Document ID: FDA-2011-N-0080-0019
Agency: fda
Document Type: Rule
Title: Amendments to Sterility Test Requirements for Biological Products
Posted Date: 2012-05-03T04:00Z

[Federal Register Volume 77, Number 86 (Thursday, May 3, 2012)]
[Rules and Regulations]
[Pages 26162-26175]
From the Federal Register Online via the Government Printing Office [www.gpo.gov]
[FR Doc No: 2012-10649]

-----------------------------------------------------------------------

DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

21 CFR Parts 600, 610, and 680

[Docket No. FDA-2011-N-0080]

Amendments to Sterility Test Requirements for Biological Products

AGENCY: Food and Drug Administration, HHS.

ACTION: Final rule.

-----------------------------------------------------------------------

SUMMARY: The Food and Drug Administration (FDA) is amending the 
sterility test requirements for biological products. This rule provides 
manufacturers of biological products greater flexibility, as 
appropriate, and encourages use of the most appropriate and state-of-
the-art test methods for assuring the safety of biological

[[Page 26163]]

products. FDA is taking this action as part of its ongoing efforts to 
comprehensively review and, as necessary, revise its regulations 
related to biological products.

DATES: This rule is effective June 4, 2012.

FOR FURTHER INFORMATION CONTACT: Paul E. Levine, Jr., Center for 
Biologics Evaluation and Research (HFM-17), Food and Drug 
Administration, 1401 Rockville Pike, suite 200N, Rockville, MD 20852-
1448, 301-827-6210.

SUPPLEMENTARY INFORMATION:

Table of Contents

I. Background
II. Summary of the Final Rule
III. Comments on the Proposed Rule and FDA's Responses
    A. General Comments and FDA's Responses
    B. Comments and FDA's Responses on Specific Topics From the 
Proposed Rule
IV. Revisions to Other Regulations
V. Legal Authority
VI. Analysis of Impacts
VII. Environmental Impact
VIII. Federalism
IX. The Paperwork Reduction Act of 1995

I. Background

    This rule revises the sterility requirements for most biological 
products under title 21 of the Code of Federal Regulations (CFR), 
subchapter F, parts 600 through 680 (21 CFR parts 600 through 680) \1\ 
and is intended to promote improvement and innovation in the 
development of sterility test methods by allowing manufacturers the 
flexibility needed for sterility testing of some novel products that 
may be introduced to the market, enhancing sterility testing of 
currently approved products, and encouraging manufacturers to utilize 
scientific and technological advances in sterility test methods as they 
become available.
---------------------------------------------------------------------------

    \1\ The sterility test provisions of this regulation do not 
apply to Whole Blood, Cryoprecipitated Antihemophilic Factor (AHF), 
Platelets, Red Blood Cells, Plasma, Source Plasma, Smallpox Vaccine, 
Reagent Red Blood Cells, Anti-Human Globulin, or Blood Grouping 
Reagents. The provisions also do not apply in cases where the 
Director of the Center for Biologics Evaluation and Research (CBER) 
or the Director of the Center for Drug Evaluation and Research 
(CDER), as appropriate, exempts a product from the requirements 
because the Director finds the manufacturer's data adequate to 
establish that the mode of administration, the method of 
preparation, or the special nature of the product precludes or does 
not require a sterility test or that the sterility of the lot is not 
necessary to assure the safety, purity, and potency of the product. 
(See 21 CFR 610.12(g)(4).)
---------------------------------------------------------------------------

    In the Federal Register of June 21, 2011 (76 FR 36019), FDA 
published a proposed rule that proposed revisions to update 
requirements for sterility testing of biological products. As described 
in the preamble of the proposed rule (76 FR 36019 at 36019 to 36020), 
any product that purports to be sterile should be free of viable 
contaminating microorganisms to assure product safety (Sec.  600.3(q) 
(21 CFR 600.3(q)). Absolute sterility of a lot cannot be practically 
demonstrated without complete destruction of every finished article in 
that lot (USP, Chapter 1211). Therefore, sterility assurance is 
accomplished primarily by validation of the sterilization process or of 
aseptic processing under current good manufacturing practice (CGMP), 
and is supported by sterility testing using validated and verified test 
methods (see e.g., USP Chapter 71, European Pharmacopeia 2.6.1.).
    In the Federal Register of November 20, 1973 (38 FR 32048), we 
reorganized and republished the biologics regulations, which included 
regulations governing sterility testing, as parts 600 through 680.
    Over the years, FDA has amended the biologics regulations, as 
necessary, to clarify and update the sterility test requirements. On 
March 11, 1976 (41 FR 10427) and March 2, 1979 (44 FR 11754), we 
updated Sec.  610.12 (21 CFR 610.12) to clarify the procedures for 
repeat testing. On December 15, 1986 (51 FR 44903), we clarified and 
updated certain requirements for sterility testing to ensure the 
reliability of the growth-promoting qualities of the sterility test 
culture media and to provide greater consistency with the test methods 
of USP XXI. Finally, on September 15, 1997 (62 FR 48174), we 
incorporated by reference into Sec.  610.12(f) the 1995 edition of the 
USP concerning the procedures for the membrane filtration test method.
    Prior to this final rule, Sec.  610.12 required that the sterility 
of most licensed biological products \2\ be demonstrated through the 
performance of tests prescribed in Sec.  610.12(a) and (b). 
Specifically, Sec.  610.12 provided that the sterility of each lot of 
each product, with the exception of certain products,\3\ be 
demonstrated by the performance of prescribed sterility tests for both 
bulk and final container material, unless different sterility tests 
were prescribed in the license (see Sec.  610.12(g)(1)) or the 
manufacturer submitted adequate data \4\ establishing that the mode of 
administration, the method of preparation, or the special nature of the 
product precluded or did not require a sterility test, or that the 
sterility of the lot was not necessary to assure the safety, purity, 
and potency of the product (Sec.  610.12(g)(4)(ii)).
---------------------------------------------------------------------------

    \2\ See list of exemptions in Sec.  610.12(g)(4).
    \3\ Whole Blood, Cryoprecipitated AHF, Platelets, Red Blood 
Cells, Plasma, Source Plasma, Smallpox Vaccine, Reagent Red Blood 
Cells, Anti-Human Globulin, or Blood Grouping Reagents (Sec.  
610.12(g)(4)(i)).
    \4\ In such an instance, the Director of CBER or CDER, as 
appropriate, would determine the adequacy of the data (Sec.  
610.12(g)(4)(ii)).
---------------------------------------------------------------------------

    The regulation also specified the test method and culture media to 
be used. For example, the prescribed sterility test methods relied upon 
culture media (either Fluid Thioglycollate Medium or Soybean-Casein 
Digest Medium) to detect growth of microorganisms (Sec.  610.12(a)(1) 
and (a)(2)). Moreover, Sec.  610.12 specified criteria, such as 
incubation conditions (time and temperature) to be used during testing, 
suitable test organisms for the evaluation of the growth-promoting 
qualities of the culture media, storage and maintenance of test 
organism cultures, and storage and condition of media.
    Since we last clarified and updated our regulations governing 
sterility testing, advances in technology in recent years have allowed 
the development of new sterility test methods that yield accurate and 
reliable test results in less time and with less operator intervention 
than the currently prescribed methods. Some examples of novel methods 
include the Adenosine Triphosphate bioluminescence, chemiluminescence, 
and carbon dioxide head space measurement. Manufacturers may benefit 
from using such sterility test methods with rapid and advanced 
detection capabilities.
    Accordingly, we have amended Sec.  610.12 to promote improvement 
and innovation in the development of sterility test methods, to address 
the challenges of novel products that may be introduced to the market 
in the future, and to potentially enhance sterility testing of 
currently approved products. This final rule provides manufacturers the 
flexibility to take advantage of methods as they become available, 
provided that these methods meet certain criteria.

II. Summary of the Final Rule

    FDA is adopting as final, without material change, the proposed 
requirements for sterility testing. Specifically, this final rule:
     Eliminates specified sterility test methods, culture media 
formulae (or formulation), and culture media test requirements;
     Eliminates specified membrane filtration procedure 
requirements for certain products;
     Eliminates specified sterility test requirements for most 
bulk material;

[[Page 26164]]

     Modifies the repeat sterility test requirements, so that 
repeat tests will occur only once for each lot. These repeat tests are 
limited to situations when the quality control unit conclusively 
determines, after conducting an investigation upon detection of viable 
microbial contamination during the initial test of the lot, that the 
contamination is the result of laboratory error or faulty materials 
used in conducting the sterility test;
     Replaces the storage and maintenance requirements for 
cultures of test organisms used to determine the ``growth-promoting 
qualities'' of culture media with: (1) Validation requirements 
specifying that any sterility test used is able to consistently detect 
the presence of viable contaminating microorganisms and (2) 
verification of ``growth-promoting properties'' or microorganism-
detection capabilities of test and test components;
     Replaces the sample size or amount requirement with a 
requirement that the sample be appropriate to the material being 
tested;
     Replaces the Interpretation of test results section under 
Sec.  610.12(c) with a requirement that manufacturers establish, 
implement, and follow written procedures for sterility testing that 
describe, at a minimum, the test method used, the method of sampling, 
and the written specifications for acceptance or rejection of each lot;
     Simplifies and clarifies the Exceptions section under 
Sec.  610.12(h); and
     Identifies the Director of CDER as one of the two Center 
directors authorized to grant an exemption under the exception 
provision at Sec.  610.12(h)(2). In the proposed rule, the Center for 
Devices and Radiological Health was erroneously identified in this 
exception, instead of the Center for Drug Evaluation and Research.
     Revises the definition of the term ``sterility'' under 
Sec.  600.3(q); and
     Eliminates certain exceptions for allergenic products 
related to sterility testing under Sec.  680.3(c).

