PQRI

PQRI
Aseptic Processing Working Group
-Final Report-
Aseptic Processing Work Group
-Final Report-
The PQRI Aseptic Processing Working Group has completed their work in accordance with the December
23rd, 2002 approved work plan. The working group completed its activities on March 10, 2003, 110 days
from the date of it’s formation. The working group, comprised of 41 members, did an outstanding job of
focusing on the specified topics and goals set forth in the work plan.
Not all areas of the FDA’s concept paper “Sterile Drug Products Produced by Aseptic Processing” were
discussed in this activity, but rather several specific areas that where identified as needing additional
scientific input.
As part of the work plan an industry survey was conducted to gather specific information regarding aseptic
filling practices. The responses from over 45 aseptic filling sites were received and collated for use by the
team. Since that date several additional surveys have been received. The complete survey results,
including the additional surveys received, will be submitted for publication in the near future.
The 8 clarifications and 10 recommendation developed by the working group are presented in this
document and were developed based on the expert knowledge of the work group members, survey results,
and publications where applicable.
As Chairman of the Aseptic Processing Working Group, I would like to recognize, on behalf of PQRI, the
outstanding dedication and contributions made by each one of the FDA, Industry, and Academia working
group members. In addition, I would like to recognize and thank the working group members from the
DMPQ in Office of Compliance at FDA, for their willingness to bring this topic into the PQRI framework
and for their efforts to make this activity a success.
Glenn E. Wright
Chairman
PQRI Aseptic Processing Work Group
Page 2 of 31 March 13,2003

-Final Report-
TABLE OF CONTENTS
WORK PLAN 4
CLARIFICATIONS 10
RECOMMENDATIONS 14

-Final Report-
PQRI
Work Plan
(Approved by the PQRI Steering Committee December 23, 2002)

-Final Report-
Aseptic Processing Working Group Work Plan

(Approved December 23, 2002)
Overview:
The PQRI Aseptic Processing working group will cover a defined list of points. The composition of the group will
be made up of experts form the FDA, industry, and academia. The points to be covered have been divided into two
categories: points for with recommendations will be made and points for which clarification comments will be
made.
Working Group Members (updated):
Mr. James P. Agalloco Nigel Halls, Ph.D. Mr. Rainer F. Newman

Agalloco & Associates GlaxoSmith Kline (ret.) Johnson & Johnson
James E. Akers, Ph.D. Mr. Karl L. Hofmann Ms. Jean I. Olsen

Akers Kennedy & Associates Brystol-Myers Squibb Co. GlaxoSmithKline
Ms. Diane Alexander David Hussong, Ph.D. Ms. Carolyn Renshaw

FDA FDA FDA
Barbara Bassler, Ph.D. Mr. Richard M. Johnson Mr. Robert Sausville

Bridge Associates International Abbott Laboratories FDA
Mr. Martyn Becker Kunio Kawamura, Ph.D. Neal J. Sweeney, Ph.D.

Merck & Co. Otsuka Pharma. Co., Ltd. FDA
Ms. Susan Bruederle Lee E. Kirsch, Ph.D. Mr. Ian Symonds

FDA University of Iowa GlaxoSmithKline
Don G. Burstyn, Ph.D. Ms. Carol M. Lampe Laura A. Thoma, Ph.D.

Alkermes Baxter Healthcare Corporation University of Tennessee
Roger Dabbah, Ph.D. Mr. Joseph Lasich Ms. Debbie Trout

USP Alcon Laboratories, Inc. FDA
Mr. Roger Deschenes John Lindsay, S.M. (NRM) Brenda Uratani, Ph.D.
Astra Zeneca Aseptic Solutions Inc. FDA
Mr. Joseph C. Famulare Mr. Russell E. Madsen Mr. Martin Van Trieste
FDA PDA Abbott Laboratories
William R. Frieben, Ph.D. Leonard Mestrandrea, Ph.D. Richard T. Wood, Ph.D.

Pharmacia Corporation Pfizer Inc. Pfizer, Inc.
Mr. Richard L. Friedman Mr. Karl Andrew Minor Mr. Glenn E. Wright
FDA Eli Lilly & Co. Eli Lilly & Co.
Mr. John Grazal Kenneth Muhvich, Ph.D. Jeff Yuen, MPH, MBA
AstraZeneca Pharmaceuticals Micro-Reliance. Jeff Yuen and Associates
Klaus Haberer, Ph.D. Mr. Terry E. Munson

Compliance Advice & Services KMI/PAREXEL, Inc.

