Lyophilization

JIHOČESCKÁ UNIVERZITA POLITEHNICA UNIVERSITY
V ČESKÝCH BUDĚJOVICÍCH OF TIMISOARA

CZECH REPUBLIC ROMANIA
ERASMUS
programme
LYOPHILIZATION
Univ.Prof. Eng. Dumitru MNERIE, PhD

“POLITEHNICA” University of Timisoara
ROMANIA
APRIL - MAY 2008

ČESKÉ BUDĚJOVICE
LYOPHLIZATION D. Mnerie
CONTENS
1. Introduction
1.1. Historical review
1.2. Evolution of process and equipment
1.3. Definition of lyophilization
1.4. General description of the process
2. Freezing Process
2.1. Primary Drying Process
2.2. Secondary Drying Process
3. Applications
3.1. Healthcare Industry
3.2. Veterinary
3.3. Food
3.4. Cereals
3.4. Other Applications
4. Traditional Lyophilization Technology
5. Lyophilization Equipment
6. Installation of laboratory
7. Conclusions
References
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1. INTRODUCTION
Freeze-drying (lyophilisation) is a widely used method for dehydrating a vast range of

materials, including foodstuffs, pharmaceuticals, biotechnology products, vaccines, diagnostics and
biological materials. It may be carried out on a range of scales, from bench top through pilot-scale
to a full-scale manufacturing process and offers a number of advantages over conventional drying
and many other processing methods. Despite its wide use, however, it is still apparent that many
regard freeze-drying as somewhat of an art. This is perhaps not surprising, given the lack of
available texts devoted to the process itself, with many published articles tending to describe
specific applications of the process. Specialised training courses are now arguably the most
effective means of learning about the various aspects of freeze-drying technology. This article seeks
to provide a brief overview of the process:
The foods conservation engineering - important part in program of human nutrition.
Interdisciplinary research, for:
- to improve human nutrition
- promote good health through new and traditional foods.
For all the people - the choices - the nutritional status of the World.
One of the best methods for foods conservation engineering is the lyophilization, (freeze-
drying)
That is a complex process that requires a careful balancing of product, equipment, and
processing techniques.
Freeze-drying (lyophilization) is a widely used method for dehydrating a vast range of
materials, including foodstuffs, pharmaceuticals, biotechnology products, vaccines,
diagnostics and biological materials.
The lyophilization process, also known as freeze-drying or sublimation, has many
advantages over other processing methods. Since freeze-drying is achieved at lower
pressures and temperatures than other methods, it is an inherently gentle process.
Lyophilization - drying, achieved by freezing the wet substance and causing the ice to
sublime directly to vapor by exposing it to a low partial pressure of water vapor.
In practice the substance may not be completely frozen, especially if no aqueous solutions
are present, and most lyophilization processes are completed by a period of desorption
drying.
The conventional drying (with hot air) has for results dehydrated products that can have a
warranty period of a year at most. Unfortunately, the quality of products dried by the conventional
method is inferior to that of the initial product. Freeze-drying is based upon the dehydration of the
frozen product through sublimation. Because this process does not require liquid water, and the
product is at a low temperature, most of the microbiological reactions and deterioration are stopped,
obtaining a final product of excellent quality. The water, being present in the freeze-drying process
as a solid entity, it protects the primary structure and the shape of the products with minimum
decrease volume. Despite the advantages, freeze-drying has been recognized as being the most
expensive method to make a dehydrated product. These processes can affect (partially or totally) the
quality of a product. Ended, a lot of changes can occur in the physical, chemical and/or biological
characteristics of the products during the process, storage and distribution.
For demonstrated outstanding qualities, the food products, preserved by drying are in great
demand on the market. Depending on the technology applied, they can finally have different
characteristics, differently appreciated qualities, and cost and sale prices also. For the producer, the
methodical concept integrated upon the production and the product can assure a technological
management suited to the requests of the consumer. To grant the product with more quality, there
can be used more expensive technologies. The dried food can be produced as the requests of the
consumer, with a decisive effect on the technical level necessary to be obtained by the producer.
The more we learn about freeze-drying, the more it reveals itself to be an exact (yet complex)
science, rather than the ‘art’ (or even scientific curiosity) it was historically thought to be. However,
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there is clearly much more to be done in order for the process to be understood more fully and
Biopharma Technology Limited strives to build upon its current position at the forefront of freeze-
drying technology to rationalize the process further using a scientific approach.
Freeze-drying (lyophilisation) is a unit operation in which a solvent, usually water, is frozen
and then sublimed in a vacuum. It is commonly used in the pharmaceutical industry when there are
stability issues with the active ingredient in solution, as is often true for proteins1. In order to
prevent processing defects during freeze-drying, active ingredients are formulated with excipients,
which may serve specific functions, such as providing bulk properties, thermal stability and activity
preservation to the product. Many groups of molecules have been shown to perform these functions,
including disaccharides, amino acids, polymers and non-ionic surfactants. The aim of this study was
to evaluate the correlation between the different analysis methods based on pre- and post-
lyophilisation properties of a range of protein-excipient mixtures. Success of these mixtures was
based on retained protein activity.
1.1. Historical review

The basic process of freeze-drying food is known to the ancient Peruvian Incas of the Andes.
They knew the basic process of freeze-drying food. Lyophilization, is the sublimation/removal of
water content from frozen food.The Incas were drying their frozen meat in the sun under the
reduced atmospheric pressure of high altitude. The dehydration occurs under a vacuum, with the
plant/animal product solidly frozen during the process. Shrinkage is eliminated or minimized, and a
near-perfect preservation results. Freeze -dried food lasts longer than other preserved food and is
very light, which makes it perfect for space travel. The Incas stored their potatoes and other food
crops on the mountain heights above Machu Picchu. The cold mountain temperatures froze the food
and the water inside slowly vaporized under the low air pressure of the high altitudes.
Freeze - drying methods have been used for over 100 years for various technical purposes.
During World War II, equipment and techniques were developed to supply blood plasma and
penicillin to the armed forces. In the late 70's freeze - drying was commonly used for taxidermy,
food preservation, museum conservation, and pharmaceutical production. It wasn't until the late 80's
the freeze - drying industry discovered the allurement and longevity of freeze -dried flowers.
There is no real invention of a freeze -dryer. It appears to have evolved with time from a
laboratory instrument that was referred to by Benedict and Manning (1905) as a "chemical pump".
Shackell took the basic design of Benedict and Manning and used an electrically driven vacuum
pump instead of the displacement of the air with ethyl ether to produce the necessary vacuum. It
was Shackell who first realized that the material had to be frozen before commencing the drying
process - hence freeze - drying. The literature does not readily reveal the person who first called the
equipment used to conduct this form of drying a "freeze -dryer". For more information on freeze -
drying or lyophilization, is referred to the book "Lyophilization - Introduction and Basic
Principles". Dr. Jennings' company has developed a number of instruments that are directly
applicable to the lyophilization process, including their patented D2 and DTA thermal analysis
instrument.
Everybody knows that, in the winter time, snow might disappear from the land, without
melting, by direct sublimation under radiant heat. It is claimed also, that, in the upper reaches of
their mountainous kingdom, the Incas were drying their frozen meat in the sun under the reduced
atmospheric pressure of high altitude. Freezing and subsequent drying from frozen state are thus
well known phenomena since time immemorial. However, only in the XXth Century this technique
has industrial applications.
During World War II, the freeze -dried process was developed commercially when it was used
to preserve blood plasma and penicillin. Freeze - drying requires the use of a special machine called
a freeze -dryer, which has a large chamber for freezing and a vacuum pump for removing moisture.
Over 400 different types of freeze -dried foods have been commercially produced since the 1960s.
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Two bad candidates for freeze - drying are lettuce and watermelon because they have too high a
water content and freeze -dry poorly. Freeze -dried coffee is the best-known freeze -dried product.
Drying is very old process used to preserve food. The term “drying” usually is to relegate to the
elimination of moisture from the substance.
1.2. Evolution of process and equipment

Freeze-drying has a great impact upon the production of dehydrated foods because of the
superior quality of the product obtained and promise continued expansion of the number of
applications.
Substances that are not damaged by freezing can usually be lyophilized so that refrigerated
storage is unnecessary.
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,
and museum specimens for display such as plants and animals, and vegetable matter for research
programs.
Ratti and Crapiste (1992) developed a lumped parameter model for hygroscopic shrinking
food systems. This model is represented by the following equation:
k g (a w p ws − p w∞ )
nw =
⎡ ⎛ φ ⎞ ⎤
⎢1 + ⎜⎜ ⎟⎟ Bimd ⎥
⎣ ⎝ X0 ⎠ ⎦ (1.1)
where Bimd is the Biot number for mass transfer defined as
Bimd=(kgLo)/(P1Drs) (1.2)
and P1 is the equilibrium relationship at the solid¯gas interface, which can be obtained from
P1=(X/pw)T. (1.3)
The parameter φ was shown theoretically and experimentally to be independent of drying

conditions and particle geometry, and only a function of moisture content
− 1, 079
⎛ X ⎞
φ = 5 ,32 ⋅ 10 −3
⎜⎜ ⎟⎟
⎝ X0 ⎠ (1.4)
The freeze-drying is analyzed under the viewpoint of the relationship between process
conditions, transport properties and drying rate.
The freezing process has a strong influence on the entire freeze-drying procedure.
Since a change in heat transfer has an effect on mass transfer and a change in pressure has
an effect on heat transfer, the relationship between the two processes is quite complicated. Thus,
differentiating between the two processes in order to determine which one is the limiting factor can
be difficult.
The transport properties – conductivity and permeability – mainly depend upon the structural
nature of the dried layer (porosity) and secondary on the operating factors such as pressure or
temperature.
Process improvements can be obtained if an accurate interpretation of the phenomena
involved is available.
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Figure 1.1. Basic Industrial Freeze Dryer
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
1.3. Definition of lyophilization

From dictionary: r.v., -dried, -dry·ing, -dries. To preserve (food, for example) is by rapid
freezing and drying in a high vacuum.
Freeze-drying, (lyophilization) means the sublimation of water content from frozen food.
Lyophilization is a process in which liquid materials are slowly frozen in a vacuum, thereby
removing water by maintaining a very low water vapor pressure. In practice the substance may not
be completely frozen, lyophilization processes are completed by a period of desorption drying.
The technology of lyophilization appear as a relatively simple process but as the practitioner
soon learns the process is deceptively complex and, as a result, is often treated as an art rather than
a science. Lyophilization - process, which extracts the water from foods and other products so that
the foods or products remain stable and are easier to store at room temperature (ambient air
temperature).
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- Drying, evaporation, desiccation, dehydration mean the processes of extracting moisture.

- Freeze, freezing are the withdrawal of heat to change something from a liquid to a solid.
Lyophilisation can be a method of drying food or blood plasma or pharmaceuticals or tissue
without destroying their physical structure; material is frozen and then warmed in a vacuum so that
the ice sublimes
Lyophilization can be a method of drying food or blood plasma or pharmaceuticals or tissue
without destroying their physical structure; material is frozen and then warmed in a vacuum so that
the ice sublimes
The fundamental principle in freeze - drying is sublimation, the shift from a solid directly into
a gas. Just like evaporation, sublimation occurs when a molecule gains enough energy to break free
from the molecules around it.
1.4. General description of the process

The technology of lyophilization appear as a relatively simple process but as the practitioner
soon learns the process is deceptively complex and, as a result, is often treated as an art rather than
a science.
Lyophilization is a process, which extracts the water from foods and other products so that the
foods or products remain stable and are easier to store at room temperature (ambient air
temperature).
The literature does not readily reveal the person who first called the equipment used to
conduct this form of drying a “freeze-dryer”.
The lyophilization (freeze drying):
a process in which water is removed from a product after it is frozen and placed
under a vacuum, allowing the ice to change directly from solid to vapor without
passing through a liquid phase.
The process consists of three separate, unique, and interdependent processes:
freezing
primary drying (sublimation),
secondary drying (desorption).
The theoretical principle of freeze drying is clearly defined in the diagram "Pressure
Temperature". 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 in this diagram,
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.
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.
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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. The temperature of the product stabilizes after time period at about 0° C. At Point
B, the ice crystals previously formed have expanded, and consist practically of pure water. At Point
C, 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. At
Point D, 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.
It has 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, Point D on the curve, represent the temperature of
the complete freezing of the product. The establishment of this eutectic zone is very important.
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 is depicted on Curve,
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
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
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evaporated). The ability of vapor release from the matrix is a function of molecular agitation inside
the matrix.
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. (Curves B, C, or D represent the
heating rate)
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.
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.
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.
The ideal curve of lyophilization is depicted, with temperature displayed on the ordinand line,
and time displayed on the abscissa.
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.
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2. FREEZING PROCESS
The primary goal of research on freeze-drying is to improve process economics by reducing
processing time. An important step in this process is determining the rate limiting factors.
Heat transfer and mass transfer are the two processes that are rate controlling processes.
Understanding the mechanisms of heat and mass transfer and their dependence on product
temperature and pressure in the drying chamber is very important for determining the rate limiting
process. If the outer surface temperature limit is encountered first as surface temperature of the
dried layer is raised, the process is considered to be heat transfer controlled; to further increase the
drying rate, in the food drying case, the thermal conductivity of the dried layer must be raised. If the
melting point temperature is encountered first, then the process is considered to be mass transfer
limited, and in order to increase the drying rate, the effective diffusivity of water vapor in the pores
of the dried layer and total mass flux must be raised. This means an increase of the convective
velocity of the vapor in the pores of the dried layer. The values of effective diffusivity and total
mass flux could be raised by decreasing the pressure in the drying chamber.
To extract water from foods, the process of lyophilization consists of:
• Freezing the food so that the water in the food become ice;
• Under a vacuum, sublimating the ice directly into water vapour;
• Drawing off the water vapour;
• Once the ice is sublimated, the foods are freeze-dried and can be removed from the machine.
Figure 2.1. Diagram pressure – temperature.
There are two major factors that determine what phase (solid, liquid or gas) a substance will
take: heat and atmospheric pressure.
For a substance to take any particular phase, the temperature and pressure must be within a
certain range. Without these conditions, that phase of the substance can't exist.
Lyophilization is carried out using a simple principle of physics called sublimation.
Sublimation is the transition of a substance from the solid to the vapor state, without first passing
through an intermediate liquid phase.
Water will sublime from a solid (ice) to a gas (vapor) when the molecules have enough energy
to break free but the conditions aren't right for a liquid to form. The chart below shows the
necessary pressure and temperature values of different phases of water. The water can take a liquid
form at sea level (where pressure is equal to 1 ATM) if the temperature is in between the sea level
freezing point (32 degrees Fahrenheit or 0 degrees Celsius) and the sea level boiling point (212 F or
100 C). But it increase the temperature above 32 F while keeping the atmospheric pressure below
06 atmospheres (ATM), the water is warm enough to thaw, but there isn't enough pressure for a
liquid to form. It becomes a gas.
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Figure 2.2. - The transformation of phases for lyophilization process.
Freeze-drying is a multiple operation which involves the following three stages:

1. the freezing stage,
2. the primary drying stage (sublimation), and
3. the secondary drying stage (desorption).
The first step in the lyophilization of a product is to convert it into a frozen state. The material
is cooled down to a temperature that is always below its freezing point, which depends on the nature
of the product. It is important that during the freezing process that solvent (water) is crystallized.
The shape of the pores, the pore size distribution, and pore connectivity of the porous network of
the dried layer formed by sublimation of frozen water during the primary stage depend on the ice
crystals that formed during freezing stage [7-10]. This dependence is very important because the
parameters that characterize the mass and heat transfer rates in the dried layer are influenced by the
porous structure of the dried layer and the performance of the overall drying process depends
significantly on this stage.
2.1. Primary Drying Process

After freezing stage, the chamber pressure is reduced to a value that would allow the
sublimation of the ice. For sublimation, the applied heat may come from warmed fluid flowing
through the shelves or walls of the chamber, or from microwave energy, or any conceivable source.
The maximum allowable temperature that the frozen layer could tolerate, without loss of product
property or stability, is denoted by convention the melting temperature of the sublimation interface
of the frozen layer.
There have been many experimental studies of sublimation rates and mechanisms of mass
transfer through the dried layer during freeze-drying.
In a study of sublimation rates for aqueous solutions, Pikal et al took measurements of weight
loss on a cylindrical micro-sample suspended from a vacuum micro balance kept at a constant
temperature. The variables in the experiments were freezing rate, product thickness, temperature,
pressure and solute concentration. An important result of this work was the conclusion that at higher
temperatures hydrodynamic surface flow is an important flow mechanism in the dried layer.
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Livesey and Rowe, in their discussion on the effect of chamber pressure, noted that the rate
limiting process in freeze-drying changes as sublimation proceeds. Initially, the process is limited
by heat transfer when the dried layer is small and an unrealistic heat flux is required to push the
sublimation rate to its maximum. After the dried layer thickens, the process becomes limited by
mass transfer since the required heat flux is easily maintained for the decreasing sublimation rate.
Wolff and Gibert determined the rate limiting factor in vial freeze-drying of milk. By fitting
their model to measured data, they were able to determine three transport parameters in their model.
These were the water vapor diffusivity in the dried layer, the external mass transfer coefficient and
the resistance to the heat transfer from the heated shelf to the ice. Since the contact resistance
between the vial and the shelf is the most significant barrier to heat transfer, it was determined to be
the overall rate controlling parameter and thus controlled the drying kinetics.
The water vapor produced by sublimation travels by diffusion and convection the porous
structure of dried layer and enters in the drying chamber of the freeze dryer. The water vapor must
be continuously removed from drying chamber in order to maintain non-equilibrium conditions for
the drying process in the system. The time at which there is no more frozen layer is taken to
represent the end of the primary stage
During the primary drying phase the pressure is lowered and enough heat is supplied to the
material for the water to sublimate. The amount of heat necessary can be calculated using the
sublimating molecules’ latent heat of sublimation. In this initial drying phase about 98% of the
water in the material is sublimated. This phase may be slow, because if too much heat is added the
material’s structure could be altered.
In this phase, pressure is controlled through the application of partial vacuum. The vacuum
speeds sublimation making it useful as a deliberate drying process. Furthermore, a cold condenser
chamber and/or condenser plates provide a surface(s) for the water vapour to re-solidify on. This
condenser plays no role in keeping the material frozen; rather, it prevents water vapor from reaching
the vacuum pump, which could degrade the pump's performance. Condenser temperatures are
typically below −50°C (-58°F).
2.2. Secondary Drying Process

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.)
• The vacuum in the drying chamber.
The secondary drying stage starts when all the ice has been removed by sublimation. In the
secondary stage the water that did not freeze (bound water) is removed. The bound water is due to
physical adsorption, chemical adsorption and water of crystallization. Since the amount of the non-
frozen water is about 10 – 35% of the total moisture contents, its effect on the drying rate and
overall drying time is very significant.
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The bound water is removed by desorption which takes place by heating the product under
vacuum. Moisture may move by vapor diffusion through the solid under a vapor pressure gradient.
Usually the pressure is also lowered in this stage to encourage sublimation. However, there
are products that benefit from increased pressure as well. After the freeze drying process is
complete, the vacuum is usually broken with an inert gas, such as nitrogen, before the material is
sealed.
Secondary drying is the process of removing the unfrozen or bound water contained in the
amorphous material. The mechanism for removal of the unfrozen moisture is primarily desorption.
This means that release of the bound moisture can be predicted by desorption isotherms. A
desorption isotherm gives the moisture content as a function of humidity for a given temperature.
Using these data one can determine a temperature-pressure protocol for secondary drying that will
bring the residual moisture content to the required level. Water will follow the same path as in
primary drying, from the material to the condenser. However, in secondary drying, diffusion of
unfrozen water from within the material to its surface and subsequent desorption of that water
become the most important mechanisms.
The primary goal of research on freeze-drying is to improve process economics by reducing
processing time. An important step in this process is determining the rate limiting factors. Since the
freeze-drying rate is limited by the rates of heat and mass transfer across a dried layer, the values of
its thermal conductivity and permeability are indispensable to determine the drying rate. These
values are mainly governed by the structure of the dried layer and operating factors such as
temperature and pressure. Since the structural nature of the dried layer is also affected by freezing
operations, the freezing condition, the structure and transport properties of the dried layer are
fundamental information to design the optimum drying cycle and to control the quality of final
products.
Although the basic concepts of freeze-drying are known, the details are not well understood.
Fundamental knowledge in some areas concerning the physical processes of freeze-drying is not
complete. Optimum parameter settings are usually found by trial and error, a lengthy and expensive
procedure. The understanding of the fundamental phenomena involved in this complex process
allows process optimization. However, fundamental knowledge of the physical and chemical
processes of freeze-drying needs to be put into a form that is useful to equipment designers and
users. Combining this fundamental knowledge with process development research can result in an
improved freeze-drying process for food industry and not only.
The freezing stage represents the first separation step in the freeze-drying process, and the
performance of the overall freeze-drying process depends significantly on this stage. The structural
nature of the dried layer is mainly affected by freezing operations. The fast freezing before freeze-
drying leads to smaller pore spacing after freeze-drying. It follows that faster freezing should lead to
lower thermal conductivities at a given pressure. If a freeze-drying process is rate limited by
internal heat transfer, the rate of freeze-drying for fast-frozen material should then be less than that
of a slowly frozen material.
The relationships among the freezing condition, structure and transport properties (thermal
conductivity and permeability) of the dried layer are fundamental information to design the
optimum drying cycle and control the quality of final products. As long as no collapse occurs,23
during the primary and secondary drying, the space previously occupied by the ice crystals becomes
the main passageway for vapour transport.24 Different morphologies of solidification texture may
results depending on the chosen solidification parameters. These different textures then lead to
different resistances against vapor transport during drying. In fact, the porous structure of the dried
product determines the values of the heat and mass transfer coefficients, and thus the drying rate.
The quality of the final freeze-dried product is greatly influenced by the freezing process. For
instance, the freezing process controls the color and the flavour of freeze-dried coffee extract or
retention of volatile compounds. Proteins cells, subjected to freeze-drying, can be damaged during
the freezing process already.
13
One important goal of the freezing step is to produce a uniform product batch, which is
difficult because of the stochastic nature of nucleation. The degree of super-cooling, defined as the
difference between the equilibrium freezing point and the temperature at which ice crystals first
form, is both a statistical or random event as well as one that depends on the solution properties and
process conditions. The degree of super-cooling is important because it determines the number of
nuclei at any time, and thus determines the number of ice crystals formed. More ice crystals from
the same amount of water means smaller crystals, which means smaller pore size and thus longer
primary drying time. Nucleation rates increases rapidly with the degree of super-cooling, yielding
ice crystal sizes that are inversely proportional to the degree of super-cooling. Thus, as super-
cooling is increased, the resulting increases in nucleation rate cause the ice crystal particle size
distribution to shift to smaller sizes. In turn, this produces pores with lower volume/surface area
rations, which results in decreased diffusive flux and slower sublimation.
Theoretically, solidification may result in different patterns: the interface may be planar, i.e.,
smooth and flat, cellular (columnar) or dendritic, i.e. needle, tonque or finger – shaped, or tree-like
with side branches.
A large portion of most freezing processes used in freeze-drying may be regard as directional
solidification, i.e. the growth direction of the solidification interface is opposite to the main heat
flow direction. The solidification pattern is governed by two parameters: velocity of solidification
interface general case and mean temperature gradient at the interface ice-liquid interface in the
direction of ice finger growth. A complete description of the geometric form of dendrites or cells in
freezing aqueous solutions is difficult to achieve because of complexity of the theoretical models.
Y. Sagara studied the influence of the freezing stage on the transport properties in the foods
freeze drying. He found a linear relationship between permeability and ice crystallization time for
mashed sample. The freezing rate of the group A was relatively larger than the group B. This
behavior is attributed to the larger ice crystals formed in the sample as the freezing rate decreased.
The effects of the freezing rate on the permeability were found to be critical for the mashed cellular
food materials. In liquid food systems the porosity of the dried layer is governed by the solid
content or concentration, and thus the porosity mainly affects the thermal conductivity of the dried
layer as shown in Fig.2.3.
Fig. 2.3 - Permeability vs. ice crystallization time. Fig. 2.4 - Thermal conductivity vs. porosity
14
In addition to concentration, the permeability is influenced significantly by freezing manner,

because the capillary type ice columns is formed by ice crystals along with the direction of heat
flow during freezing. Consequently, the morphology of capillary type void as well as grain
orientation and pore size of the dried layer are arranged during freezing process and thus the
transport properties of the dried layer is decisively governed by the structure fixed during freezing.
From these results, it would be suggested for liquid materials that under the constant solid
concentration the material should be frozen in a manner to form straight ice columns whose
orientation is parallel to that of heat transfer during drying and also make larger ice crystals by
employing the slower freezing rate.
If the higher permeability coefficient could be obtained by controlling freezing method, the
drying rate would be limited by heat transfer rate across the dried layer. Under this condition the
surface temperature of the dried layer is allowed to increase within a certain range that is decided
from viewpoint of quality control for final product.
The freezing process thus has a strong influence on the entire freeze-drying procedure.
The main processing factors in freeze-drying are product temperature and chamber pressure.
The freeze-drying process should be conducted within certain temperature and pressure limits in
order to avoid product damage during drying process.
During the primary drying stage there are two temperature constraints (limits). First, the
surface temperature of the dried layer must not become too high because of risk of thermal damage
– loss of bioactivity, color change, degradative chemical and biochemical reactions, structural
deformation in the dried layer; the second, the temperature at the interface must be kept below the
melting temperature, which may be 10ºC or more below the melting point of ice. The melting at the
sublimation interface, or any melting that occur in the frozen layer, can give rise to gross material
faults such as puffing, shrinking, and structural topologies filled with liquid solution. When melting
has occurred in the frozen layer, then the solvent at that point cannot be removed by sublimation. If
the material has a glass form and the minimum freezing temperature is exceeded during primary
drying stage then the phenomenon of collapse can occur. The product collapse causes a loss of
rigidity in the solid matrix. The maximum allowable temperature in the frozen layer is determined
by both structural stability and product stability factors.
During secondary drying the moisture concentration and temperature in the sample could vary
widely with location and time and this implies a potential for product alteration. Since very many
products are temperature sensitive, it is usual to control product stability by limiting the value of the
temperature during secondary drying process and then the final moisture content.
The pressure constraints (limits) are more complex to define because of the pressure effect on
both the heat transfer (the pressure has effect on the thermal conductivity of the dried layer) and the
mass transfer (the pressure has effect on the effective diffusivity of water vapor in the dried layer
and the total mass flux). In the dried material, the effective diffusivity varies significantly with the
total pressure and with the type of gas present. At very low pressures, independent of the
surrounding gas, the thermal conductivity reaches a lower asymptotic value. This reflects the
geometric structure of the solid matrix with no contribution from the gas in the voids of the material
because the pressure is very low. At high pressures, the thermal conductivity reaches a higher
asymptotic value and is characteristic of the heterogeneous matrix composed of solid material and
gas voids. The thermal conductivity is dependent of the gas composition and its thermal
conductivity. There is a transition between asymptotes corresponding to 0.1 and 100 mmHg
pressure range. The pressure effect on the mass transfer is explained by the fact that the effective
diffusivity of water vapor in the dried layer and the total mass flux are function of the total pressure
in the drying chamber. The magnitude of the effective diffusivity decreases if the total pressure is
increased. Furthermore, when the total pressure is increased, the gradient of total pressure in the
dried layer is reduced, and this decreases the convective velocity and total mass flux.
Above a certain pressure, the freeze drying process becomes internal mass transfer controlled.
The highest rate mass transfer control will occur at the pressure of the transition from heat transfer
control to mass transfer control (diffusivity and mass flux decrease with increasing pressure and the
15
thermal conductivity increases with pressure), and the attainable drying rate will decrease at higher
pressures.
There has been considerable interest in investigating the factors affecting the batch time of
freeze-dryers because these are most amenable to control, and efforts have been made to minimize
the batch time.
Nail studied a typical pharmaceutical freeze-drying situation: small glass vials of a frozen
product sitting on a metal tray which is placed on a heated shelf in the freeze-dryer. He states that in
some situations mass transfer may be the rate limiting factor, but that heat transfer from the heat
source to the sublimation front is usually the rate limiting process. Nail established that resistance to
heat transfer caused by air gaps between the vial and the tray and between the tray and the heated
shelf becomes significant in the low pressure environment of freeze-drying. This is explained by the
reduction of the thermal conductivity of a gas at pressures in the range where free molecular flow
occurs. Nail's theory shows that the drying rate decreases with decreasing pressure.
Pikal et al determined that the resistance of the dried layer decreases with decreasing pressure
and that a lower pressure increases the sublimation rate.
Livesey and Rowe considered a situation in which the temperature of the product was at its
optimum and pressure was increased. This would result in the decrease of the sublimation rate,
because the heat flow to the product had to be reduced to maintain the temperature at its optimum
value without overheating. Another situation considered was one in which the shelf temperature
was kept at a constant value. In this situation increasing the pressure resulted in an increased heat
flow, increased product temperature and an associated increase in sublimation rate.
In a study that focused on the secondary drying stage, Pikal et al determined that pressure
variation in the range of 0 - 0.2 Torr had no effect on drying rate during secondary drying for the
materials used. However, it was determined that temperature has a significant effect on the drying
rate and that the rate limiting process was either desorption from the solid-vapor interface or
diffusion in the solid.
The transport properties, such as the thermal conductivity and the permeability, have a great
influence on the processing factors, and finally on the drying rate. Various methods, both transient
and steady state, have been proposed to determine the transport properties of freeze dried food, the
latter gives better accuracy.
To analyze the effects of the structural parameters on transport properties and drying rate, one
physical and mathematical model is necessary.
To this end, let consider a material to be dried on a tray (Fig. 2.5) The thickness of the sides
and bottom of the tray is small and thermal conductivity of the material of tray is high, such that the
resistance of the tray to the heat transfer could be considered negligible. As a result, the temperature
of the tray can be considered equal with the temperature of the heating plate.
Fig. 2.5 - Drying of the material in a tray
16
Heat qI could be supplied to the material by conduction, convection and /or radiation.
The heat qII is supplied by a heated plate and is conducted through the bottom of the tray to
the material. The heat qIII represents the amount of heat transferred between the environment in the
drying chamber and the vertical sides of the tray. The contribution of the qIII is negligible when
compared to the contribution of qI and qII.
The study of the freeze-drying can be done in the following cases (Fig 2.5):
1. qI ≠ 0 and qII = 0,
2. qI = 0 and qII ≠ 0 and
qI ≠ 0 and qII ≠ 0.
The transport properties of the dried layer were determined by applying the drying data to a
model based on heat and mass transport in a sample, and then the effects of processing parameters on
transport properties and drying rate in connection with the freezing operations. The theoretical model
used by Y. Sagara to determine the thermal conductivity and permeability is shown in Fig. 2.6.
Fig. 2.6 - Freeze-drying model for transport properties analysis
The sample is assumed to have the geometry of a semi-infinite slab dried layer is separated
from the frozen layer by an infinitesimal sublimation interface retreating uniformly from the sample
surface. The bottom of the sample is insulated while the surface is exposed to an evacuated space at
the temperature θs and pressure ps. Since the thermal conductivity of the frozen layer is 20 - 50
times greater than that of the dried layer, the temperature of the frozen layer can be supposed to be
uniform and same as that of sublimation front.
An approximate method was developed for predicting the structural parameters of the dried
layer by assuming this layer to be a bundle of capillary tubes with the pore space having an
equivalent pore radius, porosity and tortuosity factor.
The expression of the rate of heat transfer across the dried layer according to Massey and
Sunderland, is:
θs
⋅ λ ⋅
(2.1)
q= (θ s − θ f ) − m ∫ c p dθ
m (t ) θf
The mass flux may be expressed as

