Lyophilization

Subtitle

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Lyopholization or freeze drying
• It is 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.
• This is in contrast to dehydration by most conventional methods
that evaporate water using heat.

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• In this process, the moisture content of the product is reduced
to such a low level that does not support biological growth or
chemical reactions which gives the stability to the
formulation.
• This technique useful in formulation development of drugs
which are thermolabile and/or unstable in aqueous medium.
3,4,5

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• This process is performed at temperature and pressure conditions below
the triple point, to facilitate sublimation of ice.

• The entire process is performed at low temperature and pressure, so that

useful for drying of thermolabile compounds.

• Four important Steps involved in lyophilization process to obtain the

final dried product with desired moisture content (Figure 2).

i. sample preparation

ii. freezing,

iii. primary drying (sublimation),

iv. secondary drying (desorption)

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Lyophilization process is based on the principle of sublimation of ice, without entering the
liquid phase.
The phase diagram of water (Figure 1) represent that two phases coexist along a line under
the given conditions of temperature and pressure, at the triple point .

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Figure 1- Phase diagram showing the triple point of water at 0.01°C, 0.00603 atm. Lyophilization is take place below the triple point.
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Figure 2. Steps involved in lyophilization process from sample preparation to final product formation.
Annealing is an another optional step, occasionally used to crystalline substance.

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Stages of freeze drying
Pretreatment
• Pretreatment includes any method of treating the product prior to freezing

• This may include

1. concentrating the product,

2. formulation revision (i.e., addition of components to increase stability,

preserve appearance, and/or improve processing),

3. decreasing a high-vapor-pressure solvent, or increasing the surface area

• Food pieces are often IQF treated to make them free flowing prior to
freeze drying.

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Freezing and annealing
• During the freezing stage, the material is cooled below its triple point, the
lowest temperature at which the solid, liquid and gas phases of the material
can coexist.

• This ensures that sublimation rather than melting will occur in the following
steps.

• To facilitate faster and more efficient freeze drying, larger ice crystals are
preferable. The large ice crystals form a network within the product which
promotes faster removal of water vapor during sublimation.

• To produce larger crystals, the product should be frozen slowly or can be

cycled up and down in temperature in a process called annealing.

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Cont… freezing

• The freezing phase is the most critical in the whole freeze-drying process,
as the freezing method can impact the

• speed of reconstitution,

• duration of freeze-drying cycle,

• product stability, and

• appropriate crystallization

• Amorphous materials do not have a eutectic point, but they do have a

critical point, below which the product must be maintained to prevent
melt-back or collapse during primary and secondary drying.

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Figure 3 Flowchart showing the concept of eutectic temperature and Tg, and their importance
during primary drying.

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Structurally sensitive goods
• In the case of goods where preservation of structure is required,

• like food or objects with formerly-living cells,

• large ice crystals will break the cell walls which can result in increasingly
poor texture and loss of nutritive content.

• In this case, the freezing is done rapidly, in order to lower the material to
below its eutectic point quickly,

• thus avoiding the formation of large ice crystals.[2]

• Usually, the freezing temperatures are between −50 °C (−58 °F) and
−80 °C (−112 °F).

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Primary drying
• During the primary drying phase, the pressure is lowered (few millibars), and
enough heat is supplied to the material for the ice to sublime.

• In this initial drying phase, about 95% of the water in the material is
sublimated.

• This phase may be slow (can be several days in the industry), 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 up the sublimation, making it useful as a deliberate drying

process.

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Cont…pri freezing

• Furthermore, a cold condenser chamber and/or condenser plates provide a

surface(s) for the water vapour to re-liquify and solidify on.

• During primary drying, drying temperature should not exceed the critical
temperature, which otherwise leads to ‘meltback’ or ‘collapse’
phenomenon in case of crystalline or amorphous substance respectively
(Figure 3).11

• It is important to note that, in this range of pressure, the heat is brought

mainly by conduction or radiation; the convection effect is negligible, due
to the low air density.

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Secondary drying
• The secondary drying phase aims to remove unfrozen water molecules,
since the ice was removed in the primary drying phase.

• In this phase, the temperature is raised higher than in the primary drying
phase, and can even be above 0 °C (32 °F), to break any physico-chemical
interactions that have formed between the water molecules and the frozen
material.

