Professional Documents
Culture Documents
to Freeze Drying
State of the
Performance
An Introduction
to Freeze Drying
© Jouan Nordic A/S
Text by James M. Flink and Henning Knudsen
Printed in Denmark for Center-Tryk ApS, Holbæk 2002
An Introduction to Freeze Drying
1. Introduction
Goal of booklet, whom it is for
Organisation, 2 parts- introduction and supplements
3. What is drying?
3
11. Questions to ask when choosing a freeze dryer,
or when operating a freeze dryer
Material properties
• freezing requirements, conc., comp., temp.
• stability behaviour, temp., limits, final m.c.
Capacities required
• batch capacity, loading in chamber
• rate of production
Freeze drying parameters
• heater temperatures
• chamber types, sample surface, areas in containers
12. Supplements
A) Water properties
B) Material properties
C) Mass transport
D) Heat transport
4
1. Introduction
This booklet is intended as an introduction to the freeze drying process
for persons who will be using freeze drying in the future, or are cur-
rently using a freeze dryer and wish to know more about how it works.
3. What is drying?
Drying or dehydration is a process where the solvent in the sample, in
our case water, is physically removed from the sample such that the
solids are obtained in an essentially dry (eg. water-free) condition.
Table 1:
• Product-based goals for freeze drying
• Preparation of tissue specimens suitable for structure (histological)
studies using various microscopic techniques, including electron
microscopy.
Table 2:
Some typical materials which are dried by freeze drying
It can be seen from above examples that the various end-uses for the
product will place different requirements on the process such as
retention of biological activity, retention of physical structure, preven-
tion of chemical changes in storage, etc. It is, thus, important in the
following discussion to keep in mind that it is the product which
defines which of the possible drying methods can be used and what
process conditions will be possible. It will be noted later how the prop-
erties of the product and the above mentioned usage-based require-
ments together will act to define the range of conditions which can be
used in preparing the freeze dried product.
While one can freeze dry samples without knowing much about freeze
drying, it is the aim of this booklet to give various staff levels, from the
freeze dryer operator to the project manager, a better understanding
of the freeze drying process, and especially the relationship of process
conditions to product properties so that they can obtain a fuller
understanding of both the opportunities and the limitations associated
with freeze drying.
There are 2 main differences between freeze drying and all other drying
processes; (1) the material is solidified prior to drying, and (2) the
removal of the major amount of water from the sample occurs through
direct conversion of water from the solid state to the vapour state. In
all other drying processes, water is present in the material as a liquid,
and water removal occurs by vaporisation, (eg. water is converted from
the liquid state to the vapour state). The mobility of liquid water in most
drying processes vs. the immobility of solid water (ice) during freeze dry-
ing has major consequences on the properties of the dry product ob-
tained. Furthermore, the sizable differences in temperature levels at
which water is converted to vapour is also important. The sublimation
process (in freeze drying) occurs at low temperature (generally about
-20°C to -30°C) whereas for vaporisation of liquid water, the temper-
ature for the liquid-to-vapour transition will generally occur at higher
temperatures in the range of +50°C to +100°C
That freeze drying may not be optimal for all drying situations relates
primarily to cost considerations (both capital costs and operating
costs) since freeze drying is generally more expensive than most other
drying processes. Thus, for cases where specific product-quality fac-
tors are of lesser importance, it is possible that the higher cost of freeze
drying will not be justified. (For laboratory situations, operating cost
considerations are generally not significant). As we have noted earlier
(Table 1), there are many areas where particular product-related
requirements will necessitate the use of freeze drying if a product of
satisfactory quality is to be obtained.
10
In the chapters below, the freeze drying process will be examined in
further detail and related to the properties of the materials to freeze
dry. But before doing that, we will examine the physical layout of a typ-
ical freeze dryer, as the reader may already have one in the laboratory.
On the left can be seen the vacuum pump (1), which is connected by
means of a tube and valve to a refrigerated condenser system (2) which
removes the water. On the front of the freeze dryer are a number of
controls and measurement devices (3), and lastly, there are a number
of sample chambers shown (4), some of which are connected to the
condenser via tubing and valve systems (5).
All the shown freeze dryers are built up of the same basic components,
though in most cases the vacuum pump and refrigerated condenser
are hidden in the cabinet under the chamber for aesthetic reasons. Sev-
eral types of tube and valve systems (5) can be used to connect vari-
ous chambers to the refrigerated condenser. Some of the connections
are rather long and may have a complicated geometry.
11
Figure 1.
Typical configurations of laboratory freeze dryers.
➂
➅
➃ ➁ ➄
➇
➆ ➃ ➄
➀ ➂
1. Vacuum pump 5. Tubing and valve systems
2. Refrigerated condenser system 6. Tubing and valve systems
3. Controls and measurement devices 7. Shelf controller for temp. reg.
4. Sample chambers 8. Serial printer (via RS 232C)
12
Figure 2.
Schematic diagram of a typical freeze dryer.
Figure 3.