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Process:
The working group will work on the 10 recommendations and 8 clarifications as a single group. A project kickoff
meeting will be held the first week in January. Members will be required to attend one 90-minute conference call
each week.
Listed below are the working groups deliverables and the processes to be used. Included in the deliverables is the
industry survey.
• Clarifications
Clarifications on 8 specific topics are to be worked on by the working group.
Topic leaders will be chosen from the PQRI working group for each point.
Suggested redline clarifications are to be made by the working group and sent to the topic leader. This
activity is to begin in early January.
The topic leader will collect and collate the suggested clarifications and develop a redline strikeout version
of the text that incorporates the various suggestions.
The topic leader will lead discussions on the clarifications and make the modifications needed.
A final clarification will be developed and approved by the working group.
The redline clarifications are to be completed by January 31st and sent to the PQRI steering committee for
approval.
• Survey
An industry survey will be performed to collect current industry information.
The survey will be a data collection type of survey designed to provide information on the industries
current practices.
The target date to receive completed surveys will be January 20th.
The surveys will be blinded by PQRI and data tabulated.
The tabulated data will be provided to the work group on February 4th.
• Recommendations
Recommendations on 10 specific topics are to be worked on by the working group.
Discussions relating to environmental monitoring and sterilization options will be lead by Carol Lampe
while the discussion on process simulation and aseptic processing isolators will be led by Richard Johnson.
Discussions are targeted to begin the first week in February.
The recommendations are to include the question being asked, the recommendation being provided and a
brief rational section that provides information on how the recommendation was reached.
A final recommendation will be approved by the working group.
The recommendations must be completed by February 28th and sent to the PQRI steering committee for
approval.
Timeline:
• Week of December 16th - Work Plan and Survey sent to Steering Committee for approval.
• Week of December 23rd - Survey sent to companies.
• First week in January - Work group begins work on clarifications.
• January 20th - Surveys due to PQRI.
• January 31st - Clarification work completed and sent to Steering Committee for Approval.

-Final Report-
• February 4th - Survey results compiled and provided to work group.

• First week in February - Work Group begins work on recommendations.
• February 28th - Work group completes recommendations and sends the recommendations to PQRI Steering
Committee for approval.
Clarifications:
The PQRI working group will develop suggested clarifications for the following 8 points. The clarifications will be
presented in the form of redlined revisions to the current text.
1.) Concept Paper Line Number Reference: 637
Subsequently, routine semi-annual revalidation runs should be conducted for each shift and processing line to
evaluate the state of control of the aseptic process.
Clarification: What clarification should be provided in regards to each shift to help the reader understand
that the process simulation needs to represent the various shifts but that a separate media fills
for each shift may not be required.
Written procedures should include a list of locations to be sampled. Sample timing, frequency, and location
should be carefully selected based upon its relationship to the operation performed. Samples should be taken
throughout the aseptic processing facility (e.g., aseptic corridors; gowning rooms) using appropriate,
scientifically sound sampling procedures, standards, and test limits.
Clarification: What clarification should be provided for the term “test limit” so that it is not confused with a
specification?
Upon preparation, disinfectants should be rendered sterile, and used for a limited time, as specified by written
procedures. Disinfectants should retain efficacy against the normal microbial flora and be effective against
spore-forming microorganisms. Many common sanitizers are ineffective against spores, for
example,70%isopropyl alcohol is not effective against Bacillus,spp .spores. A sporicidal agent should be
used regularly to prevent contamination of the manufacturing environment with otherwise difficult to
eradicate spore forming bacteria or fungi.
Clarification: What clarification should be suggested regarding a disninfectants efficacy against spore
forming microorganisms?
After the initial assessment of sanitization procedures, ongoing sanitization efficacy should be frequently
monitored through specific provisions in the environmental monitoring program, with a defined course of
action in the event samples are found to exceed limits.
Clarification: What type of clarification should be made in regards to expectations surrounding sanitization
efficacy being monitored by the environmental program?
b. Active Air Monitoring-

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Manufacturers should be aware of a device's air monitoring capabilities, and should determine suitability of
any new or current devices with respect to sensitivity and limit of quantification.
Clarification: What type of clarification should be made in regards to determining the suitability of new or
current devices and the comment around sensitivity and limit of quantification?
Properly operated RTPs (rapid transfer ports) are also generally considered to be an effective transfer
mechanism. The number of transfers should be kept to a minimum because the risk of ingress of
contaminants increases with each successive material transfer.
Clarification: What clarification should be suggested regarding the number of RTP transfers and the risk of
contamination?
In addition to microbial counts beyond alert and action limits, the presence of any atypical microorganisms in
the cleanroom environment should be investigated, with any appropriate corrective action promptly
investigated.
Clarification: What clarification should be suggested regarding the expectation concerning the “typical
microflora”, and the definition for an atypical microorganism?
Evaluating the quality of air and surfaces in the cleanroom environment should start with a well-defined
written program and validated methods. The monitoring program should cover all production shifts and
include air, floors, walls, and equipment surfaces, including the critical surfaces in contact with product and
container/closures.
Clarification: What clarification should be made regarding the validation of the environmental monitoring
methods?
Recommendations:
The PQRI working group will develop recommendations using data from the industry survey as well as good
scientific principals for the following 10 points (questions). The recommendation format will follow the example
given in the Attachment.
Process Simulations
Questions: What is an appropriate number of units to be filled during a process simulation (media fill)?
Question: What is an acceptable temperature range for the incubation of media fill units using TSB and
FTM? If alternative practices are used what type of justification is required?
Question: What is an appropriate limit for the contamination rate in a process simulation (media fill)?
What is an appropriate target for contaminated units in a process simulation (media fill)?