⋅ KM w
m= grad . p (2.2)
RT
Equations representing the thermal conductivity k and permeability K are presented by Sagara
and expressed as:
17
θs
k = αρ w l (ΔH + ∫ c pw dθ )
2
(2.3)
θf
K = βρ w l 2 RT f / M w (2.4)
where:
1− m
α= (2.5)
(θ s − θ f )(−dm / dt )
1− m
β= (2.6)
( p s − p f )(− dm / dt )
The thermal conductivity was found to increase with increasing the solute concentration and is
markedly affected by the porosity of the dried layer and the permeability was found to increase with
pressure and temperature of the dried layer. This behavior is in good agreement with Mellor and
Lovett’s theoretical investigations based on the collision theory, and also with their experimental
results obtained for several kinds of solutions. The expressions for the permeability coefficient
given by Mellor and Lovett are:
ε
K= DK Ω (2.7)
τ
where:
3π r π 2r / λ 1
Ω= + + (2.8)
64 λ 4 (1 + 2r / λ )) 1 + 2r / λ
Ω indicates the contributions due to Poiseule`s flow, slip flow and Knudsen`s flow.
The mean free path of water vapor molecule λ is given by:
κ ⋅T
λ= (2.9)
2 ⋅ πσ w2 p
and Knudsen diffusivity is defined in terms of the pore radius r and the average molecular
velocity as follows
2
DK = vr
3 (2.10)
where:
1/ 2
⎛ 8RT ⎞
v = ⎜⎜ ⎟⎟ (2.11)
⎝ π M w ⎠
In the Sagara`s the experimental data, the permeability was found to depend mainly on the
porosity (Fig. 2.7) of the dried layer and then other factors such as temperature or pressure of it.
One can conclude that the transport properties mainly depend upon the structural nature of the
dried layer and secondary on the operating factors such as pressure or temperature.
As previously discussed, freeze-drying is an expensive and lengthy process, but one that can
be improved. The characteristics and physical properties of the material, the mechanism of the
drying that occurs (depending on the structure of the ice crystals) and operating conditions, in
particular the way heat is supplied, determine which factor becomes rate controlling. One way to
improve the process is to initially look at the physical mechanisms of drying in the primary and
secondary drying stages, then determine the rate limiting mechanisms and subsequently alter the
characteristics of the material and/or process which are responsible for the rate limitations.
Modification of operative conditions for enhancing the transfer fluxes is used. The injection of
inert gas is sometimes used as a control variable in lyophilization process.
18
Fig. 2.7 - Permeability vs. porosity of the dried layer
In the case of the processes limited by heat transfer through dried material, which is
determined by the thermal conductivity of the porous solid, it was proposed to use nitrogen,
hydrogen or helium pulsed atmospheres. The diffusion coefficient of these gases is sufficiently high
to allow them to diffuse into the solid countercurrent to the water vapor, increasing two to threefold
the heat transfer as effect of their high thermal conductivity. The optimal pressure cycle must be
determined, depending not only on the characteristics of the product but also on the size of the
chamber and on the performances of condenser and pump, and it must be avoided that the
temperature oscillation at the interface be of such an amplitude to cause melting.
Mellor evidenced that a cyclic-pressure process can significantly reduce the time as not only
the effective thermal conductivity of the porous solid increases during the part of the cycle in which
the pressure increases, enhancing the heat transfer, but also the vapor transfer mechanism is altered
from bulk diffusive flow to a much more effective combination of viscous and free flow.
Liapis et al pointed out that an increase in pressure increases the thermal conductivity of the
solid but decreases the temperature driving force, as a consequence of the increased resistance to
mass transfer that causes the ice temperature to rise. It is important to find an optimal pressure
which maximizes the heat transfer. However, in practice was shown that an optimal pressure policy
brings only minor improvements in drying time when compared to a policy at constant pressure,
especially if the additional complexity and cost of the apparatus are considered.
Mellor discussed the effect of pressure on heat and mass transfer in freeze-drying. He noted that
low pressure increases the driving force for mass transfer, but reduces heat flow in the dried layer
because of the dependence of heat transfer coefficients on pressure. In an attempt to overcome this
problem, Mellor developed a cyclic pressure freeze-drying process. In this process, pressure is
momentarily increased to increase heat flow and then momentarily decreased to increase vapor flow.
Litchfield et al. studied a typical vial freeze-drying case using a numerical simulation that
includes a resistance to heat transfer between the bottom of the vial and the heated surface. In this
study, Litchfield et al. found that even the best of the cyclical pressure policies was not as good as
an optimal constant pressure policy previously determined by Liapis and Litchfield.
The heat required for ice sublimation is usually supplied by conduction or radiation. Different
techniques were studied for improving the efficiency of energy transfer, but technical difficulties or
poor quality control have limited their use.
19
The main driving force for the movement of water is a pressure difference. The high pressure
is at the sublimation interface where the water vapor is generated and the low pressure is on the
surface of the condenser where the water vapor condenses to form ice again. Most of the low
pressure range typically encountered in freeze-drying, this resistance is at least partially dependent
on pressure: decreasing pressure increases the resistance. Thus, when the chamber pressure is
decreased, the heat flow to the sublimation interface decreases and subsequently the drying rate
decrease.
A better design for a freeze-dryer would be one in which changes in pressure do not affect the
ability to provide heat to the sublimation interface. If heat is supplied volumetrically, as with
microwave energy, the working chamber pressure could be lowered and the drying rate increased.
Previous attempts at using microwave energy to accelerate the freeze-drying process have
focused on food applications. It is evident that incorporating microwave energy into the
pharmaceutical freeze-drying process has the potential to reduce the overall processing cost by
reducing the processing time. The most significant advantage to volumetric heating is the ability to
decrease chamber pressure without a detrimental effect on heating. Other advantages are the ability
to heat the dried layer during primary drying and improved control of heating. The benefit of
heating the dried layer during primary drying is that secondary drying will start at lower residual
moisture content, thereby significantly reducing the secondary drying time. Improved control of
heating results from the heating rate being reduced as the volume of ice is reduced and also power
can be quickly adjusted allowing for an optimized power schedule.
Process improvements can be obtained if an accurate interpretation of the phenomena
involved is available. Indeed, chemical engineering research can give a sound contribution to
freeze-drying processes by studying kinetics, mass and heat transfer.
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
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.
20
3. APPLICATIONS
As a leading multi-product freeze drying company, European Freeze Dry works with
manufacturing companies both large and small to develop appropriate drying processes for a
diverse range of products. Over the years, our company has acquired a wealth of expertise in freeze
drying in a wide range of applications:
• Specialty Chemicals
• Natural Enzymes
• Biological Products
• Pigments
• Resins
• Active Pharmaceutical Intermediates
• Food Ingredients
• Flavorings
• Flood damage/Document Recovery
The freeze drying process is particularly suitable for products which are sensitive to heat,
oxidation, or are shear sensitive. The key benefits of freeze drying are:
• Retention of structure and surface area
• Retention of morphological, biochemical and immunological properties
• High viability/activity levels maintained through the drying cycle
• Lower temperature, oxygen, and shear conditions compared to other drying methods
• High recovery of volatiles
• High yield
• Long shelf life of dried product
• Reduced weight for shipping, storage and handling
3.1. Healthcare Industry

Freeze-drying (lyophilisation) is commonly used in the pharmaceutical industry when there are
stability issues with the active ingredient in solution, as is often true for proteins.
In order to prevent processing defects during freeze-drying, active ingredients are formulated
with excipients, which may serve specific functions, such as providing bulk properties, thermal
stability and activity preservation to the product.
Many groups of molecules have been shown to perform these functions, including
disaccharides, amino acids, polymers and non-ionic surfactants. The aim of this study was to
evaluate the correlation between the different analysis methods based on pre- and postlyophilisation
properties of a range of proteinexcipient mixtures. Success of these mixtures was based on retained
protein activity.
Pharmaceutical companies often use freeze drying to increase the shelf life of products, such as
vaccines and other injectables. By removing the water from the material and sealing the material in
a vial, the material can be easily stored, shipped and later reconstituted to its original form for
injection.
Supercritical during. Freeze drying is a method of drying food or blood plasma or
pharmaceuticals or tissue without destroying their physical structure; material is frozen and then
warmed in a vacuum so that the ice sublimes
lyophilisation, lyophilization
21
3.2. Veterinary
Taxidermy is a general term describing the many methods of reproducing a life-like, three-
dimensional representation of an animal for permanent display. The word "taxidermy" is derived
from two Greek words; taxis; meaning movement; and derma; meaning skin. The modern practice
of taxidermy incorporates many crafts, such as carpentry, woodworking, tanning, molding and
casting; but it also requires artistic talent, including the art of sculpture, painting and drawing.
Because of the diversity of specimens being freeze-dried for taxidermy purposes (from
Brahma bull head mounts to minnows used for fishing bait), trial and error has often been the
method used to determine how different taxidermy specimens should be pre-frozen and freeze
dried. The success or failure often directly relates to the proper pre- freezing of the taxidermy
specimen, and the temperature at which the specimen chamber is held during the drying stage.
Figure 3.1.
When frozen, various taxidermy specimens will totally solidify at different temperatures. The
temperature of complete solidification is the taxidermy specimen’s eutectic temperature. The
American Heritage Dictionary, 1969, defines eutectic as, "the lowest possible temperature of
solidification for any mixture of specified constituents"
An example of the variance found in freezing temperatures is that of water - not all water
freezes at 32 degrees F. When water is contaminated with other elements, such as salt, nitrogen,
mercury, industrial waste, etc., it's temperature of complete solidification will change. Since one of
the basic premises if freeze dry taxidermy is animals are made up of about 70% water by weight,
the importance of eutectic temperature is readily evident.
When determining eutectic temperatures, taxidermy specimens may be divided into two
general groups: those that contain large amounts of body fats and oils (most fish, certain species of
ducks, amphibians, etc.) and those that do not (the majority of mammals, reptiles, game birds,
delicate flowers, etc.) Because of the chemical makeup, taxidermy specimens in the group (greasy
specimens) require lower eutectic temperatures than the second group (non-greasy specimens).
Because natural salts are usually found in combination with water in taxidermy specimens,
pre-freezing should be done in the shortest time possible. Slow freezing times lead to salt
concentration in the specimen resulting in lower eutectic temperatures and increased chance for
shrinkage. Rapid freezing reduces salt concentrations and also results in smaller ice crystals, which
create less tissue distortion.
Rapid freezing may be obtained by freezing taxidermy specimens in commercial freezers with
temperatures of less than -13 degrees F. (-25 degrees C.) it is generally more economical to pre-
freeze the specimens in a commercial deep freezer, than use the specimen chamber as a freezer.
Freeze dry machines are most efficient when "drying" frozen material, rather than initially freezing
the material, then drying it. The important factor in pre-freezing any type of specimen is that the
specimen is completely frozen throughout.
Once the specimen has been prepared for preservation, mounted, posed, and completely
frozen, it is ready for the drying stage. the important factor in the drying stage is that the eutectic
22
temperature is maintained. Because of the lower eutectic temperature of greasy specimens, a low
processing temperature in the specimen chamber is needed to prevent shrinkage. Generally this
temperature should be as 0 degrees F. and then raised even higher after a period of time. Using
these figures as guidelines, the taxidermist using the freeze dry machine should determine at what
temperature his work is most successful. Some factors to consider when analyzing proper
processing temperatures include:
1. Working in batch loads (all the same type specimen) may be the most economical way to
work, depending on your own situation. Batch loads allow easy raising of temperatures as the
process progresses.
2. When working with mixed loads (greasy & non-greasy specimens) one initial temperature
should be decided upon (e.g. -5 degrees F. or 0 Degrees F).
3. Some shrinkage may be traded off for speed. Mammals may be run at slightly higher
temperatures allowing slightly faster drying times. The animals fur will cover most of the shrinking
that takes place.
4. Not only does a variety of chemical make-ups exist between different species, it may also
exist between members of the same species due to different diets, and physical environments.
5. Because if the lack of air to conduct heat within the specimen chamber, some differences of
temperature will exist inside the specimen chamber itself. This will require "rotating of the stock".
6. The greatest water loss will occur early in the drying stage when the dried shell is the
thinnest and offers the least resistance to water vapor movement.
The rate of freeze drying is directly related to vapor pressure. the higher the specimen
chamber temperature is, the higher the vapor pressure is at a given vacuum, and the faster the
drying is achieved. The rate of moisture migration from the specimen chamber, to the ice bank, is a
also related to vapor pressure. The greater the temperature difference between the specimen
chamber, and the ice bank. for example, a unit with the specimen chamber set at 5 deg. F. and the
Ice bank running at -55 deg. F., resulting in a temperature difference of 60 deg. Will move moisture
faster than the same machine with the specimen chamber set a -5 deg. F., resulting in a temperature
difference of 50 deg. F.
Certain steps may be taken to raise the low eutectic temperatures of greasy specimens. This
will allow the taxidermist to operate the freeze dry unit more economically at higher temperatures.
The first step is to eliminate the extreme amounts of fats and oils that cause the low eutectic
temperatures. In the preparation stage, a 20 minute bath in a mixture of one cup of bicarbonate of
soda per gallon of water will assist in neutralizing fats and oils. Also, injections of grease
tallowfiers, such as antioxidant will help in preparing greasy and oily specimens for more efficient
freeze drying.
Some of the questions that should be addressed when considering the right temperatures to
process a particular specimen are:
• What is the Physiology of the specimen?
• What is the Chemistry of the specimen?
• What solid mass of the specimen can be removed to speed up the process, and not effect
shrinkage?
If you analyze specimens with these questions in the initial planning stages, problems can be
eliminated. There is nothing, to our knowledge, that can not be successfully freeze dried without
shrinkage, as long as it is freeze dried at the proper temperature.
23
Duck Wildcat
Bobwhite quail Fisher
Wolf Longhorn
Figure 3.2.
24
3.3. Food
Lyophilization is recommended for most of the domains in food industry, especially in meat,
milk, canned goods industry and groups of food that cannot be preserved through other methods
(dairy products, eggs whites, enzimes, lactic bacterias) or food products destined to small children.
Bean curd Green bean Garlic
Scallion Red pepper Beef
Crab meat Chicken meat Pork
Corn Dried shrimp Ham
Carrot Mushroom Goloden needle