A p-T diagram showing phase equilibrium lines, the triple point, and the critical
point.
13
Water: - Water can exist in 3 forms (called states), referred to as solid
(as ice), liquid (as the solvent in our sample), and vapour (as steam).
The state of pure water depends at any time on its temperature and
the pressure in the system. If solids are present in the water, then the
composition of the sample will also have an effect on the state of
water. The various relationships between the states of water are given
in various state diagrams. Two are of particular interest: the tempera-
ture-pressure diagram for pure water (Figure 3) and the temperature-
composition diagram (Figures 4a and 4b). These are discussed in
detail in Supplement A. It can also be mentioned now that to change
the state of water, it is necessary to either supply or remove energy to
the water molecules.
We have already noted that several changes in the state of water are
important in freeze drying. One is the freezing of the water (a change
from liquid state to solid state) while another is sublimation (a change
from solid state to vapour state). The energy transfer required for these
changes in the state of water is quite different. For example: to freeze a
gram of water requires removing 80 calories from the water, while sub-
limating a gram of ice requires supplying 680 calories to the ice.
Figure 4.
Typical temperature-composition phase diagrams for aqueous solutions
14
In the freeze drying process, freezing water occurs at atmospheric pres-
sure (perhaps in a chest freezer, etc.), and thus the temperature of the
change in state (liquid state to solid state) is the well-known freezing
point of water (0°C).
Solids: - First we can note that, when discussing the solids in a sample
and their influence on freeze drying, it is necessary to include as a part
of the »solids« - those water molecules directly associated with (eg.
»bound to«) the solids. In freeze drying, there are a number of mate-
rial-related factors which are important to determine whether a high-
quality product will be obtained. These factors are called stability fac-
tors and can be divided into 2 major groups, structural stability and chem-
ical stability. The properties of the material, relative to these stability fac-
tors, will determine how well a product can be freeze dried.
16
Table 4:
Temperature-water vapour pressure relationship
Figure 6.
Structural stability diagram
Table 5:
Temperatures for structure transitions, °C
Collapse during freeze drying
Apple juice -42 Maltose -32
Citrate buff -40 Methocel -9
Coffee extr. -20 MSG -50
Dextran -9 Orange juice -24
Fructose -48 Ovalbumin -10
Gelatin -8 Phosphate buff -80
Glucose -40 PVP -23
Inisitol -27 Sucrose -32
Lactose -32 Sorbitol -45
Lemon juice -36
18
Collapse of dry material
Fructose 37 Maltose 96
Glucose 35 Maltotriose 88
Lactose 101 Orange juice 52
Maltodextrin (DE=10) 226 Sucrose 56
Maltodextrin (DE=20) 174 Xylose 24
Maltodextrin (DE=25) 150
Table 6:
Chemical reactions which affect the chemical stability of freeze dried
materials:
Beyond the chemical composition of the sample, the other factors hav-
ing a significant influence on the chemical stability of the material are
its temperature and moisture content. It is generally known that chem-
ical reaction rates increase with increasing temperature and fall with
decreasing moisture (though some exceptions relative to the influence
of moisture are known).
During freeze drying, the sample will have widely varying temperatures
and moisture contents the magnitudes of which will depend on the
process conditions chosen. To achieve our goal of a successful freeze
drying, the process conditions chosen must take into consideration
the properties of the materials relative to sensitivity to chemical reac-
tions.
The stability factors (both structural and chemical) of the material will
be important when placing constraints on the operating conditions in
our freeze dryer. These considerations will be taken up again in chap-
ters 10 and 11. The Supplements also give further information on these
relationships.
We can see that a high transport rate can result from either a high driv-
ing force, a small resistance or a combination of intermediate values of
each. However, we should note that without a driving force, there will be
no transport even if there were zero resistance.
case, we would say that our system has high resistances to water flow.
In Figure 7 are shown 2 tube configurations for connecting the sample
chamber with the refrigerated space. The upper tube being wide-
diametered and straight will give fewer collisions between water mole-
cules and the tube walls and will thus be a low resistance pathway rel-
ative to the lower tube with its small diameter, many bends and partly
closed valve.
In freeze drying, there are limits to the pressure differences that we can
achieve. In accordance with theory, the condenser cannot have a lower
pressure than absolute zero (perfect vacuum) while the ice pressure of
the product is limited in practice by the melting properties, this defining
a maximum temperature and pressure for the sample region. Thus, it is
quite important to keep the mass transport resistances as low as possi-
ble. Removal of air and non-condensable gases is most important, hence
we have a vacuum pump attached to the system. Beyond that, the
22
dimensions of the freeze dryer should reflect the magnitude of the water
transport loads to be placed on the system. In general, connecting tubes
should be short and have relatively large diameters. The number of valves
should be kept to a minimum and those used should be wide-throat types.
In addition, the design should have a minimum of sharp bends in the con-
necting tubing. We will return to these very important considerations in
chapter 11.