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Environmental Monitoring
Question: When should critical surfaces be monitored? What are appropriate expectations in regards to
results obtained?
5.) Concept Paper Reference Line Number Reference: 1014:
Question: What data should be considered when establishing monitoring limits? What is an appropriate
frequency for re-evaluating monitoring limits?
TABLE 1- Air Classificationa
Clean Area >0.5 um >0.5 um Microbial Limitb

Classification particles/ft3 particles/m3 cfu/10 ft3 cfu/m3
100 100 3,500 <1c <3c
1000 1000 35,000 <2 <7
10,000 10,000 350,000 <5 <18
100,000 100,000 3,500,000 <25 <88
a - All classifications based on data measured in the vicinity of exposed articles during periods of activity.
b- Alternate microbiological standards may be established where justified by the nature of the operation.
c- Samples from class 100 environments should normally yield no microbiological contaminants.
Question: What is the maximum number of viable organisms allowed in air samples for the various
classifications?
Aseptic Processing Isolators
Question: What type of airflow is required in closed isolators?
8.) Concept Paper Line Number Reference:1372
Question: What is the appropriate requirement for air handling systems in isolators?
Question: What are appropriate methods for use in the development of decontamination cycles?
Sterilization Options
10) Concept Paper Line Number Reference: 57
Question: With respect to terminal sterilization and adjunct processing what flowcharts represent the most
risk-based and scientifically developed approach?

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PQRI
Clarifications
-Report-
(Approved by the PQRI Steering Committee February 5th, 2003)

-Final Report-
The PQRI Aseptic Processing Working group has completed the Clarification section of its approved Work Plan.
As detailed in the Work Plan, clarifications have been developed for specific passages of text identified in the
FDA’s concept paper “Sterile Drug Products Produced by Aseptic Processing” published in September of 2002.
The following clarifications are being recommended by the PQRI Working Group:
• Clarification #1
Concept Paper Line Number Reference: 637
Original Text
“Subsequently, routine semi-annual revalidation runs should be conducted for each shift and processing line to
evaluate the state of control of the aseptic process.”
Clarified Text
Subsequently, routine semi-annual revalidation should be conducted for each processing line to evaluate the
state of control of the aseptic process. Activities and interventions representative of each shift and shift change
over should be incorporated into the design of the semi-annual revalidation.
Concept paper Line Number Reference: 984
Original Text
“Written procedures should include a list of locations to be sampled. Sample timing, frequency, and location
throughout the aseptic processing facility (e.g., aseptic corridors; gowning rooms) using appropriate,
scientifically sound sampling procedures, standards, and test limits.”
Clarified Text
Written procedures should include a list of locations to be sampled. Sample timing, frequency, and location
throughout the aseptic processing facility (e.g., aseptic corridors; gowning rooms) using scientifically sound
sampling procedures..
Original Text
“Disinfectants should retain efficacy against the normal microbial flora and be effective against spore-forming
microorganisms. Many common sanitizers are ineffective against spores, for example, 70%isopropyl alcohol is
not effective against Bacillus spp spores. A sporicidal agent should be used regularly to prevent contamination
of the manufacturing environment with otherwise difficult to eradicate spore forming bacteria or fungi.”
Note: The text to be modified was narrowed from that present in the working plan with the first sentence in
the paragraph being removed. It was determined by the working group that this first sentence was out
of scope.

-Final Report-
Clarified Text
Routinely used disinfectants should be effective against the normal microbial vegetative flora recovered from
the facility. Many common sanitizers are ineffective against spores, for example, 70%isopropyl alcohol is not
effective against Bacillus spp spores. Therefore a sound disinfectant program should also include a sporicidal
agent, used according to a written schedule, and if environmental data suggests the presence of bacterial spores.
Original Text
“After the initial assessment of sanitization procedures, ongoing sanitization efficacy should be frequently
monitored through specific provisions in the environmental monitoring program, with a defined course of action
in the event samples are found to exceed limits.”
Clarified Text
Once sanitization procedures are established, adequacy is evaluated through the routine environmental
monitoring program.
Paper Line Number Reference: 1070
Original Text
“Manufacturers should be aware of a device's air monitoring capabilities, and should determine suitability of
any new or current devices with respect to sensitivity and limit of quantification”
Clarified Text
Manufacturers should be aware of a device's air monitoring capabilities, and the air sampler should be evaluated
for its suitability for use in an aseptic environment based on cleanability, ability to be sterilized, and disruption
of unidirectional airflow. Manufacturers should assure that it is calibrated and used according to instructions.
Because these attributes vary from device to device, the user should assess the suitability of all monitoring
devices before they are placed into service.
Original Text
“Multiple material transfers are generally made during the processing of a batch. Frequently, transfers are
performed via direct interface with a decontaminating transfer isolator or dry heat depyrogenation tunnel with
balanced airflow. Such provisions, if well designed, help ensure that microbiological ingress does not result
from the introduction of supplies. Properly operated RTPs (rapid transfer ports) are also generally considered to
be an effective transfer mechanism. The number of transfers should be kept to a minimum because the risk of
ingress of contaminants increases with each successive material transfer.”
Clarified Text
Multiple material transfers are generally made during the processing of a batch. Frequently, transfers are
performed via direct interface between manufacturing equipment and properly maintained and operated rapid
transfer ports (RTPs) are an effective mechanism for aseptic transfer of materials into and out of isolators.

