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General Principles of Freeze Drying (The Lyophilization Process)

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 1
FREEZ DRYING
General Principles of Freeze Drying (The Lyophilization Process)
By Dr. Mostafa Khater, B.Pharm.
Introduction
Application and Uses
Freeze drying, or lyophilization as it is referred
to in the Pharmaceutical and Diagnostic
Industries, is a dehydration technique, which
enables liquid or slurry products, which have
previously been frozen to be dried under a
vacuum. The applications of freeze drying are
numerous, but are generally employed when the
requirements demand:
• Preservation of temperature sensitive
products, particularly those of biological origin,
such as enzymes, blood plasma, vaccines, etc.
• To achieve a chemical balance, such as for
biological reagents.
• To provide a practical solution for certain
delivery problems, for example, the packaging
of constituents that cannot be mixed in the liquid
state, but which are solidified in successive
stages and then freeze dried.
• To implement an important stage of a product
(such as concentration).
• To improve storage life and improved
marketing of the end product.
• To resolve certain filling problems. It may be
difficult, for instance, to divide several
milligrams of powder into precise vial dosages,
due to the difficulty of measuring tiny amounts,
homogeneity, granulation, static electricity etc..
The distribution of the product from the liquid
state eliminates such production problems.
• Product temperature sensitivity and its
relation to taste.
Description of the Operation
Generally, the Freeze Drying, or Lyophilization
cycle is divided in three phases:
• An Initial freezing process carried out in
such a way that: The product exhibits the
desired crystalline structure. The product is
frozen below its eutectic temperature.
• A primary drying (sublimation) phase
during which: The partial pressure of the vapor
surrounding the product must be lower than the
pressure of the vapor from the ice, at the same
temperature. The energy supplied in the form of
heat must remain lower than the product's
eutectic temperature (the highest allowable
product temperature during the conditions of
sublimation)
• A Secondary drying aimed at eliminating
the final traces of water which remain due to
absorption, and where: The partial pressure of
the vapor rising from the product will be at its
lowest levels. At the completion of the process,
the treated product will have retained its form,
volume and original structure-as well as all its
physical, chemical and biological properties. It
can then be stored (provided packaging is
effective to the reduction of moisture migration)
for an almost indefinite period of time. As the
product is porous, it can be re-dissolved by the
simple addition of a proper solvent. From this
description of the process of freeze drying, three
facts emerge:
• The sublimation characteristics of the product
are greatly dependent of the frozen structure.
• This structure cannot be altered during the
process.
• Product temperature plays an active role in all
three phases, and in execution it is
upon this that the choice of other parameters
(vacuum, heat rate, etc.) is based.
Theoretical Basis of Freeze Drying
The theoretical principle of freeze drying is
clearly defined in "Pressure Temperature
diagram".
• In order to avoid the liquid phase, it is
absolutely essential to lower the partial pressure
of water, below the triple point pressure. A
freeze-drying cycle is shown, which has been
designed to conform to a typical example
(described below):
• Freezing of a product from 20 °C to -20 °C at
atmospheric pressure.
• Sublimation of the product at -20 °C.
• Transfer of evolved vapor to the condenser at
low temperature.
• Vacuum release.
• Defrost.
2
Freeze drying is a complex operation, and all
facets cannot be addressed in this type of
explanation. Instead, certain aspects will be
highlighted which play a part in the development
of a freeze-drying operation:
- Freezing. - Drying.
- Vacuum influence.
- The liquid shelf on which the product is
placed.
- Essential control aspects during freeze drying.
Freezing
Upon completion of product freezing, the
product will have acquired a frozen structure,
which cannot be changed during freeze drying.
Sublimation and the qualities of the finished
product are greatly dependent on this crystal
structure. In fact, it is considered the most
crucial stage of the freeze-drying process.
Speed of Freezing
On the pilot level, fast or very fast freezing is
relatively easy to achieve. However, for
industrial production settings, freezing at the
same rates is unrealistic because of the problems
of product preparation (filling, loading time) and
larger systems costs will dictate compromises in
the same process.
As soon as the product reaches 0 °C, some of the
particles transform to ice. This is the nucleation
process. Generally, biological products contain
between 80% and 95% water.
