Review of FDA warning letters for microbial bioburden issues (2001 ...

Article
Review of FDA warning letters for microbial
bioburden issues (2001-2011)
*Dr. Tim Sandle
Head of Microbiology, BPL, UK
The monitoring of bioburden, either as a product being
manufactured, or prior to sterilization (be that through a sterilizing
grade filter or through a terminal sterilization process) is an issue
of FDA concern.
A review of FDA warning letters between 2001 and 2011
indicates that seventy-nine companies have received a warning
letter observation relating to bioburden testing. With some
companies, multiple observations have been given within the same
warning letter and 111 observations have been made in relation to
bioburden testing for this period.
The majority of the observations were made during the period
2004-2007, as indicated in the graph below. However, a number of
warning letter observations continues to be issued each year with
the current rate averaging at ten per year.
Figure 1: Graph displaying the number of warning letters
which mention a bioburden issue for the period 2001-211.
The range of areas covered by the bioburden observations
varies across a number of subject categories, of which the failure to
conduct adequate bioburden testing for material prior to the point of
filtration is the largest category. The second largest area relates to
a failure to conduct adequate investigations into bioburden results
which exceed alert or action levels. The third largest grouping was
warning letter observations which relate to sampling (either an
insufficient number of samples taken or samples not being taken
at all).
Together, these three most populated categories account for
65% of the warning letter observations.
A categorization of bioburden warning letter observations is
displayed in the chart below:
Each of the categories is examined further below.
Out of limits investigations
Many 483 warning letter observation related the investigation
of out-of-limits results for viable counts. These citations were
divided into:
a) Excursions above the action level not being investigated.
Figure 2: Categorization of warning letter observations in
relation to bioburden for the period 2001-2011.
b) Investigations having taken place into bioburden excursions,
but at a level deemed inadequate in that the FDA inspectors
considered that the root cause had not been identified.
c) Investigations for which the root cause had been identified
but where the corrective and preventative actions were
considered to be inadequate in that sufficient safeguards had
not been put in place to prevent the contamination event from
re-occurring.
d) Aspects of the investigation not being sufficiently recovered.
For example, in one company (Genzyme Corporation), a
potentially pathogenic microorganism had been isolated but
this had not been commented upon by the company and no
further investigation had taken place into the likely origin of
the contamination or as a product impact assessment.
Sampling
Four comments were made in relation to samples from the
warning letters where samples had been taken according to sampling
plans but there was no evidence of the samples having been tested.
These observations probably relate to laboratory error.
There were twenty comments in relation to other sampling
related issues. These have been categorized as:
a) No rationale in place for why particular samples are taken.
b) No sampling plans in plans.
c) No clear procedures in place relating to how samples are
taken.
d) The size of the sample taken was insufficient for the type of
bioburden test required. Two of these observations related to
the assessment for the sample taken prior to final filtration and
it is assumed that this was in relation to the membrane filtration
method.
e) An insufficient number of samples were taken for bioburden
testing. This was by far the most common observation with
sixteen 483 observations.
*Email Id: tim.sandle@bpl.co.uk
Pharma Times - Vol. 44 - No. 12 - December 2012 29

Pre-sterilization bioburden
Observations relating to pre-sterilization bioburden related
to product subject to terminal sterilization processes. For such
processes there is a requirement to periodically assess the typical
bioburden levels on the product and to characterize the types of
microorganisms recovered.
The observations made in relation to such testing fell into the
following categories:
a) Bioburden levels not assessed.
b) Bioburden levels not assessed periodically (in most cases
the failure to undertaken such an assessment quarterly was
cited).
c) The only comment which slightly differed to the quarterly
required related to a failure to determine of the bioburden on
the pre-sterilized product varied seasonally.
d) Observations were made in relation to a failure to characterize
the microorganisms detected in terms of their theoretical
resistance to the sterilization process.
e) Two observations related to a failure to relate the bioburden
assessments back to the sterilization validation and a failure to
undertaken a re-validation exercise.
Test controls
Warning letters which in relation to test controls were primarily
concerned with in-process samples being tested and reported
without any alert or action levels being in place. The absence of
any acceptance criteria made the review of the intermediate product
meaningless.
A second criticism was that where alert and action levels were
in place, no rationale was available to describe how the alert and
action levels had been set. For alert and action levels, it is common
to either assign these based on one of the following:
a) Regulatory guidance values.
b) Based on some measure of risk (where a bioburden above a
certain threshold could lead to final product contamination).
c) A statistical analysis of past data.