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Note: During the review the working group concluded that the last sentence of the text to be clarified (“The
number of transfers should be kept to a minimum because the risk of ingress of contaminants increases
with each successive material transfer.”) should be revised and moved into Line 275 of the concept
paper. This location represents the best location fit for the information.
Added Clarification text (concept paper reference Line 275).
…surrounding environment. The number of transfers into an isolator or the critical zone of a clean room should
be minimized.
Original Text
In addition to microbial counts beyond alert and action limits, the presence of any atypical microorganisms in
the cleanroom environment should be investigated, with any appropriate corrective action promptly
implemented
Clarified Text
Significant changes in microbial flora should be considered in the review of the ongoing environmental
monitoring program.
Original Text
Evaluating the quality of air and surfaces in the cleanroom environment should start with a well-defined written
program and validated methods. The monitoring program should cover all production shifts and include air,
floors, walls, and equipment surfaces, including the critical surfaces in contact with product and
container/closures.
Clarified Text
Evaluating the quality of air and surfaces in the cleanroom environment should start with a well-defined written
program and scientifically sound methods. The monitoring program should cover all production shifts and
include air, floors, walls, and equipment surfaces, including the critical surfaces in contact with product and
container/closures.

-Final Report-
PQRI
Recommendations
- Report-
(Approved by the PQRI Steering Committee March 11th, 2003)

-Final Report-
RECOMMENDATION #1
Question: What is an appropriate number of units to be filled during process simulation (media fill)?
Recommendation:
• The number of units to be filled should be sufficient to accurately simulate activities that are representative
of the manufacturing process. Such activities include but not limited to:
Aseptic manipulations during setup and during production

Interventions – type and appropriate number
o Typical/Routine
o Atypical/Non-Routine
Staffing Levels
Shift Changes
Gown Changes
Multiple day fills
• A generally acceptable starting point is between 5,000 to 10,000. For batch sizes under 5,000 the number of
media filled units should equal the batch size.
• Where the technology is such that the possibility of contamination is higher (manually intensive filling
lines), a larger number of units, generally at or approaching the full batch size, should be considered.
Rationale:
• The number of units filled during the process simulation should be based on contamination risk for a given
line, and sufficient to accurately simulate manufacturing operations. For instance, a process conducted in
an isolator can have a very low risk of contamination because of the lack of human intervention, while a
manual (or highly manual) aseptic process has a higher risk of contamination. In these instances, the
isolated process may be simulated by filling a lower number of units as a proportion of the overall
operation versus a manual operation that would fill the maximum number of units.
Survey Data Summary:
• Number of units filled during the process simulation compared to production batch sizes.
Mfg. Lot Size 5,000 units and under ≥Lot size 92%
<Lot size 8%
Mfg. Lot Size 5,001 – 10,000 >75% of lot size 33%

50% to 75% of lot size 63%
<50% of lot size 4%

50% to 75% 25%
<50% 47%
Mfg. Lot Size20,001 – 40,000 >75% of lot size 1%

51% to 75% 3%
25% to50% 49%
<25% 47%

-Final Report-

51% to 75% 2%
25% to50% 18%
<25% 74%
Mfg. Lot Size >80,000 >75% of lot size 0%

51% to 75% 0%
25% to50% 9%
<25% 90%
• Distribution for the total number of media units filled.
Media Fill Lot Size <5,000 23%
Media Fill Lot Size 500110,000 38%
Media Fill Lot Size 10,00120,000 28%
Media Fill Lot Size >20,000 11%
• Average number of media units filled; 12,981

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RECOMMENDATION #2
Question: What is an acceptable temperature range for the incubation of media fill units using TSB and FTM? If
alternative practices are used what type of justification is required?
Recommendation:
• Incubation temperatures should be suitable for recovery of bioburden and environmental isolates.
• Incubation conditions should be not less than 14 days at either a temperature or temperatures between 20-
35°C. If two temperatures are used for the incubation of the media filled samples, these filled samples
should be incubated for at least 7 days at each temperature.
• The incubation temperature should be maintained within 2.5oC of the target temperature. The temperature
should at no time be below 20oC or above 35oC.
Rationale:
• The temperature range provided is suitable for the recovery of wide range of mesophilic bacteria (the
largest portion of bacteria are classified as mesophilic). The temperature range is similarly suitable for a
wide range of yeast and molds.
• The recommendation is consistent with current USP, PDA, and PIC/S guidance.
• Regarding the second part of the rationale question. No specific temperature was selected as a set
temperature to allow flexibility for the individual firms to select the temperature range(s) appropriate for
their operation. The justification for all temperatures is specified in bullet one.
• The survey data indicates that industry currently uses various incubation conditions that can recover a wide
range of mesophilic flora. The recommendation provides for the use of these different approaches.
References:
• PDA Technical Report No. 22, “Process Simulation Testing for Aseptically Filled Products” 1996.
• PIC/S, “Recommendation on The Validation Of Aseptic Processes,” April 2000.
• What is temperature range does your firm currently use for the incubation of media fill units?
Number of respondents =49
A 20-25 o C 1 2%
B 25-30 o C 1 2%
C 30-35 o C 3 6%
D 20-25 then 30-35 o C 24 49%
E 30-35 then 20-25 o C 7 14%
F other (specify) ______________ 13 27%