The driving force for heat transport is the difference in temperature from
one location to another. Energy moves from locations of higher tem-
perature towards locations of lower temperature.
not too important. As the space has so few molecules, we can con-
sider it to be almost empty, and thus radiation can be an important
mode of heat transport. In actuality, combinations of conduction and
radiation heat transport are quite common, either by design or where
good thermal contact required for conduction is not achieved. Typi-
cal configurations are shown in Figure 8.
For heat transport, the driving force can theoretically have any mag-
nitude wished, but in practice, there are temperature limits above which
the product should not be subjected. These temperature limits give a
maximum level for the heat transport driving force. Some of the heat
transport resistances are under our control (such as how good the
sample contacts its holder, this determining the magnitude of one of
the conduction resistance) while other heat transfer resistances are less
controllable (eg. the resistance regarding radiation).
First, even though the freeze drying process has 3 steps, it is generally
very common to conduct the freezing step in a chest freezer, in a
refrigerated liquid bath or with liquid nitrogen, using equipment not
directly associated with the freeze drying apparatus. In these cases,
only the sublimation and desorption steps take place in the freeze
dryer itself.
Secondly, while we can say that all locations in the sample have com-
pleted the freezing step before we begin the sublimation step, the same
situation is not precisely true when describing the relationship between
the sublimation and desorption steps. While at each location in the
sample sublimation proceeds desorption, if we consider the whole
sample at one time, some regions are in the sublimation step while
other regions, which have already been through sublimation, are in the
desorption step. Thus, even though we discuss sublimation and des-
orption as separate steps, it is important to realize that they are
occurring simultaneously in the freeze dryer at different locations in
the sample.
26
In general, we can note that while slower freezing gives smaller num-
bers of larger ice crystals which tend to be more orientated in one
direction, rapid freezing gives large numbers of small ice crystals which
are not oriented in one direction (Figure 9).
Figure 9.
Influence of freezing rate on ice crystals morphology in the frozen sample.
In order to remove the water vapour formed, a pathway from the ice
crystal to the sample surface will be required. This means that subli-
mation always starts at an open surface and thereafter moves inwards
into the sample. After sublimation of some of the ice, we will find that
the sample can be divided into 2 regions, the dry layer (from which the
ice crystals have sublimated) and the frozen layer (where the ice crystals
are still present). The meeting place of these 2 regions is called the ice
interface, sublimation interface, freeze drying interface or simply the interface.
At the interface, the water content makes a sharp change from that
nominally in the frozen region (that of the original material) to that in
the dry layer (starting at about 25-30% water and falling to 0-3%
water).
The frozen layer has a much lower heat transport resistance than the
porous dry layer, so much lower temperature gradients are required
for heat transport through the frozen layer than through the dry layer.
Since the dry layer surface must be open so water vapour can be
removed from the interface, it follows that heat transport to this sur-
face from the heat source must occur by radiation.
Heat transport from the surface to the interface through the dry layer
will occur primarily by conduction along the solids.
29
Figure 11.
Heat and massflows together with corresponding temperature and moisture gra-
dients for conduction and radiation heating.
After the vapour has been formed at the interface, it must be trans-
ported out of the sample to the refrigerated condenser. This requires
mass transport, and thus there must be a pressure difference, called a
pressure gradient, from the interface to the refrigerated condenser sur-
face. As noted above under sample morphology, the mass transport
resistances in the sample are established in the freezing process. As all
the vapour which is formed (this being determined by the magnitude
of the heat transport) must be removed, the water pressure at the
interface will establish itself at a level relative to the refrigerated con-
denser pressure so that the resultant mass transport of vapour away
from the interface will balance the vapour formation rate at the inter-
face as determined by the heat transport. Herein lies a potential prob-
lem since, in accordance with the equilibrium conditions given in Fig-
ure 3, an increase in the water pressure at the interface will also result
in an increase in the ice temperature, and the structural stability of the
material depends very much on this interface temperature. It is in this
way that an increase in heat transport to the sample can eventually
result in a loss of sample structure.
It has been noted earlier that the stability factors of a material place
limitations on the freeze drying conditions which may be used. In the
above, it is noted that to conduct the sublimation step, it is necessary
to transport energy to the ice crystals and to remove the vapour formed.
To accomplish this requires establishing temperature and water pres-
sure gradients, which if not properly chosen with consideration for the
stability properties of the material to be dried, will result in an alter-
ation of product properties. In particular, if one wishes to retain prod-
uct properties, it is usually necessary to place limitations on the rate
of heat transport to the interface.
There are diagrams (Figures 12a and 12b), called sorption isotherms,
which give the relationship between the water pressure and water con-
tent of a material at a given temperature (Figure 12a). The water pres-
sure is often expressed in a normalised form that is relative to the water
(vapour) pressure over pure water at the same temperature. The nor-
malised water pressure is termed the water activity (Aw= p/po) and sorp-
tion isotherms generally use this representation (Figure 12b). A materi-
al will have different sorption isotherm values as temperature changes
such that at each sample moisture level, the water pressure increases as
the temperature of the sample increases.