Observe that the temperature of the product
stabilizes after time period at about 0 °C.
The ice crystals previously formed have
expanded, consist practically of pure water. The
crystals have grown larger, and now occupy
80% to 90% of the initial volume of the solution.
The crystallization of the free water is nearly
complete. These crystals seem to be contained in
an interstitial state, still liquid, but which
constitutes the principal active element of the
solution.
The interstitial component itself has reached
freezing temperature, and the amorphous
appearance is even more apparent, and a barely
visible “skin” has formed on the surface. This
structure is ideal for sublimation.
We now have a paradoxical situation: a slow
cooling which can lead to a rapid coagulation of
the constituent water. In many cases, freezing
induced by these conditions may be necessary to
achieve successful freeze drying of a sensitive
product.
Freezing Temperature
In these examples, the temperature represents
the complete freezing of the product. The
establishment of this eutectic zone is very
important. In between, the concentration and
consistency of the liquid phase is increased, and
in the case of biological products, may produce a
change in the bacteria as a result of the hyper
concentration of the active ingredient, and the
mechanical effect of the ice crystals.
During the sublimation phase, signs of melting
appear in the product, which induces a
temperature higher than that corresponding to
the eutectic point. Temperature analysis permits
determination of the onset of the fusion
temperature. This acts as an indicator to help
prevent “melt back”, or other such accidents
during the course of freeze drying.
• Generally, a structure of large crystals
presents more difficulties with regard to freeze
drying, in that a thick crust forms at the surface.
The appearance of the freeze-dried product is
heterogeneous. This very often makes
dissolution difficult.
• A product structure of fine crystals freezes
more easily. The freeze-dried product has an
amorphous appearance, and re-dissolves more
quickly. Obtaining the desired crystalline
structure is not always easy, as the formation of
this type of crystal depends on several factors:
• The nature of the product.
• The processing.
• Freezing speed.
• Type of freezing
Given the diverse range of products, which may
be treated in the dryer, the desired freezing rate,
and the type of final packaging that may used,
the manufacturer must consider:
• The quantity of product per container.
• The form of the container.
• The type of freezing.
• And, plan for a freeze dryer that is flexible to
these different demands.
Freeze Drying
Once the product is properly frozen, it must be
sublimated (evaporated) at a low temperature
under reduced pressure.
The ideal curve of lyophilization with
temperature displayed on the ordained line, and
time displayed on the abscissa.
With the product maintained at a constant
temperature, it will be necessary to supply the
3
energy of sublimation, a combination of the
latent heat of fusion (which supplies the
transformation of the liquid to the ice state) and
the sublimation energy (about 700 calories per
gram of ice evaporated). The ability of vapor
release from the matrix is a function of
molecular agitation inside the matrix.
Ideally, the temperature of the frozen product
should be brought to the highest temperature
compatible with the frozen condition, without
exceeding it, which will lead to the irreparable
production of “foam” (commonly known as melt
back) and product deterioration. On the other
hand, if the heat energy is insufficient, the
product will sublime at too low a temperature,
and the length of the freeze-drying cycle will
become abnormally long.
After the disappearance of the final ice crystals,
the temperature of the product rapidly increases,
and must be maintained at the most maximum
permissible temperature to liberate the lowest
residual moisture embedded in the matrix
(secondary drying). The liquid shelves on which
the product is loaded transfer the required
energy of sublimation.
A chilled surface known as the ice condenser
collects the vapor from the evolving product.
During lyophilization, the pressure in the drying
chamber follows the fluctuations identified by
Curves P or P1.
Importance of Vacuum in the Freeze-Drying
Process
Freeze drying can only take place if the partial
pressure of the vapor in the drying chamber is
lower than the water vapor pressure above the
product. By strict definition, the vacuum in the
chamber is not essential, as sublimation can take
place at atmospheric pressure by passing
dehydrated air above the product. By artificially
lowering the pressure in the drying chamber the
time can be reduced.
For example, a product sublimated at –20 °C at
the given vapor pressure shown as 0.8 torr. As
soon as the value in the chamber reaches below
0.8 torr, the ice begins to sublime.