-Final Report-
2 30-32C
5 A+C
1 28C
1 28-32C
1 A+D
1 C+E
1 C+D
1 NA (small batch)

-Final Report-
RECOMMENDATION #3
Question: What is an appropriate limit for the contamination rate in a process simulation (media fill)? What is an
appropriate target for contaminated units in a process simulation (media fill)?
Recommendation:
• The target is zero contaminated units.
• Any contaminated unit indicates a potential sterility assurance problem, regardless of run size. All
contaminated units should result in a thorough, documented investigation.
• Clear acceptance criteria should be established for media fills.
• The following criteria are generally acceptable:
When filling less than 5000 units no contaminated units should be detected.
When filling from 5,000 to 10,000 units

1 contaminated unit requires an investigation and a determination if any further action is needed
such as a repeat of the media fill.
2 contaminated units are considered cause for revalidation following investigation.
When filling more than 10,000 units

1 contaminated unit requires an investigation
2 contaminated units are considered cause for revalidation following investigation.
• Recurring incidents of contaminated units from media fills for an individual line, regardless of the set
acceptance criteria, should be a signal that action should be taken.
Rationale:
• With older technology, standard manufacturing operations were normally easily simulated in small media
fills. In 1973, the World Health Organization recommended demonstrations of less than 0.3%
contamination, which could be demonstrated with a fill of as low as 1,000 or 2,000 containers. By 1980,
PDA Technical Monograph #2 suggested demonstration of less than 0.1% contamination, which was also
adopted by the FDA in their 1987 Guideline on ‘Sterile Drug Products Produced by Aseptic Processing’,
requiring media fills of 3,000 units or more (using the 95% confidence interval statistical approach). This
size was considered a minimum because the absence of growth was evidence of contamination at a rate of
less than 0.1%.
• In these examples, the expression of acceptance based on a percent contamination was meant to provide a
standard based on the realistic limitations of media fills of these sizes. However, the use of percent
contamination in these examples was not meant to provide a tolerance for greater contamination in larger
media fills. The 0.1% tolerance was linked to the detectable limit of the test using 3,000 units. For this
reason, larger media fills should be subject to acceptance criteria that are not linked to percent
contamination.
• Limits based on statistical calculations have been demonstrated to be flawed for aseptic operations, in part
because it is not appropriate to apply the statistics of large-scale populations to smaller ones. A statistical

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approach makes faulty assumptions that the distribution and frequency of potentially contaminated units are
the same in these populations. Statistically-derived contamination rates are therefore not relevant in setting
acceptance criteria for process simulations.
• The target for contaminated units should be zero, which is consistent with the definition of sterile products.
This is commensurate with data indicated in the PQRI survey.
• A phased approach is appropriate for simulations above the minimum number of 5,000 units, as likelihood
of detection will increase with simulation size. An isolated incident (no trend) in which one contaminated
unit beyond the 5000 unit level is detected may be considered a random event. This should however,
require an investigation and assessment for repeat, whereas more than one contaminated unit may be more
indicative of an issue that needs to be resolved.
References:
• FDA, “Sterile Drug Products Produced By Aseptic Processing”, 1987

• EU GMP Annex 1, “Manufacture Of Sterile Medicinal Products”
• PIC/S, “Recommendation on The Validation Of Aseptic Processes,” April 2000.
• Percentage of Media fills with contamination.(606 Media Fills Total)
Contaminated
9%
Not
Contaminated
91%

-Final Report-
• Breakdown of Contaminated Media Fills (54 of 606 runs contaminated).
1200
Contaminations 2%
5 Contaminations
4%
4 Contaminations
15%
3 Contaminations
7%
2 Contaminations
6% 1 Contamination
66%

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Recommendation #4
Question: When should critical surfaces be monitored? What are appropriate expectations in regards to results
obtained?
Recommendation:
• The selection of sample sites should be strategic in an environmental monitoring program. This should
include consideration as to when, or if a critical site should be monitored.
• Each manufacturer should review each type of process and the points of risk for product contamination.
Consideration should be given to the level of contamination risk based on factors such as: difficulty of set-
up; length or processing time; impact of interventions, etc.
• It is well understood that the sampling and incubation methods used in surface monitoring are manual
operations that, due to personnel involvement, results in a low rate of false positives. For this reason, the
detection of microorganisms on a critical site should not necessarily result in batch rejection, but should be
investigated. The other EM data and procedures, that support the operation, should be reviewed to
determine if the positive result is supported. If this review does not support the positive result and there is
no negative trend for the critical surface site there is a strong case for not rejecting the lot due to the
positive result.
• PQRI strongly supports the concept discussed on line 993 of the concept paper; that, when performed,
“Critical surface sampling should be performed at the conclusion of the aseptic processing operation to
avoid direct contact with sterile surfaces during processing.”
Rationale:
• The strategic monitoring of critical surfaces can provide useful data on the state of environmental control
within the aseptic processing area. It can allow adverse trends to be detected, investigated and corrected
promptly.
• Environmental monitoring is performed in an environment that does not afford the same controls as that
present in sterility testing. For this reason the testing cannot be used to determine the sterility of the
product. Occasional positive results are expected due to the involvement of EM monitoring personnel
during sampling and as a result of sample handling and incubation within the laboratories.
• The sampling process requires contact with the critical surface (e.g., Rodac sample). To reduce the
possible negative impact this activity has on the critical areas, it should be performed at the conclusion of
the processing operation.
• When does your firm perform environmental monitoring on critical surfaces (product contact surfaces)?
Number of respondents = 49
A at beginning of operations 0 0%
B during operations 7 14%
C conclusion of operations 26 53%
D not monitored 4 8%
E other (add specific comment) 12 24%
1 B+C
4 A+B+C