The water pressure exerted by the sorbed water molecules is one of the
two pressures determining the mass transport driving force required
for removing the sorbed water. The other pressure is the water pres-
32
sure at the refrigerated condenser. Theoretically, our product will cease
to lose water (ie. mass transport stops) when the water pressure at the
sample is equal to the water pressure at the condenser since the driv-
ing force will then be equal to zero.
Figure 12.
Typical water sorption isotherms, Ts > T2 > T1.
Using a series of sorption isotherms for our material, we can for each
sample temperature find the moisture content of the material for which
the corresponding water pressure is equal to the condenser water pres-
sure. This will then be the theoretical lowest final moisture content
possible for the sample at that temperature. In actuality, the final mois-
ture content will be higher, as the rate of water loss decreases sub-
stantially as the pressure differences between sample and condenser
get smaller and smaller.
If the final moisture content is not low enough to ensure product sta-
bility, it will be necessary to alter the freeze drying conditions so that
additional water transport from the sample occurs. This additional
water loss can be achieved in 2 ways (See Figure 13).
(1) One can lower the condenser temperature (from say -50°C to
-80°C). This will result in a lower water pressure at the condenser, and
thus the water pressure gradient will become zero (ie. mass transport
ceases) at a lower water pressure in the sample, which will correspond
to a lower sample moisture content. Since, in reality, it is the reduc-
33
Figure 13.
Effect of changing sample temperature and/or condenser temperature on the min-
imum possible sample moisture content.
tion in driving force (pressure gradient) as the sample dries which lim-
its the final moisture level, this approach has a limited effectiveness for
most normal materials.
We can further note that as in the other 2 freeze drying steps, it is also
necessary in the desorption step to input energy to form water vapour
from the bound water molecules. As heat transport resistances of the
dried material are high, to have the required energy input, there will
exist temperature gradients in the sample. The fact that temperature
varies in the sample will mean that the sample will have a distribution
of moisture contents in the dry layer. These gradients are shown in
Figure 11. It is essential that at the end of the freeze drying process, all
locations of the sample have moisture contents which are below the
level required for product stability. It is not adequate that the average
moisture content for the sample is below the level required for stability.
This means that after the last ice crystals disappear and the sublima-
tion step is completed everywhere in the sample, sufficient time must
be allowed for all locations to desorb their remaining bound water.
The last region to desorb to the desired level will be the region adja-
cent to the last disappearing ice crystals.
Summary of chapter 10
Since chapter 10 covers an extensive amount of material, it may be
worthwhile making a short summary here. In chapter 10, we have seen
that the freeze drying process can be divided into 3 steps, the first (freez-
ing) in which the liquid water is divided into 2 groups, ice (solid water)
and solute-associated water. The extent of this division depends on the
sample temperature and the initial composition.
The third step (desorption) also requires the input of energy. Following
the sublimation of the ice crystals at a given location, the solid-
associated water diffuses from the concentrated solution region to the
vapour pathways.
The ultimate final moisture content of the sample is attained when the
water pressure of the solid-associated water is equal to the water pres-
sure of the condenser.
The envisaged end-use of the product can also have a practical signif-
icance with respect to the best configuration of the freeze drying
apparatus, especially regarding the choice of chambers and sample
holders. This is discussed further below.
After one knows why the sample is being considered for freeze drying,
the next most important questions are related to the properties of the
material to be freeze dried. It is important to have information about
the chemical composition of the sample and the concentration of the
solids. This information will be important in determining the freezing
conditions that are required to adequately freeze the product.
Condenser-related considerations
Capacity-related questions: How big a freeze dryer do I need? The
refrigerated condenser as a capacity measure.
When buying a freeze dryer, there is always the question of how big a
dryer is required. While a number of different size criteria could be
used, laboratory freeze dryers are usually rated by the capacity of the
condenser to hold ice (ie. water from the sample). This capacity says
nothing about how fast the vapour can be transported to the con-
denser, only how much ice it can hold before defrosting is necessary.
Once we know the sample concentration, this capacity information
39
will allow us to calculate how much product can be processed before
it is necessary to defrost the condenser.
As the dryer cannot be used during the defrost period (generally 2-4
hours, longer if the condenser does not have built-in defrost heating),
frequent defrosting is not desirable. On the other hand, the condens-
ing capacity costs money, so an overly large condenser is not neces-
sarily the answer either.
It is possible to add more product to the dryer than one has condens-
ing capacity for - for example if one keeps adding manifold elements to
a freeze drying system with external manifolds. A similar situation can
arise by inserting an extra tray or two in a bulk sample chamber. Such
overloading of the system will result in incomplete drying of the sam-
ple, no matter how long the freeze drying continues.
The condenser size can be larger than that required for a single batch.
In this case, several sequential freeze dryings can be conducted before
the condenser need be defrosted. In a laboratory setting, this can have
some value as much of the working day can be used to observe the
sample in the initial stages of the drying process instead of this time
being used for defrost.
A manifold system can also be used together with larger jars or flasks.
