BASIC prInCIpleS OF lYOpHIlIZATIOn prOCeSS

ABOUT AUTHOR
MR.AMOL P.MANE

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ABSTRACTS :
The main purpose of this article is to clear the basics concepts about the principles , operation , and
validation procedure of the deep freezers in the pharmaceuticals.The successful working of the lyophilizer
are depends upon the scalability , process modeling by using statistical models and finally trial and error
model . Now days trial and error model is very popular. The critical process parameters and critical
quality attributes are plays an important role in the process .The statistical method to be incorporated for
the cycle development in which the heat and the mass transfer are directly involved in lyophilization
process. Different stages to be monitored as an critical such as primary stage , cooling stage and the
secondary phase.The critical parameter such as the temperature , time and camber pressure are directly
affect in the process. The lyophilization is carried out by the simple principle of physics by
sublimation .Sublimation is the process in which conversion of the substance from solid to vapour state
without first passing through an intermediate liquid state.
INTRODUCTION:
The stabilization of the drug products through their shelf life is a challenge to the researcher . The
physical , chemical nature of the drug products remains same as per standard specifications is very
important. Most of the pharmaceutical drug products are unstable in its original forms. The products like
dry powder injectable , lyophilized products are very difficult to store at their original form due to their
drug degradation nature .The same drugs are used as and when required by reconstitution with the sterile
water for injection.The manufacturing of the lyophilized product is the big challenge . If failure in the
batch happens it gets huge impact on the financial / costing, hence the selected batches are very small
and taken on the trial and error base.While manufacturing of the batches ICH Q8 guidelines for
pharmaceutical development to be taken into consideration and the batches are manufacture by using
quality by design tool.The potential risk are identified , mitigated , investigated and control as per ICH Q9
guideline as Quality risk management and the quality of the product shall be maintained through the
entire shelf life of the manufactured drug product as per ICH Q10 pharmaceutical quality system.
OVERVIEW ON LYOPHILIZATION :
The Lyophilization is also called as an freeze drying process . Lyophilization is the process of removal of
ice or other frozen solvents from a material through the process of sublimation and the removal of bound
water molecules through the process of desorption.This process is called sublimation, transforms the ice
directly into water vapor, without first passing through the liquid state. The water vapor given off by the
product in the sublimation phase condenses as ice on a collection trap, known as condenser. The typical
drying cycle is consist of the three different stages: (1) freezing stage, (2) primary drying stage, and (3)
secondary drying stage as described in the diagram.

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 DIAGRAM -A

BASIC PRINCIPLES OF FREEZE DRYER:

The steps requirement to lyophilize a product in a batch process are summarized as follows:

 Pre-treatment / Formulation

 Loading in Freeze Dryer (vials)

 Freezing (Thermal Treatment) at below atmospheric Pressure

 Eutectic temperature

 Primary Drying (Sublimation) under Vacuum

 Secondary Drying (Desorption) under Vacuum

 Pre-aeration and Stoppering (For Product in Vials) under Partial Vacuum followed by aeration

 Removal of Dried product from Freeze Dryer

The schematic diagram A represents the red and green lines shows the product and actual shelve
temperature . The diagram B represents the actual difference between the lab scale , pilot scale and
industrial production scale lyophilizers .

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 DIAGRAM -B

The batch is manufactured under stated conditions and in the manufacturing tanks . The same batch is
filtered through the 0.22µ filter . The same batch is taken for filling activity . During filling activity all the
environmental conditions are to be taken into consideration such as temperature , relative humidity ,
viable and non viable particulate monitoring, pressure differential . All the primary packing materials like
vials , rubber stoppers shall be sterilized through the autoclave. In filling stage half stoppering or semi
stoppering to be done . This is very important step at filling stage of lyophilized products . The filled vials
shall be transferred to the deep freezing chamber shelves. The operation shall be performed in the clean
room class of A/B.Before going to lyophilization process there is need to set the defined parameters or
working process some times its referred as an the Recipe.Individual time , pressure and temperature are
specified in each step . For every new product there is requirement of development of product recipe.
In lyophilization the first stage is the deep freezing in which the water shall be converted in to the ice in
vials by super cooling mechanism.The temperature is changes time to time.This stage is not more lengthy
as the operation shall be completed in to the few hours.
The primary drying is the phase in which the bulk water removed from the product via sublimation of all
of the free ice crystals . Organic solvents are also removed during primary drying.
Primary drying is a slow process conducted at cooler temperatures, safely below the product’s critical
collapse temperature. All methods of heat transfer- conduction, convection and radiation are employed
during primary drying.The heat transfer induces the sublimation of ice in vials . The heat lost from the
product as a latent heat of sublimation will be supplied again from the chamber shelf .The primary drying
stage required more time and economical patience s.

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In addition to the free ice that is sublimated during primary drying, there remains a substantial amount of
water molecules that are bound to the product. This water is removed by desorption in secondary drying.
Secondary drying is done at increased temperature because all of the free ice has been removed during
primary drying hence there are no chances of melting or product collapse. Some freeze dryer have the
facility to remove the samples for moisture content determination in secondary drying step based on
which the drying step can be terminated. In this stage the product temperature elevates higher temperature
than the primary drying stage.The objective of secondary drying is to reduce the final residual water
content to an acceptable level .
There are three different steps involved in the freeze drying as ,
 Partial Aeration (Pre- Aeration)
Once the secondary drying step is completed, partial aeration-also called pre- aeration is being performed
usually using inert gas like nitrogen so as to leave some vacuum inside the vials.The compressed air is
avoided in this operation.
 Full Stoppering
Once the Partial Aeration is completed, the vials are full stoppered by lowering the freeze dryer shelves
one by one. This can be automatic step in the freeze dryer recipe or can be performed in manual mode by
pushing the button.Once the full stoppering is completed, shelf position is restored to original position.
 Aeration
Final aeration is done usually with compressed air to bring the chamber to the atmospheric pressure.
Condenser aeration is done after chamber aeration. Door of the freeze dryer is opened at aseptic area side
to unload vials.The vials are then sent to capping machine for further operation. These above mentioned
steps are interdependent on each other one of the steps failure impacted on further operation.
OVERVIEW ON THE QUALIFICATION OF THE LYOPHILIZER:

Generally qualification of the lyophilizer shall be performed as per below mentioned stages
URS : (User requirements specification) The URS comprises the basic user requirement aspect with
respect to the product manufacturing and as per cGMP requirement.
DQ: (Design qualification) it comprises the basic design consideration as per user requirement .
FAT: ( Factory acceptance test )This can be done at the manufacturer site , i.e. Pre shipment critical test
to be preformed and verified.
SAT: (Site acceptance test )This can be done on site . The verification of actual shipment can be done
with respect to the any damage during the shipment.
Installation Qualification: The installation qualification shall be performance to verify the actual
machine is installed in dedicated place in which the basic component verification , identification ,
leveling , alignment can be done.
Operational Qualification: in which the operational parameters shall be verified and critical calibration
component verified . Draft standard operating procedure shall be prepared in this stage.
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Performance Qualification: in which the actual performance of the machine shall be verified .
 Equipment Qualification:
 Shelf Temperature Performance Verification
Ultimate shelf heating and cooling rates (+20°C to -40°C in ≤ 60 minutes) Ultimate high and low
temperatures (minimum temp below -55°C after 30 min cooling and max temp above +80°C).
 Condenser Cooling Verification
Maximum condenser cooling rate (Ambient to -40°C in ≤30 minutes) Ultimate condenser cooling test
(Temperature ≤ -75°C after 60 minutes of cooling).
 Vacuum Performance Verification
Time to achieve 100 µbar should be less than 40 minutes Ultimate vacuum of 5 µbar.
 Main Valve Leak Rate Test (0.01 mbar/Liter / Sec)
Main valve leak rate of 2 µbar in 30 minutes, Allowable rise = 0.05x1800 seconds /4360 liters =
0.0021 mbar.
 Shelf Temperature Set point Control at -40°C, 0°C and +40°C ± 2°C.
 Shelf Temperature Uniformity Verification -40°C, 0°C and +40°C ± 2°C.
 Vacuum Set point Control at 500, 100 and 50 µbar ± 5 µbar.
 CIP Coverage Verification (Riboflavin Test).
 SIP Effectiveness Verification (Sterilization and cooling).
 System Leak Rate Test (0.01 mbar /Liter / Sec).
Allowable rise = 0.01x3600 seconds / 8140 liters = 0.0044 mbar
 Shelf Ram Bellows Integrity Test.
Verify the pressure rise test of the bellows was performed to confirm the leak tight to 0.01 mbar
Liter / Sec. Record the pump down time to 10 μbar here as the pass criteria.
 Main Valve Bellows Integrity Test.
Verify the pressure rise test of the bellows was performed to confirm the leak tight to 0.01 mbar
Liter / Sec. Record the pump down time to 10 μbar here as the pass criteria.
 Condenser Capacity Verification
 Condenser Defrosting Test
 Freeze Drying Cycle with Placebo
 CIP and SIP

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 Advantages of Freeze Drying
1. To dry heat labile and water-unstable materials.
2. To get accurate, sterile dosing into the final product container.
3. To extend the shelf life or stability.
4. To preserve the biological activity of a product.
5. To eliminate the need for refrigerated storage.
 Disadvantages of Freeze Drying
1. To Very complex process requires high degree of automation and process control and men power with
special skills and training for routine operation.
2. The total Capital cost of the Freeze Dryer equipment is very high.
3. Operating cost, regular maintenance, utilities and spare parts cost of the freeze dryer is very high
resulting in higher cost of end product.
4. Freeze Drying is very long process running for several days.
5. A least scope of saving the product in condition of equipment failure during running process.

REFERENCES:
1. S. L. Nail, S. Jiaang, S. Chongprasert, and S. A. Knopp, “Fundamentals of freeze-drying,”
in Development and Manufacture of Protein Pharmaceuticals, S. L. Nail and M. J. Akers, Eds.,
Kluwer Academic/Plenum Publisher, New York, NY, USA, 2002.View at: Google Scholar
2. V. M. Zavala and L. T. Biegler, “Optimization-based strategies for the operation of low-density
polyethylene tubular reactors: nonlinear model predictive control,” Computers & Chemical
Engineering, vol. 33, 2009.View at: Publisher Site .
3. A. A. Barresi, V. Rasetto, and D. L. Marchisio, “Use of computational fluid dynamics for
improving freeze-dryers design and process understanding. Part 1: modelling the lyophilisation
chamber,” European Journal of Pharmaceutics and Biopharmaceutics, vol. 129, 2018.View
at: Publisher Site .
4. D. L. Marchisio, M. Galan, and A. A. Barresi, “Use of computational fluid dynamics for
improving freeze-dryers design and process understanding. Part 2: condenser duct and valve
modelling,” European Journal of Pharmaceutics and Biopharmaceutics, vol. 129, 2018.
5. Recent Development of Optimization of Lyophilization Process Hidenori
awasaki,1,2 Toshinori Shimanouchi and Yukitaka Kimura Graduate School of
Environmental and Life Science, Okayama University, 3-1-1, Tsuhima-naka, Kitaku, Okayama,
Japan .
6. Lyophilization basic principle slide share by Dhaval surti.