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1 C+D
2 end of media fill
1 C (UK), D (US)
1 do not test product contact surface, sample surface nearby not monitored routinely, only in case of
failure investigation
• How does your firm respond when positive results are obtained from critical surface monitoring? (If your
firm does not monitor these surfaces go to next question)
A automatic rejection 0 0%
B investigation 40 81%
C no follow-up necessary 0 0%
D other (add specific comment) 9 18%
1 not monitored (BFS)

2 less than alert, no action; > alert, investigation
5 NA
1 media fill monitoring of critical surface, if exceed, investigation

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RECOMMENDATION #5
Question: What data should be considered when initially establishing monitoring limits? What is an appropriate
frequency for re-evaluating monitoring limits?
Recommendation:
• Initially, published data and/or historical data from similar operations should be used to set action and alert
levels
• Historical data may be derived from areas of similar aseptic operations or represent a homogenization of a
company’s monitoring levels, by room class, across lines and facilities.
• For aseptic areas where the allowable levels are less than 1 cfu, consideration should be given to the use of
count incidence rates as an indicator of an unfavorable trend.
• Alert and action levels are generally re-evaluated (and re-set if deemed necessary) on an annual basis using
primarily the previous years data for setting monitoring levels for the upcoming year. Published data
should be considered when re-evaluating the action level.
Rationale:
• There are no US regulations stipulating environmental action levels. However, published data should be
considered when setting action levels to assure that historical data-based action levels do not slowly creep
beyond generally accepted levels.
• Remedial action to exceeding alert or actions levels in all room classes should be taken based on
unfavorable trends as opposed to individual data point excursions. This is particularly true (and difficult to
accurately measure) for ISO Class 5 aseptic areas where the action level is 1cfu/m3.
• The frequency at which monitoring levels are re-evaluated is somewhat dependant on the frequency at
which the line is used. It may take additional time to gather sufficient data to re-evaluate lines that are
seldom used.
• What data does your firm consider when initially establishing monitoring limits for a new area?
# Respondents Yes No
A. Historical databases 49 39 (80%) 10 (20%)
B. Cleanroom qualification 45 35 (78%) 10 (22%)
C. Sanitization studies 46 15 (33%) 31 (67%)
D. Publications 49 45(92%) 4 (8%)
E. Media fills 47 16 (34%) 31 (66%)
F. Relationship of monitoring
location to overall operation 46 33 (72%) 13 (28%)
G. Other (add specific comment) 0

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• What data does your firm consider when reevaluating monitoring limits for an existing area?
# Respondents Yes No
A. Historical databases 47 45 (96%) 2 (4%)
B. Cleanroom qualification 45 21 (47%) 24 (53%)
C. Sanitization studies 45 15 (33%) 30 (67%)
D. Publications 44 35 (80%) 9 (20%)
E. Media fills 44 19 (43%) 25 (57%)
F. Relationship of monitoring
location to overall operation 45 26 (58%) 19 (42%)
G. Other (add specific comment) 0
• What frequency does your firm use for re-evaluating environmental monitoring limits? Answer in number of
months between reevaluations, e.g., 6, 12, 18, 24, etc.
Frequency
6 mo. 1 (2%)
12 mo. 33 (70%)
18 mo. 0 (0%)
24 mo. 5 (11%)
Other: 8 (17%)
2 5 yr.
1 12-18 mo.
1 6 and 12 mo.
1 12 and 24 mo.
3 no SOP

-Final Report-
Recommendation #6
Question: What is the maximum number of viable organisms allowed in air samples for the various classifications?
Recommendation:
• The document should be standardized to the ISO designations.
• The air classification table should only use metric units for the microbial action levels.
• Replace the term “limits” in the table with “Action Levels”.
• Add Microbial Settling Plates to Harmonize with EU Annex 1
TABLE 1- Air Classificationsa
Clean Area ISO > 0.5 um Microbiological Microbiological

Classification Designation particles/m3 Action Levelsb Settling Plates
cfu/m3 Action Levelsb,c
(diam. 90mm) cfu/4
hours
100 5 3,520 1d 1d
1000 6 35,200 7 3
10,000 7 352,000 10 5
100,000 8 3,520,000 100 50
a- All classifications based on data measured in the vicinity of exposed articles during periods of activity.
b- Alternate microbiological standards may be established where justified by the nature of the operation.
c- The use of settling plates are optional.
d- Samples from class 100 environments should normally yield no microbiological contaminants.
• Remedial action when exceeding actions levels in all room classes should be taken based on unfavorable
trends (intra and inter day) as opposed to individual data point excursions. For example, in the Class 100
areas a single action may not require a corrective action after a review of EM trend data for the area.
Rationale:
• The recommended modifications to the table harmonize it with international standards and reflect the most
current published standards.
• The incorporation of the term “action levels” clearly convey that the numbers provided are not product
related specifications but levels that when exceeded must be investigated.
• Note: ISO class 5 is approximately equal to EU Grade A.
References:
• <1116> Microbiological Evaluation of Clean Rooms and Other Controlled Environments,