In this case, the jar or flask acts as a small freeze drying chamber. Mod-
ern freeze drying flasks have greaseless connections (based on rubber
seals) and are available in either plastic (polycarbonate) or glass. For
particulate sample, the wide-mouth type freeze drying flask will be pre-
ferable, as it is easier to insert and remove the sample from this type
of flask than from the standard conical-neck, round-bottom labo-
ratory flask. For liquid samples, the preferences between wide-mouth
and conical-neck flasks are less clear, as wide-mouth flasks are easier
to empty while conical-neck flasks are easier to use with shell freezing.
In some cases, the manifold system can also be utilised for freeze dry-
ing different materials at the same time. These possibilities for greater
handling flexibility could be useful in a chemical laboratory where freeze
42
drying involves a variety of small sample lots as part of an analysis pro-
cedure.
43
chambers, however, are simpler, as the usual freeze drying goals do not
make such high requirements.
After having presented the pros and cons of manifold and chamber
systems, it is perhaps best to conclude that it would be most ideal if
the freeze dryer is designed so it can utilise either system. This is espe-
cially true when the freeze dryer is used in the laboratory environment,
as this will give the highest degree of flexibility.
Sample Thickness - Sample thickness affects both the freeze drying time
and the amount of material that may be present in the sample hold-
er. It is obvious that the thicker the layer of material in an ampoule or
on a tray, the more sample we can load into the dryer. This would give
44
us a higher loading capacity. However, thicker samples also take longer
to dry.
Figure 14.
Freezing configurations which give increased sample surface area.
During drying, a very thick sample will eventually get a large dry layer
thickness, which has high resistances to heat transfer and mass trans-
port. To transport energy to the interface across such a thick, high-
resistance dry layer requires a sizable temperature difference (driving
force) and since most products will have a quality-related upper tem-
perature limit, it will generally be difficult to maintain the same drying
rate with a thick sample that was possible for a thinner sample of the
same material. Thus, for thick samples, it is also necessary to accept
a further slowing of the drying process.
45
Thus, if drying is to be completed in a reasonable time, one should try
to observe the practical rule of thumb for maximum sample thickness
(generally given as 15-20 mm). In some cases, this is impossible (for
example when freeze drying whole animals), and in this case it is nec-
essary to expect that freeze drying may take days or weeks.
The surface can be increased in various ways such as by wall coating dur-
ing freezing (called spin or shell freezing), by using larger freeze drying
flasks, by granulation, or by using more flasks or trays (See Figure 14).
If freeze drying flasks are filled with a given amount of liquid sample,
the liquid sample will sit in the bottom of the flask in a pool which,
when frozen, will have a given thickness. The larger the radius of the
flask, the more area the liquid will cover, and the thinner the sample
layer will be. The combination of increased transport area and reduced
thickness will give a sample which dries more rapidly.
If the flask is tilted at an angle and rotated during the freezing step,
the liquid can be distributed on the wall surface of the flask, which will
result in a sample having a much higher transport surface and a much
lower thickness.
The above approaches only apply to liquid materials. Granulation
(grinding) of the frozen material to small chunks (called granules) is
applicable to both liquids or solids. Here one increases the specific sur-
46
face of the sample by decreasing the particle size. Granulation can be
simulated by freezing the liquid as droplets, either by spraying into an
immiscible chilled liquid, into liquidified gas or by impingement on a
very cold metal plate.
One can say that there exist maxima for the amount of sample and
sample surface that can be placed on trays in a chamber or in flasks
on a manifold. These maxima are flexible as materials that are inher-
ently more structurally stable can tolerate higher loadings than the
more sensitive materials. A further factor entering into this considera-
tion will be sample heating, which is discussed immediately below.
Sample heating - We have earlier discussed that to sublimate ice and des-
orb bound water (i.e. to conduct the last 2 steps of the freeze drying
process), it is necessary to supply energy to the sample. Even when we
make no conscious effort to supply energy during freeze drying, nature
is in actuality assuring that the required energy is being transferred.
The heat will be transferred by conduction through the wall of the flask
towards the sample, and after crossing the flask wall/ frozen layer
interface, is conducted further to the interface in the sample where the
frozen layer and dry layer meet.
At the interface, the energy is used to sublimate some ice crystals so that
the product becomes a little dryer. For samples on a tray in a drying
chamber, the course of the heat transfer will be a little different, but
in effect we would note that even if we make no effort to supply heat,
nature will do it.
We have now noted that heat is both required for freeze drying and will
be supplied by nature (if we don’t do it), and we can immediately
remark that the rate at which heat is supplied to our sample by nature
is quite arbitrary and will most likely be either too fast or too slow. As
we have noted earlier, it will be the properties of the sample which will
determine which rate of heat transfer can be tolerated. Thus, if we have
a sample which is sensitive to structural change, the natural heat trans-
48
fer rate may be too fast, while if we have a relatively insensitive sam-
ple, the natural rate will be too slow. In most cases, the natural rate is
slow enough so that freeze drying can occur with what is called »no
external heating«.
