Regarding autoclave cycle development

  1. Sir how can we develop sterilization cycle by having a Dvalue of 0.5 mins
  2. how can we recommend freezing temperature in lyophilization process

Hello and welcome.

  1. To develop a sterilization cycle, first you need to establish the level of lethality (F0) that you want to achieve. This is done by selecting either an overkill approach (F0 ≥ 12min) or a product specific approach. I will assume you’ll use a product specific approach since you’re already considering to develop a cycle for a Dvalue of 0.5 min. Now you need to establish the level of bioburden you want to sterilize, worst case scenario (overkill) assumes a 10^6 CFU/unit. If that is the case, you use the following formula to calculate the lethality needed to achieve a PNSU (or SAL) of 10^-6:

F0 = [Log(N0) - Log(Nf)] * Dvalue
F0 = [Log(10^6)-Log (10^-6)] * 0.5 = 6 min

Next, you need to know at what temperature you can sterilize the product, since lethality rate is a function of temperature. I will assume you can use the typical sterilization temperature of 121.1 °C. The lethality rate can be calculated with the following formula:

L = 10^[(Tb-Tr)/z]
L = 10^[(121.1 - 121.1)/10] = 1

At 121.1 °C, each minute the product is exposed at 121.1°C, the product is accumulating 1min fo lethality (F0). Thus, you will need a sterilization cycle that ensures the product is kept at 121°C for a total of 6 min, in order to achieve a lethality (f0) of 6 min needed to obtain a steirlization process with a PNSU or SAL of 10^-6 as required by regulation.

  1. The freezing temperature ina lyophilization process is a bit harder to specify since it is excplusive to the product and solution characteristics. The main idea is that the freezing temperature achieved at the end of the freezing stage must be lower than the glass transition of the product, wich will allow you to have a better lyophilized structure during the primary and secondary drying process, avoiding cake collapse or other defects that could impact the final moisture of the product. That being said, a common temperature coul be -40 °C but it is higle dependant on each product.

I hope this information is helpful. If you want me to elaborate further, please let me know.