Flosdorff has shown that as soon as the pressure
in the chamber is reduced, evaporation increases,
but the rate of evaporation is not without limit,
and reaches a maximum when the pressure in the
chamber has a value equal to about 50% of the
vapor pressure above the product.
It was represented that the changes in
evaporation rate for a product which is
undergoing sublimation at –20 C, as a function
of the pressure in the chamber; the rate reaching
a maximum of 0.4 torr.
Contrary to widely held opinion, it is not
necessary to have a very low vacuum during the
sublimation period, because below the limit
defined, the evaporation rate is not improved,
and that too low a pressure acts as a barrier to
effective heat transfer.
Freeze Drying
Once the product is properly frozen, it must be
sublimated (evaporated) at a low temperature
under reduced pressure. With the product
maintained at a constant temperature, it will be
necessary to supply the energy of sublimation, a
combination of the latent heat of fusion (which
supplies the transformation of the liquid to the
ice state) and the sublimation energy (about 700
calories per gram of ice evaporated). The ability
of vapor release from the matrix is a function of
molecular agitation inside the matrix.
Ideally, the temperature of the frozen product
should be brought to the highest temperature
compatible with the frozen condition, without
exceeding it, which will lead to the irreparable
production of “foam” (commonly known as melt
back) and product deterioration.
On the other hand, if the heat energy is
insufficient, the product will sublime at too low
a temperature, and the length of the freeze-
drying cycle will become abnormally long.
After the disappearance of the final ice crystals,
the temperature of the product rapidly increases,
and must be maintained at the most maximum
permissible temperature to liberate the lowest
residual moisture embedded in the matrix
(secondary drying). The liquid shelves on which
the product is loaded transfer the required
energy of sublimation.
A chilled surface known as the ice condenser
collects the vapor from the evolving product.
(Curve F) During lyophilization, the pressure in
the drying chamber follows the fluctuations
identified by plotting the relations as curves.
4
Conclusion:
• By controlling chamber pressure, you can
reduce the heat transfer coefficient between the
warm shelves and the product. This simple
control will greatly improve the energy transfer
and reduce primary drying times. The curve of
shelf temperature is shown in schematic form in
B of Fig. 3, and the vacuum curve in P.
• The control of vacuum in the lyophilization
process can become a useful means of
controlling heat transfer, and the means of
getting energy to the product. A laboratory
example illustrates the influence of pressure in
heat transfer:
• A specially fitted freeze dryer equipped with
the necessary measurement and control
equipment was used to simulate these
sublimation phenomena. Freeze drying of the
product was carried out at –20 °C, with a
controlled pressure in the chamber in the range
of 0.4 torr, and a shelf temperature of 30 °C.
• When the injection of non-condensable gas
was terminated, the product cooled rapidly, and
the vapor removal rate slowed. To regain the
sublimation temperature of –20 °C, it was
necessary to bring the shelf temperature to 125
°C, a difference of 95 °C in the heating source to
produce the same sublimation temperature of the
product, and virtually the same evaporation rate.
• The chamber pressure acts as a thermal
regulator, which can, in the space of a few
moments can produce the same effect as raising
the temperature 95 °C.
Rate of Vapor Transfer to the Condenser:
Thermodynamic formulae enable the calculation
of values of the volume occupied by vapor as a
function of chamber pressure. For one gram of
water, this volume is in the region of 100m3 at a
pressure of 10-2 torr, and 1000m3 at a pressure
of 10-3 torr.
Consider a product with a eutectic of about –
20°C, freeze dried under the following
conditions:
• 500 ml bottle filled with 300ml liquid
product.
• A water content of approximately 250 grams.
• An evaporation surface of about 0.02 sq.
meters.
• A chamber pressure of 2 x 10-2 torr.
• The length of primary drying (sublimation of
the ice) around ten hours.
