Exclusion of Objectionable

Microorganisms from Non-Sterile

Pharmaceuticals, Medical Devices
and Cosmetics
Tony Cundell, Ph. D.
Microbiological Consultant,
Scarsdale, New York

September, 2015 bioMerieux Customer Meeting 1

 Presentation Overview
• Introduction to the PDA Technical Report on the
exclusion of objectionable microorganisms from non-
sterile drug products.
• Regulatory definition of objectionable microorganisms.
• The role of microbial testing in detecting objectionable
microorganisms
• Decision trees for objectionable organisms in different
dosage forms
• Review of clinical literature, product recalls and
infection outbreaks
• Conclusions
September, 2015 bioMerieux Customer Meeting 2
 PDA Technical Report
• As the result of the work of a broad-based industry
task force, PDA TR No. 67 Exclusion of Objectionable
Microorganisms from Non-Sterile Pharmaceuticals,
Medical Devices and Cosmetics published October,
2014 in conjunction with the PDA Global
Pharmaceutical Microbiology Conference,
Bethesda, Maryland.

September, 2015 bioMerieux Customer Meeting 3

 Other Notable Publications
• USP <1115> Bioburden Control in Non-sterile
Drug Substances and Drug Products that
emphasizes the frequency of monitoring
should reflect the potential risk associated
with the dosage form was published in
Second Supplement to USP 37 official on
December 1, 2014.

September, 2015 bioMerieux Customer Meeting 4

 Technical Report
The objective of the technical report was to define
industry best practices on how to mitigate the risk of
microbial contamination in non-sterile products
• Emphasis on:
– Risk-based decision criteria

– Assessing whether microorganisms if found in a non-sterile product

were objectionable or not.

– No list of objectionable microorganisms was provided

September, 2015 bioMerieux Customer Meeting 5

 Task Force Members
• Anil Sawant, Ph.D. J&J (Co-chair) • Michael Long, Consultant
• Tony Cundell, Ph.D. Consultant (Co- • Patrick Murray, Ph. D. Becton
chair) Dickson
• Donald G. Ahearn, Ph.D. Georgia • Judith Noble-Wang, Ph.D. CDC
State University • Per Arne Parment, MD. Ph.D.
• Matthew Arduino, Ph.D. CDC Consultant
• Julie Barlasov, Perritt Laboratories • Dona Reber, Pfizer
• Mark Dato, MD, Ph.D. P&G • David Roesti, Novartis
• Andrew Dick, J&J • Frank Settineri, Consultant
• Donald English, Avon • Linda Skowronsky, GlaxoSmithKline
• Rhonda Ezell, Qualitest • Donald Singer, GlaxoSmithKline
Pharmaceuticals • John Stone, Ph.D. Kao, USA
• Dennis Guilfoyle, Ph.D. FDA • Scott Sutton, Ph.D. Consultant
• David Hussong, Ph.D, FDA • Edward Tisdale, Ph.D. Baxter
• Mark Kaiser, Lancaster Laboratories • Myriam Sosa, Novartis

September, 2015 bioMerieux Customer Meeting 6

 Scope of the Task Force
• The contamination of marketed products with
potential objectionable microorganisms continues
to be an infrequent but chronic problem. i.e.
around 20 U. S. recalls annually.
• The U.S., Japanese and European pharmacopeias
have harmonized the microbial test methods for
enumeration and the detection of specified
microorganisms that would be the basis of testing.
• Mycotoxins, viruses and sterile dosage forms were
out of scope of this Technical Report.

September, 2015 bioMerieux Customer Meeting 7

 US Recalls of Non-sterile Products
• A recent U.S. survey of 144 reported recalls of non-
sterile pharmaceutical drug products (5%), over-
the-counter drug products (42%), cosmetics (31%),
medical devices (14%) and dietary supplements (8%
of the total recalls) for microbiologically-related
issues for the 7-year period from 2004 through
2011.
• Publication highlighted that the majority of these
recalls (72%) were associated with objectionable
microorganisms and not for exceeding microbial
enumeration limits (Sutton and Jimenez, 2012).

September, 2015 bioMerieux Customer Meeting 8

 Industry Challenge
• An absence of objectionable
microorganisms requirement for
a non-sterile product is a Critical
Quality Attribute without a
defined test method and
acceptance criteria making it a
unique product specification.
• Furthermore, there is no
consensus amongst
manufacturers and regulators
how to approach this issue.