The easiest way to increase the capacity of the freeze dryer is to increase
the rate of heat transfer to the sample by means of artificial, control-
lable heating. With the presence of artificial, controllable heaters, we
can regulate the rate of heat transfer to our sample in accordance with
the sensitivity of the sample to physical or chemical changes. Increa-
sing the rate of heat transfer to the sample by using heating at tem-
peratures above ambient is the most usual method to increase the rate
of freezing drying and thus, the capacity of the freeze dryer.
As has been noted, with each method to increase freeze dryer capacity,
there are limits to the possible increase, and this observation is especially
true for the heating of the sample. We have noted earlier that the sam-
ple will have temperature limits which should not be exceeded in regard
of either loss of structural stability or chemical changes.
This will result in a water pressure rise at the sample, which is neces-
sary so that the pressure difference for mass transport is large enough
to give the required rate of water transport away from the sample. As
noted earlier, the pressure rise at the sample gives a concurrent tem-
perature rise at the sample, and if this is allowed to become large
enough (for example, by a steady increase of the heat transfer rate),
49
Figure 15.
Sample heating configurations in freeze drying.
We can note that if contact between the sample and the inside wall of
the flask is not very good (for example, due to breakage of the sam-
ple), the rate of heat transfer will become quite low and the drying time
will become long. In this case the use of a radiant heat source, such as
a heat lamp, can improve the drying rate as it can transfer heat directly
to the sample surface. But, such a direct transfer can be dangerous
and quickly lead to overheating if not carefully controlled.
In fact, it is similar to the case of the flasks on the manifold except that
due to the vacuum in the freeze drying chamber, convection is not too
significant as a heat transfer mechanism. This means that if the con-
51
tact between heating plate and sample holder surfaces is not good,
the conduction heat transfer will not be good, and the drying rate will
fall significantly. This is especially important for the cases of contact
between trays and heaters (the trays often become warped due to
rough handling) or ampoules/ flasks and trays (most ampoules have
a slighty concave bottom). Eventual heat transport control must
account for these non-ideal situations.
The second heat transport mode for drying samples on trays in the
freeze drying chamber is through the use of radiant heating to the sam-
ple and the tray surface. To have a reasonable heat transfer rate, it is
necessary to use much higher heater temperatures than with the
conduction-based heating systems.
Radiant heating has its advantages (it tends to be more even across a
tray, as well as less sensitive to tray mishandling), and disadvantages
(it is more energy-demanding due to heat losses from the dryer cham-
ber associated with the higher temperatures). In addition, since it is
usual to provide radiant heat from 2 sides, part of the heat transport
will go via the dry layer.
If the throughput rate is adequate, one can then be satisfied with the
52
natural heat input. On the other hand, if experience shows that the
capacity of the dryer when using natural heating is becoming inade-
quate relative to the amount of material to be dried, it could be
suggested to first examine the possibility of obtaining improved
drying rates by increasing the sample surface or decreasing the sample
thickness, though taking care not to exceed the overloading limitations
noted above. If further capacity increases are required, there is little
alternative to utilising artificial heating to accelerate the drying process.
Again, the limitations noted above must be respected so that the
required product quality will be maintained.
The how of heating partly depends on the type of system involved. For
a manifold system, heating can occur by means of heater-blowers, elec-
tric resistance heaters or heat lamps, or by special arrangements which
allow the use of a circulating liquid from a heating bath. In this latter
case, the freeze drying flask could have a built-in jacket, or the
manifold and flasks could be arranged so that the flasks could be
dipped into an open trough through which the liquid was circulated.
In general, a circulating liquid heating method for a manifold freeze
drying unit will be awkward to use if there are more than a few flasks
on the manifold.
Yet, a liquid circulation system with both heating and cooling will, in
general, be preferred when drying heat-sensitive materials as temper-
ature control can be more precise.
53
Supplement A:
Water properties
Equilibrium phase diagrams for water and aqueous solutions show the
various states in which water can exist under defined conditions of tem-
perature (T), pressure (P) and composition (X). By states, we mean the
physical condition of water, e.g. solid water (ice), liquid water and
vaporous water (water vapour). The basis for representing states in
terms of temperature, pressure and composition stems from these
being somewhat controllable parameters. Lastly, by equilibrium, we
mean that if we do not change the environment conditions (T, P or X),
the state(s) present will then remain unchanged for all time.
Since there are 3 controllable variables (T, P, X), the phase diagram for
water exists as a 3-dimensional diagram, with axes for T, P and X. As
the 3-dimensional diagram is quite complicated, it is usual to discuss
water properties in a series of 2-dimensional diagrams. The two of
most interest to freeze drying are the pressure-temperature relation-
ship for pure water (called the P-T diagram at X=0) and the tem-
perature-composition diagram at atmospheric pressure (T-X diagram
at P = I atm).