III. Comments on the Proposed Rule and FDA's Responses

    We received 17 letters of comments on the proposed rule. These 
comments were received from biologics manufacturers, industry 
associations, and other interested persons. A summary of the comments 
received and our responses follow. We first respond to general comments 
and then respond to comments on the specific topics set forth in the 
preamble of the proposed rule.
    To make it easier to identify the comments and our responses, the 
word ``Comment,'' in parentheses, will appear before the comment's 
description, and the word ``Response,'' in parentheses, will appear 
before our response. We have also numbered each comment to help 
distinguish between different comments. The number assigned to each 
comment is purely for organizational purposes and does not signify the 
comment's value or importance or the order in which it was received. 
Certain comments were grouped together because the subject matter of 
the comments was similar.

A. General Comments and FDA's Response

    (Comment 1) Thirteen of the letters of comments supported the 
proposed rule. Many of the comments agreed that the proposed amendments 
would provide manufacturers of biological products greater flexibility 
and would promote improvement and innovation in the development of 
sterility test methods. Several comments agreed that the proposed 
amendments would allow manufacturers to use the most appropriate and 
state-of-the-art test methods for assuring the safety of biological 
products. Several comments applauded FDA's effort to amend sterility 
test requirements to permit the use of new methods and systems in 
assessing microbiological contamination in sterile products. Another 
comment was pleased to see FDA's commitment to advancing the principles 
of innovation in product development for public health.
    (Response) FDA acknowledges and appreciates the supportive 
comments. As stated previously, the rule provides needed flexibility 
and encourages manufacturers to benefit from scientific and 
technological advances in sterility test methods as they become 
available.
    (Comment 2) One comment noted an error in the reference to the 
European Pharmacopeia 2.6.2. provided in the first paragraph in section 
I of the preamble to the proposed rule. The comment pointed out that 
European Pharmacopeia 2.6.2. is the chapter for Mycobacteria testing.
    (Response) We agree with this comment. The reference should have 
been to European Pharmacopeia 2.6.1. Sterility testing.
    (Comment 3) One comment concurred with the preamble statement that 
``* * * sterility assurance is accomplished primarily by validation of 
the sterilization process or by the aseptic processing procedures under 
CGMP, and is supported by sterility testing using validated and 
verified test methods,'' (76 FR 36019 at 36019). However, the commenter 
went on to state that ``* * * the regulations would be better suited by 
ensuring that the aseptic manufacturing processes follow strict GMP, 
further leveraging the requirements for aseptic environments, media 
fill programs, and strict oversight of the aseptic process as opposed 
to the perceived assurance that sterility testing of samples provides. 
This is best illustrated through existing verbiage in Sec.  211.113(b) 
(21 CFR 211.113(b)) but should be further expanded upon to provide 
improved guidance to industry and investigators.''
    (Response) We acknowledge that product sterility testing does not 
provide absolute assurance of product sterility. However, we believe 
validation of aseptic processes,\5\ using process simulations or media 
fills, together with operational controls and product sterility 
testing, provide a sufficient level of assurance that products 
purported to be sterile are in fact sterile. Therefore, we do not agree 
that additional requirements are necessary because the existing CGMP 
requirements under parts 210 and 211 (21 CFR parts 210 and 211) and the 
other applicable regulations in parts 600 through 680 already address 
the concerns raised by the commenter. We believe this final rule, 
together with the other applicable regulations and Agency guidance, 
provide manufacturers appropriate latitude to determine how to achieve 
the level of control necessary for compliance.
---------------------------------------------------------------------------

    \5\ See the applicable requirements in parts 210, 211, and 600 
through 680, and FDA's guidance document entitled ``Guidance for 
Industry: Sterile Drug Products Produced by Aseptic Processing--
Current Good Manufacturing Practice,'' dated September 2004.
---------------------------------------------------------------------------

    (Comment 4) One comment expressed a concern that an environmental 
requirement is not part of the proposed rule. The commenter stated, 
``Environmental conditions are important to avoid cross-contamination'' 
and proposed the addition of the following wording described in 
European Pharmacopeia 2.6.1. ``The test for sterility is carried out 
under aseptic conditions. In order to achieve such conditions, the test 
environment has to be adapted to the way in which the sterility test is 
performed. The precautions taken to avoid contamination are such that 
they do not affect any microorganisms which are to be revealed in the 
test. The working conditions in which the tests are performed are 
monitored regularly by appropriate sampling of the working area and by 
carrying out appropriate controls.''

[[Page 26165]]

    (Response) In discussing ``environmental conditions,'' we 
understand the comment to mean environmental controls. We have 
considered the issue, including the points raised in this comment and 
have decided not to adopt the suggested language or revise the rule in 
light of the suggested language because the concerns expressed by the 
commenter are currently addressed in the CGMP requirements in parts 210 
and 211 and the applicable regulations in parts 600 through 680. In 
addition, manufacturers may turn to relevant Agency guidance documents 
for additional guidance. Furthermore, as the commenter states, the 
proposed wording regarding environmental controls under which the 
sterility test is to be performed is already described in European 
Pharmacopeia 2.6.1., and USP Chapter 71, both of which are additional, 
valuable resources for manufacturers.
    (Comment 5) One comment noted that while Sec.  610.12 addresses 
aspects of sterility, the current theme of the section is specific to 
sterility testing. The commenter therefore suggested either renaming 
the title of Sec.  610.12 as ``Sterility Test,'' or broadening Sec.  
610.12 so that the regulation addresses all critical elements in the 
content area of sterility.
    (Response) We decline to adopt either recommended change because we 
believe that the current title of Sec.  610.12 remains appropriate and 
that the suggested title change is unnecessary. In response to the 
comment expressing a desire to broaden Sec.  610.12 to address all 
critical elements in the content area of sterility, FDA notes that this 
comment is outside the scope of this final rule.

B. Comments and FDA's Response on Specific Topics From the Proposed 
Rule

    The following are comments and FDA's responses, as identified by 
the specific topic in the proposed rule to which the comment and FDA's 
response applies.
1. When is sterility testing required?
    For the reasons discussed in the preamble to the proposed rule (76 
FR 36019 at 36020 to 36021), we proposed amending Sec.  610.12 to 
eliminate the sterility test requirement for most bulk materials. We 
have determined that, in most cases, for purposes of sterility testing, 
the most appropriate test material is the final container material. We 
recognize that due to the nature of some biological products, testing 
the final container material may not always be feasible or appropriate. 
Thus, as finalized, Sec.  610.12 requires that prior to release, 
manufacturers of biological products must perform sterility testing of 
each lot of each biological product's final container material or other 
material (e.g., bulk material or active pharmaceutical ingredient 
(API), in-process material, stock concentrate material), as 
appropriate, and as approved in the biologics license application (BLA) 
or BLA supplement. For example, as discussed in the preamble to the 
proposed rule (76 FR 36019 at 36021), certain allergenic and cell and 
gene therapy products may need to be tested for sterility at an in-
process stage or some other stage of the manufacturing process (e.g., 
intermediate, API, bulk drug substance) instead of the final container 
material because the final container material may interfere with the 
sterility test. Likewise, as discussed in the preamble to the proposed 
rule, some cell therapy products and cell-based gene therapy products 
may need to be tested for sterility at an in-process stage or some 
other stage of manufacturing process because low production volumes may 
result in an insufficient final container material sample for sterility 
testing or a short product shelf-life may necessitate administration of 
the final product to a patient before sterility test results on the 
final container material are available.
    (Comment 6) Three comments were particularly supportive of FDA's 
proposal to eliminate the sterility test requirements for bulk 
material. One comment noted this change will be particularly helpful 
for cellular therapy products.
    (Response) We appreciate the supportive comments. We agree that the 
elimination of specified sterility test requirements for most bulk 
materials will provide manufacturers with greater flexibility and in 
most cases, for purposes of sterility testing, the most appropriate 
test material is the final container (76 FR 36019 at 36021). We also 
acknowledge that due to the nature of some biological products, this 
change could result in the need for some manufacturers to modify their 
testing procedures to eliminate testing for bulk materials. However, we 
note that these modifications to eliminate testing for bulk materials 
would be made following existing change control procedures and a 
submission to FDA to report the change would not be required.
    If it is determined that sterility testing needs to be performed on 
material other than the final product, due to the nature of the final 
product, we would expect the manufacturer, as required under Sec. Sec.  
601.2 and 601.12, to include in its BLA or BLA supplement: (1) A 
description of the details of the sterility test method used, including 
the procedure for testing the alternate material instead of the final 
container material; and (2) the scientific rationale for selecting the 
specific test material instead of the final container material.
    As discussed in the preamble to the proposed rule (76 FR 36019 at 
36021), a manufacturer who desires to utilize an alternate sterility 
test method other than the one approved in its BLA must submit a BLA 
supplement in accordance with Sec.  601.12(b).
    (Comment 7) One comment asserted that upon finalization of the 
rule, a manufacturer who desires to utilize an alternative sterility 
test other than the one approved in its BLA should be permitted to 
submit the change to FDA in its annual report in accordance with Sec.  
601.12(d), as opposed to a prior approval supplement to an approved 
application under Sec.  601.12(b).
    (Response) We consider changes that may affect the sterility 
assurance level of a product to have substantial potential to affect 
the safety, purity, or potency of a product and have consistently 
identified this change as one that requires prior approval. Therefore, 
a manufacturer who desires to utilize an alternate sterility test 
method other than the one approved in its BLA must submit a prior 
approval supplement to an approved application in accordance with Sec.  
601.12(b). We note that approval of the supplement will be based on the 
determination that the data submitted with the request establishes a 
regulatory basis for approval.
2. What are the sterility test requirements?
    a. Test methods--We proposed amending Sec.  610.12 to eliminate 
references to specific test methods and culture media for sterility 
testing and to instead require that the sterility test be appropriate 
to the material being tested such that the material does not interfere 
with or otherwise hinder the test. As discussed in the preamble to the 
proposed rule (76 FR 36019 at 36021), we believe this revision 
recognizes current practices and provides manufacturers the flexibility 
to take advantage of suitable modern sterility test methods and keep 
pace with advances in science and technology.
    As also discussed in the preamble to the proposed rule (76 FR 36019 
at 36021), because we are expanding potentially acceptable sterility 
test methods to include non-culture-based methods in addition to 
culture-based methods, we also have removed the definition of ``a lot 
of culture medium.'' Previously, Sec.  610.12(e)(2)(i) defined this 
term as ``* * * that quantity of uniform material identified as having 
been