USP 24, United States Pharmacopeial Convention, Inc., Rockville, MD, 1999.
• PDA Technical Report No. 13 (Revised), “Fundamentals of an Environmental Monitoring Program”
Sept/Oct 2001.
• ISO 13408-1, “Aseptic Processing of Health Care Products- Part 1: General Requirements,” August 1,
1998.

-Final Report-
• ISO 14644-1 “Cleanrooms and associated controlled environments – Part 1: Classification of air
cleanliness”, 1999.
• EU Annex 1

• What air microbial action level does your firm use for your Class 1,000 area? Answer in number of CFU
per cubic meter.
facility has a class 1000 area 10 21%

no class1000 area 37 79%
• What action level is used (CFU/m3 ) for class 1000 area:
(<0.0042, 1, 3, 4, 6, 7, 10, 15, 18)

-Final Report-
RECOMMENDATION #7
Question: What type of airflow is required in a closed isolator?
Recommendation:
• The section needs to recognize, up front, that there are two major ‘types’ of isolators; open and closed.
• The term “Unidirectional” in 1369 should be split out for open isolators.
• Within a closed isolator turbulent flow is normally acceptable.
• The terms open isolator and closed isolators should be better defined. Recommend that the definitions be
based on those contained in PDA Technical Report No. 34.
Rationale:
• It is important that open and closed isolators be differentiated between in the document since they have
different operational expectations. Open isolators are characterized by an interface between the external
and internal environments (i.e., to allow ingress of sterile vials and/or egress of filled vials), and therefore,
uses elements of conventional clean room technology (i.e., pressure differentials and unidirectional flow
air) to provide assurance of environmental quality. Closed isolators have no such openings and therefore
generally do not require unidirectional airflow.
• What type of airflow is required at your firm for closed isolators?

A unidirectional 8 (57%)
B turbulent 6 (43%)
NOTE: Some question was raised, in regards to the survey results, if those reporting unidirectional airflow in
close isolators truly had unidirectional flow.

-Final Report-
RECOMMENDATION #8
Question: What is the appropriate recommendation for air handling systems in isolators?
Recommendation:
• Redundant HEPA (or ULPA) filters, in series, are not necessary.
• The document should not specify the type of filter to be used to reduce the microbial load of the air
entering isolator.
• Suggest the following wording... “The air handling system should be capable of maintaining the
requisite environmental conditions within the isolator”
Rationale:
• Redundant filters HEPA (or ULPA) are also not required for traditional aseptic processing areas; therefore,
there should be no a priori requirement to use them to maintain the environment of an isolator.
• Multiple types of filters and potentially new technologies can be used for this purpose. The purpose of
these filters is to insure air entering the isolator meets environmental requirements.
• The important point is that we control ingress and egress of everything entering and exiting the isolator (air,
components, etc.) to insure maintenance of a suitable environment for assurance of sterility of the products
of aseptic processing. How we do it is less important.
• Does your firm use ULPA and HEPA filters installed in series on its isolator system?
Yes 6 (43%)
No 8 (57%)
• If you answered "no" to the last question, do you have multiple pre-filters, including one that is at least
95% ASHREA-rated, preceding the single terminal HEPA or ULPA?
Yes 9 (75%)
No 3 (25%)

-Final Report-
RECOMMENDATION #9
Question: What are appropriate methods for use in the development of decontamination cycles?
Recommendation:
• Isolators should be decontaminated with a sporicidal agent. The decontamination cycle needs to be
qualified.
• Normally, a 4 – 6 log reduction can be justified depending on the application. The specific BI spore titer
used and the selection of BI placement sites should be justified.
• All product contact surfaces, within the isolator should be sterilized, i.e., demonstration of 6 log reduction
of suitable BIs.
• Chemical indicators and fraction negative studies can be used to help develop a decontamination cycle.
However, demonstration of suitable kill of BIs is the ultimate standard.
• It is important to ensure uniform distribution of the decontaminating agent during cycle development.
• Recommend that no change is made in the statement regarding fraction negative studies.
• Clarify various materials statement to stress texture and porosity rather than composition.
Rationale:
• The environment within an isolator is not purported to be sterilized. Sterilization, in this instance, would be
an imprecise technical term. The internal environment is decontaminated to ensure a suitable environment
for aseptic processing.
• Requirements for isolators should not be greater than for conventional areas used for the same function.
Just as with conventional filling areas the use dictates the acceptable decontamination requirements and
environmental expectations.
• What method/s does your firm use in the development of decontamination cycles?
total kill analysis Number of respondents =21 Yes 16 (76%) No 5 (24%)

half-cycle Number of respondents =18 Yes 10 (55%) No 8 (45%)
fraction negative Number of respondents =17 Yes 7 (41%) No 10 (59%)