mushroom
Figure 3.4 – Lyophilization products
The process is used for drying and preserving a number of food products, including meats,
vegetables, fruits, and instant coffee products. The dried product will be the same size and shape as
the original frozen material and will be found to have excellent stability and convenient
reconstitution when placed in water. Freeze-dried products will maintain nutrients, colour, flavour,
and texture often indistinguishable from the original product.
25
Prunes lyophilization (freeze-drying) consists of freezing below -30°C and later removing the
ice formed by sublimation in a chamber with controlled temperature under vacuum. The water
vapor produced is removed by condensation at temperatures around -60°C. Different lyophilised
prunes derived from many kind of these have been developed and can be used to improve existing
method, create new ones, or to extract compounds of interest in a fruits matrix almost free from
water. The know-how on lyophilization and the problems during the process have been acquired by
using pilot equipment. The optimization of the process can be also addressed by calculating the
curves of weight loss during drying and the electrical consumption. Quality and safety parameters
in fresh and lyophilised prunes can be also determined compared with the fresh prunes. The main
fruits are freeze-dried samples obtained from a near isogenic line collection. This collection
rendered products with different taste, flavor, color, etc.
It made some comparisons between the prunes resulted through heating using the roaster
(shown in figure 1) with those after lyophilization.
Lyophilised (freeze-dried) prunes are with long shelf life under storage temperatures above
the regular used for freezing. The manufacture of lyophilised prunes requires knowledge about the
optimum conditions of processing in each fruit including the right format for any of them. Usually,
lyophilization, a non-thermal treatment, preserves most of the nutrients and aroma compounds from
the original prunes, particularly when compared with other treatments. Vacuum is a critical step of
the process in order to avoid melting ice. Optimization of freeze-drying requires a pilot plant and
knowledge on the curves of water loss from the product, though this information is not easy to find
in fruits. In the laboratory the research group has experience in many fruits. Also there is a pilot
freeze-dryer with heated trays able to condense up to 18 kg ice. With this equipment the weight loss
of the products can be modeled in order to select the optimum conditions of the process and to
know the electrical consumption of the operation. It is interested in contacting companies able to
optimized lyophilization conditions to be used later for developing new fruits. Quality parameters
from all the dried prunes produced can be determined compared with the quality of fresh product in
the laboratories.
a. b.
Figure 35. - Dried prunes: a.- lyophilizated; b. – by heating (roaster).
It can realize o lot of researches about the prunes behavior in different conditions. For
example X-, K- and Q-band EPR (Electron paramagnetic resonance) studies on lyophilized
whole pulp parts of prunes before and after X–ray irradiation are reported. Before irradiation
show in X band a weak singlet EPR line with g=2.0030±0.0005. Immediately after irradiation
all samples exhibit complex fruit-depending spectra, which decay with time and change to give,
in ca. 50 days, an asymmetric singlet EPR line with g=2.0041±0.0005. Singlet EPR lines
recorded after irradiation in X -band are K- and Q-band resolved as typical anisotropic EPR
spectra with g||=2.0023±0.0003 and g =2.0041±0.0005. In addition, K- and Q-band EPR
spectra of all samples show a superposition with the six EPR lines of Mn2+ naturally present in
the prunes. The differences in g factors of samples before and after X-ray irradiation might be
used for the identification of radiation processing of fruits in the case of prunes pulp .
The degradation of vitamin C [ascorbic acid] in prunes as affected by processing
temperature and moisture content of samples (0.05, 0.11, 0.18, 0.38 or 1.40 g water/g dry
26
matter) was modeled using the Bigelow equation. Prunes were freeze-dried and conditioned in
desiccators with various salt solutions at 4°C. After reaching equilibrium, the temperature of the
desiccators was adjusted to 40-80°C. Vitamin C degradation was analyzed for 5 days by
titration method using a solution of 0.01% 2,6-dichlorophenolindophenol. The results showed
that the samples subjected to 40-60°C exhibited slower degradation of vitamin C than those
subjected to higher temperatures. The parameters z-value and D<sub>T</sub> showed a linear
and quadratic dependence, respectively, with the moisture content of the samples. In general,
vitamin C degradation was faster in samples with higher moisture content, due to the high
solubility of vitamin C.
As a member of the largest freeze drying group in the world, European Freeze Dry offers an
extensive range of high quality freeze dried ingredients. These are ideal for inclusion in dried ready
meals, pot snacks, dried soups, breakfast cereals and bars, drinks, dairy items, confectionery and
bakery products.
The freeze dried meat, seafood and vegetable products are perfect for dried savory applications
such as instant pot and sachet products. With rapid rehydration within 3 to 5 minutes in boiling water
they meet customer expectations of "instant" food. When the customer then tastes the freeze dried
ingredients, they will recognize all the textural and flavor attributes of a freshly cooked product.
It can also offer an extensive range of freeze dried culinary herbs. Chives and culinary herbs
have wide application possibilities. Examples of use are dressings and dips, soups and sauces, dairy
product applications and dry meals based on rice, potatoes and pasta. For catering and retail purposes,
herbs and herb mixes give both colour and taste in restaurants as well as in private households.
For manufacturers of breakfast cereals and bars, bakery ingredient suppliers, cake
manufacturers and confectioners we present a wide range of freeze dried fruits. These products are
natural, wholesome, and contain no artificial additives or added sugar. Our fruit range includes both
berries and orchard fruits - please check our product list to see the range available. Alternatively, if
you can't see what you require, contact us, and we will be happy to discuss your requirements.
Here at European Freeze Dry we take pride in the service that we offer to all our customers.
Whether your requirement is for small quantities or for large scale manufacture, we have the
flexibility to fulfil your order promptly ands efficiently.
A selection from our product list is shown below. Please remember to contact us if you can't
see what you need.
Vegetables and Pulses Meats

Broccoli, beads Bacon, diced various dimensions
Courgette Beef, diced, strips, mince
Grilled Mixed Bell Pepper Chicken Powder. Pure chicken meat.
Kidney Beans, red Chicken Pressed, diced to various
Leek dimensions
Lentils, brown Chicken, Whole muscle breast meat
Olives, sliced, wedges or granulated (no additives)
Peas, whole and crushed Ham, diced various dimensions
Red Bell Peppers, diced
Spinach Seafood
Sun Dried Tomato, granules
Sweetcorn Alaskan Pollock, diced (uncooked)
Arctic Prawns cooked and peeled
Clams / Vongole
Herbs COD, diced (uncooked)
Basil Hoki, diced (uncooked)
Chervil Langostino
Chives Mussels
27
Dill Smoked Salmon

Green Peppercorns Surimi (Crabstick)
Juniper berries Tuna
Lemon Grass Warm water shrimps
Marjoram *All of the above are also available in
Mint powdered form
Oregano
Parsley
Rosemary Dairy and Egg
Sage
Savory Cheddar Cheese
Tarragon Mozzarella Cheese
Thyme Omelette
*Blends of the above herbs are available Red Leicester Cheese
upon request Ricotta Cheese
Stilton Cheese
Fruits and Berries
Bananas, sliced or diced
Bilberries
Blackberries
Blackcurrants, whole or powder
Blueberries, whole or powder
Cherries, whole, sliced or diced
Cranberries, sliced
Fig Pieces
Freeze Dried Fruit Blends
Grape slices
Lingonberries, whole or powder
Mango, diced
Pineapple, diced
Pomegranate Kernels
Raspberries, whole, pieces or
crumble
Redcurrants
Strawberries, whole, sliced, powder
or diced
*Kosher available upon request
Foods for space: Mercury astronauts following John Glenn were forced to endure bite-sized
cubes, freeze dried foods, and semi-liquids in aluminum toothpaste-type tubes. They found the food
unappetizing, had trouble rehydrating the freeze-dried foods, and disliked squeezing the tubes.
In the Gemini missions eating in space became more normal.
In the Apollo program, food packages were similar to those used on Gemini missions but the
variety of food was considerably greater. Apollo astronauts had the added luxury of heated water
for hot drinks and foods at a temperature of 67 degrees C (154 degrees F) and chilled water at 7
degrees C (45 degrees F). Water temperatures from the dispenser of the Gemini spacecraft hovered
at the 21 degrees C (70 degrees F) ambient spacecraft temperature. With hot water available, food
was easier to rehydrate and much improved in taste.
28
In 1973 and 1974, the Skylab spacecraft was occupied by three teams of astronauts. Space
food systems there were much improved over systems used in Apollo, Gemini, and Mercury.
Unlike previous space vehicles for astronauts, Skylab featured a large interior volume and space
was available for a dining room table. The table was a pedestal where food trays were mounted.
When dining, the three-astronaut teams would "sit down" in the air by means of foot and thigh
restraints and eat in an almost normal fashion. The food trays not only held the food in place but
also served as warming devices. Underneath three of eight cavities in the trays were warmers that
could raise temperatures of foods needing heating to 66 degrees C
In 1975, the last Apollo flights took place with the Apollo-Soyuz docking mission. The
Apollo spacecraft did not have the freezer that Skylab featured but many of the food advances from
Skylab and the earlier Apollo missions were incorporated. Because of the short duration of the
flight (nine days), many short shelf-life items were added to the foods carried. Fresh breads and
cheese were included as a part of 80 different varieties of food dined upon by the Apollo while
others were placed in spoon-bowl packages or plastic drinking bags. To make eating easier, a food
tray was carried on the mission. The tray did not warm the food as the Skylab tray did, but it held
the food in place with springs and Velcro fasteners. The tray was secured to the crewmember's leg
during meal time.
The process of lyophilization consists of:
- freezing the food so that the water in the food becomes ice;
- under a vacuum, sublimating the ice directly into water vapor;
- drawing off the water vapor;
- once the ice is sublimated, the foods are freeze-dried and can be removed from the machine;
- lyophilization has many advantages compared to other drying and preserving techniques.
Lyophilization maintains food quality.
The use of lyophilization is particularly important when processing lactic bacteria, because
these products are easily affected by heat.
Foods, which are lyophilized, can usually be stored without refrigeration, which results in a
significant reduction of storage and transportation costs.
Lyophilization greatly reduces weight, and this makes the products easier to transport. For
example, many foods contain as much as 90% water. These foods are 10 times lighter after
lyophilization.
Because they are porous, most freeze-dried foods can be easily rehydrated. Lyophilization
does not significantly reduce volume; therefore water quickly regains its place in the molecular
structure of the food.
Ratti and Crapiste (1992) developed a lumped parameter model for hygroscopic shrinking
food systems.
Lyophilization, is like “suspended animation” for food. It can store a freeze-dried meal for years
and years, and then, when that is finally ready to eat it, it can completely revitalize it with a little hot
water. Even after all those years, the taste and texture will be pretty much the same. It can to explore
the basic idea behind freeze-drying, and that can look at the different steps involved in the process.
Also it can see how freeze drying is different from ordinary dehydration, and it find out.
Figure 3.7. Ice cream after freeze dried and rehydratation process
29
A package of freeze-dried ice cream, sold as a novelty item.

The process has been popularized in the forms of freeze -dried ice cream, an example of
astronaut food. It is also popular and convenient for hikers because the reduced weight allows them
to carry more food and reconstitute it with available water. Instant coffee is sometimes freeze dried,
despite high costs of freeze dryers. The coffee is often dried by vaporization in a hot air flow, or by
projection on hot metallic plates. Currently, the freeze –drying process is used more commonly in
the pharmaceutical industry.
It is healthier to eat raw grains or vegetables without heating because living enzymes are
consumed. A plant embryo cannot sprout when heated, and that occurs because the plants enzymes
have been destroyed by the heat. Only live food can create life; dead food cannot create life.
Through instant lyophilization natural color, aroma and flavor are preserved. The vitamins,
minerals, enzymes and chlorophyll are kept intact which has the same effect in the body as eating
natural raw vegetables.
3.4. Cereals
The composition of cereals has five food substances:
• Water
• mineral matter
• Protein
• fat
• carbohydrate
Mineral Matter or
Poz Protein Fat Carbohydrate Cellulose
Ash
1 Wheat Oats Rice Oats Oats
2 Rye Corn Rye Buckwheat Barley
3 Oats Barley Corn Barley Buckwheat
4 Barley Buckwheat Wheat Wheat Rye
5 Corn Rice Barley Rye Wheat
6 Buckwheat Wheat Buckwheat Corn Corn
7 Rice Rye Oats Rice Rice
The cereals are a very good vegetarian resource for eating, with important nutrients like
vitamin B12-(fortified soy milk and cereals), iron-fortified cereals and breads, especially whole-
wheat, zinc from whole grains (especially the germ and bran), whole-wheat bread. Also, the cereals
are the most important source of dietary fiber in many countries.
For example, after the freezing, 100 g of uncooked maize was introduced in the dried room of
a lyophilizer for lab (Christ, Alpha 1-2). The grains was put in a tray be Stainless Steel, (After DIN
symbolized X6CrNiTi18.10). The time necessary for lyophilization was 18 hours and 32 minutes.
After that it rested 85 g maize dried. The grains were reconstituted with warm water and the weight
was 98.8 g. (In figure 3 it show the lyophilizer, in the figure 5 is the maize lyophilized and the
figure 6 is the maize reconstituted after lyophilization).
For establish the optimal solution for foods conservation it must to analyze the specific
situation, in according with the destination. Also, it must to study the advantages and disadvantages
of foods lyophilizeted.
One experiment put in evidence a very quality of dried grains, also after easy milling.
30
The maize lyophilized The maize reconstituted after lyophilization
Figure 3.8.
Advances in freeze drying equipment, such as automatic loading, has to some degree reduced
the need for the use of a tray. But this is only for products that are contained in vials or bottles and
these advance systems are generally not used to freeze dry bulk materials. Trays are generally
constructed out of three basic materials: metal, glass and plastic. As a result of the composition,
each material will offer its own unique advantage and disadvantage.
The lyophilization of cereals is not so used because the cereals contain very little water in
their composition.
This absence of water is a distinct advantage, for it makes their nutritive value proportionately
high and improves their keeping quality.
The strength of a beverage is lowered by the addition of water. So the nutritive value of foods
decreases when they contain a large amount of water.
The keeping quality of cereals could scarcely be improved, since the germs that cause foods to
spoil grow only in the presence of water. This low proportion of water also permits them to be
stored compactly, whereas if water occurred in large amounts it would add materially to their bulk.
That dried grains was very porous, since it occupies the same volume as the original and so
rehydrates rapidly.
There is less loss of flavor and texture than with most other methods of drying.
The experiment put in evidence a very good quality of dried grains, also after an easy milling
Another product from cereals lyophilizated:
corn oil,
wheat flour,
maltodextrin (from corn),
juices hydrolyzed corn gluten,
dried whey,
autolyzed yeast extract,
modified cornstarch,
flavorings (hydrolyzed wheat gluten, partially hydrogenated soybean and cottonseed oil,
autolyzed yeast extract, lactic acid and bonito fish extract),
hydrolyzed vegetable protein (soybean and wheat),
soy sauce (wheat, soybeans, salt),
instant rice,
hydrolyzed vegetable protein (hydrolyzed corn torula, brewers yeast, wheat gluten, soy protein),
monosodium glutamate (a flavor enhancer),
hydrolyzed soybean protein.
3.4. Other Applications
In chemical synthesis, products are often lyophilized to make them more stable, or easier to
dissolve in water for subsequent use.
31
In bioseparations, freeze drying can also be used as a late-stage purification procedure,