[[Page 26166]]

thoroughly mixed in a single vessel, dispensed into a group of vessels 
of the same composition and design, sterilized in a single autoclave 
run, and identified in a manner to distinguish one lot from another.'' 
Although we have deleted this term from Sec.  610.12, we believe (as 
stated in the preamble to the proposed rule) that this concept is 
captured by the definition of ``lot'' in Sec.  600.3(x). We note that 
this change is also consistent with our understanding that prepared 
culture media may be purchased, in which case a lot may be 
predetermined by the vendor.
    (Comment 8) Two comments opposed the elimination of the specified 
sterility test methods and culture media because eliminating the 
specific requirements may lead to different interpretations by 
industry, as well as FDA investigators. One comment stated that the 
current text on acceptable culture media, reference organisms, and 
incubation temperatures for sterility testing represents essential 
guidance for industry. The comments suggested that either the current 
regulations be retained in addition to the proposed amendments or 
retained as guidance.
    (Response) We reiterate that the purpose of this rule is to provide 
manufacturers of biological products greater flexibility and to 
encourage use of the most appropriate and state-of-the-art test methods 
for assuring the safety of biological products. Accordingly, at this 
time, we decline to retain the current specified sterility test 
methods, culture media, reference organisms, and incubation 
temperatures in regulation or guidance. Furthermore, we disagree that 
this rule may lead to inconsistent interpretations by industry and FDA 
staff because sterility test methods for biological products are 
approved in the manufacturer's BLA or BLA supplement, and hence, the 
data submitted with the request are reviewed in a consistent manner in 
accordance with review management procedures. Therefore, we believe the 
commenters' concerns about inconsistencies in interpretation are 
unfounded.
    (Comment 9) One commenter expressed concern about the applicability 
of the proposed changes in the global regulatory market in that the use 
of approved alternative sterility methods would not be globally 
applicable in the absence of compendial harmonization. The commenter 
inquired whether FDA has plans to harmonize the use of alternative 
sterility methods with the three main global compendia.
    (Response) We do not agree that the final rule and the use of a 
suitable modern sterility test method will interfere with the global 
regulatory market. The purpose of the rule is to provide for greater 
flexibility and to encourage use of the most appropriate and state-of-
the-art test methods for assuring the safety of biological products. We 
believe this final rule will foster the adoption of novel methods and 
that alignment with global pharmacopeial methods will occur over time. 
With respect to FDA's future plans to harmonize the use of alternative 
sterility methods with the three main global compendia, we note that 
any such discussion is outside the scope of this rule.
    (Comment 10) One comment proposed adding a reference in the 
regulations to a compendial method and allowing for the implementation 
of alternative methods. The commenter expressed concern that, in the 
global marketplace, implementation of a novel method different from USP 
Chapter 71 would not be harmonized with other compendia and might pose 
risks to approval of marketing authorizations if new tests are not 
recognized or accepted by foreign health authorities.
    (Response) We do not agree with the comment and note that 
incorporating such a reference would be inconsistent with the intent of 
this rule. We reiterate that we do not agree that this final rule will 
interfere with the global marketplace. Rather, we believe that 
facilitating flexibility and encouraging the use of the most 
appropriate and state-of-the-art test methods will foster the adoption 
of novel method technologies and that alignment with pharmacopeia 
methods will occur over time. Furthermore, as we have explained in the 
preamble to the proposed rule, FDA considers established USP compendial 
sterility test methods to already have been validated using an 
established validation protocol; therefore their accuracy, specificity, 
and reproducibility need not be reestablished to fulfill the validation 
requirements under the final rule. Only a manufacturer who desires to 
utilize an alternative method other than the one approved in its BLA 
must submit a BLA supplement in accordance with Sec.  601.12(b). This 
rule does not require manufacturers to utilize an alternative method 
other than the one approved in their BLA.
    (Comment 11) One comment stated that the absence of references to 
standards such as USP Chapter 71 within Sec.  610.12 may lead to 
confusion and suggested that a general disclaimer that FDA is not 
endorsing any particular standard or the provision of specific examples 
within the regulation may provide an important point of reference for 
compliance. Two comments stated that USP Chapter 71 and European 
Pharmacopeia 2.6.1. should be listed within Sec.  610.12 as a baseline 
or standard for sterility testing. Two other comments recommended 
referring to the USP Chapter 71 as the ``referee'' method instead of 
referring to it as an example.
    (Response) The concerns expressed in the comments are unfounded. We 
reiterate that we consider the current sterility test methods in a 
manufacturer's BLA or BLA supplement to already have been validated. In 
contrast, newer methods (for example, non-culture-based methods that 
have not been validated according to an established protocol) or those 
that deviate from the official compendial sterility test methods will 
require validation.
    Moreover, the final rule requires that a novel method be validated 
in accordance with an established protocol to demonstrate that the test 
is capable of consistently detecting the presence of viable 
microorganisms. We believe methods validation is a well recognized 
activity and can be performed without comparison to a ``referee'' test 
method.
    Furthermore, we note that there is no single ``referee'' test 
method that would work for all products and that some novel methods 
cannot be easily compared to culture-based methods such as USP Chapter 
71 because these testing methods do not measure microbial growth. 
Therefore, we believe that it is neither necessary nor appropriate to 
add a reference to a standard or ``baseline'' in this final rule.
    (Comment 12) We received two comments regarding growth-promotion 
testing. One comment asserted that the proposal to eliminate the 
requirements to test culture media with specific test organisms, to 
eliminate the number of organisms that must be used to demonstrate 
growth-promoting qualities of culture media, and to eliminate specific 
incubation conditions and visual examination requirements may lead to 
different interpretations on which organisms can and should be used. 
The comment proposed that a reference to a ``referee'' method be added 
to the regulation including requirements for growth promotion and the 
strains and number of organisms to be used. The other comment supported 
the elimination of the list of specified organisms, while also stating 
that providing a list of organisms for manufacturers to consider would 
be a benefit to facilities that do not have the necessary expertise or 
staffing.
    (Response) Because we are providing manufacturers the flexibility 
to use

[[Page 26167]]

sterility test methods that are either culture-based or non-culture-
based, which may necessitate different verification activities, we 
decline to retain the existing requirements for specified sterility 
test reference organisms. For similar reasons, we do not believe a 
reference to a ``referee'' method is necessary or appropriate and we 
decline to adopt the recommended change.
    Instead of specifying the number and type of test organisms, under 
Sec.  610.12(b) of the final rule, we require that: (1) The sterility 
test must be appropriate to the material being tested such that the 
material does not interfere with or otherwise hinder the test; (2) the 
sterility test must be validated to demonstrate that the test is 
capable of reliably and consistently detecting the presence of viable 
contaminating microorganisms; and (3) the sterility test and test 
components must be verified to demonstrate that the test method can 
consistently detect the presence of viable contaminating 
microorganisms.
    Due to the variety of currently available and potential future 
sterility test methods, we have eliminated specified incubation 
conditions (time and temperature) and visual examination requirements 
previously prescribed in Sec.  610.12. Since we are allowing any 
validated sterility test method that is appropriate to the material 
being tested, rather than specifying the test and the media used, we 
have also eliminated the Fluid Thioglycollate Medium incubation 
temperatures previously prescribed in Sec.  610.12(a)(1)(ii) for the 
final container material containing a mercurial preservative.
    (Comment 13) One comment recommended that, with respect to 
validation, a definition for the terms ``reliably'' and 
``consistently'' be added to the regulation for greater utility in 
understanding expectations when validating a method. The commenter 
offered, for example, ``* * * that a validated method, though 
performing consistently and reliably, may still not be centered on the 
true value of the specific parameter being tested. Consequently, when 
this method would be used during testing the results may be in a 
statistical state of control, but not necessarily statistically capable 
of measuring the true value.'' The commenter asked FDA to consider ``* 
* * that the use of the terms `reliably and consistently' may infer 
that the validation of a test for non-sterility does not require proof 
of performance at least equivalent to the USP referee method.'' The 
comment therefore asked that Sec.  610.12(b)(2) be revised to require 
that the sterility test be validated to demonstrate an equivalent or 
superior detection of viable contaminating microorganisms compared to 
the USP compendial or like method.
    (Response) FDA has considered the issues raised by these comments 
and has determined that making the suggested changes would be 
inconsistent with the intent of this rule. With respect to the comment 
that the rule should be revised to require that the sterility test be 
validated to demonstrate an equivalent or superior detection of viable 
contaminating microorganisms compared to the USP compendial or like 
method, we reiterate that some novel methods cannot be easily compared 
to culture-based methods such as USP Chapter 71 because they do not 
measure microbial growth. Moreover, we note that the final rule 
requires that a novel method be validated in accordance with an 
established protocol to demonstrate that the test is capable of 
consistently detecting the presence of viable microorganisms. With 
respect to the comment that the terms ``reliably'' and ``consistently'' 
should be defined, we note that these terms are already well understood 
in the industry.
    b. Validation--As discussed in the preamble to the proposed rule 
(76 FR 36019 at 36021 to 36022), the International Conference on 
Harmonisation (ICH) publication entitled ``Validation of Analytical 
Procedures: Text and Methodology Q2(R1)'' dated November 2005, states 
that ``The objective of validation of an analytical procedure is to 
demonstrate that it is suitable for its intended purpose.'' \6\ 
Similarly, USP General Chapter 1223, ``Validation of Alternative 
Microbiological Methods,'' states ``Validation of a microbiological 
method is the process by which it is experimentally established that 
the performance characteristics of the method meet the requirements for 
the intended application.'' For sterility testing, this means that the 
test can consistently detect the presence of viable contaminating 
microorganisms.
---------------------------------------------------------------------------

