. However the link between them that determines
stability has not been identified. Karel et al.[54] attempted to relate water activity and glass
transition by plotting equilibrium water content and glass transition as a function of water
activity. By drawing a vertical line on the graph, stability criteria could be determined
from the moisture isotherm curve and the glass transition line. At any temperature (say
25oC), stability moisture content determined from the glass transition line was much
higher than the stability moisture from the isotherm. Similarly Sablani et al.[55] predicted
stability criteria (solids content) from an isotherm model (GAB equation), considering
water activity 0.20, and then estimate the solids content from glass model (Gordon-Taylor)
considering storage temperature as Tg. They found a surprising gap between the predic-
tions of the stable solids contents using the two different approaches for a number of food
products. At present it is a challenge to link them in a meaningful way.
As a first attempt, Rahman[22] plotted the BET-monolayer value as the LNO line in
the state diagram shown in Fig. 6. It intersects at point N with the glass line EQFS, which
shows that at least in one location (point N), glass and water activity concepts provide the
same stability criteria. This also justifies the variability of deteriorations observed by
Sablani et al.[55] This approach forms more micro-regions, which could give different
degrees of stability in the state diagram. More studies regarding stability need to be done
on both the left (above and below glass) and right sides (above and below) of the line
LNO. It should be mentioned here that the BET-monolayer could be achieved by mainly
removing water from a system (since the isotherm is relatively unaffected by tempera-
ture), but glassy state could be achieved by removing water through drying, as well as by
decreasing the temperature of the system. A successful combination of water activity and
glass transition could develop a more in depth knowledge on stability criteria. In addition,
how other factors, such as pH, and preservatives act could be linked with these concepts.
At present the scientific community far from developing unified theory.
It is evident from the literature that stability below or above glass transition varies
even based on specific cases, indicating that applying only the glass transition temperature
for developing the stability rule is not enough. Samples with freezable water are more
complex and four characteristic temperatures were defined by Rahman et al.[52] In current
paper a total of five temperatures are defined as Tgiv > Tm′ > Tg′′′ > Tg′ > Tg′′. In addition,
736 RAHMAN

the stability below or above Tgiv, Tm′, Tg′′′, Tg′, and Tg′′ should also be explored. There are
only a few references available that include all four characteristic temperatures with their
moisture content. It is important to know how these temperatures affect the stability of
foods. It would be interesting to explore the differences in stability in product within these
different ranges.[22]

VALIDITY OF GLASSY STATE

Applications of glassy state concept in food systems are thoroughly reviewed by
Kalichevsky-Dong[3] and Rahman.[22] They grouped the applications of glassy state in the
processes of controlling diffusion process, structure, crystallization, stickiness, grain
damage, pore formation, microbial stability, seed stability, oxidation, non-enzymatic
browning, enzymatic reaction, protein denaturation, hydrolysis, and enzyme inactivation.
Most of the literature used two criteria to test the validity specifically: (i) whether food is
stable if it is in the glassy state or unstable if it is above the glass transition; and/or
(ii) whether the change in attributes above glass is related to the (T-Tg) or T/Tg. It is clear
from the literature that all experimental results could not be explained by the above rules,
thus it is now important to identify when it fails and why. It is also a challenge to combine
other concepts with glass transition.
The water activity concept is based on the binding nature of water molecules in the
matrix. When water is bound (i.e., unavailable to take part in reactions) to the solid matrix
or non-solvent, then no deterioration reactions could be expected. The glass transition
concept is based on the molecular mobility of the reacting components in the matrix, thus
diffusion of the reactants through the system to take part in reactions is very slow and
stability is achieved. Although combining both the concepts could be a powerful tool for
stability determination. A successful combination of water activity and glass transition
could open more precise and unified determination of stability criteria. Attempts should be
made to provide alternative solutions to combine both concepts. In fact there are other
factors which play a role and in many instances it is incorrect to say even combining both
would give complete stability. However, other building blocks could be added. Stability
could be determined from boundary modeling and then kinetics prediction models.