A third area to which the warning letters related was where
action levels had been set but had then been increased without
any explanation or justification provided for the increase.
A fourth area, which related to one warning letter, was that
the company (Genzyme Corporation), had used the term “Too
Numerous To Count” (TNTC) for test data without defining what
TNTC meant and what the implications of a result which was
deemed to be TNTC meant.
Test method validation
Five of the 483 observations related to test method validation.
In four of the cases the warning letters related to absence of test
method validation for different bioburden samples in that the test
methods used did not show recovery of microorganisms.
The fifth observation related to a company which had used a
contract test facility to conduct bioburden testing. Here, there was no
evidence of method transfer or with the contract laboratory having
conducted validation studies.
Test procedures
There were four 483 observations relating to test procedures.
These concerned either the lack of procedures in place (that is,
there was no description of how the bioburden test was conducted)
or that the procedure was unclear. The lack of clarity related to
poor method descriptions and to no indication of how frequently
the bioburden test was to be conducted.
Pre-sterilisation filtration
Comments relating to the pre-sterilization bioburden related to
aseptically filled products. In the case of the 483s, these applied to
the pre-sterilization bioburden not being conducted or to the sample
not being taken on the day of the filtration (the sample of the bulk
product was taken the day before the filtration took place).
A related observation was made to aseptic technique not being
followed in relation to the sampling of the bulk product.
Batch review
The observations relating to batch review concerned the release
of batches of final product where there was no clear evidence that
the in-process bioburden tests had been included as part of the
batch release process.
Raw materials
Three warning letters made reference to in-coming raw
materials not being assessed for bioburden prior to being released
for processing. These observations probably related to the tests for
microbial enumeration described in USP .
Environmental connection
Two warning letters made reference to high bioburden results
not being linked, in the course of batch assessment or investigation,
to the microorganisms commonly found in the manufacturing
environment. The inference here was that the contamination
to which the bioburden related to may have arisen from crosscontamination
from the process area environment.
In-process monitoring
Observations relating to in-process monitoring related to stages
of the manufacturing process where samples for bioburden were
not taken, which the FDA inspector considered should be sampled.
A process flow or risk assessment had not been conducted.
Process hold times
The 483 relating to process hold times related to a process
where the intermediate product was held for a period of time prior to
the next stage of processing. The FDA inspector made a comment
that the hold time should be assessed in order to determine if the
bioburden increased during the hold time period. Such a test would
have involved sampling the product prior to the start of the hold
time and at the end of the hold time.
Microbial identification
There were two 483 observations which discussed the
characterization of microorganisms. In both cases in-process
bioburden counts had exceeded the alert level and the counts had
not been sent for microbial identification. This was considered to
be a lack of process control.
Process cleanliness
There was one 483 which made specific reference to an
observed lack of process hygiene and that this had not been
adequately captured through testing samples for bioburden.
Training
There was one 483 observation which related to the person
tasked with taking the sample for bioburden testing not having been
trained in sampling procedures.
Pharma Times - Vol. 44 - No. 12 - December 2012 30

Minutes of CEC Meeting
Minutes of the CEC meeting held on 18 th August 2012 at IPA Conference
room Mumbai at 2.30 pm
Members Present
Office Bearers
Dr. J.A.S. Giri, President, Dr. C. Gopalakrishna Murty, Immediate
Past President, Mrs. Manjiri Gharat, Vice President, Chairman IPA-
CPD, Dr. R.N. Gupta, Vice President, Chairman IPA - HPD, Mr. S.
D. Joag, Hon. Gen. Secretary, Mr. S.R. Vaidya, Hon. Treasurer,
Dr. Rao V.S.V. Vadlamudi, Editor – I.J.P.S., Dr. Alka Mukne, Editor
- Pharma Times
Associate Secretary
Dr. S.C. Mandal, Mr. Nitin Maniar
Council Members
Dr. Hemant Mondkar, Dr. Anand Shedge
Permanent Invitees
Dr. Ajit Dangi, Mr. Kaushik Desai, Mr. Subodh Priolkar, Mr. P.D.
Sheth
Special Invitees
Prof. V.B. Desai, Mr. Ranga Iyer, Dr. A. K. Kapadia, Mr. A.I. Mehta,
Dr. S.P. Manek, Mr. S.M. Mudda
Secretariat
Mr. T. B. Nair, Executive Secretary
Leave of Absence
Dr. Tulsi Chakrabarti, Dr. Divakar Goli, Dr. Sunil K. Jain, Prof. T.V.