    \6\ This guideline for industry was previously named ``Text on 
Validation of Analytical Procedures'' (ICH-Q2A), dated March 1995 
(approved by the Steering Committee in October 1994). An 
accompanying guideline entitled ``Validation of Analytical 
Procedures: Methodology (Q2B),'' dated November 6, 1996, was 
subsequently developed and approved by the Steering Committee in 
November 1996. The parent guideline is now renamed ``Validation of 
Analytical Procedures: Text and Methodology Q2(R1)'' and was revised 
in November 2005. At that time, the guideline on methodology (Q2B) 
was incorporated into the parent guideline.
---------------------------------------------------------------------------

    We have eliminated the prescribed sterility test methods found in 
Sec.  610.12 and instead will allow the use of sterility test methods 
that are validated in accordance with established protocols to be 
capable of consistently detecting the presence of viable contaminating 
microorganisms. If an established USP compendial sterility test method 
is used, a manufacturer must verify that this established method is 
suitable for application to the specific product (see Sec. Sec.  
211.165(e) and 211.194(a)); however, FDA considers established USP 
compendial sterility test methods to already have been validated using 
an established validation protocol, so their accuracy, specificity, and 
reproducibility need not be reestablished to fulfill the validation 
requirement under the final rule. In contrast, novel methods and any 
methods that deviate from the USP compendial sterility test methods 
require the detailed validation discussed in this document and 
elsewhere in this preamble.
    We again note that Sec.  610.12 requires the use of a material 
sample that does not interfere with or otherwise hinder the sterility 
test from detecting viable contaminating microorganisms. This 
requirement is crucial because the material itself or substances added 
to the material during formulation may make some sterility tests 
inappropriate for use. A validated sterility test method is a critical 
element in assuring the safety, purity, and potency of the product. USP 
General Chapter 1223, as well as the ICH guideline referenced earlier 
entitled ``Text on Validation of Analytical Procedures,'' dated March 
1995 (ICH-Q2A), provide general descriptions of typical validation 
parameters, how they are determined, and which subset of each parameter 
is required to demonstrate validity, based on the method's intended 
use. Validation of each test method should be performed on a case-by-
case basis to ensure that the parameters are appropriate for the 
method's intended use. In the context of reviewing sterility test 
methods as part of BLAs and BLA supplements, FDA may decide, as 
appropriate, to encourage the use of the compendial method as a 
benchmark or starting point for validation of novel methods and certain 
other methods.
    (Comment 14) One comment requested clarification regarding 
validation of novel methods and any methods that deviate from the USP. 
This commenter stated that to validate novel test methods, ``the 
sponsor not only has to test the matrix effects'', but also has to 
validate the new method against the USP compendial method. The

[[Page 26168]]

commenter also stated that this would impede the use of innovative 
technologies and increase the risk and cost to the sponsor. In 
addition, the commenter recommended that duplicative testing 
requirements be avoided and that the manufacturer of the technology or 
a third party be allowed to perform the validation of new methods.
    (Response) The commenter misinterpreted the validation requirements 
under the proposed (and final) rule. The revisions we are adopting in 
the final rule do not require duplicative validation of novel methods 
against the USP compendial method or testing under a separate 
validation procedure. Instead, novel methods and any methods that 
deviate from the USP compendial sterility test methods will require a 
single, detailed validation study to be conducted, which may include 
the use of the compendial method as a benchmark or starting point. We 
disagree that such validation will impede the use of innovative 
technologies and will increase the risk and cost to the sponsor. 
Instead, we believe that, as discussed elsewhere in this document and 
in the preamble to the proposed rule, that this final rule will 
encourage the use of innovative technology.
    (Comment 15) One comment referenced the preamble statement that ``* 
* * FDA may decide, as appropriate, to encourage the use of the 
compendial method as a benchmark or starting point for validation of 
novel methods and certain other methods.'' (76 FR 36019 at 36022) and 
suggested that the use of the compendial method as a benchmark or 
starting point should be more strongly encouraged.
    (Response) While FDA may decide, as appropriate, to encourage the 
use of the compendial method as a benchmark or starting point for 
validation of some novel or other methods, we also may decide not to 
encourage such use for some (for example, non-culture-based) methods 
that cannot easily be compared to culture-based methods such as the USP 
compendial method. Therefore, we disagree that the use of the 
compendial method as a benchmark or starting point should be more 
strongly encouraged or required.
    (Comment 16) We received two comments in response to our request in 
the proposed rule for comments on whether the proposed requirements are 
sufficient to ensure adequate validation of novel sterility test 
methods or whether additional criteria or guidance is needed. One 
comment recommended that any guidance to accompany the final rule be 
developed to include such things as a list of organisms for 
manufacturers to consider in the development of their validation and 
verification plans, including examples of when verification is 
required. One comment suggested that such additional guidance include 
information related to a determination of the panel of relevant 
organisms in the sample matrix used in challenging the sterility test 
during validation.
    (Response) We appreciate the interest in additional guidance for 
validation of novel sterility test methods and will consider the need 
to develop future guidance in accordance with the good guidance 
practices set out in 21 CFR 10.115.
    As discussed in the preamble to the proposed rule, it is important 
to consider validation principles, such as limit of detection, 
specificity, ruggedness, and robustness, while developing the 
validation protocol and performing validation studies. These terms are 
defined as follows:
     The ``limit of detection'' reflects the lowest number of 
microorganisms that can be detected by the method in a sample matrix. 
This is necessary to define what is considered contaminated.
     ``Specificity'' is the ability of the test method to 
detect a range of organisms necessary for the method to be suitable for 
its intended use. This is demonstrated by challenging the sterility 
test with a panel of relevant organisms in the sample matrix.
     ``Ruggedness'' is the degree of reproducibility of results 
obtained by analysis of the same sample under a variety of normal test 
conditions, such as different analysts, different instruments, and 
different reagent lots.
     ``Robustness'' is the capacity of the test method to 
remain unaffected by small, but deliberate, variations in method 
parameters, such as changes in reagent concentration or incubation 
temperatures.
    (Comment 17) One comment stated that for the detailed validation of 
a novel method, the validation principles should be restricted to the 
limit of detection, specificity, and robustness (i.e., to not include 
ruggedness).
    (Response) We agree that the validation principles of limit of 
detection, specificity, and robustness are important to consider when 
developing protocols and performing validation studies. However, we 
understand the comment to suggest excluding ruggedness. We view 
ruggedness as an important validation principle to be considered, and 
we do not agree with excluding it from the scope of this rule. We note 
that the final rule does not include prescriptive details on how to 
conduct validation studies; it simply codifies our longstanding policy 
that the sterility test must be validated to demonstrate that the test 
is capable of reliably and consistently detecting the presence of 
viable contaminating microorganisms.
    (Comment 18) One comment objected to the requirement in existing 
Sec.  211.160(b) as to the establishment of sampling plans because ``* 
* * it is not practical or feasible to develop a scientifically sound 
sampling plan to ensure a product conforms to standards of sterility.'' 
The comment recommended as a solution to either remove the requirement 
for scientific sampling plans with respect to sterility testing or to 
provide a clarification of ``scientifically sound'' versus 
``appropriate.''
    (Response) The suggested revisions go beyond the scope of the 
proposed changes to the sterility test requirements. Furthermore, Sec.  
211.160(b) is an existing current good manufacturing practice 
requirement for finished pharmaceuticals, which states that laboratory 
controls must include the establishment of scientifically sound and 
appropriate specifications, standards, sampling plans, and test 
procedures designed to assure that components, drug product containers, 
closures, in-process materials, labeling, and drug products conform to 
appropriate standards of identity, strength, quality, and purity. We 
consider such laboratory controls to be needed for both culture-based 
and non-culture-based sterility test methods. As stated in the preamble 
to the proposed rule (76 FR 36019 at 36022), the manufacturer must 
establish and document the test method's accuracy, sensitivity, 
specificity, and reproducibility (Sec.  211.165(e)), as specified in 
the BLA or BLA supplement (Sec. Sec.  601.2, 601.12). For sterility 
tests, FDA believes that a validation protocol that would meet these 
standards would, at a minimum, include samples of the material to be 
marketed and incorporate appropriate viable contaminating 
microorganisms to demonstrate the sterility test's growth-promoting 
properties or the method's detection system capabilities, depending on 
the type of test method used. In addition, validation protocols for 
culture-based methods should include both aerobic and anaerobic 
microorganisms when selecting test organisms and include microorganisms 
that grow at differing rates so that manufacturers can establish that 
the test media are capable of supporting the growth of a wide range of 
microorganisms.