September, 2015 bioMerieux Customer Meeting 9

 Regulatory Requirements
• FDA GMPs: The FDA CGMP regulations 21 CFR
211.113 Control of microbiological contamination
states: a) Appropriate written procedures, designed
to prevent objectionable organisms in drug
products not required to be sterile, shall be
established and followed.
• Furthermore, 21 CFR 211.165 Testing and release
for distribution (b) states: There shall be
appropriate laboratory testing, as necessary, of
each batch of drug product required to be free of
objectionable microorganisms.
September, 2015 bioMerieux Customer Meeting 10
 TGA Requirements
• ICH Q4B Annex 4 A Microbiological Examination of
Non-Sterile Products: Microbial Enumeration Tests
(General Test Chapter); Test for Specified
Microorganisms (General Test Chapter) and
Acceptance Criteria for Pharmaceutical
Preparations and Substances for Pharmaceutical
Use (General Informational Chapter).
• Note: These are harmonized with USP <61>, <62>
and <1111>

September, 2015 bioMerieux Customer Meeting 11

TGA 17.3.2 Objectionable Microorganisms
• In addition to being free from contamination with
specified microorganisms, a non-sterile medicine
should also be free from contamination with other
microorganisms that might be objectionable in the
dosage form.
• For example, pseudomonad-type bacteria are
considered to be objectionable in aqueous dosage
forms that are intended for inhalant, cutaneous,
nasal, auricular, oromucosal, gingival or vaginal use
and in transdermal patches.
• These dosage forms are expected to be free from
contamination with these types of bacteria.
September, 2015 bioMerieux Customer Meeting 12
 TGA Recommendations
Evaluation of the significance of, and risk from, other
objectionable microorganisms should consider:
• the formulation of the medicine
• its route of administration
• its method of application
• the population for which the medicine is intended,
including:
• the possibility of underlying illness in the user of
the medicine
• the possible concurrent use of immunosuppressive
agents or corticosteroids.
September, 2015 bioMerieux Customer Meeting 13
What are Objectionable Microorganisms?
CGMP regulations, i.e. CFR 211.113 does not define
the term objectionable microorganisms, but they can
be broadly defined as:
1) Microorganisms that can proliferate in a product
adversely affecting the chemical, physical, functional
and therapeutic attributes of that pharmaceutical
product.
2) Microorganisms that due to their numbers in the
product and their pathogenicity can cause infection in
the patient in the route of administration when
treated with that pharmaceutical product.
September, 2015 bioMerieux Customer Meeting 14
 Microbiological Testing
Product-release and shelf-life testing
• Three levels of testing are required:
1) microbial enumeration,
2) testing for the absence of specified
microorganisms, and
3) screening for objectionable microorganisms.

September, 2015 bioMerieux Customer Meeting 15

 Regulatory Requirements
• The inclusion of the phrase "as necessary“ in many
regulations implies a risk-based approach to
product testing and decisions about which products
will or will not be routinely tested.

• Drug manufacturers cannot rely solely on finished

product testing to comply with regulation but must
ensure the quality of their products from the
receipt of production materials to the end of the
manufacturing process by following current GMPs.

September, 2015 bioMerieux Customer Meeting 16

 Regulatory Requirements
Bioburden control is achieved by:
• Procuring pharmaceutical ingredients of high
microbiological quality.
• Formulating robust products with low water activities
and effective preservative systems that resist microbial
contamination.
• Good bioburden control through sound equipment
cleaning, disinfectant programs, utility management
and personnel hygiene.
• Emphasis on cGMP compliance.
• Risk-based microbial testing programs.

September, 2015 bioMerieux Customer Meeting 17

 Recommended Microbiological
Quality Requirements
• The recommended microbiological quality
requirements by pharmaceutical dosage form can
be found in the harmonized general informational
chapter USP <1111> Microbiological attributes of
non-sterile pharmaceutical products.
• Specific microbial requirement may be found in
individual USP product monographs.