MACRO-MICRO REGION CONCEPT

BET-monolayer line as LNO in the state diagram (shown in Fig. 6) makes four
regions: below BET-monolayer, one above and one below; and above BET-monolayer,
one above and another below.[22] This approach forms more micro-regions, which could
give different stability in the state diagram and could explain the limitations of each
concept. The present hypothesis proposed thirteen micro-regions having highest to lowest
stability based on the locations from glass and BET-monolayer lines. For example, the
region-1 (relatively non-reacting zone, below the BET-monolayer line and glass line) is
the most stable and region-13 (highly reacting zone, far from BET-monolayer line and
glass line) is least stable. The stability decreased as the zone number increased. Each
micro-region could be studied for specific deterioration separately, considering different
characteristics of a medium.
It is clear that the stability based on macro-regions in the state diagram considering
below and above BET-monolayer or glass transition would not be enough to determine
stability. The literature showed that many systems were stable above glass transition and
 MACRO-MICRO REGION CONCEPT IN THE STATE DIAGRAM 737

unstable below glass transition. One of the arguments was proposed that mobility of water
could also occur below glass transition. Based on the variability in the stability, micro-
regions were identified within the macro-regions. In the state diagram, 3 factors are
clearly identified or mapped: (i) how far it is from the BET-monolayer line; (ii) how far it
is from the glass transition line; and (iii) how low is the temperature. In addition to the
binding nature of water (water activity) and mobility of reactants (glass transition), sample
temperature could be identified as a separate factor in the state diagram, considering
temperature dependent reaction rates. The reactivity does not increase only based on a
function of (T-Tg) or T/Tg and variation in increasing or decreasing behavior could be
observed, for example micro-region 10 is below the BET-monolayer and above the glass
transition and could be very reactive. The micro-region 1 is considered most stable since it
is below the glass transition and BET-monolayer, and low temperature storage. The most
unstable micro-region is region 13 since it is the most reactive. In this case, there is no
point in applying the concepts of glass transition or water activity alone and other preser-
vation hurdles must be used. The micro-region could also be sub-divided further to
explain the variability of stability, for example regions between 3 to 9 based on Tu, Tm′,
Tg′′′, Tg′′, or Tg′. This could open another dimension to the food stability map with more
micro regions. Similarly regions 10 and 11 could be divided based on Tg and Tgiv for the
samples containing no freezable water. Another important aspect in stability is to study the
types of glass formed in foods along with their characteristics, such as fragile/strong
glass,[56, 57] degree of plasticization,[57] strength and density of hydrogen bonds [58], vari-
ability in molecular mobility and/or relaxation below glass,[59,60] and complex matrix of
crystalline/semi-crystalline/amorphous regions. There is a definite need to clarify the
validity of each concept by identifying the valid and invalid conditions and systems.

COMBINING MULTICONCEPTS
IFT[6] presented the factors that influence the microbial growth in foods. In this
report the multi-factor nature of microbial stability was identified, and how two factors,
pH and water activity interact each other were described. The USDA pathogen-modeling
program was used to identify critical limits of water activity and pH when both factors are
interacting. They also identified that a general model for foods to cover all interactions of
atmospheric gases and/or preservative combinations with pH and water activity does not
currently exist. In reality the problem of stability determination could not be solved by
identifying macro-regions or micro-regions, but it could help in developing scientific,
systematic and rational approaches to determine the stability. The next task is to test the
stability in each micro region and there may be a possibility to explore more generic rules
for stability of individual stability criteria in the micro-region. A knowledge-based
approach could be used to identifying how other factors, such as pH, preservatives, and
types of solutes affect the stability in each micro-region.

KNOWLEDGE BASE OR DATA MINING

The author believes that developing a knowledge base of the stability for each
macro and micro region would be a valued approach for exploring further generic rules for
stability. He is confident that a data mining approach and/or a boundary modeling technique
could be used to explore and to develop the generic rules in the future. This database, if
developed, could be the foundation of new theoretical progress.
738 RAHMAN

CONCLUSION
The state diagram could be used to combine water activity and glass transition con-
cepts by including BET-monolayer and glass lines. In addition different macro- and
micro-regions could also be drawn to identify food stability. In this paper 13 micro regions
are proposed for determining food stability. A library of knowledge needs to be developed
for each macro- and micro-region for developing further generic rules.