[[Page 26169]]

    When utilizing culture-based methods, where appropriate, validation 
protocols should require that challenge organisms be added directly to 
the product prior to membrane filtration or direct inoculation. If this 
is not possible due to inhibition by the product, then validation 
protocols should require that the challenge organism be added to the 
final portion of sterile diluent used to rinse the filter, if a 
membrane filtration test method is used, or directly to the media 
containing the product if a direct inoculation test method is used.
    For non-culture-based methods, the feasibility of identifying 
microorganisms from a contaminated sample should be evaluated during 
validation. If a method does not have the capability to identify 
microorganisms to the species level, the validation protocol should 
require that an additional method for species identification be 
utilized for investigation of detected contaminants. The test organisms 
selected should reflect organisms that could be found in the product, 
process, or manufacturing environment.
    (Comment 19) Two comments sought clarification of the following 
statement in the preamble to the proposed rule: ``When utilizing 
culture-based methods, validation protocols should require that 
challenge organisms be added directly to the product prior to membrane 
filtration or direct inoculation. If this is not possible due to 
inhibition by the product, then validation protocols should require 
that the challenge organism be added to the final portion of sterile 
diluent used to rinse the filter if a membrane filtration test method 
is used, or directly to the media containing the product if a direct 
inoculation test method is used.'' (76 FR 36019 at 36022)
    One commenter stated that this language is inconsistent with the 
harmonized compendial method suitability test which states, ``After 
transferring the content of a container or containers to be tested to 
the membrane, add an inoculum of small number of viable microorganisms 
(not more that 100 colony-forming units) to the final portion of 
sterile diluents used to rinse the filter.'' Another comment sought 
clarification of the suggested limits for the density of the inoculum 
of challenge organisms added directly to the product.
    (Response) The intent of these statements was to clarify that for 
certain biological products utilizing culture-based methods, method 
suitability testing necessitates adding the challenge organism directly 
to the product prior to membrane filtration or direct inoculation. 
Therefore, we are now clarifying that when utilizing culture-based 
methods, where appropriate, validation protocols should require that 
challenge organisms be added directly to the product before membrane 
filtration or direct inoculation. If this is not possible due to 
inhibition by the product, then validation protocols should require 
that the challenge organism be added to the final portion of sterile 
diluent used to rinse the filter if a membrane filtration test method 
is used or directly to the media containing the product if a direct 
inoculation test method is used.
    (Comment 20) One comment addressed the selection of organisms to be 
used. The comment suggested that with respect to validation protocols, 
for consistency, the wording regarding the selection of organisms 
should specifically include wild-type isolates that have been recovered 
from the controlled manufacturing environment and past contaminants of 
the product or any of its sterile components. The comment also 
suggested that this requirement should extend beyond culture-based 
methods. Further, the comment suggested that the statement in the 
preamble that `` `The test organisms selected should reflect organisms 
that could be found in the product, process, or manufacturing 
environment (emphasis added) [76 FR 36019 at 36022],' should be 
tightened to require use of strains actually isolated from the product, 
process, or manufacturing environment, as the word `reflect' probably 
implies use of relevant species that might be sourced from culture 
collections rather than explicitly requiring use of wild-type strains 
(plant isolates).''
    (Response) Our intention with respect to this statement was to 
include those organisms recovered both from the controlled 
manufacturing environment and from the product. Furthermore, the 
preamble statement was intended to refer to validation protocols in 
general, where appropriate, to both culture-based and non-culture-based 
test methods.
    The validation study design should contain the appropriate controls 
to evaluate the product sample's potential to generate false-positive 
and false-negative results. Validation of the sterility test should be 
performed on all new products, and repeated whenever there are changes 
in the test method or production method that could potentially inhibit 
or enhance detection of viable contaminating microorganisms.
    (Comment 21) One comment recommended the addition of ``or 
production method'' to the statement in the preamble so that it would 
now read, ``Validation of the sterility test should be performed on all 
new products, and repeated whenever there are changes in the test 
method or production method that could potentially inhibit or enhance 
detection of viable contaminating microorganisms.'' (See original 
statement 76 FR 36019 at 36022.) The commenter stated that the 
additional language is appropriate because the production process may 
influence the matrix of the test article, which may in turn influence 
the sterility test verification.
    (Response) We agree that changes in the production method or 
manufacturing process could affect the results of testing conducted on 
the product. Therefore, we agree that validation of the sterility test 
should be performed on all new products and repeated whenever there are 
changes in the test method or production method that could potentially 
inhibit or enhance detection of viable contaminating microorganisms.
    c. Verification--As stated in the proposed rule (76 FR 36019 at 
36022), verification is the confirmation that specified requirements 
have been fulfilled as determined by examination and provision of 
objective evidence. While validation of a sterility test method is the 
initial process of demonstrating that the procedure is suitable to 
detect viable contaminating microorganisms, verification occurs over 
the lifetime of the sterility test method and is the process of 
confirming that the sterility test and test components continue to be 
capable of consistently detecting viable contaminating microorganisms 
in the samples analyzed. This verification activity may be necessary on 
a periodic basis or each time a sample is tested, depending upon the 
test method used. Under Sec.  610.12(e) of the final rule, we require 
that the sterility test and test components be verified, as 
appropriate, to demonstrate that they can continue to consistently 
detect viable contaminating microorganisms.
    (Comment 22) One comment maintained that the section of the 
preamble to the proposed rule regarding verification was not totally 
clear and should be reworded to explain the intended purpose. 
Specifically, the comment suggested, in order to clarify the goal of 
verification, adding the following sentence, ``The intended purpose of 
the verification is to confirm that all the reagents utilized in the 
sterility test are qualified.'' The commenter also noted that 
validation is to be done using the product to be tested and proposed 
adding the phrase ``in the product to be tested'' to the following 
statement in the preamble ``While

[[Page 26170]]

validation of a sterility test method is the initial process of 
demonstrating that the procedure is suitable to detect viable 
contaminating microorganisms, verification occurs over the lifetime of 
the sterility test method and is the process of confirming that the 
sterility test and test components continue to be capable of 
consistently detecting viable contaminating microorganisms in the 
samples analyzed.'' (76 FR 36019 at 36022 to 36023)
    (Response) To the extent that the commenter is arguing that our 
explanation is unclear, we disagree. As stated in the preamble to the 
proposed rule at section III.E (76 FR 36019 at 36022 to 36023), we 
believe that in order to verify the sterility test, verification 
activities are necessary to demonstrate that sterility test methods can 
continue to reliably and consistently detect viable contaminating 
microorganisms and that verification is the process of confirming that 
the sterility test and test components continue to be capable of 
consistently detecting viable contaminating microorganisms in the 
samples analyzed. In addition, we acknowledge that method suitability 
testing using the product is an important part of a validation protocol 
for a sterility test method.
3. What information is needed in written procedures for sterility 
testing?
    We have finalized, as proposed, the replacement of the requirements 
found in current Sec.  610.12(c) entitled Interpretation of test 
results, with the requirements that manufacturers must establish, 
implement, and follow written procedures for sterility testing. Written 
procedures are essential to ensure consistency in sampling, testing, 
and interpretation of results and to provide prospective acceptance 
criteria for the sterility test. Written procedures should include all 
steps to be followed in the sterility test method for initial and 
repeat tests and be detailed, clear, and unambiguous. Under the current 
good manufacturing practice regulations, manufacturers are required to 
document that a drug product satisfactorily conforms to final 
specifications for the drug product (Sec.  211.165(a)). As such, 
scientifically sound and appropriate specifications, standards, 
sampling plans, and test procedures must be designed and written to 
ensure that materials conform to appropriate standards of sterility; 
and written procedures must include a description of the sampling 
method and the number of units per batch to be tested (see Sec.  
211.165(c)).
    Under the final rule, manufacturers may use either culture-based or 
non-culture-based sterility test methods to evaluate material for 
sterility. There are marked differences between culture-based and non-
culture-based sterility tests. Section 610.12(c) provides the minimum 
critical considerations that must be included in the written procedures 
for culture-based and non-culture-based sterility tests.
    For culture-based sterility test methods, the written procedures 
must include, at a minimum, a description of the composition of the 
culture media, growth-promotion test requirements, and incubation 
conditions (time and temperature). For non-culture-based sterility test 
methods, the written procedures must include the composition of test 
components, test parameters, including the acceptance criteria, and the 
controls used to verify the test method's ability to consistently 
detect the presence of viable contaminating microorganisms.
4. What is an appropriate sample for sterility testing?
    Selection of an appropriate sample of a lot is critical for 
purposes of sterility testing. Under Sec.  610.12(d) as finalized, due 
to the variety of products covered under Sec.  610.12, the regulation 
requires that the sample be appropriate to the material being tested.
    (Comment 23) Five comments requested clarification of the proposed 
requirement that the sample be ``appropriate to the material being 
tested,'' with respect to the size or volume of the final product lot. 
The comments asserted that the example provided in the preamble of the 
proposed rule, ``For example, a final product lot size of 100,000 units 
would necessitate a greater number of samples to be evaluated than a 
final product lot size of 5,000 units,'' (76 FR 36019 at 36023), 
conflicts with USP Chapter 71 regarding the minimum number of articles 
to be tested in relation to the number of articles in the batch.
    (Response) We acknowledge that the example provided in the preamble 
of the proposed rule erroneously compared a final product lot size of 
100,000 units to one of 5,000 units. We had intended to compare a final 
product lot size of 100,000 to one of 500 units. We recognize that this 
error may have caused confusion among some readers, and that the 
example was inconsistent with the USP Chapter 71 methods for the 
minimum number of articles to be tested in relation to the number of 
articles in the batch. It was not our intent to suggest that 
established USP compendial sterility test methods, including the 
minimal number of articles to be tested in relation to the number of 
articles in the batch, were unacceptable under the new requirements in 
Sec.  610.12(d).
    In order to clarify the new requirement that the sample be 
``appropriate to the material being tested,'' we reiterate that in 
selecting an appropriate sample size, Sec.  610.12(d) requires that the 
following minimal criteria be considered:
     The size or volume of the final product lot. For example, 
a final product lot size of 100,000 units would necessitate a greater 
number of samples to be evaluated than a final product lot size of 500 
units;
     The duration of manufacturing of the drug product.\7\ For 
example, it is important that samples be taken at different points of 
manufacture, which, at a minimum, should include the beginning, middle, 
and end of manufacturing, in an effort to provide evidence of sterility 
of the drug product throughout the duration of the manufacturing 
process; \8\
---------------------------------------------------------------------------

    \7\ See Sec.  210.3(b)(4) for the definition of the term ``drug 
product.''
    \8\ See Sec.  211.160(b) for general requirements for laboratory 
controls.
---------------------------------------------------------------------------