September, 2015 bioMerieux Customer Meeting 18

 Microbiological Quality Requirements
Dosage Form TAMC TCYMC Absence of Specified Microorganisms (in 1 g or 1 mL)

Non-aqueous preparations for oral use (e.g., E. coli Salmonella spp. (unrefined plant or animal
tablets and capsules) 103 102 material only)

Aqueous preparations for oral use (e.g., oral E. coli

liquids, syrups and suspensions) 102 101

Rectal products 103 102 -

Preparations for oromucosal, gingival and S. aureus and
auricular use 102 101
P. aeruginosa
Preparations for cutaneous use (e.g., topical S. aureus and
liquids, ointments, gels and creams 102 101 P. aeruginosa

Nasal products (e.g. drops and sprays) S. aureus and

102 101
P. aeruginosa
Preparations for vaginal use (e.g., suppositories, S. aureus, P. aeruginosa and
ointments and creams) 102 101 C. albicans

Inhalants (e.g., dry powder inhalants and aerosol S. aureus, P. aeruginosa and
inhalants) 102 101 bile-tolerant, Gram-negative bacteria
* TAMC/ TCYMC counts in cfu/g or cfu/mL

September, 2015 bioMerieux Customer Meeting 19

 Specified Microorganisms
• Tests for the Absence of Specified
Microorganisms that are applied to different
dosage forms are found in USP <62> while USP
<1111> contains the acceptance criteria.
• The screening tests are absence and quantitative
of bile salt-tolerant gram-negative bacteria,
absence of E. coli, P. aeruginosa, S. aureus, C.
albicans, and Clostridium spp (in 1g) and
absence of Salmonella spp (in 10g).

September, 2015 bioMerieux Customer Meeting 20

 Specified Microorganisms
These USP <62> tests generally consist of three steps:
• General enrichment in soybean – casein digest or
Sabouraud dextrose broth to increase the number
of microorganisms.
• Selective enrichment using specialized broth and
incubation conditions to select for the target
specified microorganisms.
• Growth on solid diagnostic media for isolation and
presumptive identification of the specified
microorganisms.
• Confirmatory identification to species.
• Note: These tests are too selective to screen for
objectionable microorganisms

September, 2015 bioMerieux Customer Meeting 21

 Test for Absence of P. aeruginosa
• After a general enrichment in TSB incubated at 30-
35°C to 24-48 hours streak out on Cetrimide Agar
• P. aeruginosa ATCC 9027 is a growth-promotion
organism & E. coli ATCC 8739 inhibitory organism
for Cetrimide Agar used for the indicative medium
of P. aeruginosa
• Note: Related pseudomonads like B. cepacia or P.
fluorescens would not be isolated

September, 2015 bioMerieux Customer Meeting 22

 Test for Absence of P. aeruginosa
• Cetrimide (quaternary ammonium compound) is
highly selective for P. aeruginosa. The water
soluble blue pigment pyocyanin is stimulated by
magnesium chloride and potassium sulfate in
the medium. Colonies with pyocyanin
production that fluoresces under ultra violet
light is indicative of P. aeruginosa.
• The objectionable organism isolation rating is
poor as CET agar is highly selective for P.
aeruginosa inhibiting other pseudomonads,
enterics, and gram-positive bacteria especially at
higher incubation temperatures.

September, 2015 bioMerieux Customer Meeting 23

 Risk-based Microbial Testing

September, 2015 bioMerieux Customer Meeting 24

 Risk-based Microbial Testing

Low Risk Moderate Risk High Risk

Products pass the test if Products pass the test if Products pass the test if
the microorganisms the microorganisms the microorganisms
identified in USP <1111> identified in USP <1111> identified in USP <1111>
and in the relevant and in the relevant and the relevant
monographs were not monographs are not monographs are not
isolated and isolated, other colonies isolated, other colonies
enumeration counts are observed on TSA are not observed on TSA or
below the specified objectionable (see risk blood agar or LMA are
limit. decision tree in Section not objectionable (see
9) and enumeration risk decision tree in
counts are below the Section 9) and
specification limit. enumeration counts are
below the specification
limit.

September, 2015 bioMerieux Customer Meeting 25

 Decision Tree
Objectionable Organism Decision Tree
Failed Microbial Organism of
Fail Pass No Accept
Specifications 1 Specifications Concern

Yes Preservative
Activity 3 Immunoco
mpromised
Infant
Geriatric
Above Below
Failed Internal Number of Growth in Route of High Intended 5 High
Internal Internal Dosage Form No Risk to Patient No Accept
Specifications Organisms product administration risk Population risk
Limit Limit

Water >.06 Low risk

Yes Yes
Activity2

Objectionable Objectionable
AW Accept
Organism Organism
Measurement Low risk
Definitions

Growth in Product
1. Water Activity Immunocompromised
2. Antimicrobial effectiveness test <.06
3. Challenge Infant
Geriatric
Intended 5 High
Organisms of Concern 4 Accept
Population risk
Risk to Patient No Accept

1. Associated with outbreaks

2. Produced recalls
3. Clinically significant infections
Low risk Yes

Objectionable
Accept
Organism

September, 2015 bioMerieux Customer Meeting 26

 Hierarchy of Risk
• Aerosol and dry powder inhalants
• Nasal sprays
•
Higher risk

