A workshop with limited number of participants

1. [Case Study (B) : 6-1]
Sterile Operations for Parenteral Preparations
Roohi B. Obaid
23 Jun 2018 at Karachi

2. Mycoplasm
Opinion & Perspective
FDA
Challenge continues … A probable issue

3. A firm has multiple media fill failures
Investigation did not show obvious causes
What could be the source of contamination
?

4. Please list down what can be the potential causes
Firstly focus on main causes

5. Human Being
Flaws in
Maintenance
Design &
Facilities

6. Begin to resolve …

7. They conducted media fills
using Tryptic Soy Broth
(TSB) prepared by filtration
through 0.2 um sterilizing
filter
Any obvious cause could
not be identified in the
investigation
Background

8. They conducted media fills
using Tryptic Soy Broth
(TSB) prepared by filtration
through 0.2 um sterilizing
filter
Any obvious cause could
not be identified in the
investigation
Background

9. They conducted media fills
using Tryptic Soy Broth
(TSB) prepared by filtration
through 0.2 um sterilizing
filter
Any obvious cause could
not be identified in the
investigation
Background

10. Chain of Event
Media fill run simulating
the filling process during
production were
conducted inside an
isolator
1

11. Chain of Event
Media fill run simulating
the filling process during
production were
conducted inside an
isolator
Used TSB (non-
sterile bulk powder)
from a commercial
source
1
2

12. Chain of Event
Investigation
conducted to detect
contamination
3

13. Chain of Event
Did not succeed in
isolation or recovery of
the contaminant (by
conventional
microbiological
techniques)
4
Investigation
conducted to detect
contamination
3

14. Findings
Eventually, it was identifed as Acholeplasma laidlawii
Firm conducted studies to confirm its presence in the lot
of TSB used
Thus, it was not the contaminant from the process, but
from the media source used

15. Take Home Message
Acholeplasma laidlawii belongs to an order of mycoplasma
They are not susceptible to beta-lactams and do not take up
Gram stain
Individual organisms vary in size from 0.2 to 0.3 microns or
smaller

16. Take Home Message
Acholeplasma laidlawii is capable of penetrating a 0.2 micron
filter, but is retained by a 0.1 micron filter
It is known to be associated with animal-derived material, and
microbiological media is often from animal sources
Environmental monitoring of mycoplasma requires
selective media

17. 1
• Firm has decided to filter prepared TSB, for use in media
fills, through a 0.1 micron filter
2
• For future, they will use sterile, irradiated TSB
3
• The firm will continue monitoring for mycoplasma and has
revalidated their cleaning procedure to verify its removal
Resolution