     The final container configuration and size. We believe 
this will ensure appropriate representation of the lot;
     The quantities or concentrations of inhibitors, 
neutralizers, and preservatives, if present, in the test material;
     For a culture-based test method, the volume of test 
material that results in a dilution of the product that was determined 
not to be bacteriostatic or fungistatic; and
     For a non-culture-based test method, the volume of test 
material that results in a dilution of the product that does not 
inhibit or otherwise hinder the detection of viable contaminating 
microorganisms.
    (Comment 24) Two comments stated that the proposed changes related 
to sample size are vague and leave too much room for interpretation by 
industry as well as investigators or auditors when determining an 
appropriate sample size.
    (Response) We disagree that requiring the sample to be appropriate 
to the material being tested is vague and leaves too much open to 
interpretation. Our intent in requiring that the sample be 
``appropriate to the material being tested,'' with consideration of a 
list of minimal criteria, is to provide manufacturers flexibility to 
retain their existing procedures for sterility testing using culture-
based methods, or to take

[[Page 26171]]

advantage of modern methods as they become available, provided that 
these modern methods meet certain criteria, as described in our 
response to Comment 23. In addition, as noted previously, sterility 
test methods are approved by FDA in either a manufacturer's BLA or BLA 
supplement, thereby alleviating concern that the final rule leaves too 
much room for interpretation.
    (Comment 25) One comment asked FDA to clarify whether the 
quantities or concentrations of inhibitors, neutralizers, and 
preservatives, if present in the test material, have an impact on 
sample size and selection. The comment also asked about the 
relationship between the impact of preservatives and any increase in 
the sample size.
    (Response) In selecting an appropriate sample size, Sec.  610.12(d) 
requires consideration of certain minimal criteria, including the 
quantities or concentrations of inhibitors, neutralizers, and 
preservatives, if present in the test material. The consideration of 
the quantities or concentrations of inhibitors, neutralizers, and 
preservatives, if present in the test material, will depend upon the 
product and the test method utilized. This provides both manufacturers 
of future innovative products, as well as manufacturers of currently 
approved products, the flexibility to take advantage of modern methods 
or to retain the sterility testing method as approved in the BLA or BLA 
supplement.
5. What is required to verify the sterility test?
    As discussed in the preamble to the proposed rule (76 FR 36019 at 
36023), verification activities are necessary to demonstrate that 
sterility test methods can continue to reliably and consistently detect 
viable contaminating microorganisms. The degree of verification that is 
necessary depends upon the sterility test method employed. Depending 
upon the sterility test method, verification of each individual test 
might be appropriate. On the other hand, some sterility test methods 
may only need verification activities performed on the selected culture 
media or test organisms. Under Sec.  610.12(e), a manufacturer must 
perform verification activities appropriate for the sterility test 
method chosen, as set forth in the final rule.
    (Comment 26) In the proposed rule (76 FR 36019 at 36020, footnote 
6), we proposed to refer to ``growth-promoting properties'' rather than 
``growth-promoting qualities'' and requested comments on which term is 
most appropriate. We received two comments in response to our request. 
Both comments support the use of ``growth-promoting properties'' and 
agree that ``growth-promoting properties'' reflects more accurate and 
current terminology.
    (Response) We appreciate and agree with these comments and have 
retained the term ``growth-promoting properties'' in the final rule.
    (Comment 27) Two comments requested clarification of the 
requirements for verification of culture-based test methods. One 
comment asked if, for culture-based test methods, all media must 
undergo growth-promotion testing over their shelf-life, and if 
validation were performed for three lots, whether it is acceptable to 
perform growth-promotion testing on the media only when it is initially 
received. One comment acknowledged that each media lot would have to be 
tested for growth-promotion at least at the beginning and the end of 
its use; however, the comment sought clarification whether companies 
would be expected to keep performing the test at regular intervals.
    (Response) For culture-based methods, it is important that each lot 
of all culture media undergo growth-promotion testing at regular 
intervals over the shelf-life of the media, not just when the media is 
initially received. The final rule requires that the sterility test and 
test components be verified, as appropriate, to demonstrate that they 
can continue to consistently detect viable contaminating 
microorganisms. The degree of verification depends upon the sterility 
test method employed.
    For culture-based test methods, studies must be conducted to 
demonstrate that the performance of the test organisms and culture 
media are suitable to consistently detect the presence of viable 
contaminating microorganisms, including tests for each lot of culture 
media to verify its growth-promoting properties over the shelf-life of 
the media and not only at the beginning and end of use. Growth-
promotion testing is important to demonstrate that the culture media 
are capable of supporting the growth of microorganisms.
    (Comment 28) One comment recommended that with the proposal to 
remove the definition of a lot of culture medium currently defined in 
Sec.  610.12(e)(2)(i), revisions to the rule should clearly state that 
each delivery of each vendor lot of media be ``QC tested'' by the end 
user to verify its ability to detect viable microorganisms. The comment 
states, ``It must be made clear that the vendor cannot be totally in 
control of the product once it has been shipped from the distribution 
centre.'' Further, the comment states it is the user's responsibility 
to test each delivery of each vendor lot to ensure that undetected 
mistreatment of the testing product during its shipment and delivery to 
the end-user has not caused deterioration in its efficacy.
    (Response) We agree that the user of the culture media must verify 
that each lot can continue to consistently detect viable contaminating 
microorganisms. For the reasons noted previously, we do not believe the 
suggested changes are needed because the rule, as proposed and now 
finalized, already reflects this requirement.
    (Comment 29) One comment stated that usually validation data 
provided by the media suppliers are used to cover the shelf-life of the 
media and proposed adding the following text ``or media supplier 
validation data must be available'' after the text ``over the shelf-
life of the media'' in proposed Sec.  610.12(e)(1) to capture the fact 
that the supplier of the media may also supply this parameter.
    (Response) We do not agree that reliance on media supplier 
validation data alone, in lieu of testing by the manufacturer, would be 
acceptable. Under Sec.  610.12(e)(1) of the final rule, for culture-
based test methods, manufacturers must conduct tests to demonstrate 
that the performance of the test organisms and culture media are 
suitable to consistently detect the presence of viable contaminating 
microorganisms, including tests for each lot of culture media to verify 
its growth-promoting properties over the shelf-life of the media. 
Therefore, reliance on media supplier validation data alone, in lieu of 
testing by the manufacturer, would not be acceptable.
6. Can a sterility test be repeated?
    For the reasons discussed in the preamble to the proposed rule (76 
FR 36019 at 36023 to 36024), we have amended the regulations in Sec.  
610.12(b) for repeat testing. Therefore, we have eliminated the 
reference to repeat testing of bulk material because, under the final 
rule, sterility testing is no longer required on bulk material in most 
instances. We also have finalized the proposal to eliminate the use of 
a second repeat test for final container material to harmonize our 
regulatory expectations with current scientific understanding of 
quality manufacturing controls.\9\ Under the final rule,

[[Page 26172]]

consistent with USP Chapter 71, if the initial test indicates the 
presence of microorganisms, then the product being examined does not 
comply with the sterility test requirements, unless a thorough 
investigation by the quality control unit can conclusively ascribe the 
initial evidence of microbial presence to a laboratory error or faulty 
materials used in conducting the test.
---------------------------------------------------------------------------

    \9\ See also Barr D., A. Celeste, R. Fish, et al., Application 
of Pharmaceutical CGMPs; FDLI (1997) at p. 146 (``In the case of a 
clearly identified laboratory error, the retest results substitute 
for the original test results. * * * If, on the other hand, no 
laboratory error could be identified in the first test, then there 
is no scientific basis for discarding the initial out-of-
specification results in favor of passing retest results.'').
---------------------------------------------------------------------------

