LowTemperatureSterilizationProcess(115C)andItsValidation

Learn how sterilization is done for heat sensitive material at lower temperature below 121 C and its validation using Bacillus subtilis (Bacillus atrophaeus) )
ATCC 5230 as biological indicator.
Sterilization of the media and material is generally done at 121 C but sometimes it is required to do at low temperature 115 C because of heat sensitivity
of the material. This sterilization below 121 C is called Low Temperature Sterilization.

Reinforced Medium, Rappaport Vassiliadis Medium, Wilson and Blair's BBS Agar, GN Broth, Triple Sugar Iron Agar, Bismuth Sulphite Agar Medium etc.
are sterilized at 115C and 10 psi pressure. These media are sterilized separately at this temperature.

These culture media contains some heat sensitive ingredients that can degrade at 121C losing their nutritive quality. This media shall not support the
growth of specified microorganisms. When these media are sterilized at high temperature, the microorganism can grow but shall not develop the
characteristic colonies due to the degradation of the heat sensitive ingredients.

Related: Terminal Sterilization of Sterile Pharmaceutical Preparations

121 C sterilization cycle is run for 15 minutes but when temperature is decreased to 115 C, time of sterilization is increased for effective killing of the
microorganisms. It is recommended to run the 115 C sterilization cycle for 30 minutes or it can be calculated by the D value. Suppose D value at 115 C is
3.75 and we want 8 log reduction then we have to run the sterilization cycle 3.75x8=30 minutes.

All sterilization processes must be validated. Low temperature sterilization validation is done by using Bacillus subtilis Bacillus atrophaeus ATCC 5230.
Strips and ampoules containing the spores of B. subtilis are sterilized with the media load similarly as the normal autoclave validation. Every strip contains
1x106 spores.

Related: SOP for Qualification of Biological Indicator

Spore strips are incubated in soybean casein digest medium at 35C for seven days. Growth should not be observed for a valid sterilization process. Growth
of the biological indicator is indicated by the turbidity.
Turbidity Improper sterilization
No turbidity Proper sterilization

If ampoules of Bacillus subtilis are used as biological indicator then media should released from the bulb for the growth of the spores. Ampoules are also
incubated at 35C for seven days and observed for the change of color of the ampoule.

Also see: Performance Evaluation of Biological Indicators

Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widelyread pharmaceutical
blog since 2008. Signup for the free email updates for your daily dose of pharmaceutical tips.
Need Help: Ask Question
Offline

20082017 All rights reserved to Ankur Choudhary and data may not be reproduced on other websites. Designed by Pharmaceutical Guidelines.
Privacy Policy Disclaimer