One gram of water occupies, under this pressure,
a volume of 50m3, and the amount of water
vapor which sublimates from the product per
second can be calculated from the following
formula:
• 50m3 x 250 = 0.35 m3 per/second
• 10 hrs x 3600
• The surface area inside the bottle being
around 0.02m2, the vapor leaves the product at a
speed of:
• 0.350 m3/s = 17.5 m/s0.02 m2
Inside the neck of the bottle, which has a
diameter of about 3 mm, or a section of 7 cm2 (7
x 10-4m2), the speed is calculated at:
0.350m3/s = 500 m/s, 7 x 10-4 m3
• This illustrates a very high rate advantageous
to shorter drying times. But, as the product
becomes lighter (the kinetic energy of the vapor
is proportional to the square rate), the crossing
of an already dry boundary carries product in the
flow (as much as 10 to 20%), and deposits itself
on the condenser where it is virtually
irrecoverable. In addition, these light product
particles can pollute the chamber area, where
sensitive sensing equipment may be located.
• When a non-condensable gas is introduced to
raise the pressure level, say to 0.2 torr, the
volume occupied by one gram of water (about
5m3), the speed through the neck of the
container is about 50 m/s. In short, the kinetic
energy of the vapor is divided by 100, and the
amount of product entrained in the vapor flow is
reduced to negligible amounts.
With Pressure x Volume Constant:
For one (1) gram of water at:
Atmospheric pressure (760 torr) we derive: 760
x 22.4 (l)1000
• At P=1 torr, V=1000 l or 1m3
• At P=10-1, V=1000, = 10,000 l, or 10m3
• At P=10-2 V=100m3
• At P=10-3 V=1000m3
• Pressure Vapor Velocity, Vapor Velocity,
Kinetic Energy
• Product container neck vapor (1/2mV2)
• 2x10-2 torr 17.5 m/s 500 m/s 100X
• 2x10-1 torr 1.75 m/s 50 m/s 1000X
• At the end of the sublimation phase, there is
sufficient time to re-establish very low pressure
to create the pressure differential conditions
needed for terminal drying.
The Role of the Condenser:
Given the thousands of cubic meters of water
vapor emitted from the product in a freeze dryer,
5
no vacuum pump, regardless of rate, could
remove these volumes directly. Therefore, a
vapor trap, or condenser is essential to condense
the evolving water molecules.
As the role of the vacuum pump is limited to
eliminating the tiny traces of non-condensable
gases, one could conclude that the real vacuum
effect is derived from the ice condenser (via the
pressure differential created by the surface
temperature of the product and the condenser).
You can note that each temperature value for
condensed ice corresponds to a vapor pressure,
the value of which is always lower than the
chamber pressure.
For example, if a machine is equipped with an
ice condenser for which the lowest temperature
limit is –40 °C, it would be impossible to lower
the pressure below the vapor pressure
corresponding to –40 °C, i.e. 10-1 torr.
This value is irrespective to the type or design of
the vacuum pump. A failure in the refrigeration
system leads to a chain reaction:
• A rise in condenser temperature.
• A rise in chamber pressure.
• A rise in product temperature.
• An irreversible eutectic melting and the boiling
of liquid fractions.
• Product failure.
Because of the importance of the refrigeration
system, a freeze dryer must be equipped with a
condenser designed and constructed with the
ability to:
• Condense all vapors from the product.
• Provide a vapor route of minimum distance to
avoid hindering vapor flow.
• Permit easy defrosting after the run.
• Prevent vapors from contaminating the oil in
the vacuum pump.
• Provide a simple cleaning operation.
• Provide the necessary BTU output under load
to condense vapor at a maximum rate without
disturbing the product’s selected primary
sublimation temperature.
• Insure the necessary low temperature
(saturated suction) during the secondary drying
to deliver the lower vacuum levels needed for
this phase.
• Provide a high degree of reliability.
Heating Control in the Freeze-Drying
Process:
The transmission of energy to the product needs
to be carefully controlled for three important
considerations:
• To avoid transferring too much heat and
passing the safe primary drying temperature.
• To avoid supplying insufficient heat, hence
prolonging the sublimation period.
• To achieve a homogeneous temperature in the
total batch, as the lyophilization cycle will be
determined by the temperature of the “coldest”
product area.
• The higher the sublimation temperature, the
faster the drying cycle. The conditions of low
temperature and high vacuum, can have, if they
are unnecessarily prolonged, an effect on living
organisms, which form the great bulk of freeze-
dried products.