    If the test of the initial sample is conclusively found to be 
invalid, due to laboratory error or faulty test materials, the 
sterility test may be repeated one time. If no evidence of 
microorganisms is found in the repeat test, the product examined 
complies with the test requirements for sterility. If, however, 
evidence of microorganisms is found in the repeat test, the product 
examined does not comply with the test requirements for sterility.
    Further, as discussed in the preamble to the proposed rule, both a 
comparable product that is reflective of the initial sample in terms of 
sample location and the stage in the manufacturing process from which 
it was taken, and the same sterility test method must be used for both 
the initial and repeat tests. This is intended to ensure that the same 
volume of material is used for the initial test and each repeat test, 
and that the interpretation of the results is conducted in the same 
manner.
    (Comment 30) One comment supported FDA's proposal to modify the 
provision for repeat testing to harmonize regulatory expectations with 
current scientific understanding of quality manufacturing controls by 
eliminating the use of a second repeat test of final container material 
and agreed with FDA that the proposed modification of the provision for 
repeat testing is in accordance with the USP and the European 
Pharmacopeia. However, the commenter noted that FDA's proposed 
requirement to take repeat test samples that are reflective of the 
initial samples may be difficult to fulfill. For instance, the 
commenter states, ``* * * at the time when the sterility test might 
show a positive result (after a few days), it could be that it is no 
longer possible to distinguish which vials were filled at which point 
in time.'' The comment suggested deleting the requirement in proposed 
Sec.  610.12(f)(3) that the repeat test must be conducted with 
``comparable product that is reflective of the initial sample in terms 
of sample location and the stage in the manufacturing process from 
which it was obtained.''
    (Response) We appreciate the supportive comments. However, we do 
not agree with the recommended change to Sec.  610.12(f)(3). We believe 
the final rule is consistent with current scientific understanding of 
quality manufacturing controls. If a repeat test is conducted, the same 
test method must be used for both the initial and repeat tests, and the 
repeat test must be conducted with comparable product that is 
reflective of the initial sample in terms of sample location and the 
stage in the manufacturing process from which it was obtained.
    As discussed in the preamble to the proposed rule, we appreciate 
that this final rule could result in the need for some manufacturers to 
modify their repeat test procedures. We continue to consider these 
modifications to be minor changes in accordance with Sec.  601.12(d) 
and to have a minimal potential for an adverse effect on the identity, 
strength, quality, purity, or potency of the product as they may relate 
to the safety or effectiveness of the product. Therefore, such changes 
must be reported in the annual report within 60 days of the anniversary 
date of approval of the BLA.
7. What records must be kept relating to sterility testing?
    Previously, Sec.  610.12(h) incorporated by reference the record 
keeping and maintenance requirements contained in Sec. Sec.  211.167 
and 211.194. We continue to maintain these requirements. As discussed 
in the preamble to the proposed rule (76 FR 36019 at 36024), this is 
intended to assure that data derived from sterility tests comply with 
established specifications. This includes describing the samples 
received for testing, stating the method used to test the samples, 
identifying the location of relevant validation or verification data, 
recording all calculations performed, and stating how the results of 
tests performed compare to set specifications.
8. Are there any exceptions to sterility test requirements?
    In the proposed rule we invited comments on whether any of the 
current exceptions should be removed (76 FR 36019 at 36024). We 
specifically requested comments on whether to remove the exemption for 
platelets. Bacterial contamination of platelets is a recognized public 
health risk, and the blood collection industry has already called for 
and implemented methods to detect and limit or inactivate bacteria in 
platelet components. Requiring testing for platelets would be 
consistent with these industry practices.
    (Comment 31) In response to our request for comment, a joint 
comment from industry groups recommended that FDA continue to except 
Whole Blood, Cryoprecipitated Antihemophilic Factor (AHF), Platelets, 
Red Blood Cells, and Plasma from the sterility test requirements in 
Sec.  610.12. The comment acknowledged that the blood industry has 
called for and implemented methods to detect and limit or inactivate 
bacteria in platelet components and that some culture-based methods are 
in wide use as a quality control tool. However, there are currently no 
available tests that will ensure the sterility of platelet products. In 
addition, the joint comment noted that if the current exception for 
platelets would be removed, manufacturers of blood and blood components 
would not be able to satisfy the new requirement. Further, the comment 
recommended that FDA vigorously support applications for pathogen 
inactivation processes for platelet components. Moreover, the joint 
comment noted that any sterility test requirement tied to a BLA is too 
narrow an approach to ensure optimal bacterial testing of platelet 
products, as any platelet collected or manufactured by a facility that 
does not have a BLA would not be subject to the sterility test 
regulation. Accordingly, the joint comment recommended that FDA use a 
different mechanism to require testing of all platelet products for 
bacterial contamination when testing becomes technologically feasible.
    (Response) We appreciate these comments and we generally agree. We 
recognize that blood establishments have begun to take steps to test 
for bacterial contamination in platelet components. We welcome the 
acknowledgement of the importance of bacterial testing and pathogen 
inactivation processes for platelet components and believe that 
appropriate microbial testing of platelet components may be necessary 
to assure product quality. However, while these technologies are 
developing, we have retained the exception from this rule for these 
products. Instead, we will continue to review these issues and 
available technologies and will take appropriate steps at another time 
to address microbial testing of blood components.
    (Comment 32) One comment recommended adding an exception stating 
that a manufacturer with parametric release programs is not required to 
comply with the sterility test requirements. The comment noted that 
parametric release for articles sterilized

[[Page 26173]]

with moist heat has been recognized by FDA since 1987, and that many 
companies have adopted this approach.
    (Response) We disagree with the proposed change and decline to add 
an exception for drug products terminally sterilized by moist heat 
processes and subject to parametric release because the exception under 
Sec.  610.12(h) (previously under Sec.  610.12(g)) already provides for 
an exception for such parametric release programs. As noted in FDA's 
guidance document entitled ``Guidance for Industry: Submission of 
Documentation in Applications for Parametric Release of Human and 
Veterinary Drug Products Terminally Sterilized by Moist Heat 
Processes,'' dated February 2010, FDA approval of parametric release 
must be requested either in an original application submission under 21 
CFR 314.50 or 601.2, or in a prior approval supplement under 21 CFR 
314.70 or 601.12.
    (Comment 33) Two comments recommended adding other exceptions to 
the sterility test requirements. One comment recommended adding 
granulocytes to the exception, and one comment recommended adding in 
vitro diagnostic devices regulated as biological products, which do not 
purport to be sterile.
    (Response) We decline to adopt the suggested changes because 
neither granulocytes nor in vitro diagnostic devices, which do not 
purport to be sterile, are subject to the sterility test requirements 
in Sec.  610.12. Therefore, we believe the recommendations are beyond 
the scope of this rule.
    (Comment 34) One comment recommended that the exceptions provision 
be revised to ``specifically include or exclude various biological 
product types such as Bioequivalent/Biosimilars and combination 
products.''
    (Response) We do not believe the suggested change is needed. 
Biological products must comply with the applicable requirements in 
parts 600 through 680, in addition to other applicable regulations.
    For the reasons discussed in the preamble to the proposed rule (76 
FR 36019 at 36024), we have finalized the proposed minor modifications 
to the current exception in Sec.  610.12(g)(4)(ii), under which the 
Director of CBER or CDER, as appropriate, determines that data 
submitted adequately establish that the mode of administration, the 
method of preparation, or the special nature of the product precludes 
or does not require a sterility test or that the sterility of the lot 
is not necessary to assure the safety, purity, and potency of the 
product. Specifically, the minor modification that we refer to is the 
``route of administration'' rather than the ``mode of administration'' 
and to ``any other aspect of the product'' rather than ``the special 
nature of the product'' in finalized Sec.  610.12(h)(2) so as to 
account for novel products that may be introduced to the market in the 
future. This exception allows the Director of CBER or CDER, as 
appropriate, to exempt biological material from the sterility test 
requirements of this section if, based upon the scientific evidence 
presented in the BLA or BLA supplement, the data adequately establish 
that the route of administration, method of preparation, or any other 
aspect of the product precludes or does not necessitate a sterility 
test to assure the safety, purity, and potency of the product. We note 
that in the proposed rule, the Center for Devices and Radiological 
Health was erroneously identified in this exception, instead of CDER. 
In the final rule, we have correctly identified CDER in the exception 
provision at Sec.  610.12(h)(2).
    In addition to comments regarding exceptions as stated in this 
document, we have also eliminated, as proposed, the current exceptions 
under Sec.  610.12(g)(1) and (2) because they are no longer necessary 
given the flexibility now built into the final rule. In addition, we 
have eliminated, as proposed, the current exceptions in Sec.  
610.12(g)(5) through (g)(9) because they are no longer necessary and 
because the revised rule now requires manufacturers to determine the 
appropriate sample volume and size for the material being tested and 
requires that the sterility test be ``appropriate to the material being 
tested.'' (See 76 FR 36019 at 36024 to 36025 for more information.)

IV. Revisions to Other Regulations

    In addition to the revisions to the sterility regulation in Sec.  
610.12, we have also revised, as proposed, two other FDA regulations in 
this final rule. These revisions are as follows:
     Section 600.3(q): Previously, Sec.  600.3(q) defined 
``sterility'' to mean ``freedom from viable contaminating 
microorganisms, as determined by the tests prescribed in Sec.  610.12 
of this chapter.'' As proposed, we have reworded this definition to 
eliminate the term ``prescribed'' since Sec.  610.12 no longer 
prescribes specific test methods. Thus, we have amended Sec.  600.3(q) 
to define ``sterility'' as ``freedom from viable contaminating 
microorganisms, as determined by tests conducted under Sec.  610.12 of 
this chapter.''
     Section 680.3(c) (21 CFR 680.3(c)): As proposed, we have 
amended Sec.  680.3(c) to eliminate the term ``prescribed.'' Section 
680.3(c) now states that ``A sterility test shall be performed on each 
lot of each Allergenic Product, as required by Sec.  610.12 of this 
chapter.'' Additionally, we have eliminated Sec.  680.3(c)(1) through 
(c)(4) because these exceptions are no longer necessary under the 
revisions to Sec.  610.12. (See 76 FR 36019 at 36025 for more 
information.)

V. Legal Authority

    FDA is issuing this regulation under the biological products 
provisions of the Public Health Service Act (the PHS Act) (42 U.S.C. 
262 and 264) and the drugs and general administrative provisions of the 
Federal Food, Drug, and Cosmetic Act (the FD&C Act) (sections 201, 301, 
501, 502, 503, 505, 510, 701, and 704) (21 U.S.C. 321, 331, 351, 352, 
353, 355, 360, 371, and 374). Under these provisions of the PHS Act and 
the FD&C Act, we have the authority to issue and enforce regulations 
designed to ensure that biological products are safe, effective, pure, 
and potent, and to prevent the introduction, transmission, and spread 
of communicable disease.