Otics
• Vaginal suppositories, ointments and
creams
• Topical lotions, gels, ointments,
transdermal patches and creams
• Oral liquids (aqueous)
Lower risk

• Oral liquids (non-aqueous)

• Rectal suppositories, ointments and creams
• Liquid-filled capsules
• Oral tablets and powder-filled capsules
September, 2015 bioMerieux Customer Meeting 27
 CDC Investigated Outbreaks
• 1,022 nosocomial outbreaks were reported in the
clinical literature from 1966 to 2002, i.e. 28
annually.

• The most frequent species implicated in clusters of

hospital patient infection were S. aureus (151
outbreaks, 15% of all outbreaks), P. aeruginosa (91
outbreaks, 9%), K. pneumoniae (73 outbreaks, 7%)

• It is notable that in the vast majority of outbreaks in

which drug products were implicated, the products
were sterile, not non-sterile products.
September, 2015 bioMerieux Customer Meeting 28
 Product Recalls
• The majority of these recalls (72%) were
associated with objectionable
microorganisms and not for exceeding
microbial enumeration limits (Sutton and
Jimenez, 2012).
• B. cepacia was the microorganism most
frequently implicated in the contamination of
multiple-use liquid products.

September, 2015 bioMerieux Customer Meeting 29

 Microorganisms Implicated
Incidence of Infection

12
10
8
6
4
2
0

September, 2015 bioMerieux Customer Meeting 30

 Products Implicated
Incidence of Infection

6
5
4
3
2
1
0

September, 2015 bioMerieux Customer Meeting 31

 Most Significant Microorganisms Associated with the Ten Most
Common Product Recalls, Major Outbreaks Related to
Nonsterile Products and Nosocomial Infections, in Descending
Order
Product Recalls Major Infection Outbreaks Hospital-related Infection
(N=144) (N = 23) (N = 1,022)
B. cepacia (34 B. cepacia complex (12) S. aureus (151)
occurrences)
Unspecified fungi (19) P. aeruginosa (3) P. aeruginosa (91)
B. cereus (9) S. marcescens (2 K. pneumoniae (73)
P. aeruginosa (6) R. mannitolilytica (2) S. marcescens (67)
E. meningoseptica (5) B. cereus (2) E. cloacae (34)
E. gergoviae (5) K. pneumonia (1) E. coli (27)

P. putida (3) E. cloacae (1) A. baumannii (24)

Pseudomonas spp. (2) S. liquefaciens (1) B. cepacia (21)
Salmonella spp. P. lilaceous (1) C. albicans (20)
N/A Enterobacter spp. (1) N/A
1
September, 2015 bioMerieux Customer Meeting 32
 Sources of Microbial Contamination
• Based on the analysis of drug product recalls,
the origin of microorganisms isolated from
non-sterile drug products, in descending
order, is pharmaceutical ingredients ≥
ingredient water >process equipment >>
manufacturing environment >>
manufacturing personnel.
• Cundell, A.M. Risk-based Approach to Pharmaceutical Microbiology In
Encyclopedia of Rapid Microbiological Methods. Edited Michael J. Miller Davis
Harwood/PDA 2005

September, 2015 bioMerieux Customer Meeting 33

 Conclusions
• The frequency of product recalls for objectionable
microorganisms is low, and the incidence of
infections traced back to the use of contaminated
products is extremely low and primarily associated
with the recipient’s health condition.
• Non-sterile product manufacturers share customer
and regulatory agency concerns about the serious
health hazards that objectionable microorganisms
can pose and wants to continuously improve
manufacturing controls, detection and decision
making to exclude objectionable microorganisms
from its products

September, 2015 bioMerieux Customer Meeting 34

 Conclusions
• A risk-based approach must be used for
determining the level and type of testing to be
conducted to identify potentially objectionable
microorganisms in different products.
• Standard microbiological testing procedures may be
used with slight modifications to screen products
for objectionable microorganisms.
• Organisms of concern must be identified based on a
review of the published literature, such as the
reference materials cited in this technical report.

September, 2015 bioMerieux Customer Meeting 35

 Conclusions
• The task force developed technically sound policies,
procedures and training that ensure the exclusion of
objectionable microorganisms from non-sterile
products.
• Policies and procedures developed by each
company must result in consistent decision making
that complies with the applicable regulations.

September, 2015 bioMerieux Customer Meeting 36