Narayana, Mr. Devinder Pal, Dr. M.K. Raina, Dr. T.K. Ravi, Dr.
Suresh Sarvadekar, Dr. Premnath Shenoy, Dr. B. Suresh, Mr. B.N.
Thakore, Mr. Raj Vaidya,
1) Welcome address by the Hon. Gen. Secretary
i. The Hon. Gen. Secretary welcomed all CEC members and
thanked them for attending this meeting. He informed the
members about the sad demise of Mr. Ton Hoek, CEO of FIP
on 20 th July 2012. Condolence message has been sent to his
family and FIP Secretariat. Dr. S.C. Mandal also informed about
the sad demise of Mr. Dipankar Dutta on 7 th August 2012, who
was a Life member of IPA and served IPA Bengal State branch
in various positions for almost three decades. The members
observed two minutes silence, as a mark of respect to the
departed souls.
ii. The Hon. Gen. Secretary stated that the IPA Building project
will be taken up on priority and it would be completed in next
two years.
iii. EDQM Conference - The EDQM Conference will be held on
September 28 – 29, 2012 at Sofitel Hotel, Bandra Kurla Complex.
The e. brochure was ready and two or more speakers from Dr.
Reddy’s and Ranbaxy still had to be identified. The President
requested the Vice Presidents to work towards the success of
the program. Mr. Ranga Iyer advised that IPA EDQM WHO
Conference program announcement should be inserted in
IDMA Bulletin to ensure better reach and visibility.
iv. The Hon. Gen. Secretary requested Dr. Rao V.S.V. Vadlamudi
to give an update on the discussions during the IPA Awards
Committee. Dr. Rao Vadlamudi mentioned that while it is a
very good initiative to globalize the Eminent Pharmacist award,
one needs to first revisit the main objective of the award, which
is to recognize the achievements and services rendered by
pharmacy professionals from IPA platform. If the organization
wishes to change the objective now or in near future to include
recognition to global pharmacy professionals of Indian origin as
well, it may not be acceptable to many in IPA who are serving
the profession from IPA platform and might very well be aspiring
to be a recipient of the Highest award of IPA.
2) Opening remarks by the President
i. The President, Dr. J.A.S. Giri welcomed all CEC members and
informed that few important decisions were taken in the Office
Bearers meeting held in the morning.
ii. The President emphasized on the need to boost the image of
IPA with the help of senior members, state and local branches
Presidents, industry and regulatory bodies. He requested Mr.
Subodh Priolkar, Dr. C. Gopalakrishna Murty and Mr. P.D.
Sheth to give their valuable guidance for boosting IPA’s
image and promised complete support from the Office
Bearers in this exercise.
iii. The President expressed disappointment about the fact that IPA
branches have not been as active and functional as expected.
The branches have been lax in submitting their statement of
accounts and annual reports and many of the branches have
not been holding regular elections as well.
The House decided that henceforth IPA pan card and 80G
certificate would not be given to any of the branches and the
President requested the Hon. Treasurer to enforce the same.
In cases, where the IPA pan card has been given to any of the
branches earlier, audited balance sheets of these branches
should be collected by the Hon. Treasurer before 30 th of August
2012 and amalgamated with the IPA Centre balance sheet.
The Hon. Gen. Secretary remarked that it was difficult to
garner participation from the branch representatives even for CEC
meetings. In spite of giving support in terms of travel allowance to
these representatives for attending CEC meetings, there has been
no positive response. Dr. Ajit Dangi inquired about the organogram
of IPA; the reporting hierarchies and so on. Mr. Ranga Iyer added
that if IPA Center’s Charity Commissioner’s approval is being used
by the branches, then Center becomes liable for the branch actions.
Mr. Subodh Priolkar remarked that in the past, two or three branches
has been disqualified on grounds of non - submission of annual
reports and audited account statements and/or not holding regular
elections. The branches’ share of membership fees had also been
withheld in the past. The Council now had to take suitable decision
regarding the non active branches. Mr. Kaushik Desai pointed out
that there is a communication gap between Center and Branches.
We need to look at and try and understand the problems that the
branches may have been facing in complying with the expected
requirements. It was decided that the Office Bearers and other
senior members be appointed as Mentors for the branches,
in order to supervise and guide the branch functionaries and
establish a good rapport with them. The state branches would
be responsible for statutory report and accounts submissions
from the local branches within their states. The President
requested state branch Office Bearers to follow up with their
respective local branches and submit report of the local branch
activities to the center.
Pharma Times - Vol. 44 - No. 12 - December 2012 31