• Usually the pressure is also lowered in this stage to encourage desorption

• After the freeze-drying process is complete, the vacuum is usually broken

with an inert gas, such as nitrogen, before the material is sealed.

• At the end of the operation, the final residual water content in the product
is extremely low, around 1% to 4%.

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Advantages of lyophilization:
1-It is a ideal drying technique for heat sensitive
products, 6-lyophilized products sensitive to
oxidation can be stoppered and sealed
2- It can be stored at ambient temperature over a 2 within an inert atmosphere (i.e. nitrogen)
year shelf life, enhanced product stability in a dry to minimize detrimental effects.
state.
3. Removal of water without excessive heating of
the product
3- Easy reconstitution greatly reduces weight and
makes the products easier to transport,
5- It is not limited to products for parenteral use,
but can also be used for fast dissolving sublingual
tablets. Tablets can have very low disintegration
time and have great mouth feel due to fast melting
effect. 18
 DISADVANTAGES-

1. It is a long and cost intensive process,

2. requires sterile diluents for reconstitution,
3. it should only be used when product is unstable and
heat-liable,
4. the limited amount of vials processed in each run
restricts the overall production capacity. 12
5. Cost and complexity of equipment

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 MATERIAL THAT CAN BE LYOPHILIZED-
The major type of material that can be lyophilized:
Primary applications of freeze drying include biological (e.g.,
bacteria and yeasts), biomedical (e.g., surgical transplants),
food processing (e.g., coffee) and preservation.

1-Non-living bio products this comprises the major areas of

application and include:
Enzyme, hormones, antibiotics, vitamins, blood products, inactivated
vaccines etc.
Foodstuffs, Industrially useful bio-products.
Bone and other body tissue for medical and surgical use.

2-Non-biological where the process is used to dehydrate and

concentrate reactive and heat labile chemicals.
3- Living organism- where reconstituted cells after drying
must be able to grow and multiply to produce new progency.
4- miscellaneous- Flood damaged books, museum, artifacts
etc. 20
 Equipment and types of freeze dryers
Caption There are many types of freeze-dryers available,
1. vacuum chamber,
2. process condenser,
3. shelf-fluid system,
4. refrigeration system,
5. vacuum system, and
6. control system

A benchtop manifold freeze-drier Unloading trays of freeze-dried material from a small cabinet-type
freeze-dryer
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Function of essential components
• Chamber

• The chamber is highly polished and contains insulation, internally. It is

manufactured with stainless steel

• A hydraulic or electric motor is in place to ensure the door is vacuum-tight

when closed.

• Process condenser

• The process condenser consists of refrigerated coils or plates that can be

external or internal to the chamber.

• During the drying process, the condenser traps water. For increased
efficiency, the condenser temperature should be 20 °C (68 °F) less than the
product during primary drying[29]

• and have a defrosting mechanism to ensure that the maximum amount of

water vapor in the air is condensed.
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• Shelf fluid

• The amount of heat energy needed at times of the primary and secondary drying
phase is regulated by an external heat exchanger. Usually, silicone oil is use

• Refrigeration system

• This system works to cool shelves and the process condenser by using compressors or
liquid nitrogen.

• Vacuum system

• During the drying process, a vacuum of 50-100 microbar is applied, by the vacuum
system, to remove the solvent.[29] This system compresses non-condensable gases
through the condenser.

• Control system

• Finally, the control system sets up controlled values for shelf temperature, pressure
and time that are dependent on the product and/or the process. [30][31] The freeze-dryer
can run for a few hours or days depending on the product. [29]

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Pharmaceuticals and biotechnology
• Pharmaceutical companies often use freeze-drying to increase the shelf
life of the products, such as live virus vaccines,[13] biologics[14] and other
injectables., tablets or wafers.

• Examples of lyophilized biological products include many vaccines such

as live measles virus vaccine, typhoid vaccine, and meningococcal
polysaccharide vaccine groups A and C combined.

• By removing the water from the material and sealing the material in a
glass vial, the material can be easily stored, shipped, and later
reconstituted to its original form for injection.

• Dry powders of probiotics are often produced by bulk freeze-drying of

live microorganisms such as lactic acid bacteria and bifidobacteria.[18]

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• References
• https://www.pharmatutor.org/

• GUIDE TO INSPECTIONS OF LYOPHILIZATION OF PARENTERALS

(FDA)

• wikipedia

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THANK YOU

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