Production Application:
In industrial freeze-drying applications it is
essential to reproduce identical drying protocols
for a wide range of products. To accomplish this
goal, it is vital to insure the same parameters for
each operation. The essential criteria are:
• The temperature of the product.
• The duration of the primary and secondary
drying phases.
• Heat input.
• Chamber pressure.
By a series of reproducible runs on a scaleable
the optimum cycle can be established. If the
process is automated it is essential that:
• Cycle is performed with a guarantee of
functional security.
• Validation is simple.
• Allows maximum flexibility for a variety of
product drying protocols.
In the pilot development phase, it is critical that
the favorable conditions for freezing and the
optimum parameters of primary and secondary
drying be established.
Secondary Drying:
At the end of the sublimation phase (primary
drying), all the ice will have disappeared. The
product will begin to rise in temperature and will
tend to approach the control temperature of the
shelf. However, at this stage the product is not
sufficiently dry for long term storage. For most
products, the residual moisture is in the region of
5% to 7%.
The product now enters the desorption phase,
during which the last traces of water vapor are
removed, along with traces of the “bound” water
6
within the product matrix. This phase is
identified as secondary drying. The aim of this
final phase is to reduce the product to the
acceptable moisture levels needed for long term
storage (3% to 1%).
The reasons for drying the product to these
levels are desirable for several benefits:
• When the water content is higher than these
levels, the product will denature.
• When the residual moisture is forced lower
than these levels, many products may undergo
chemical or enzymatic changes.
• Residual moisture in the product is generally
dependent on four factors:
• The product matrix (both in frozen and
sublimation mode.)
• The vacuum in the drying chamber.
• The duration of secondary drying (isothermic
phase).
• The maximum temperature allowed in the
product during secondary drying.
• Note: Products with excipients may exhibit
tendencies to retain water in the matrix and are
hygroscopic in ambient air. The final residual
moisture obtainable is dependent on more
conditions than the vacuum and moderate
temperature generally employed for this
secondary phase.
Good vacuum is essential so that the mean free
path of water molecules from the product matrix
is not impeded by pressure. The lowest possible
pressure is not necessarily desirable (below 10-2
torr) because below this pressure, desorption
produces an asymptotic rate, and the
phenomenon of “backstreaming” the vacuum
pump becomes more significant, thus risking
pollution the product matrix.
Factors affecting secondary (terminal) drying
phase:
By a series of reproducible runs on a scaleable
the optimum cycle can be established. If the
process is automated it is essential that:
• Cycle is performed with a guarantee of
functional security.
• Validation is simple.
• Allows maximum flexibility for a variety of
product drying protocols.
• In the pilot development phase, it is critical
that the favorable conditions for freezing and the
optimum parameters of primary and secondary
drying be established.
• Secondary Drying:
At the end of the sublimation phase (primary
drying), all the ice will have disappeared. The
product will begin to rise in temperature and will
tend to approach the control temperature of the
shelf. However, at this stage the product is not
sufficiently dry for long term storage. For most
products, the residual moisture is in the region of
5% to 7%.
The product now enters the desorption phase,
during which the last traces of water vapor are
removed, along with traces of the “bound” water
within the product matrix. This phase is
identified as secondary drying. The aim of this
final phase is to reduce the product to the
acceptable moisture levels needed for long term
storage (3% to 1%).
The reasons for drying the product to these
levels are desirable for several benefits:
• When the water content is higher than these
levels, the product will denature.
• When the residual moisture is forced lower
than these levels, many products may undergo
chemical or enzymatic changes.
• Residual moisture in the product is generally
dependent on four factors:
• The product matrix (both in frozen and
sublimation mode).
In considering a freeze dryer for production use,
the following general guidelines should be a part
of your determinations:
• The product’s lyophilization profile. (A vital
stage in making a determination of the
production freeze dryer’s performance
characteristics).
• This work should be carried out on scalable
pilot equipment.
Laboratory units, especially those equipped with
air cooled refrigeration systems cannot provide
the depth of data needed to determine a
production cycle. If the user does not have
access to an industrial pilot unit, this work can
be accomplished on an outsource basis at
relatively modest cost.