Some of these points are well known to us; for example point E is
located at +100°C and 1013 mbar (atmospheric pressure). We know
that at +100°C we can convert liquid water to vapour water, a process
we call boiling at 1 atmosphere pressure. The »boiling« we see is the
water molecules exerting a pressure of 1 atm, this equaling the total
air pressure.
If we lived at the top of Mt. Everest where the local atmospheric pres-
sure is only 0.31 atm (pressure falls with increasing altitude), our
water would boil at +71°C (shown at point G). We should note that
even though it is not +100°C, we still have a transition of liquid to
vapour water.
The last line, AB, which separates the solid water and vapour water
regions, is probably less well-known than the first two lines, which is
unfortunate because it is here that freeze drying occurs. The line AB
gives the corresponding pressures and temperatures at which solid wa-
ter is converted directly to vapour water (called sublimation), or vapour
water converted to solid water (called condensation). In these regions,
there is no liquid involved in the transition.
cous liquid even though it resembles a solid, and thus cooling below
the glass transition temperature does not give a real liquid-solid tran-
sition.
We can see from these 2-component diagrams (water and solute) that
the composition axis is a »mirror image« for each component (ie. 100%
water means 0% solute, etc). Thus the phase transition temperature
at 0% solute is the crystallisation (freezing)/ melting temperature of
pure water (0°C) as discussed under the P-T diagram. This point is
shown in Figures A 2 and A 3 as Tm. Similarly, if we have pure solute
(100% solute), the crystallisation/ melting temperature is shown as Tc.
We can see that if we add solute to water, or water to solute, the freez-
ing or crystallisation temperature (ie. the temperature at which the sol-
id and liquid interconvert) is decreased.
Using the water side (line AB - of most interest relative to freeze dry-
ing) we can describe what happens as we cool a solution which is ini-
tially at temperature T, and composition Xs. To avoid complicating the
discussion, the system is assumed to always be at equilibrium; in real
58
life we can have non-equilibrium conditions such as subcooling or
supercooling arising, which will give a more complicated behaviour.
Initially, cooling the solution lowers its temperature with no change in
composition. We remain in the liquid region until the temperature falls
to Tfs, which is the freezing temperature corresponding to Xs. This is
shown in Figures A 2 and A 3 by the line SR. When the solution reach-
es temperature Tfs, continued cooling will cause the system to cross
the phase boundary and the first ice crystals are forming. Ice crystals
are the solid-structure form of water molecules and are pure (ie. 100%
water). With this crystallisation of some of the water molecules, there
are now fewer liquid water molecules remaining in the solution, and
thus the composition changes in the direction of increased % solute.
If we continue down the SR line to point U, we find that we are in the
phase region, liquid plus solid water. At point U, we have a mix of ice crys-
tals (X=0) and a more concentrated solution (X=Xp) at temperature
Tfp.
Supplement B:
The material
We can note some general factors which we have called stability factors
are important for obtaining high-quality freeze dried products. The
properties of a material relative to these stability factors influence the
»freeze dryability of the material«, in particular the constraints under
which the freeze drying process must be conducted.
These stability factors are divided into two major groupings: (1) structural
stability and (2) chemical stability.
sidering the stability of the final freeze dried product, while the other
end of the moisture content scale (up to 25- 30% moisture and above)
is of most interest in the freeze drying process itself.
Figure B 2.
Influence of material molecular weight on the structural stability boundary.
The overall structural stability of the sample depends on its overall stiff-
ness, which comprises the stiffness of the ice phase and the stiffness of
the concentrated solute phase. Pure ice crystals are quite stiff even at a
temperature close to 0°C, so the ice phase present in a sample will make
an important contribution to the overall sample stiffness.
Figure B 3.
Relationship of material molecular weight and maximum temperature giving
structural stability for a given sample moisture content.
Besides the brown colour developed, these reactions also result in many
breakdown compounds having a strong aroma, and which can serve
as reactants for other reactions. In addition, Maillard browning
involving proteins can give a loss of biological activity.
Factors influencing the potential for protein changes are the own
internal bonding of the protein, the side chain reactivity, and the tem-
perature and moisture content in freeze drying.
Nutrient destruction: A number of nutrients in foodstuffs (especially
Vitamin C) are labile when subjected to heat, and as such their reten-
tion depends on the temperature conditions in the freeze drying
process. The role of moisture is less well understood, but it appears
that as the moisture level falls, the other temperature sensitivity of the
reaction also decreases.
67
Lipid oxidation: As the most important reactant, oxygen, is not pres-
ent in the chamber during freeze drying, this reaction is most impor-
tant during storage. Freeze dried material containing unsaturated lipids
is especially susceptible, as the porous structure of the freeze dried cake
gives oxygen a good access to the internal pore surface of the materi-
al. This means that lipid-containing materials must be packaged so as
to hinder the contact of lipid and oxygen, in chosen high-quality, low
transmittance packaging materials, and by closing with either vacuum
or inert gases in the package headspace.
There are also indications that free radicals, which can initiate lipid
oxidation, may be generated in the freeze drying process. In the dry
state the free radicals are quite stable, and thus their presence places
even higher requirements on the fulfillment of the packaging goals.