VI. Analysis of Impacts

    FDA has examined the impacts of the final rule under Executive 
Order 12866, Executive Order 13563, the Regulatory Flexibility Act (5 
U.S.C. 601-612), and the Unfunded Mandates Reform Act of 1996 (Pub. L. 
104-4). Executive Orders 12866 and 13563 direct Agencies to assess all 
costs and benefits of available regulatory alternatives and, when 
regulation is necessary, to select regulatory approaches that maximize 
net benefits (including potential economic, environmental, public 
health and safety, and other advantages; distributive impacts; and 
equity). The Agency believes that this final rule is not a significant 
regulatory action under Executive Order 12866.
    The Regulatory Flexibility Act requires Agencies to analyze 
regulatory options that would minimize any significant impact of a rule 
on small entities. While the rule restricts retesting when sterility 
tests are failed, the change codifies an approach for retesting that is 
similar to the approach prescribed by the USP. The rule does not 
otherwise add any new regulatory responsibilities and generally 
increases flexibility for sterility testing. Therefore, the Agency 
certifies that the final rule will not have a significant economic 
impact on a substantial number of small entities.
    Section 202(a) of the Unfunded Mandates Reform Act of 1995 requires 
that Agencies prepare a written statement, which includes an

[[Page 26174]]

assessment of anticipated costs and benefits, before proposing ``any 
rule that includes any Federal mandate that may result in the 
expenditure by State, local, and tribal governments, in the aggregate, 
or by the private sector, of $100,000,000 or more (adjusted annually 
for inflation) in any one year.'' The current threshold after 
adjustment for inflation is $136 million, using the most current (2010) 
Implicit Price Deflator for the Gross Domestic Product. FDA does not 
expect this final rule to result in any 1-year expenditure that would 
meet or exceed this amount.
    These amendments would generally provide manufacturers of 
biological products with more flexibility as to how they evaluate the 
sterility of their products and reduce the number of evaluations 
required. The net effect would be to reduce costs.
    One part of these amendments might impose some additional costs on 
manufacturers, however. Under the current regulations, if a biological 
product fails a sterility test, the test may be repeated. If the 
product passes a subsequent test, it is inferred that the first test 
was flawed and only the latter results are used. Under the new 
regulations, the test may be repeated only if it is possible to 
``ascribe definitively'' the initial failure to ``a laboratory error or 
faulty materials used in conducting the sterility testing.''
    This change could increase costs for manufacturers because 
additional products could be discarded. The size of the increase, if 
any, would be determined by the number of additional lots discarded, 
the lot sizes, and the production costs per unit. Some or all of the 
costs of this change, could, in turn, be mitigated by the reduction in 
losses associated with the provision of contaminated products.
    This change is expected to affect few manufacturers. The method for 
sterility testing described in USP Chapter 71 already limits the 
repetition of tests to circumstances similar to those described in 
these amendments. It is anticipated that, in the absence of these 
amendments, the majority of manufacturers would limit the repetition of 
sterility tests in order to comply with USP Chapter 71.
    The benefit of limiting retests would be fewer illnesses caused by 
contaminated biological products. We are unable to quantify the value 
of the reduction in illnesses because we do not have an estimate of the 
risk of illness from contaminated biological products or the decline in 
that risk associated with limiting retests.

VII. Environmental Impact

    The Agency has determined under 21 CFR 25.31(h) that this action is 
of a type that does not individually or cumulatively have a significant 
effect on the human environment. Therefore, neither an environmental 
assessment nor an environmental impact statement is required.

VIII. Federalism

    FDA has analyzed this final rule in accordance with the principles 
set forth in Executive Order 13132. FDA has determined that the rule 
does not contain policies that have substantial direct effects on the 
States, on the relationship between the National Government and the 
States, or on the distribution of power and responsibilities among the 
various levels of government. Accordingly, the Agency has concluded 
that the rule does not contain policies that have federalism 
implications as defined in the Executive order and, consequently, a 
federalism summary impact statement is not required.

IX. The Paperwork Reduction Act of 1995

    This final rule contains collections of information that were 
submitted for review and approval to the Director of the Office of 
Management and Budget (OMB), as required by section 3507(d) of the 
Paperwork Reduction Act of 1995 (44 U.S.C. 3501-3520). The collections 
of information in Sec. Sec.  211.165 and 610.12 have been approved and 
assigned OMB control number 0910-0139.

List of Subjects

21 CFR Part 600

    Biologics, Reporting and recordkeeping requirements.

21 CFR Part 610

    Biologics, Labeling, Reporting and recordkeeping requirements.

21 CFR Part 680

    Biologics, Blood, Reporting and recordkeeping requirements.

    Therefore, under the Federal Food, Drug, and Cosmetic Act, the 
Public Health Service Act, and under the authority delegated to the 
Commissioner of Food and Drugs, 21 CFR parts 600, 610, and 680 are 
amended as follows:

PART 600--BIOLOGICAL PRODUCTS: GENERAL

0
1. The authority citation for 21 CFR part 600 continues to read as 
follows:

    Authority: 21 U.S.C. 321, 351, 352, 353, 355, 360, 360i, 371, 
374; 42 U.S.C. 216, 262, 263, 263a, 264, 300aa-25.

Sec.  600.3  [Amended]

0
2. Section 600.3 is amended in paragraph (q) by removing ``prescribed 
in'' and by adding in its place the phrase ``conducted under''.

PART 610--GENERAL BIOLOGICAL PRODUCTS STANDARDS

0
3. The authority citation for 21 CFR part 610 continues to read as 
follows:

    Authority:  21 U.S.C. 321, 331, 351, 352, 353, 355, 360, 360c, 
360d, 360h, 360i, 371, 372, 374, 381; 42 U.S.C. 216, 262, 263, 263a, 
264.

0
4. Section 610.12 is revised to read as follows:

Sec.  610.12  Sterility.

    (a) The test. Except as provided in paragraph (h) of this section, 
manufacturers of biological products must perform sterility testing of 
each lot of each biological product's final container material or other 
material, as appropriate and as approved in the biologics license 
application or supplement for that product.
    (b) Test requirements. (1) The sterility test must be appropriate 
to the material being tested such that the material does not interfere 
with or otherwise hinder the test.
    (2) The sterility test must be validated to demonstrate that the 
test is capable of reliably and consistently detecting the presence of 
viable contaminating microorganisms.
    (3) The sterility test and test components must be verified to 
demonstrate that the test method can consistently detect the presence 
of viable contaminating microorganisms.
    (c) Written procedures. Manufacturers must establish, implement, 
and follow written procedures for sterility testing that describe, at a 
minimum, the following:
    (1) The sterility test method to be used;
    (i) If culture-based test methods are used, include, at a minimum:
    (A) Composition of the culture media;
    (B) Growth-promotion test requirements; and
    (C) Incubation conditions (time and temperature).
    (ii) If non-culture-based test methods are used, include, at a 
minimum:
    (A) Composition of test components;
    (B) Test parameters, including acceptance criteria; and
    (C) Controls used to verify the method's ability to detect the 
presence of viable contaminating microorganisms.
    (2) The method of sampling, including the number, volume, and size 
of articles to be tested;

[[Page 26175]]

    (3) Written specifications for the acceptance or rejection of each 
lot; and
    (4) A statement of any other function critical to the particular 
sterility test method to ensure consistent and accurate results.
    (d) The sample. The sample must be appropriate to the material 
being tested, considering, at a minimum:
    (1) The size and volume of the final product lot;
    (2) The duration of manufacturing of the drug product;
    (3) The final container configuration and size;
    (4) The quantity or concentration of inhibitors, neutralizers, and 
preservatives, if present, in the tested material;
    (5) For a culture-based test method, the volume of test material 
that results in a dilution of the product that is not bacteriostatic or 
fungistatic; and
    (6) For a non-culture-based test method, the volume of test 
material that results in a dilution of the product that does not 
inhibit or otherwise hinder the detection of viable contaminating 
microorganisms.
    (e) Verification. (1) For culture-based test methods, studies must 
be conducted to demonstrate that the performance of the test organisms 
and culture media are suitable to consistently detect the presence of 
viable contaminating microorganisms, including tests for each lot of 
culture media to verify its growth-promoting properties over the shelf-
life of the media.
    (2) For non-culture-based test methods, within the test itself, 
appropriate controls must be used to demonstrate the ability of the 
test method to continue to consistently detect the presence of viable 
contaminating microorganisms.
    (f) Repeat test procedures.--(1) If the initial test indicates the 
presence of microorganisms, the product does not comply with the 
sterility test requirements unless a thorough investigation by the 
quality control unit can ascribe definitively the microbial presence to 
a laboratory error or faulty materials used in conducting the sterility 
testing.
    (2) If the investigation described in paragraph (f)(1) of this 
section finds that the initial test indicated the presence of 
microorganisms due to laboratory error or the use of faulty materials, 
a sterility test may be repeated one time. If no evidence of 
microorganisms is found in the repeat test, the product examined 
complies with the sterility test requirements. If evidence of 
microorganisms is found in the repeat test, the product examined does 
not comply with the sterility test requirements.
    (3) If a repeat test is conducted, the same test method must be 
used for both the initial and repeat tests, and the repeat test must be 
conducted with comparable product that is reflective of the initial 
sample in terms of sample location and the stage in the manufacturing 
process from which it was obtained.
    (g) Records. The records related to the test requirements of this 
section must be prepared and maintained as required by Sec. Sec.  
211.167 and 211.194 of this chapter.
    (h) Exceptions. Sterility testing must be performed on final 
container material or other appropriate material as defined in the 
approved biologics license application or supplement and as described 
in this section, except as follows:
    (1) This section does not require sterility testing for Whole 
Blood, Cryoprecipitated Antihemophilic Factor, Platelets, Red Blood 
Cells, Plasma, Source Plasma, Smallpox Vaccine, Reagent Red Blood 
Cells, Anti-Human Globulin, and Blood Grouping Reagents.
    (2) A manufacturer is not required to comply with the sterility 
test requirements if the Director of the Center for Biologics 
Evaluation and Research or the Director of the Center for Drug 
Evaluation and Research, as appropriate, determines that data submitted 
in the biologics license application or supplement adequately establish 
that the route of administration, the method of preparation, or any 
other aspect of the product precludes or does not necessitate a 
sterility test to assure the safety, purity, and potency of the 
product.

PART 680--ADDITIONAL STANDARDS FOR MISCELLANEOUS PRODUCTS

0
5. The authority citation for 21 CFR part 680 continues to read as 
follows:

    Authority:  21 U.S.C. 321, 351, 352, 353, 355, 360, 371; 42 
U.S.C. 216, 262, 263, 263a, 264.

0
6. Section 680.3 is amended by revising paragraph (c) to read as 
follows:

Sec.  680.3  Tests.

* * * * *
    (c) Sterility. A sterility test shall be performed on each lot of 
each Allergenic Product as required by Sec.  601.12 of this chapter.

    Dated: April 27, 2012.
Leslie Kux,
Assistant Commissioner for Policy.
[FR Doc. 2012-10649 Filed 5-2-12; 8:45 am]
BILLING CODE 4160-01-P