because it can effectively remove solvents. Furthermore, it is capable of concentrating substances
with low molecular weights that are too small to be removed by a filtration membrane.
Freeze – drying is a relatively expensive process. The equipment is about three times as
expensive as the equipment used for other separation processes, and the high energy demands lead
to high energy costs. Furthermore, freeze drying also has a long process time, because the addition
of too much heat to the material can cause melting or structural deformations. Therefore, freeze
drying is often reserved for materials that are heat-sensitive, such as proteins, enzymes, freeze, and
blood plasma. The low operating temperature of the process leads to minimal damage of these heat-
sensitive products.
Floral Freeze Dry - How To freeze dry flowers. After years of research and development,
Freeze Dry Company introduced the amazing concept of freeze-drying flowers in 1981. Producing
a dried flower with the beautiful color and shape of fresh flowers has been a phenomenon that has
taken the industry by storm. Endless creative and business applications for the freeze drying
process have been discovered by floral entrepreneurs world-wide, making freeze-dry the hottest
business opportunity in the floral and crafts industry. Statistics indicate the freeze dry floral
industry is growing at an impressive rate. Industry professionals believe that the potential of the
floral freeze dried industry is just getting started. Wedding flowers, valentine bouquets, or any
special event flowers can be freeze dried for a lifetime of beauty.
1. Allow flowers to mature to the state you would like them at when completed. (If you would
like flowers such as roses to open up more, warm water will cause them to open quicker.)
2. Allow flowers to absorb moisture naturally (e.g. in a container with water, vase, etc.) for 12
hours. Ensure that you use fresh blade-cut stems. Fluff heads if bunched together.
3. Strip foliages off stem and cut stem to desired length. The stems will dry slower than the
head, therefore the less stem, the faster the drying.
4. Pre-cool the specimen chamber to -5 Degrees F. Turn the vacuum on for 10 seconds to seal
the door (to hold refrigeration).
5. Load flowers in/on rack as you prepare them, filling one rack at a time and then transferring
into the pre-cooled flower chamber.
6. Place blackout cover on door. Leave interior light off. freeze at -5 degrees F for 24 hours.
7. Turn vacuum pump on.
8. Every two days increase temperature in flower chamber by -5 degrees F. (In ten days you
will have ranged from -5 degrees F to +20 degrees F.)
After 10 to 12 days, open equipment and check to ensure flowers are dried. Inspect by
checking if flowers are cold to the touch (a dried flower will not be cold because it does not contain
any moisture) and by dissection of selected samples.
Another’s applications:
Cereal allergen preparations
Allergenic extracts from T. putrescentiae bodies
The incorporation of bio-active ingredients in soy products
Extracts of proteins from cereals a wide range of applications:
Speciality Chemicals
Natural Enzymes
Biological Products
Pigments
Resins
Active Pharmaceutical Intermediates
Food Ingredients
Flavourings
Flood damage/Document Recovery
32
Scanning Micrograph of an Aphid, Freeze dried using the K750 system and Sputter Coated
with Gold. Aphids secret wax ribbons of which show good preservation, normally 'Lost' in Drying
methods involving solvents.
Cells of the petiole assumed their normal isodiametric shape after dying.
Recently, some taxidermists have begun using freeze drying to preserve animals.
Organizations such as the Document Conservation Laboratory at the United States National
Archives and Records Administration (NARA) have done studies on freeze drying as a recovery
method of water-damaged books and documents. While recovery is possible, restoration quality
depends on the material of the documents. If a document is made of a variety of materials, which
have different absorption properties, expansion will occur at a non-uniform rate which could lead to
deformations. Water can also cause mold to grow or make inks bleed. In these cases, freeze drying
may not be an effective restoration method.
In high altitude environments, the low temperatures and pressures can sometimes produce
natural mummies by a process of freeze –drying.
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.
Generally, one looks for the final freeze-dried product to fulfill a number of set criteria, which
for biotechnology products and pharmaceuticals might typically include maintenance of cosmetic
acceptability, good reconstitution characteristics, appropriate moisture content and good shelf-life.
Due to the complex interrelationship between the numerous product and process variables, some
developmental work may be required to achieve these criteria. However, the advent of new
technologies is enabling such work to be approached on a more rational basis.
33
4. TRADITIONAL LYOPHILIZATION TECHNOLOGY

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.
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:
3 3
• 50m x 250 = 0.35m 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:
3
• 0.350m /s = 17.5 m/s
0.02 m2
Inside the neck of the bottle, which has a diameter of about 3mm, or a section of 7cm2 (7 x 10-
4m2), the speed is calculated at:
3
• 0.350m /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
3
• At P=1 torr, V=1000 l or 1m
3
• At P=10-1, V=1000, = 10,000 l, or 10m
3
• At P=10-2 V=100m
3
• At P=10-3 V=1000m
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,
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).
34
If you examine the curve, 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.
The heat transfer rate (Qs) from shelf through the bottom wall of the tray is given by the
Fourier`s law:
∂T
Q1 = − Aλ
∂l
Since the heating or cooling takes place in an unsteady state, the amount of heat (Q) will be:
∂T
Q = − Aλ t
∂l
In the case of the bottom of the tray, the relation (2) becomes:
QS = Aλ
(Ts − Tp )
δ
35
Where:
A is the total surface area of the tray, m2;
- the thermal gradient, 0C/m, or Kelvin/m
∂T Ts − Tp
=
∂l δ
It can consider the role of composition and configuration of the tray can have on the heat
transfer properties. For analyse on put the conditions:
• All the trays have the same general rectangular configuration and the only variables
will be their total mass and thickness of bottom and walls.
• That all trays contain the same volume of water.
In each case, the shelf-surface temperature is uniform and chamber pressures are equal.
Neglecting the contact resistance between shelf and tray, the external surface temperature of
the bottom of the tray is equal to that of the shelf.
It can consider that the temperature gradient has a single direction, perpendicular on the
surface of the bottom of the tray.
In the regions with a good contact area of the bottom of the tray with shelf surface there is also
a good heat transfer. In the regions where the contact is not perfect it appears a new thermal
impedance to heat transfer between shelf and bottom of the tray.
36
5. LYOPHILIZATION EQUIPMENT
A lyophilizer consists of a vacuum chamber that contains product shelves capable of cooling
and heating containers and their contents. A vacuum pump, a refrigeration unit, and associated
controls are connected to the vacuum chamber.
Chemicals are generally placed in containers such as glass vials that are placed on the shelves
within the vacuum chamber.
Cooling elements within the shelves freeze the product. Once the product is frozen, the
vacuum pump evacuates the chamber and the product is heated. Heat is transferred by thermal
conduction from the shelf, through the vial, and ultimately into the product.
Llyophlization Container Requirements:
The container in which a substance is lyophilized must permit thermal conductivity, be
capable of being tightly sealed at the end of the lyophlization cycle, and minimize the amount of
moisture to permeate its walls and seal.
The enclosed reagents can only remain properly lyophilized if the container in which they are
processed meets these requirements.
Lyophlization Heat Transfer:
Successful lyophlization is heavily dependent on good thermal conductivity. For this reason,
containers used in the lyophlization process must be capable of meeting a number of heat-transfer
requirements.
Such containers should be made of a material that offers good thermal conductivity; should
provide good thermal contact with the lyophilizer shelf, which is the source of heat during
processing; and should have a minimum of insulation separating the source of heat from the product
requiring heating.
Poor thermal conductivity often results from the use of containers made of materials with low
coefficients of heat transfer. It can also be caused by the shape, size, or quality of the container.
It may come from thermal barriers, such as excessive amounts of material, which can act as
insulation, preventing energy from being transferred to the point at which the frozen ice and dried
product interface.
Poor thermal conductivity often results in a product that is not successfully lyophilized. In a
serum vial, the surface of the frozen cake must sublime first to allow the ice vapor to escape.
There after, the sublimation front moves as the drying proceeds. Generally, the sublimation
front simultaneously moves downward toward the bottom of the serum vial and inward toward the
center of the frozen cake (the core).
If sublimation is not controlled - and it cannot be controlled when significant thermal barriers
exist - then portions of the chemicals may actually be vacuum-dried rather than freeze-dried.
The processed product will then not possess the defined and reproducible characteristics of a
properly lyophilized material, such as maximized retention of activity, optimized shelf life, rapid
reconstitution, and a consistent finished cake.
Sealing Lyophilized Products:
A properly lyophilized product must be sealed within its container prior to removal from the
ultra dry atmosphere that exists at the end of the lyophlization cycle.
A product that has been dried to less than 3% residual moisture will, when exposed to an
environment with greater than its own moisture level, absorb as much moisture as it can. The
product's quality will immediately be degraded.
All of the desirable characteristics of a lyophilized product - such as increased shelf life,
enhanced chemical performance, and rapid reconstitution - will be compromised. This
reintroduction of moisture can lead to loss of product, product failures in the field, false results, and
even product recalls.
The most common mistake that companies make is to use packages that cannot be sealed
inside the lyophilizer prior to repressurization. For example, the manufacturing process for some
diagnostic products may require lyophilizing the product inside a large number of screw-top tubes.
37
There is no practical way to seal these tubes inside of a lyophilizer prior to terminating the
batch, so the company will assemble a large production crew to apply the tops manually—often in a
room incompatible with lyophlization.
The recently stabilized chemistry will be jeopardized by exposure to unacceptably high and
variable moisture levels during the manual sealing process. Exposing lyophilized material to
atmospheric moisture in this way may result in an unstable product.
Installation of lyophilization for liquid food products: For liquid food products it is more
frequently used the separate freezing alternative, followed by a breaking up and a continuous
criodesication in small volume apparatuses, in which the product remains for a short period of
time. The advantages to this kind of installation come from the fact that the products that are going
to be lyophilized are granulated; it has a large surface of mass exchange:
the removal of water from the product is very fast, due to the movement that homogenize
the drying;
the duration of the process is short, the product remains in the installation for 20-30
minutes;
it easily allows the passing from the lyophilization of a product to that of another;
in case of technological accidents, due to their loading capacity, the economical loses are
smaller considering other types of installations.
Installation of lyophilization for solid food products: For solid food products there can be
used continuous functioning installations or discontinuous functioning. The lyophilization
equipments with continuous functioning are made out of a tunnel that contains standard
lyophilization units, coupled in line or ”U” shaped.
At the same time, considering the constructive structure, the lyophilization instalations with
discontinuous functioning can be: with interior freezinf and interior condenser (inside the same
precinct in wich the process takes place), or with interior freezing and exterior condenser.
Installation of lyopilization with interior freezing an interior condenser
Are installations with discontinuous functioning (figure 1) and comprise in their structure
components that are meant to assure the freezing and condensing of vapours inside the
lyophilizator.
The major advantaje of this installations is avoiting the installation of the frozen product and
so the posibility to make some large volumed precincts. Beside these advantages, this type of
installations presents a series of desavantages: if the instalation is made for severalgroups of
products, then the sizes of the freezing installation andvoiding group are important, which leads to
high costs of investment; the total duration of the lyophilization process and of the immobilization
of the installation is very long, due to the fact that in the same apparatus, beside the lyophilization,
are realised the other phases of the lyophilization process.
Installation of lyopilization with interior freezing and exterior condenser
Are installations with discontinuous functioning. The freezing, sublimation and lyophilization
can be done in the same installation, ussing a plate condenser. The plates are used alternativelly for
freezing and for heating, as they succesively connected to a refrigerating installation and a thermic
one.
To assure the necessary vacuum for sublimation and drying, a vacuum pump is used. The
vavapoures resulted from sublimation and drying condensate on the surface of the exterior
condenser that is also the vaporizers for a refrigerating installation. The condense collected in the
collecting container is evacuated through the evacuation tap for condense
38
Figure 5.1. – The basic scheme of the lyophilizer with interior freezing
The most important mechanical engineering elements are listed below:

- separated drying chamber and ice condenser to reduce cross-contamination;
- provision of an isolation valve between chamber and ice condenser can 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 0C 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.
Advances in freeze drying equipment, such as automatic loading, has to some degree reduced
the need for the use of a tray. But this is only for products that are contained in vials or bottles and
these advance systems are generally not used to freeze dry bulk materials.
For bulk products, it still must rely on a large container that we generally classify as a tray.
We will certainly point out the advantages, disadvantages and perhaps pose some new questions
39
that we need to address. The trays will be considered range from the traditional metal (stainless
steel) to the more recently introduced.
The trays can be generally classified as plastic trays, they do have unique features and we felt
they deserved their own category in this discussion.
The basic reason being that they are covered with a membrane barrier that will allow a
formulation prepared in an aseptic environment to be safely lyophilized or freeze dried in a non
aseptic environment.
This feature is of major importance to those working with small freeze dryers, which were not
designed to withstand the harsh rigors of steam sterilization.
However, given such an advantage there are also some disadvantages in that they tend not to
be reusable and as a result of their unique feature to be more costly than those trays fabricate from
the above materials.
Trays are generally constructed out of three basic materials: metal, glass and plastic. As a
result of the composition, each material will offer its own unique advantage and disadvantage. The
trays can be making from:
Cold Roll Steel used in the manufacture of flowers. If its are either galvanized or electroplated
especially it would be easier to construct and may represent the least expensive of all the trays.
Stainless Steel is most commonly used trays in the lyophilization or freeze-drying of products,
because that is corrosion resistant, can be depyrogenated at temperatures of 250oC, are durable,
require no special handling, are reusable which in time can off-set their initial higher cost.
Aluminum trays are used in the freeze-drying of bulk materials. They are certainly lightweight
when compared to the cold roll and stainless steel trays and are corrosion resistance. In addition, its
can be anodized to give them a black surface, which will increase their emissivity aid somewhat to
the energy transfer resulting from radiant energy. Also its can be steam sterilized. Because
aluminum is a relatively soft metal, one must be concerned with the effects of “galling” (the
removal of a surface layer generally in the form of a dust) should the product have to be removed
by scraping.
Glass is good for all bulk products that can be lyophilized or freeze-dried come into contact
with a metal surface. As a result of their composition, these trays have similar advantages as those
constructed from stainless steel, except they are by their very nature fragile and are subject to
breakage that may occur over time from stress strains resulting from age and thermal shock.
Plastic Trays can be used in some applications which not requiring high temperature
depyrogenation. It tends to be lightweight when compared to the trays made from the previous
materials and easy to clean and quite durable and reusable.
While these trays can also be generally classified as plastic trays, they do have unique features
and we felt they deserved their own category in this discussion.
The basic reason being that they are covered with a membrane barrier that will allow a
formulation prepared in an aseptic environment to be safely lyophilized or freeze dried in a non
aseptic environment. This feature is of major importance to those working with small freeze dryers,
which were not designed to withstand the harsh rigors of steam sterilization.
However, given such an advantage there are also some disadvantages in that they tend not to
be reusable and as a result of their unique feature to be more costly than those trays fabricate from
the above materials
A lyophilizer consists of a vacuum chamber that contains product shelves capable of cooling
and heating containers and their contents.
A vacuum pump, a refrigeration unit, and associated controls are connected to the vacuum
chamber.
The conception about lyophilizers was materialized in different constructive solution, in
concordance with the destination.
For experiment it was used a lyophilizer like these from the figure 5.1. For industrial
production can use a complex installation (figure 5.2), with similar principle but with a big capacity,
and different form and dimension.
40
For each application it must to realize an integrated conception based on the optimal
parameters for a good economic efficiency.
Basically, the highest optimum temperature at which the product can be lyophilized should be
between -20 C and -30 C, and often lower. The critical product temperature for primary drying is
much lower, generally.
Figure 5.1. Figure 5.2
This is the product tem-perature employed for opti-mum sublimation, and when exceeded may
induce "meltback" which results in the loss of the batch.
41
6. INSTALLATION OF LABORATORY
A suggested experimental apparatus for conducting a lyophilization process is illustrated in
the figure 6.1. As seen by this figure, the apparatus consists mainly of parts that can be found in
most laboratories with the possible exception of the vacuum gauge and vacuum pump, which one
may have to obtain from an outside supplier. While a capacitance manometer would be the desired
vacuum gauge, in this instant a more inexpensive thermal conductivity gauge such as a
thermocouple gauge or Pirani gauge can be use. When using the thermal conductivity gauge one
should bear in mind that when water vapour is present in the Sample Chamber, the gauge will tend
to read a false high pressure. Note that it is important that the Sample chamber also contains at least
one temperature sensor, preferably a thermocouple, to measure the product temperature during the
lyophilization process. The need for measuring the product temperature was already previously
considered.
Figure 6.1. - Freeze drying system for laboratory (Thomas A. Jennings)
It is recommended that the thick vacuum rubber hose between the Sample Chamber and the
Condenser Chamber have the largest possible bore diameter and be kept as short as possible to
maximise the throughput of the system.
Since a liquid cold bath [example - mixture of acetone and dry ice (solid CO2 )] will be used
to refrigerate and heat the sample and also refrigerate the condenser, its strongly advised that the
apparatus be used in a fume hood so that one is not exposed to the vapors. Acetone is a highly
flammable substance and one should make certain that there are no sparks, arks or open flames
being used in the vicinity where the apparatus is being used. The fume hood will also remove any
exhaust fume that may stem from the vacuum pumping system. It is suggested that the blank-off
pressure of the vacuum pump should be lower than 5 mTorr.
A typical machine consists of a freeze-drying chamber with several shelves attached to
heating units, a freezing coil connected to a refrigerator compressor, and a vacuum pump.
With most machines, can be placed the material to be preserved onto the shelves when it is
still unfrozen. When it seats the chamber and begins the process, the machine runs the compressors
to lower the temperature in the chamber. The material is frozen solid, which separates the water
from everything around it, on a molecular level, even though the water is still present.
42
Figure 6.2. – The laboratory scheme of lyopilizer
The machine turns on the vacuum pump to force air out of the chamber, lowering the
atmospheric pressure below .06 ATM. The heating units apply a small amount of heat to the
shelves, causing the ice to change phase. Since the pressure is so low, the ice turns directly into
water vapor. The water vapor flows out of the freeze-drying chamber, past the freezing coil. The
water vapor condenses onto the freezing coil in solid ice form, in the same way water condenses as
frost on a cold day.
This continues for many hours (even days) while the material gradually dries out. The process
takes so long because overheating the material can significantly change the composition and
structure. Additionally, accelerating the sublimation process could produce more water vapor in a
period of time then the pumping system can remove from the chamber. This could rehydrate the
material somewhat, degrading its quality.
Figure 6.3.
43
Once the material is dried sufficiently, it's sealed in a moisture-free package, often with an oxygen-
absorbing material. As long as the package is secure, the material can sit on a shelf for years and
years without degrading, until it's restored to its original form with a bit of water (a very small
amount of moisture remains, so the material will eventually spoil). If everything works correctly,
the material will go through the entire process almost completely unscathed.
Figure 6.4. - A simplified freeze-drying machine
This lyophiliser consists of a vacuum chamber that contains product shelves capable of
cooling and heating containers and their contents. A vacuum pump, a refrigeration unit, and
associated controls are connected to the vacuum chamber.
Chemicals used in IVD products are generally placed in containers such as glass vials that are
placed on the shelves within the vacuum chamber. Cooling elements within the shelves freeze the
product. Once the product is frozen, the vacuum pump evacuates the chamber and the product is
heated. Heat is transferred by thermal conduction from the shelf, through the vial, and ultimately
into the product.
Fig 6.5. Lyophilization machine
A simplified freeze dryer system is presented in Fig.6.6.

The plant consists particularly of a drying chamber (1), temperature controlled shelves (2), a
condenser (3) and a vacuum pump (4). The major purposes of the shelves are either to cool and
freeze or to supply heat to the product. This is supported by the shelf heater and refrigerator system
44
(5). On those shelves the vials or the trays (10) are placed. The chamber can be isolated from the
condenser by a valve (6). Some freeze dryers possess a mechanism for sterilizing the interior (7).
Fig. 6.6. - Simplified freeze-dryer
The vacuum system is placed after the condenser. After a freeze drying process is completed,
the condenser will be heated by means of the condenser heater and refrigerator system (8) in order
to remove all the frozen ice at the condenser walls. The water is then drained (9).
Because the process parameters of lyophilization treatment alternate in type function of agri-
food product, there is the opinion that lyophilization plant constructions are different. Exists two
category of lyofilization plant: - destined for food product liquids or viscous (milk, cream etc.); -
destined for food products with texture (meat, fish etc.). For liquid food products on utilize separate
freeze, crumbling continuation in small equipment; that is food product one short period de time.
For food products with texture on utilize continuous or discontinuous equipment. Continuous
plant has in the structure one tunnel that contains one standard lyophilization unit, coupled in line or
in U form. Discontinuous plant can be like an internal freeze and condenser; internal freeze and
external condenser; external freeze and condenser.
For experiments it was used an instalation with the lyophilizer Crist Alpha 1-4, (figure 2.).
Freeze dried agri-food when blended with the gel compositions of the invention can be a tasty treat
for many meal prepared with some lyophilized components. For example, it was study:
maltodextrin (from corn), modified cornstarch, mushrooms, nonfat dry milk, corn oil extract, spice
extract, wheat flour, dried whey, cottonseed oil, onion powder, vegetable protein (soybean and
wheat), monosodium glutamate (a flavor enhancer), garlic powder, paprika, tomato powder,
calcium caseinate, etc.
The cereals are a very good vegetarian resource for eating, with important nutrients like
vitamin B12-(fortified soy milk and cereals), iron-fortified cereals and breads, especially whole-
wheat, zinc from whole grains (especially the germ and bran), whole-wheat bread. Also, the cereals
are the most important source of dietary fiber in many countries. For example, another experiment:
after the freezing, 100 g of uncooked maize was introduced in the dried room of a lyophilizer for
lab (Christ, Alpha 1-2, figure 2). The grains was put in a tray be Stainless Steel, (After DIN
symbolized X6CrNiTi18.10). The time necessary for lyophilization was 18 hours and 32 minutes.
After that it rested 85 g maize dried. The grains were reconstituted with warm water and the weight
45
was 98.8 g. After lyophilization that is very porous, since it occupies the same volume as the
original and so rehydrates rapidly. There is less loss of flavor and texture than with most other
methods of drying.
Figure 6.7.. – Lab installation with lyophilizer Christ Alpha 1-4.
In the lab was making some comparations between the quality of lyophilized prunes and
prunes dried by heating. (Anghel, G.V., Mnerie, D., Sito, S., Tucu, D., 2007). Innovations and
advantages of the lyophilization solution are: unavailability of lyophilized prunes on the market; the
lyophilized prunes can be stored at -18°C for long periods of time, keeping most of the flavor and
taste; the lyophilized product can be added as natural prunes aroma; lyophilized prunes are less
sensitive to thawing problems and reduce the cost of transportation because of the lack of water.
The atmospheric spray-freeze-drying process is an alternative to traditional vacuum freeze
drying technology. Here, a liquid feed containing a dissolved solid is first atomized to form small
droplets which are rapidly frozen in a cold air stream. The air is then also used to dry the frozen
particles by sublimation. This is possible if the partial pressure of water vapor (not necessarily the
total pressure) is below the saturation vapor pressure of water at that temperature. Air at
atmospheric pressure can thus be used, but it must be of extremely low humidity. This project is
aimed at investigating and modeling the rates of heat and mass transfer, and looking at process
modifications to improve economic viability. It is envisaged that smaller particle sizes will
significantly accelerate drying rates. Potential products include those currently made by spray-
drying or freeze drying liquid feeds. Examples are instant tea and coffee, pharmaceuticals, ceramics
and specialty chemicals.
Figure 6.8.- The atmospheric spray-freeze-drying process
46
Spray-freeze-drying liquid feed containing a dissolved solid is first atomised to form small
droplets which are rapidly frozen in a cold air stream. Potential products include those currently
made by spray-drying or freeze drying liquid feeds. Examples are instant tea and coffee,
pharmaceuticals, ceramics and specialty chemicals.
Figure 6.9.- Freeze-Drying Microscope System
A typical machine consists of a freeze-drying chamber with several shelves attached to