Determine the total length of the process cycle,
including all of the following factors:
• Preparation of the equipment.
• Dryer loading time.
• Freezing time.
• Freeze Drying time.
• Unloading time.
• Defrosting time.
7
• Clean up time required to make the area ready
for the next batch.
Heating Power, General Rules
• A standard industrial freeze dryer should
permit sublimation of approximately
Production Cycle Duration, General Rules
• Assuming the optimum rate of flow, an
estimate for a dryer of approximately 100 square
foot capacity would be:
• 100 kg of product in bulk (liquid loaded on
trays).
• 1000 kg to 1500kg of stainless steel (product
shelves, hoses, etc).
• 300kg to 500 kg of thermal medium (heated
and cooled).
• All of the above to be chilled or heated during
the drying cycle.
If the dryer is equipped with 10kw heating
employed 80% of the cycle, the shelves and heat
exchangers represent:
• 1000kg of stainless steel with a specific heat
of 12cal/kg per degree °C
• 500kg of thermal fluid with a specific heat of
0.36kcal/kg per degree °C (Silicone)
• A set point freezing temperature of –40 °C
and a terminal temperature of 40 °C (Shelf)
The amount of energy required would be:
Q=100kg x 800kcal + 1000kg x 0.12kcal x 80°
C + 500kg x 0.36kcal x 80= 100,000kcal
The time of sublimation would be
T= 100,000 / 10kw x 860kcal x 0.8= 15 hours
The secondary drying time must be added to this
total to deduce the total cycle time.
Secondary drying time is dependent on the
desired residual moisture content.
The energy of sublimation is about 800 kcal/kg
of water sublimated. The heating value of kw/hr
=860 kcal/hr. Therefore, the freeze dryer should
be furnished with heating capacity equivalent to
1 kw/sq. meter of surface area.
Refrigeration Power, General Rules
The freeze dryer’s condensing system should be
capable of condensing the sublimated vapor
liberated from the product at a temperature of –
55 °C to –65 °C, at a rate of 800cal/kg of water,
having a theoretical refrigeration capacity of
about 1000kcal/hr at -50 C/sq. meter of surface
area.
It is important that at the end of secondary
drying, the refrigeration system must be capable
of lowering the temperature of the ice condenser
to a temperature low enough to obtain the
necessary residual moisture in the product
Production Cycle Duration, General Rules
Assuming the optimum rate of flow, an estimate
for a dryer of approximately 100 square foot
capacity would be:
• 100 kg of product in bulk (liquid loaded on
trays).
• 1000 kg to 1500kg of stainless steel (product
shelves, hoses, etc).
• 300kg to 500 kg of thermal medium (heated
and cooled).
• All of the above to be chilled or heated during
the drying cycle.
If the dryer is equipped with 10kw heating
employed 80% of the cycle, the shelves and heat
exchangers represent:
• 1000kg of stainless steel with a specific heat
of .12cal/kg per degree °C
• 500kg of thermal fluid with a specific heat of
0.36kcal/kg per degree °C (Silicone)
• A set point freezing temperature of –40° C
and a terminal temperature of 40° C (Shelf)
The amount of energy required would be:
• Q=100kg x 800kcal + 1000kg x 0.12kcal x
80° C + 500kg x 0.36kcal x 80= 100,000kcal
The time of sublimation would be:
T= 100,00 /10kw x 860kcal x 0.8= 15 hours
The secondary drying time must be added to this
total to deduce the total cycle time.
Secondary drying time is dependent on the
desired residual moisture content
Bulk Drying, General Rules
As the interface separating the dried and frozen
regions increases; the frozen portion of the
matrix moves toward the lower surface of the
frozen material. This upper dry layer is highly
insulating to heat flux but is less sensitive at low
pressures to the phenomena of mass transfer
(pressure difference between product and
condenser induced by a temperature
differential).
• Heat transfer is increased by a pressure rise,
but at the expense of an increased resistance to
mass transfer.
• Water vapor migrating through the upper
surface of the matrix and the thermal
conductivity of the ice layer is two times higher
than that of the dry layer.
• In this condition, water vapor flux is
decreased as pressure is increased.
 8

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