ACKNOWLEDGMENTS
This paper was presented in the 18th International Congress of Chemical and Process Engineering
(CHISA 2008), 24-28 August 2008, Prague, Czech Republic as a keynote lecture. The author would
like to acknowledge the support of Sultan Qaboos University towards this research in the area of food
structure and stability. He is grateful to all members of his research group for their continued support
and encouragement. Special thanks to Dr. Ann Mothershaw for checking the clarity of the paper.

REFERENCES
1. Food Preservation. http://www.answers.com/topic/food-preservation (accessed May 2009).
2. Rahman, M.S. 2007a. Food preservation: overview. In Handbook of Food Preservation, 2nd ed;
Rahman, M.S.; Ed.; CRC Press: Boca Raton, FL, 2007; 3–18.
3. Kalichevsky-Dong, M.T. The glass transition and microbial stability. In The Stability and Shelf
Life of Food; Kilcast, D.; Subramaniam, P.; Eds.; Woodhead Publishing: Cambridge, England;
CRC Press: Boca Raton, FL; 25–53.
4. Beales, N. Adaptation of microorganisms to cold temperatures, weak acid preservatives, low
pH, and osmotic stress: a review. Comprehensive Reviews in Food Science and Food Safety
2004, 3, 1–19.
5. Le Meste, M., Roudaut, G., Rolee, A. The physical state and quality of cereal-based foods. In
Food Engineering 2000; (Fito, P; Ortega-Rodriguez, E.; Barbosa-Canovas, G.V.; Eds.; pp.
Chapman & Hall: New York, 1997; 97–113.
6. IFT. IFT/FDA report on task order 4. Factors that influence microbial growth. Comprehensive
Reviews in Food Science and Food Safety 2003, 2 (Supplement), 21–32.
7. Scott, W.J. Water relations of Staphylococcus aureus at 30oC. Australian Journal of Biological
Science 1953, 6, 549–564.
8. Csonka, L.N. Physiological and genetic response of bacteria to osmotic stress. Microbiological
Review 1989, 53 (1), 121–147.
9. Gould, G.W., Christian, J.H.B. Food preservation by moisture control. In Characterisation of
the State of Water in Foods—Biological Aspects. In: Food Preservation by Moisture Control.
Seow, C. C.; Teng, T.T.; Quah, C.H.; Eds.; Elsevier: London, 1988.
10. Beuchat, L.R. Detection and enumeration of microorganisms in hurdle technology foods, with
particular consideration of foods with reduced water activity. In Food Preservation by Moisture
Control. Fundamentals and Applications; Barbosa-Canovas, G.V.; Welti-Chanes, J.; Eds.;
Technomic Publishing Company: Lancaster, PA, 1995; 603–612.
11. Lang, F.; Waldegger, S. Regulating cell volume. American Scientist 1997, 85, 456–463.
12. Russell, N.J. 1989. Functions of lipids: structural roles and membrane functions. In Microbial
Lipids; Ratledge, C.; Wilkinson, S.G.; Eds.; Academic Press: London; 279–365.
13. Russell, N.J.; Evans, R.I.; Tersteeg, P.F.; Hellemons, J.; Verheul. A.; Abee, T. Membranes as a
target for stress adaption. International Journal of Food Microbiology 1995, 28, 255–161.
14. Galinski, E.A. Osmoadaptation of bacteria. Advances in Microbiol Phys. 1995, 37, 273–328.
15. Gutierrez, C.; Abbe, T.; Booth, I.R. Physiology of the osmotic stress response in microorganisms.
International Journal of Food Microbiology 1995, 28, 233–244.
 MACRO-MICRO REGION CONCEPT IN THE STATE DIAGRAM 739

