VALIDATION OF STERILIZATION

Presented By:
Mr. Kiran D. Baviskar,
Assist. Professor
Dept. of Pharmaceutics,

Smt. Sharadchandrika Suresh Patil College of Pharmacy, Chopda.

VALIDATION is defined as a documented procedure for recording &
interpreting data required to show that any process (machine/method) will
consistently comply with predetermined specifications

Sterilization refers to the complete absence or destruction of all micro-organisms

from a surface, equipment, medications (parenterals or ophthalmics), or
biological culture medium.
Various methods of sterilization include thermal methods (dry or moist heat) and
non-thermal methods (physical – filtration or radiation and chemical - gaseous).
 105 Non-
Site REJECT
Sterile
Aseptic
Microbial
manufacture Challenge Container 103 Non-
of products REJECT
Sterile
(106) in Test Solution
suitable 1 Non- ACCEPTED
container- Sterile DESIRED SAL
closure Closure
system
Monitoring
IDENTIFICATION
sterility level
OF STERILIZATION
CYCLE
PARAMETERS

Therefore, to be operated at conditions less than the conditions

stated in the sterilization process specifications.

IN-PROCESS VALIDATION NECESSARY

• Standardized preparation of viable microorganisms (bacterial spores), of
known sterilization resistance to the sterilization mode.
•Microbial preparations are carried either directly by some of the items to be
sterilized or by carriers such as filter papers, porcelain cylinders.
•Some biological indicators may also contain two different species and
concentrations of microorganisms
•Biological indicators (BI) therefore, serve as a challenge to the effectiveness
of a given sterilization cycle.

P
U
R
P
O
S
E
The stock suspension should contain predominantly dormant
(non-germinating) spores that are held in a non-nutritive liquid.

The finished product (microbial suspension, inoculated carriers,

or biological indicators) supplied by commercial manufacturers
shall have no microorganisms, other than the test
microorganism, present in sufficient numbers to adversely affect
the product.

The system to minimise the presence of microorganisms other

than the biological indicator microorganism in the product will
itself also be validated, monitored, and recorded.
Biological indicator survival is predicated upon both resistance and
population. Therefore, a 106 biological indicator population is not
always required to demonstrate a 106 SAL.

The appropriate use for biological indicators is to employ them

to confirm that the developed process parameters result in the
desired SAL.

Parameters used for thermal calculations and to

define the rate of thermal lethality:

D - Value and Z - Value

 ↑ the temperature (hot) ↑ the microorganisms die --- FASTER
PROCESS
The speed of the process is expressed with the D-Value
D-value refers to decimal reduction time
D Value is time in minute required to kill 1 log
(90%)of the microorganisms.
Thus, after an organismis reducedby 1 D,only 10% of the
originalorganisms remain.

For example: A hypothetical organism is reduced by 90% after exposure

to temperatures of 300 degrees Fahrenheit for 2 minutes, thus the D-value
would be written as D300F = 2 minutes. If the initial population was
100cfu/ml, then 10cfu/ml would remain after a log cycle reduction.

The decimal reduction time is dependent on the temperature, the type of

microorganism and the composition of the medium containing the
microorganism
 DT =
t 2 - t1
log N0 - log N1
Log Survivors

t1= Initial Time t2= Final Time

N0= Initial Population N1= Final Population
D-value

Therefore, from the graph-

D121 = 45 – 30 / 5 - 4
D121 = 15/1
D121 = 15 sec
Time (min) at 121°C

Biological indicators with substantive D values and populations substantially

less than 106 are adequate to validate sterilization and decontamination
processes.
For example : Bacillus stearothermophilus spores on a strip has a normal
population of 1.4 * 105 and a D value of 1.5 min at 121°C.
 It is the measure of change in death
rate with a change in temperature.
The Z-Value gives an indication of the
Thermal Death Time (min) (D-value)

relative impact of different

temperatures on a microorganism,
with smaller values indicating greater
sensitivity to increasing heat.
Z-value
Z=
t 2 - t1
log a - log b
t1= Initial Temperature a= Upper D-value
t2 = Final Temperature b=Lower D-value
Therefore, from the graph-
Z = 240 – 220 / log 100 – log
10
Z = 20 / 2 – 1
Temperature (°F)
 TERMINAL
STERILIZATION

FINAL
PACKAGE

S
ASPETI PRODUCT ESING

For sterile preparations (parenterals and ophthalmics), it is often necessary to

reach a compromise between the most effective sterilization procedure that will
kill microoragnisms and the one that will not have any adverse effect upon the
material to be sterilised.
Validation of sterilization process with biological indicator will ensure that
only the most effective sterilization procedure will ensure sterility of raw
materials to be used in aseptic processing and also terminally sterilised finished
products with desired sterilization assurance level.
1) U.S. Pharmacopoeia-National Formulary [USP 32 NF 27]. Roydan,
Md: United States Pharmacopeial Convention, Inc; 2009. [1035]
Biological indicators for sterilization; 1: 416.

2) N.K.Jain; Sterilization. Pharmaceutical Microbiology; 2nd edition,

7th reprint; page no. 100-113; Vallabh Prakarshan, Delhi; 2013.

3) Roop.K.Khar, SP Vyas, Farhan J Ahmad, Gaurav K Jain;

Sterlization. Lachman/Lieberman’s The Theory and Practise of
Industrial Pharmacy; 4th edition; page no. 804-827; CBS
Publications & Distributors Pvt. Ltd., New Delhi, 2013.