Nucleic acid breaks: There have been indications in the scientific lit-
erature that genetic mutations can occur in freeze dried microbial cul-
tures. It is proposed that the genetic changes result from nucleic acid
breaks. At this stage in time, it is necessary to note that many micro-
bial species can be dried successfully without undergoing genetic
changes, and thus genetic mutation is not a necessary consequence of
freeze drying. At present not much is known about the cause of genetic
mutations or about the influence of freeze drying parameters.
68
Supplement C:
Mass transport
Mass transport (or mass transfer) is the movement of matter from one
location to another. In freeze drying we are most interested in the move-
ment of water, especially from our sample to some other location. The
driving force for mass transport in freeze drying is pressure difference
(actually, in the most general case, it is a concept called chemical
potential, which in our case can be approximated by pressure), while
the resistances are a number of friction flow elements. A simple example
can be used to clarify the relationship.
Figure C 1
Basic system for mass transport discussion.
Figure C 2
Basic system with unequal distribution of molecules before
mass transport.
In Figure C 1, we have 2 equal volume chambers, left (L) and right (R)
connected by a tube (T). To separate the two chambers, we place a
69
valve (V) in the tube line. If we have an empty chamber, then we will
have no pressure in the chamber since pressure is a measure of the number
of molecules present in a given volume. The more molecules in a fixed vol-
ume, the higher the pressure. Temperature is also important; higher
temperatures give increased pressures since each molecule has more
energy, but for our discussion here, we will let the temperature be con-
stant. If in Figure C 1, we put (with the valve closed) 100 units of
water molecules in the left chamber (NL=100) and 50 units in the right
chamber (NR= 50), we will find that each chamber shows its respec-
tive pressure, PL and PR. This is shown in Figure C 2. If we now open
the valve, the system will tend to equalise itself so that the number of
molecules is the same in each chamber (ie. the pressure will become
equal in each side). This equalization is accomplished by the net phys-
ical movement of 25 units of water molecules from the left chamber,
Figure C 3
Basic system with equal distribution of molecules after mass transport.
Figure C 4
Variant of basic system with higher mass transport resistances.
70
through the tube and valve to the right chamber (Figure C 3). Thus we
have conducted a mass transport due to the pressure difference driving force.
Figure C 5.
Dynamic mass transport in the basic system.
72
Supplement D:
Heat transport
To first consider the static case where the energy from the lower sur-
face is not allowed to escape at the upper surface, then an energy trans-
fer occurs only until the upper surface reaches the same temperature
as the lower surface. Thus, when the driving force disappears, there is
no further energy transport. The rate at which this occurs will depend
on the resistances to the heat transport, and we can generally say that
the resistances increase in the order conduction, convection and radiation.
74
Figure D 2.
Boiling system for heat transport discussion.
The energy transported to the water molecules in the pot raises their
energy and thus we note a rise in the water temperature. The rate at
which the temperature rises partly depends on the size of the resist-
ances to heat transfer which exist. If we have our pot open so that the
external pressure is 1013 mbar (1 atm) we can see, from Figure A 1,
that when the water reaches a temperature of +100°C (ie. the water
molecules reach the corresponding energy level), further transfer of
energy to the water molecules results in a change in state from liquid
to vapour, which we call boiling (ie. we cross the line at point E).
76
General references on freeze drying
In this booklet, we have started our discussion on freeze drying at a
very basic level and then, from this base, we have developed the sub-
ject through more detailed examinations of the properties of materi-
als and the steps of the freeze drying process. It is, of course, not pos-
sible in a booklet such as this to cover all the possible freeze drying-
related subjects (such as engineering calculations, or additive use when
freeze drying microbial cultures, to name two topics which could not
be covered). We therefore give here a limited list of general references
on freeze drying which can be used to supplement the information pre-
sented in this booklet.
77
Rey, L. 1960.
Traité de Lyophilisation
Herman, Paris (French and English Articles)
Fisher, F. J. 1962.
Freeze Drying of Foods
National Academy of Science - National Research Council
Washington, D. C.
Rey, L. 1964.
Aspects théoriques et industriels de la lyophilisation
(Research and Developments in Freeze Drying)
Herman, Paris (French and English Articles)
Burke, R. F. and Decareau, R. V. 1964.
Freeze Dehydration of Foods
Advances in Food Research, Vol. 13
Academic Press
Rey, L. 1966.
Advances in Freeze Drying (Lyophilisation. Recherches et Applications
Nouvelles)
Herman, Paris (French and English Articles)
King, C. J. 1971.
Freeze Drying of Foods
CRC Press
Mellor, J. D. 1978.
Fundamentals of Freeze Drying
Academic Press
78
In addition to the above books, there are the Proceeding of Congress-
es and Meetings of the International Institute of Refrigeration (espe-
cially Commission 10 (now C1)-Freeze Drying, Cryobiology, Medical
Applications). These are generally referred to as bulletins, Proceedings
of Annex of the IIR.
79
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