16. Amezega, M.R.; Davidson, I.; McLaggan, D.; Verheyul, A.; Abbe, T.; Booth, I. The role of
peptide metabolism in the growth of Listeria monocytogenes ATCC 23074 at high osmolarity.
Microbiology 1995, 141, 41–49.
17. Furmaniak, S.; Terzyk, A.P.; Gauden, P.A. The general mechanism of water sorption on
foodstuffs- importance of the multitemperature fitting of data and the hierarchy of models
Journal of Food Engineering 2007, 82, 528–535.
18. Labuza, T.P.; Tannenbaum, S.R.; Karel, M. Water content and stability of low-moisture and
intermediate-moisture foods. Food Technology 1970, 24 (5), 543–544.
19. Labuza, T.P.; McNally, L.; Gallagher, D.; Hawkes, J.; Hurtado, F. Stability of intermediate
moisture foods. I. Lipid oxidation. Journal of Food Science 1972, 37, 154–159.
20. Rahman, M.S.; Labuza, T.P. Water activity and food preservation. In Handbook of Food
Preservation, 2nd ed.; Rahman, M.S.; Ed.; CRC Press: Boca Raton, FL, 2007; 447–476.
21. Rahman, M.S. Dried food properties: challenges ahead. Drying Technology 2005, 23 (4), 695–715.
22. Rahman, M.S. State diagram of foods: Its potential use in food processing and product stability.
Trends in Food Science and Technology 2006, 17, 129–141.
23. Chirife, J. Specific solute effects with special reference to Staphylococcus aureus. Journal of
Food Engineering 1994, 22, 409–419.
24. Hardman, T.M. Interaction of water with food component. In Interaction of Food Components;
Birch, G.G.; Lindley, M.G.; Eds.; Elsevier Applied Science Publishers: London, UK, 1986; 19–30.
25. Sperber, W.H. Influence of water activity on foodborne bacteria—a review. Journal of Food
Protection 1983, 46 (2), 142–150.
26. Ferry, J.D. Some reflections on the early development of polymer dynamics: viscoelasticity,
dielectric dispersion, and self-diffusion. Macromolecules 1991, 24, 5237–5245.
27. Angell, C.A. Perspective on the glass transition. Journal of Physics and Chemistry of Solids
1988, 49, 863–871.
28. Buitink, J.; Leprince, O. Glass formation in plant anhydrobiotes: survival in the dry state.
Cryobiology 2004, 48, 215–228.
29. Kasapis, S. Glass transition phenomena in dehydrated model systems and foods: a review.
Drying Technology 2005, 23 (4), 731–758.
30. Rey, L.R. Etude Physiologique et Physico-chimique de I’Action des Basses Temperatures sur
Tissus Animaux Vivants. PhD. thesis, 1958, 122 pp.
31. White, G.W.; Cakebread, S.H. The glassy state in certain sugar-containing food products.
Journal of Food Technology 1966, 1, 73–82.
32. Luyet, B.; Rasmussen, D. Study by differential thermal analysis of the temperatures of instability
of rapidly cooled solutions of glycerol, ethylene glycol, sucrose and glucose. Biodynamica 1968,
10, 167–191.
33. Levine, H.; Slade, L. A polymer physico-chemical approach to the study of commercial starch
hydrolysis products (SHPs). Carbohydrate Polymer 1986, 6, 213–244.
34. Slade, L.; Levine, H. Non-equilibrium behavior of small carbohydrate-water systems. Pure
Applied Chemistry 1988, 60, 1841–1864.
35. Slade, L.; Levine, L. A food polymer science approach to structure property relationships in
aqueous food systems: non-equilibrium behavior of carbohydrate-water systems. In Water
Relationships in Food; Levine, H.; Slade, L.; Eds.; Plenum Press: New York, 1991; 29–101.
36. Rahman, M. S. Glass transition and other structural changes in foods, In Handbook of Food
Preservation, 2nd ed.; Rahman, M.S.; Ed.; CRC Press: Boca Raton, FL, 2007; 75–93.
37. Roos, Y. Characterization of food polymers using state diagrams. Journal of Food Engineering
1995, 24, 339–360.
38. Rahman, M. S. Food Properties Handbook; CRC Press: Boca Raton, FL, 1995. pp. 87–177.
39. Allen, G. A history of the glassy state. In ‘The Glassy State in Foods; Blanshard, J.M.V.;
Lillford, P.J.; Eds.; Nottingham University Press: Nottingham, UK, 1993; 1–12.
40. Rahman, M.S. State diagram of date flesh using differential scanning calorimetry (DSC).
International Journal of Food Properties 2004, 7 (3), 407–428.
740 RAHMAN