-Final Report-
Recommendation #10
Question: With respect to terminal sterilization and adjunct processing what flowcharts represent the most risk-
based and scientifically developed approach?
Recommendation:
• No detail should be added to the current text present in the concept paper.
• The comment beginning on line #56 regarding adjunct processing should be reworded to clearly indicate
that adjunct processing is not an expectation at this point.
• The group strongly recommends to PQRI that a group be formed within PQRI or another organization to
further discuss and develop this topic.
Rationale:
• This topic involves adjunct processing being used in conjunction with aseptic processing as a general
concept to increase sterility assurance. As indicated by the survey the topic of adjunct processing has value
and should be explored. However, it will need to be further developed before it can be included, on a
scientific basis, in a guidance document. Added scientific discussion, research, and the establishment of
new standard methods will be needed to understand how it might be used and what expectations from a
regulatory perspective should be considered.
• Since terminal sterilization is far better understood, a firm should not default automatically to aseptic
processing but should explore terminal sterilization during product development.
• What of the following flowcharts represent the most risk-based and scientific development approach? [Pick
only one.]
A terminal sterilization (explore heat methods only) -> aseptic processing 12 27%
B terminal sterilization (explore heat and irradiation methods) -> aseptic
Processing 7 16%
C terminal sterilization (explore heat methods only) -> consider adjunct
process -> aseptic processing 5 11%
D terminal sterilization (explore heat and irradiation methods) -> consider
adjunct process-> aseptic processing 14 32%
E automatically default to aseptic process without any studies 6 14%
• What percentage of sterile products at your firm are aseptically filled and could be terminally sterilized
(capable of withstanding Fo > 8 minutes without change in current packaging and formulation)? [Enter a
number as a percentage.]
0% 36
2% 2
5% 2
10% 4
20% 1
30% 2
don’t know 2

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Aseptic Processing Working Group

Page 2 of 31 March 13,2003

Page 3 of 31 March 13,2003

Aseptic Processing Working Group

Page 4 of 31 March 13,2003

Aseptic Processing Working Group Work Plan

Working Group Members (updated):

Mr. James P. Agalloco Nigel Halls, Ph.D. Mr. Rainer F. Newman

James E. Akers, Ph.D. Mr. Karl L. Hofmann Ms. Jean I. Olsen

Ms. Diane Alexander David Hussong, Ph.D. Ms. Carolyn Renshaw

Barbara Bassler, Ph.D. Mr. Richard M. Johnson Mr. Robert Sausville

Mr. Martyn Becker Kunio Kawamura, Ph.D. Neal J. Sweeney, Ph.D.

Ms. Susan Bruederle Lee E. Kirsch, Ph.D. Mr. Ian Symonds

Don G. Burstyn, Ph.D. Ms. Carol M. Lampe Laura A. Thoma, Ph.D.

Roger Dabbah, Ph.D. Mr. Joseph Lasich Ms. Debbie Trout

William R. Frieben, Ph.D. Leonard Mestrandrea, Ph.D. Richard T. Wood, Ph.D.

Klaus Haberer, Ph.D. Mr. Terry E. Munson

Page 5 of 31 March 13,2003

Page 6 of 31 March 13,2003

• February 4th - Survey results compiled and provided to work group.

1.) Concept Paper Line Number Reference: 637

2.) Concept Paper Line Number Reference: 984

3.) Concept Paper Line Number Reference: 1047

4.) Concept Paper Line Number Reference: 1055

5.) Concept Paper Line Number Reference: 1070

b. Active Air Monitoring-

Page 7 of 31 March 13,2003

6.) Concept Paper Line Number Reference: 1419

7.) Concept Paper Line Number Reference: 1033

8.) Concept Paper Line Number Reference: 981

1.) Concept Paper Line Number Reference: 661

2.) Concept Paper Line Number Reference: 730

3.) Concept Paper Line Number Reference: 787

Page 8 of 31 March 13,2003

4.) Concept Paper Line Number Reference: 981

5.) Concept Paper Reference Line Number Reference: 1014:

6.) Concept Paper Line Number Reference: 82

TABLE 1- Air Classificationa

Clean Area >0.5 um >0.5 um Microbial Limitb

Aseptic Processing Isolators

7.) Concept Paper Line Number Reference: 1369

Question: What type of airflow is required in closed isolators?

8.) Concept Paper Line Number Reference:1372

9.) Concept Paper Line Number Reference: 1448

10) Concept Paper Line Number Reference: 57

Page 9 of 31 March 13,2003

Aseptic Processing Working Group

Page 10 of 31 March 13,2003

Concept Paper Line Number Reference: 637

Concept paper Line Number Reference: 984

Concept Paper Line Number Reference: 1048

Page 11 of 31 March 13,2003

Concept Paper Line Number Reference: 1055

Paper Line Number Reference: 1070

Concept Paper Line Number Reference: 1419

Page 12 of 31 March 13,2003

Added Clarification text (concept paper reference Line 275).

Concept Paper Line Number Reference: 1033

Concept Paper Line Number Reference: 981

Page 13 of 31 March 13,2003

Aseptic Processing Working Group

Page 14 of 31 March 13,2003