SHORT AND BREIFE

Principles of Lyophilization
The material is first frozen and transferred to a drying chamber. During the drying stage, the
material in the chamber is subjected to a high vacuum. Heat is applied carefully to the material,
and a condenser is used in the chamber to collect the water. When water is leaving rapidly, its
heat of vaporization is taken from the material and helps to keep it cool and safe. As the material
dries, this cooling diminishes so that it is possible to overheat and damage the material.

Heat supplies the energy necessary for sublimation of the water. An ice crystal is composed of
pure water that is rather rigidly confined in a crystal lattice. The molecules have natural
vibrations, however, so that extra thermal energy increases the probability of breaking free.
When the water molecule breaks free, it diffuses through the already dried surface of the solid
and sublimes. As the water molecules diffuse and sublime, the thickness of the dry outer surface
of the specimen increases, and thus more energy is required to transport the molecules through
the dry shell. The actual force driving water vapor from the drying boundary, through the dry
shell and to the specimen surface, is a concentration gradient, and not, as some would assume,
the vacuum sucking on the sample.

As the molecules sublime and use up the latent thermal energy, the thermal reserves in the
specimen are depleted and thus the probability of further sublimation decreases. Rate of transfer
through the dried solids is low. The rate of drying of the specimen decreases until such time that
so much external thermal energy would have to be supplied that the specimen may be harmed.
Sublimation can continue safely if no heat is supplied, but drying time is greatly extended.

The removal of the water vapor that reaches the specimen surface is critical to completion of the
drying process. The water molecules that have successfully sublimed must be removed from the
free space of the vacuum. The molecules move through the vacuum-induced free space and are
trapped by condensation. Some condensers are plates, but a device known as a 'cold finger' is
common. The 'cold finger' is a long thin condenser.
Wet samples can be frozen by placing them in a vacuum. The more energetic molecules
escape, and the temperature of the sample falls by evaporative cooling. Eventually it freezes.
About 15% of the water in the wet material is lost.

The simplest form of lyophilizer would consist of a vacuum chamber into which wet sample
material could be placed, together with a means of removing water vapor so as to freeze the
sample by evaporative cooling and freezing and then maintain the water-vapor pressure below
the triple-point pressure. The temperature of the sample would then continue to fall below the
freezing point and sublimation would slow down until the rate of heat gain in the sample by
conduction, convection, and radiation was equal to the rate of heat loss as the more energetic
molecules sublimed away were removed.

This simple approach creates numerous difficulties. When a material is frozen by evaporative
cooling it froths as it boils. This frothing can be suppressed by low-speed centrifugation.
Centrifugation also helps to dry faster by reducing material thickness and exposing a greater
surface area.

An alternative is to freeze the material before it is placed under vacuum. This is commonly done
with small laboratory Lyophilizers where material is frozen inside a flask. The flask is then
attached to a manifold connected to the ice condenser. To speed the process the material can
be shell-frozen by rotating the flask in a low-temperature bath, giving a large surface area and
small thickness of material.

For larger-scale equipment it is usual to place the material on product-support shelves inside the
drying chamber, which can be cooled so that the material is frozen at atmospheric pressure
before the vacuum is created. Without a controlled heat in [put to the sample its temperature
would fall until drying was virtually at a standstill. For this reason it is usual to arrange a heat
supply to the product-support shelves so that, after their initial use for freezing the product, they
can be used to provide heat to replace the energy lost with the subliming water vapor and
maintain the product at a constant low temperature.
One milliliter of ice produces more than 1,000,000 ml. of water vapor at typical lyophilization
cycle pressures. The more energy-efficient vacuum pumps cannot handle large quantities of
water vapor. For this reason it is usual to fit a refrigerated trap (called the ice condenser)
between the lyophilization chamber and the vacuum pump. Modern lyophilizers incorporate
refinements.