heating units, a freezing coil connected to a refrigerator compressor, and a vacuum pump.
still unfrozen. When it seal the chamber and begin the process, the machine runs the compressors to
lower the temperature in the chamber. The material is frozen solid, which separates the water from
everything around it, on a molecular level, even though the water is still present.
Figure 6.10.
The machine turns on the vacuum pump to force air out of the chamber, lowering the
atmospheric pressure below .06 ATM. The heating units apply a small amount of heat to the
shelves, causing the ice to change phase. Since the pressure is so low, the ice turns directly into
water vapor. The water vapor flows out of the freeze-drying chamber, past the freezing coil. The
47
water vapor condenses onto the freezing coil in solid ice form, in the same way water condenses as
frost on a cold day.
This continues for many hours (even days) while the material gradually dries out. The
process takes so long because overheating the material can significantly change the composition
and structure. Additionally, accelerating the sublimation process could produce more water vapor in
a period of time then the pumping system can remove from the chamber. This could rehydrate the
material somewhat, degrading its quality.
Figure 6.11.- Structure regained on cooling
Once the material is dried sufficiently, it's sealed in a moisture-free package, often with an
oxygen-absorbing material. As long as the package is secure, the material can sit on a shelf for
years and years without degrading, until it's restored to its original form with a bit of water (a very
small amount of moisture remains, so the material will eventually spoil). If everything works
correctly, the material will go through the entire process almost completely unscathed.
In this paper the authors propose one technical system for lyophilization process in laborator.
It is designed with minimal conditions for all processes.
This lyophiliser consists of a vacuum chamber that contains product shelves capable of
cooling and heating containers and their contents. A vacuum pump, a refrigeration unit, and
associated controls are connected to the vacuum chamber.
Chemicals used in IVD products are generally placed in containers such as glass vials that are
placed on the shelves within the vacuum chamber. Cooling elements within the shelves freeze the
product. Once the product is frozen, the vacuum pump evacuates the chamber and the product is
heated. Heat is transferred by thermal conduction from the shelf, through the vial, and ultimately
into the product.
No doubt that tray played a major role in the early days of lyophilization or freeze drying of
products and continues to this day to be a major vessel in conducting these drying processes.
Advances in freeze drying equipment, such as automatic loading, has to some degree reduced the
need for the use of a tray. But this is only for products that are contained in vials or bottles and these
advance systems are generally not used to freeze dry bulk materials. So for bulk products, we still
must rely on a large container that we generally classify as a tray.
48
It intends to generally discuss the various materials used in fabricating trays that have been
and continue to be used in the lyophilization or freeze drying of products. We will certainly point
out the advantages, disadvantages and perhaps pose some new questions that we need to address.
The trays will be considered range from the traditional metal (stainless steel). It should be clear to
the reader that we have neither preference nor bias for or against any given type of tray but just
wish to present an objective evaluation and make it clear that the selection of the tray will no doubt
be very product dependent. So what may work for one product may not be applicable for another.
Composition: Trays are generally constructed out of three basic materials: metal, glass and
plastic. As a result of the composition, each material will offer its own unique advantage and
disadvantage.
Cold Roll Steel: To my knowledge, we know of no application today where this material is
used in the fabrication of trays. Certainly, they would not be applicable in the food or health care
industry because of the potential for rust contamination. However, we do not see any objections for
their use in the manufacture of flowers and especially if they are either galvanized or electroplated.
They would perhaps be easier to construct and may represent the least expensive of all the trays.
Stainless Steel: Trays made from stainless steel are perhaps the most commonly used trays in
the lyophilization or freeze drying of products. They have numerous advantages which would
include and not be limited to (a), they are corrosion resistant: (b), such trays can be depyrogenated
at temperatures of 250 oC; (c), these trays are also durable and require no special handling and (d),
they are reusable which in time can off-set their initial higher cost.
Aluminum: Aluminum trays are used in the freeze drying of bulk materials. They are certainly
light weight when compared to the cold roll and stainless steel trays and are corrosion resistance. In
addition, they can be anodized to give them a black surface which will increase their emissivity aid
some what to the energy transfer resulting from radiant energy. They can be steam sterilized but we
do not have any knowledge where they have been depyrogenated at 250 0C. Because aluminum is a
relatively soft metal, one must be concerned with the effects of “galling” (the removal of a surface
layer generally in the form of a dust) should the product have to be removed by scraping.
Glass: Not all bulk products that can be lyophilized or freeze dried come into contact with a
metal surface. As a result of their composition, these trays have similar advantages as those
constructed from stainless steel, except they are by their very nature fragile and are subject to
breakage that may occur over time from stress strains resulting from age and thermal shock.
Plastic Trays: Molded trays are useful in those applications not requiring high temperature
depyrogenation. They tend to be (a), light weight when compared to the trays made from the
previous materials; (b), easy to clean and (c), quite durable and reusable.
Lyoguard Trays: While these trays can also be generally classified as plastic trays, they do
have unique features and we felt they deserved their own category in this discussion. The basic
reason being that they are covered with a membrane barrier that will allow a formulation prepared
in an aseptic environment to be safely lyophilized or freeze dried in a non aseptic environment [1].
This feature is of major importance to those working with small freeze dryers which were not
designed to withstand the harsh rigors of steam sterilization. However, given such an advantage
there are also some disadvantages in that they tend not to be reusable and as a result of their unique
feature to be more costly than those trays fabricate from the above materials.
Heat Transfer Properties: It have discussed the impact and need to understand heat transfer
properties in previous and references. In this, we shall just consider the role that composition and
configuration of the tray can have on the heat transfer properties. In making such a comparison,
allow me to make a number of assumptions and conditions so as to avoid possible confusion.
o All the trays have the same general rectangular configuration and the only variables
will be their total mass and thickness of bottom and walls.
o That all trays contain the same volume of water.
o In each case, the shelf-surface temperature is uniform and chamber pressures are
equal.
49
The heat transfer rate (Qs) from shelf through the bottom wall of the tray is given by the Fourier`s
law:
∂T
Q1 = − Aλ
∂l (6.1)
Since the heating or cooling takes place in an unsteady state, the amount of heat (Q) will be:
∂T
Q = − Aλ t
∂l (6.2)
In the case of the bottom of the tray, the relation (2) becomes:
Q S = Aλ
(T s
− Tp )
δ (6.3)
Where:
A is the total surface area of the tray, m2;
∂T Ts − Tp
=
∂l δ - the thermal gradient, 0C/m, or Kelvin/m
- ∂ - the thickness of the bottom of the tray, m;
- Ts - the temperature of the shelf surface, given in 0C;
- Tp - the temperature of the internal bottom surface of the tray, 0C;
- t - time, sec.
Neglecting the contact resistance between shelf and tray, the external surface temperature of
the bottom of the tray is equal to that of the shelf.
One can consider that the temperature gradient has a single direction, perpendicular on the
surface of the bottom of the tray. In the regions with a good contact area of the bottom of the tray
with shelf surface there is also a good heat transfer. In the regions where the contact is not perfect it
appears a new thermal impedance to heat transfer between shelf and bottom of the tray.
This impedance is given by the existing medium (gas at very low pressure and low density),
between shelf and bottom of the tray and will increase as the pressure is lowered. Consequently, the
heat transfer is reduced in these regions. But the heat transfer is more complicated.
To the normal temperature gradient (perpendicular on shelf) new gradients can be added on
the other directions, anywhere a temperature difference exists between different points (position).
Cold Roll Steel: For purposes of this we shall assume that the thickness of the walls and
bottom are 0,051 inches (1,2 mm). The thermal conductivity (lambda) cold roll steel is about 45,6
W/(m 0C) and the specific heat over the normal temperature range for lyophilization or freeze
drying is 0,4609 kJ( kg· 0C) [4].
Now it is well known that it is rather difficult to fabricate a steel tray in which the bottom
surface is in full Thermal contact with the shelf surface. Thus one can only expect a fraction of the
bottom surface to be in direct contact with the shelf surface. In the worse cases, contact may be only
at three corners or rest on a small area in the middle. For our comparative model (which we will use
for all trays except for the Lyoguard trays) we will assume only 10% of the bottom surface will be
in contact with the shelf. Given that (Ts - Tp), is 1 0C, the value of Qs now becomes 3,875·A (in
kW). One can see that as A increases, the thermal conductivity across the tray becomes more of a
real factor. Energy will flow from the edges towards the center so one can expect a temperature
gradient across the bottom of the tray as the temperature of the shelf is varied. This temperature
50
gradient will be directly proportional to the thickness of the tray and inversely proportional to the
length of the conduction path so the configuration of the tray is an important consideration.
If the height of the walls of the tray are 1 inch (2,54 cm) and the density of the steel is taken as
7850 kg/m3 [4], the mass of the tray is defined as “abd”, where “a” is the width of the tray and “b”
is the tray length in m such that the mass (M) can be expressed as:
M = V m ⋅ 7850 ( kg ) (6.4)
where Vm is defined as the volume of metal used to fabricate the tray.
The amount of energy (Q) required just to change the tray temperature by 1 0C (∂T =1) can be
expressed as:
Q = M ⋅ c p ⋅ ∂T = M ⋅ 0,460911 ⋅ 1 = 0,460911 ⋅ M ( kJ )
(6.5)
Stainless Steel: Given the same thickness as the cold roll steel, the thermal conductivity
(lambda) is given as 15,11 W/(m 0C). A comparison of the thermal conductivity shows that the cold
roll steel is three (3) times more thermally conductive than that of the stainless steel. For a tray of
the same dimensions as that of the cold roll steel and with only 10% of the bottom of the tray in
contact with the shelves, the heat transfer rate (Q) between the shelf surface and the tray when (Ts -
Tp) is 1 0C will be 1,259·A (kW). As a result, for a tray constructed out of stainless steel, it will only
transfer 33 % of the energy of that for the cold rolled tray.
The specific heat of the 304 stainless steel is 0,5028 kJ/(kg 0C), the amount of energy one [5]
while the density is given as being 8020 kg/m3. The mass (M) of the tray having the same
dimensions as above, will then be given as:
M = V m ⋅ 8020 ( kg ) (6.6)
The amount of energy (Q) required just to change the tray temperature by 1 0C can be
expressed as:
Q = M ⋅ c p ⋅ ∂T = M ⋅ 0,5028 ⋅ 1(kJ )
(6.7)
Since the mass of the tray will be greater than that of the cold roll steel, the amount of energy
to change the temperature of the tray by 1 0C will be more than that for the cold roll steel.
Aluminum: Aluminum trays (alloy 2017) having the same thickness as the cold roll steel, the
thermal conductivity (lambda) is given as about 104 W/(m 0C). A comparison of the thermal
conductivity shows that this alloy of aluminum is 2,23 times more thermally conductive than that of
the cold roll steel and 6,88 times more thermally conductive than the 304 stainless steel tray. For a
tray of the same dimensions as that of the cold roll steel and with only 10% of the bottom of the tray
in contact with the shelves, the heat transfer rate (Qs) between the shelf surface and the tray when
(Ts - Tp) is 1 0C will be 8,66·A (kW). Thus the heat transfer rate for an aluminum tray would be
2,23 twice that for a cold roll steel tray and more than 6,88 times that for one constructed out of 304
stainless steel.
The specific heat of the of aluminum alloys is 0,8799 kJ/(kg 0C) [6], while the density is given
as being 2800 kg/m3 [4]. The mass (M) of the tray having the same dimensions as above, will then
be given as:
M A1 = Vm ⋅ 2800(kg )
(6.8)
The amount of energy (Q) required just to change the tray temperature by 1 0C can be expressed as
51
Q A1 = M A1 ⋅ c p ⋅ ∂T = M A1 ⋅ 0,8799 ⋅ 1(kJ )
(6.9)
Or:
Q A1 = Vm ⋅ 2800 ⋅ 0,8799 = 2463,72 ⋅ Vm (kJ )
While for cold roll steel we have:
Q = Vm ⋅ 7850 ⋅ 0,460911 ⋅ 1 = 3618 ⋅ Vm (kJ )
Although the specific heat for aluminum is about twice that for the cold roll steel and the
stainless steel, it will require less energy to increase its temperature 1 0C, in fact, when compared to
stainless steel it will only require 68 % as much energy.
Glass: Because glass trays are more fragile and subject to breakage the tray thickness will be
greater and be of the order of 0,20 inches (5 mm) or about four times thicker than the metal trays.
For a borosilicate glass the thermal conductivity is about 0,83 W/(m0C) [4] while the specific heat is
0,75 kJ/(kg 0C) [7]. The heat transfer rate (Qs) for this type of tray will be 0,166 ·A (kW). The
density of borosilicate glass is about 2300 kg/m3.
The mass (M) of the tray having the same dimensions as above, will then be given as:
M glass = Vmglass ⋅ r = Vm ⋅ 2300(kg )

(6.10)
The amount of energy (Q) required just to change the tray temperature by 1 0C can be expressed as:
Q glass = M glass ⋅ c p ⋅ ∂T = M glass ⋅ 0,75 ⋅ 1 = 0,75 ⋅ M glass (kJ )

(6.11)
Even though it will require less energy to increase the temperature of the tray by 1 0C, because
of the poor thermal conductivity, one could expect at times large temperature differentials that can
exist across such trays as a result of the much lower thermal conductivity.
Plastic Trays: Assume that the plastic material that we have used to fabricate the tray is PETG
[8] and has a gauge thickness of 18 (1,2 mm) and so will be similar in dimensions to that of the
above metal trays. The thermal conductivity of the PETG is 0,1675 W/m0C and the heat transfer
rate (Qs) for the tray when there is a 1 0C differential would be just 0,166 ·A (kW). This “Qs” value
is considerably lower than that for a thicker layer of glass which is surprising.
The specific heat of the PETG plastic is 1,089 kJ/(kgoC) [8], while the density is given as
being 1270 kg/m3 [8]. The mass (M) of the tray having the same dimensions as above, will then be
given as:
M PETG = VmPETG ⋅ 1270(kg )

(6.12)
The amount of energy (QPETG) required just changing the tray temperature by 1 0C can be
expressed as:
Q PETG = M PETG ⋅ c p ⋅ ∂T = 1,089 ⋅ 1 ⋅ M PETG (kJ )

(6.13)
While it will require little energy to change the temperature of the tray, the poor thermal
conductivity could, at times, result in large temperature differentials across such trays during the
lyophilization or freeze drying process.
52
Lyoguard Trays:
The thickness of the Lyoguard tray is 0,008 inches (0,2 mm) and according to Steve Delrosso
of W. L. Gore & Associates, Inc.[1], the material is flexible enough to conform to the shelf surface.
In view of this information, we shall assume that at least 90% of the bottom of the tray is in contact
with the shelf surface.
Mr. Delrosso also informed me that the bottom of the tray was fabricated from polyethylene.
The thermal conductivity (lambda) of polyethylene ranges from 0,32 - 0,5028 W/(m 0C). As a result
the heat transfer rate (Qs) (assuming the l for this tray is estimated to be 0,5028 W/(m 0C) will be
2,514·A (kW) for a temperature differential of 1 0C. A comparison of the (Qs) for this tray shows
that its heat transfer rate will be 15 times to 18 times higher than that expected for either the glass or
plastic tray. However, when compared to that of the metal trays it is considerably lower, e.g., the q
will be 1/6 th that for the stainless steel tray.
The specific heat of the polyethylene is 1,88 kJ/(kg 0C) [8], while the density is given as being
940 kg/m3 [8].
The mass (M) of the bottom of the tray will be:
M Lyoguard = VmLyoguard ⋅ 940 (kg )

(6.14)
The amount of energy (Cp) required just to change the tray temperature by 1 0C can be expressed as:
Q Lyoguard = M Lyoguard ⋅ c p ⋅ ∂T = 1,88 ⋅ M Lyoguard (kJ )

(6.15)
Because of the relative low mass of the bottom of the tray, the amount of energy required to
increase the temperature of the polyethylene by 1 0C is considered to be insignificant.
The above calculations are presented in the following table:
Heat transfer Amount of heat to
Specific
Thermal rate/1 degree change the tray
Density heat
Tray material conductivity temperature temperature by
ρ [kg/m3] cp
λ [W/m0C] difference 10
C
[kJ/kg0C]
Qs [kW/10C] Q [kJ/10C]
Cold Roll Steel
0,4609-
(thickness = 7850 46,5 38,75·A 0,46091·M
0,48
1,2mm)
Stainless Steel
15,119-20,93
(thickness = 8020 0,5028 12,59·A 0,5058·Mst.steel
(15.11)
1,2mm)
Aluminium
Alloy 2017
2800 104 0,8799 86,66·A 0,8799·MAl
(thickness =
1,2mm)
Glass (thickness
2300 0,83 0,75 0,166·A 0,75·Mglass
= 1,2mm)
PETG
(thickness = 1270 0,1676 1,089 0,139·A 1,089·MPETG
1,2mm)
Lyoguard trays
(thickness = 910-940 0,32-0,5028 1,88 2,514·A 1,88·MLyoguard
0,2mm)
53
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.
Schematic of a Laboratory Freeze Dryer with Shell Frozen Flasks Attached to a Manifold
Figure 6.12.
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.
54
Freeze Dryer with Shelf
Figure 6.13.
Typical Capacities
Product Type Vegetables Shrimps Boiled Tea Granulate Coffee
Beef Granulate
Dry Matter % 10.0 22.0 35.0 25.0 40.0
Load kg/batch 6.0 6.6 7.8 5.5 6.9
Batch time h 8.0 8.0 6.0 6.0 6.0
Sublimation kg 5.4 5.1 5.0 4.1 4.1

H2O/batch
Production 0.6 1.5 2.8 1.4 2.8

kg/batch
(Capacity and freeze-drying time depend on e.g. product type, bulk density, product layer thickness
and heat sensitivity)
Figure 6.14- Pilot Freeze Drying Plant - RAY™ 1.
55
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.
Laboratory Lyophilizer Model WR 1: "As simple as an oven as reliable as a household
refrigerator"
Performs all these tasks:

• Bulk Drying in Trays
• Vial Drying with Stoppering under full vacuum
or partial pressure
• Monitoring of all essential parameters:
o Product Temperature
o Tray Temperature
o Condenser Temperature
o System Vacuum
• Controlled Temperature rise for Product
• System Vacuum Control (optional)
Success in Freeze Drying depends a great deal on simplicity and reliability, and most
importantly, in the technician's comfort in operating the Freeze Dryer.
As in most mechanical equipment, simplicity is the key to both understanding and reliability.
Quite simply, the WR 1 Freeze Dryer is the simpliest and most reliable Freeze Dryer you can buy---
regardless of cost!
The WR 1 Freeze Dryer is designed with over thirty years experience acquired in servicing all
brands of Freeze Dryers, both laboratory and production models. The WR 1 Freeze Dryer reflects
all the advantages and the correction of most of the disadvantages of conventionally manufactured
freeze dryers.
The differences between this lyophilizer and all others are too detailed to list here.
SPECIFICATIONS
Refrigeration CFC Free
Cabinet Caster mounted, welded steel frame cabinet, finished in bonderized
Appliance enamel
Weight 600 lbs
Product Trays One (1) to Four (4) Stainless Steel Trays
Product Tray
14" wide, 14" deep
Dimensions
Stoppering Adjustable height trays that move upward within the chamber.
Stoppering mechanism is air driven.
Overall Dimensions 48" wide, 34" deep, 35" high
56
Figure 6.15. - THE LYOLAB 984 MICRO-CONTROLLER: 21 CFR Part 11 Compliant
Upgrade your freeze dryer's control system and improve product quality. The Lyolab984
micro-controller was designed to control and monitor the main lyophilization parameters on a
variety of machines. The Lyolab984 combines the functions of separate temperature and pressure
controllers together with the cycle sequencer to integrate the automation of the lyophilization cycle
on a Windows 2000 ® PC. The Lyolab984 comes with a moderate price tag and targets the retrofit
market.
It believe that a control system must facilitate the inter-relationship between the freeze dryer, the
product, the recipe and the operator. Therefore, a control design must be evaluated in its ability to:
• minimize operator error
• clearly define recipe parameters
• provide a synoptic of the machine for maintenance
• automate intervention (AI) to alert possible malfunctions and product related
behavior deviations
• facilitate cycle modifications before and during a cycle
• document a cycle in both data log and graph modes
• support cGMP operation
• be user-friendly
The LyoLab984 Micro-controller was designed with these functional characteristics in mind.
57
One of the most important functions of a fully integrated control system is to simplify recipe
definition. With our unique EZ-n-Tree screens, recipe definition is a breeze. Creating a cycle profile
for lyophilization is simplified by a series of prompting messages, help screens and warnings. Once
a recipe is outlined, the temperature profile can be plotted, providing a visual representation of the
recipe prior to running a cycle.
Since readable and accessible data is important in cycle development, the cycle batch log can
be printed in a variety of formats, including graphical and numerical modes. Cycle data can also be
imported into most spreadsheets, databases, and can even be appended to a document with a word
processor. Our software is among the most flexible on the market.
Lyolab 984 features include:
• Automated lyophilization cycle
• Automated performance cycle
• Manual synoptic mode
• Alarms and safeties annunciation
• Offline analysis of logs and graphs
• Password access security
• Multiple data presentation (event log, graphic)
• Graphic recipe utility
• PLC-OMI Smart Link monitoring
• Recipe conversion utility
• Printing of logs/graphs in full color
• Simulation mode for training
• Easy to learn interface
At Lyotech we understand that a control system is only as good as its supporting
documentation. In the cGMP environment of most pharmaceutical manufacturing, documentation is
critical for effective qualification of the control system. The LyoLab984 Micro-controller, which
includes the PLC logic and OMI software, was designed for such a cGMP environment. The PLC
ladder logic and the OMI code are fully documented including an operator interface manual, an
interface technical supplement, and electrical schematics. In addition, a Control Qualification (CQ)
package is offered, providing static I/O testing, safety interlock checks, sequencing checks and
software test scripts (black box testing), for complete GMP readiness.
Figure 6.16. Screen shots
58
Figure 6.17.
A typical machine consists of a freeze-drying (lyophilization) chamber with several shelves

attached to heating units, a freezing coil connected to a refrigerator compressor, and a vacuum
pump.
still unfrozen. When it seal the chamber and begin the process, the machine runs the compressors to
lower the temperature in the chamber. The material is frozen solid, which separates the water from
everything around it, on a molecular level, even though the water is still present.
Benchtop manifold freeze dryer.
There are essentially three categories of freeze dryers: rotary evaporators, manifold freeze
dryers, and tray freeze dryers.
Rotary freeze dryers are usually used with liquid products, such as pharmaceutical solutions
and tissue extracts.
Figure 6.18. Unloading trays of freeze -dried material from a small cabinet-type freeze dryer
Manifold freeze dryers are usually used when drying a large amount of small containers and
the product will be used in a short period of time. A manifold dryer will dry the product to less than
5% moisture content. Without heat only primary drying (removal of the unbound water) can be
59
achieved. A heater needs to be added for secondary drying, which will remove the bound water and
will produce a lower moisture content.
Production freeze dryer. Tray freeze dryers are more sophisticated and are used to dry a
variety of materials. A tray freeze dryer is used to produce the driest product for long-term storage.
A tray freeze dryer allows the product to be frozen in place and performs both primary (unbound
water removal) and secondary (bound water removal) freeze drying, thus producing the driest
possible end-product. Tray freeze dryers can dry product in bulk or in vials. When drying in vials,
the freeze dryer is supplied with a stoppering mechanism that allows a stopper to be pressed into
place sealing the vial before it is exposed to the atmosphere. This is used for long term storage, such
as vaccines.
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.
• 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 one kilogram
of water per square meter of surface per hour, considering that 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:
60
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
--------------------------- = 15 hours
10kw x 860kcal x 0.8
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.
Collapse temperature (Tc): Where a frozen amorphous solution starts to lose structure whilst
freeze-drying. Can occur over several degrees before total loss of structure.
Eutectic temperature (Teu): Where a frozen crystalline solution will no longer freeze-dry
with any structure. A discrete temperature.
Tm: The melting temperature of the frozen mass
61
7. CONCLUSIONS
The advantages of lyophilizeted products are: very lightweight; food retains it original shape,
with good nutritional value, and flavor; readily reconstitutes, requiring a minimum of fuel in
preparation; keeps for years; can be repackaged; many products are delicious in their dried state, for
example whole freeze dried berries; freeze dried products combine well into premixed package
meals, they do not readily exchange flavors, meaning the broccoli will still taste like broccoli, the
chicken like chicken, and the rice like rice, even if they have been in the same package for two
years; excellent for creating drink crystals, the most popular being freeze dried instant coffee.
Lyophilization has many advantages compared to other drying and preserving techniques:
Ease of processing a liquid, which simplifies aseptic handling
Enhanced stability of a dry powder
Removal of water without excessive heating of the product
Enhanced product stability in a dry state
Rapid and easy dissolution of reconstituted product
maintains food quality because the food remains at a temperature that is below the freezing
point during the process of sublimation.
Foods, which are lyophilized, can usually be stored without refrigeration, which results in a
significant reduction of storage and transportation costs.
Lyophilization greatly reduces weight, and this makes the products easier to transport.
For example, many foods contain as much as 90% water. These foods are 10 times lighter
after lyophilization.
Because they are porous, most freeze-dried foods can be easily rehydrated, with friable
structure.
Lyophilization does not significantly reduce volume, therefore water quickly regains its
place in the molecular structure of the food.
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
But in the present it is known some disadvantages of lyophilizated: expensive to produce

requiring large scale equipment; some freeze dried products can be brittle and tend to powder or
crush easily if knocked about; limited to products that freeze well. For example leafy produce such
as lettuce does not freeze well and, therefore, is unsuitable for freeze drying.
Increased handling and processing time
Need for sterile diluent’s upon reconstitution
Cost and complexity of equipmente
xpensive to produce requiring large scale equipment;
some freeze dried products can be brittle and tend to powder or crush easily if knocked
about;
limited to products that freeze well. For example leafy produce such as lettuce does not
freeze well and, therefore, is unsuitable for freeze drying.
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)
The future for application of the lyophilization in the study of foods is in the development of
that through increasing a good method of freeze-drying for make some experiments with many kind
of different foods, for the improvement that in food production.
It is necessary and is possible to study the corn oil extract, wheat flour, the maltodextrin from
corn, modified cornstarch, spice extracts, soy sauce, hydrolyzed wheat gluten, partially
62
hydrogenated soybean and cottonseed oil etc. That is very porous, since it occupies the same
volume as the original and so rehydrates rapidly. There is less loss of flavor and texture than with
most other methods of drying.
• Food is not the only material that is freeze-dried. Pharmaceutical products such as
antibiotics and vaccines are often preserved this way. Specialty chemicals, pigments, and
ceramics powders are also produced using freeze-drying. Currently, there is development
work on freeze-drying various aerosol sprays.
• One of the most interesting applications is freeze-drying flowers to produce bouquets that
can be stored for many months before being reactivated to make "fresh" flowers. This would
be especially beneficial for those who want flowers that bloom only during a short season.
• Lyophilization integrated can be a very important part in foods conservation engineering,
special for a high food quality.
• For the future the lyophilizated food products can make a revolution in human nourishment
rational and scientific, because this solution is good for form whatever composition without
water reaction, in quantities well determined, with a maximum precision.
• Also, it can make many study and/or experiments about the compatibility between the meal
and the human body who used that.
Thus, full–scale the integrated engineering principles, for each human organism can determine
the adequate meal.
• Freeze-drying process is not widely used in the food industry due to its high operation cost.
• Although new improvements such as adsorption, fluidisation, and microwaves have been
researched in the last decade in order to reduce costs, vacuum freeze-drying is, up to now,
the only technology used in an industrial scale to dry coffee, spices, meats, food ingredients
and other high-value foods.
• However, with the increasing concern about food quality, this process could be considered
as a valuable alternative to preserve other foods.
• The high progress of the science and the techniques in this domain are created many
possibilities for partial or total elimination all the disadvantages
63
REFERENCES
[1]. Gabriela ANGHEL, Dumitru MNERIE, Dumitru ŢUCU - OPPORTUNITY OF

RESEARCH BY FREEZE DRYING THE FOOD -, International Conference Integrated
Systems for Agri-food Production ISAP’03, 20-22.11.2003, Timişoara, Romania
[2]. A. Bacaoanu and S. Picos, Rev. Roum. de Chim., 2005, no.6
[3]. R. Bruttini, “Freeze Drying, Pharmaceuticals Encyclopaedia of Bioprocess Technology
Fermentation, Biocatalysis and Bioseparation”, Ed. John Wiley and Sons Inc., New York,
1999, 1276 – 1289.
[4]. M.N. Estiaghi, R. Stute and D. Knorr, J. Food Sci., 1994, 59, 1168-1170
[5]. T. A. Jennings, “Lyophilization - Introduction and Basic Principles”, Interpharm Press,
Buffalo Grove, IL 1999.
[6]. M. Kochs, Ch. Körber, B. Nunner and I. Herschel, Int. J. Heat Mass Transfer, 1991, 34,
2395-2408
[7]. A. I. Liapis and R. Bruttini, in A.S. Mujumdar ed., “Handbook of industrial Drying” vol.1,
2nd ed., Marcel Dekker, New York and Basel, 1995, 309 – 343.
[8]. R.J. Litchfield and A.I. Liapis, Che . Eng. Sci,. 1982, 37, 45-55
[9]. R.G. Livesey and T.W.G. Rove, J. Parenteral Sci. Technol., 1987, 41, 169-171
[10].
[11]. A.P. MacKenzie, “Collapse during freeze drying-qualitative and quantitative aspects. in
Freeze-Drying and Advanced Food Technology”, edited by S.A. Goldblith , L. Rey and W.
Rothmayr, Academic Press, New York, 1975
[12]. D.W. McPeak, E. Idziak and T.G. Smyrl, Can. Inst. Food. Sci.Technol. J., 1987, 20, 176-
178
[13]. M.J. Millman, A.I. Liapis and J.M. Marchello, J. Food Technol., 1985, 20, 541
[14]. Dumitru MNERIE, Gabriela Victoria ANGHEL - SOME APPLICATIONS OF
LYOPHILIZATION IN AGRICULTURE, - Opatia08LyophAgriculture
[15]. M.J. Pikal, S. Shah and J.E. Lang, J. Pharm. Sci., 1983, 72, 635-650
[16]. Y. Sagara, Drying Technnol., 2001, 19, 281 – 296
Contact:
Univ.Prof. eng.Ec. Dumitru MNERIE PhD

University POLITEHNICA of Timisoara, Romania
dumitru_mnerie@yahoo.com
64

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JIHOČESCKÁ UNIVERZITA POLITEHNICA UNIVERSITY

V ČESKÝCH BUDĚJOVICÍCH OF TIMISOARA

Univ.Prof. Eng. Dumitru MNERIE, PhD

APRIL - MAY 2008

Freeze-drying (lyophilisation) is a widely used method for dehydrating a vast range of

1.1. Historical review

1.2. Evolution of process and equipment

where Bimd is the Biot number for mass transfer defined as

The parameter φ was shown theoretically and experimentally to be independent of drying

Figure 1.1. Basic Industrial Freeze Dryer

The most important are listed below:

1.3. Definition of lyophilization

- Drying, evaporation, desiccation, dehydration mean the processes of extracting moisture.

1.4. General description of the process

Figure 2.1. Diagram pressure – temperature.

Figure 2.2. - The transformation of phases for lyophilization process.

Freeze-drying is a multiple operation which involves the following three stages:

2.1. Primary Drying Process

2.2. Secondary Drying Process

In addition to concentration, the permeability is influenced significantly by freezing manner,

Fig. 2.5 - Drying of the material in a tray

Fig. 2.6 - Freeze-drying model for transport properties analysis

The mass flux may be expressed as

Fig. 2.7 - Permeability vs. porosity of the dried layer

3.1. Healthcare Industry

Bobwhite quail Fisher

Bean curd Green bean Garlic

Scallion Red pepper Beef

Crab meat Chicken meat Pork

Corn Dried shrimp Ham

Carrot Mushroom Goloden needle

Figure 3.4 – Lyophilization products

Figure 35. - Dried prunes: a.- lyophilizated; b. – by heating (roaster).

Vegetables and Pulses Meats

Dill Smoked Salmon

A package of freeze-dried ice cream, sold as a novelty item.

1 Wheat Oats Rice Oats Oats

2 Rye Corn Rye Buckwheat Barley

3 Oats Barley Corn Barley Buckwheat

4 Barley Buckwheat Wheat Wheat Rye

5 Corn Rice Barley Rye Wheat

6 Buckwheat Wheat Buckwheat Corn Corn

7 Rice Rye Oats Rice Rice

The maize lyophilized The maize reconstituted after lyophilization

3.4. Other Applications

In bioseparations, freeze drying can also be used as a late-stage purification procedure,

In industrial freeze drying applications it is essential to reproduce identical drying protocols

4. TRADITIONAL LYOPHILIZATION TECHNOLOGY

Installation of lyopilization with interior freezing an interior condenser

Installation of lyopilization with interior freezing and exterior condenser

The most important mechanical engineering elements are listed below:

Figure 5.1. Figure 5.2

Figure 6.1. - Freeze drying system for laboratory (Thomas A. Jennings)

Figure 6.2. – The laboratory scheme of lyopilizer

Figure 6.4. - A simplified freeze-drying machine

Fig 6.5. Lyophilization machine

A simplified freeze dryer system is presented in Fig.6.6.

Fig. 6.6. - Simplified freeze-dryer

Figure 6.7.. – Lab installation with lyophilizer Christ Alpha 1-4.

Figure 6.8.- The atmospheric spray-freeze-drying process

Figure 6.9.- Freeze-Drying Microscope System

A typical machine consists of a freeze-drying chamber with several shelves attached to