41. Van den Berg, C. Water activity. In Concentration and Drying of Foods; MacCarthy, D.; Ed.;
Elsevier Applied Science: London, 1986; 11–36.
42. Levine, H.; Slade, L. The glassy state in applications for the food industry, with an emphasis on
cookie and cracker production. In The Glassy State in Foods; Blanshard, J.M.V.; Lillford, P.J.;
Eds.; Nottingham University Press: Nottingham, UK, 1993. pp. 333–373.
43. Slade, L.; Levine, H. Structure stability of intermediate moisture foods- a new understanding?
In Food Structure—its Creation and Evaluation; Blanshard, J.M.V.; Mitchell, J.R.; Eds.;
Butterworths: London,1988; 115–147.
44. Slade, L.; Levine, H. Water and the glass transition- dependence of the glass transition on com-
position and chemical structure: special implications for flour functionality in cookie baking.
Journal of Food Engineering 1994, 22, 143–188.
45. Slade, L.; Levine, H. Glass transitions and water-food structure interactions. Advances in Food
and Nutrition Research 1995, 38, 103–179.
46. Roos, Y.H.; Karel, M.; Kokini, J.L. Glass transitions in low moisture and frozen foods: Effects
on shelf life and quality. Food Technology 1996, 50 (11), 95–108.
47. Slade, L.; Levine, H.; Ievolella, J.; Wang, M. The glassy state phenomenon in applications for
the food industry: Application of the food polymer science approach to structure-function rela-
tionships of sucrose in cookie and cracker systems. Journal of the Science and Food Agriculture
1993, 63 (2), 133–176.
48. Kokini, J.L.; Cocero, A.M.; Madeka, H.; De Graaf, E. The development of state diagrams for
cereal proteins. Trends in Food Science and Technology 1994, 5, 281–288.
49. Kokini, J.L.; Cocero, A.M.; Madeka, H. State diagrams help predict theology of cereal proteins.
Food Technology 1995, 49 (10), 74–82.
50. Roos, Y.; Karel, M. Applying state diagrams to food processing and development. Food
Technology 1991, 44 (12), 66–71.
51. Cuq, B.; Abecassis, J.; Guilbert, S. State diagrams to help describe wheat bread processing.
International Journal of Food Science and Technology 2003, 38, 759–766.
52. Rahman, M.S.; Sablani, S.S.; Al-Habsi, N.; Al-Maskri, S.; Al-Belushi, R. State diagram of
freeze-dried garlic powder by differential Scanning Calorimetry and cooling curve methods.
Journal of Food Science 2005, 70 (2), E135–E141.
53. Roos, Y.; Karel, M. Plasticizing effect of water on thermal behavior and crystallization of
amorphous food models. Journal of Food Science 1991, 56, 38–43.
54. Karel, M.; Anglea, S.; Buera, P.; Karmas, R.; Levi, G.; Roos, Y. Stability-related transitions of
amorphous foods. Thermochimica Acta 1994, 246, 249–269.
55. Sablani, S.S.; Kasapis, S.; Rahman, M.S. Evaluating water activity and glass transition concepts
for food stability. Journal of Food Engineering 2007, 78, 266–271.
56. Angell, C.A.; Bressel, R.D.; Green, J.L.; Kanno, H.; Oguni, M.; Sare, E.J. Liquid fragility and
the glass transition in water and aqueous solutions. Journal of Food Engineering 1994, 22, 115–142.
57. Champion, D.; Le Meste, M.; Simatos, D. Towards an improved understanding of glass transition
and relaxations in foods: molecular mobility in the glass transition range. Trends in Food Science
and Technology 2000, 11, 41–55.
58. Wolkers, W.F.; Oliver, A.E.; Tablin, F.; Crowe, J.H.A Fourier-transform infrared spectroscopy
study of sugar glasses. Carbohydrate Research 2004, 339 (6), 1077–1085.
59. Moates, G.K.; Noel, T.R.; Parker, R.; Ring, S.G. Dynamic mechanical and dielectric character-
ization of amylase-glycerol films. Carbohydrate Polymers 2001, 44, 247–253.
60. Lin, X.; Ruan, R.; Chen, P.; Chung, M.; Ye, X.; Yang, T.; Doona, C.; Wagner, T. NMR state
diagram concept. Journal of Food Science 2006, 71 (9), R136–R144.