The most important are listed below:

• Separated drying chamber and ice condenser to reduce cross-contamination

• Provision of an isolation valve between chamber and ice condenser to allow for end-point
determination and simultaneous loading and defrosting
• Construction of the chamber and ice condenser as pressure valves to allow for steam
sterilization at 121 C or higher
• Cooling and heating of the product -support shelves by a circulating intermediate heat-
exchange fluid to give even and accurate temperature
• Additional instruments to control, monitor, and record process variables
• Movable product-support shelves to close the slotted bungs used in vials and to facilitate
cleaning and loading
• Automatic control system with safety interlocks and alarms, duplicated vacuum pumps,
refrigeration systems, and other moving parts to enable drying to proceed without
endangering the product in the event of mechanical breakdown

Comparison with Liquid-Phase Drying

Lyophilization gives the opportunity to avoid denaturation caused by heating the product, by
maintaining it frozen throughout drying. This is the most obvious advantage over liquid-phase
drying.

Equally important is that in liquid-phase drying there is an undesirable shrinkage and

concentration of active constituents that causes damage as well as a movement of these
constituents to the surface of evaporation, where they form a dense, impermeable skin that
inhibits drying and later, rehydration. Such effects can be avoided by spray drying, but this
requires brief exposure to temperatures around 100 C.

Further advantages of lyophilization for parenterals products are that the wet material can be
dispensed accurately and can be sterile filtered just before filling into final containers so that
particulate and bacterial contamination is reduced.

Thus, the principle advantages of lyophilization as a drying process are:

• Minimum damage and loss of activity in delicate heat-liable materials

• Speed and completeness of rehydration
 • Possibility of accurate, clean dosing into final product containers
• Porous, friable structure

The principle disadvantages of lyophilization are:

• High capital cost of equipment (about three times more than other methods)
• High energy costs (2 - 3 times more than other methods)
• Long process time (typically 24-hour drying cycle)

Lyophilization should be used when the product meets one or more of the following criteria:
unstable; heat liable; minimum particulates required; accurate dosing needed; quick; complete
Rehydration needed; high value .
Some other less common applications of Lyophilization are recovery of water-damaged books
and manuscripts and preservation of archaeological specimens, tissue for spare-parts surgery,
museum specimens for display such as plants and animals, and vegetable matter for research
programs.

Lyophilization: Freeze-Drying
A Downstream Process
Reference: Snowman, John W. Downstream Processes: Equipment and Techniques, pages 315-351,
1988 Alan R. Liss, Inc.

term project by Lisa Menyhart

Integrated with an earlier term project by K. Theriault, INTRODUCTION TO FREEZE

DRYING.

This was a BASIC program that was published in

BINARY - Computing in Microbiology, 6: 19-20 (1994)

Freeze-drying, technically known as lyophilization, is a process of sublimation: water molecules

in a solid phase specimen are directly converted to free water molecules in the vapor phase.
The free water molecules are then trapped and removed from the process. Porous dried
specimens usually are easily rehydrated. The purpose of freeze-drying is to increase the shelf
life, or preserve a specimen, be it food, microbial organisms, or, in some circumstances to
decrease the size of the product. Different temperatures and drying times are used to freeze and
sublime, different specimens, depending on the nature of the specimen. Because lyophilization
is the most complex and expensive form of drying, its use is usually restricted to delicate, heat-
sensitive materials of high value.

A slightly different definition of lyophilization

Substances that are not damaged by freezing can usually be lyophilized so that refrigerated
storage is unnecessary. (Important exceptions are mammalian cells, nearly all of which are
destroyed by lyophilized.) Many microorganisms and proteins survive lyophilization well, and it is
a favored method of drying vaccines, pharmaceuticals, blood fractions, and diagnostics. Some
specialist food products are also lyophilized. They rehydrate easily and quickly because of the
porous structure left after the ice has sublimed. (The word lyophilized is derived from the Greek
"made solvent-loving")

Occasionally materials are lyophilized to achieve a porous, friable structure rather than for
preservation. Lyophilizers are sometimes used for concentration of delicate materials.
The form of the product and the type of container it is to be freeze-dried in influence the type of
lyophilizer needed and how it should be operated.