美国药典35版USP 1226 VERIFICATION OF COMPENDIAL PROCEDURES

882?1226? Verification of Compendial Procedures / General Information USP 35

terial to which the procedure is applied. Although complete ?1226? VERIFICATION OF revalidation of a compendial method is not required to ver-ify the suitability of v a procedure v USP35 under actual condi-COMPENDIAL PROCEDURES

tions of use, some of the analytical performance characteris-tics listed in chapter ?1225?, Table 2, may be used for the verification process. Only those characteristics that are con-sidered to be appropriate for the verification of the particu-The intent of this general information chapter is to pro-lar v procedure v USP35 need to be evaluated. v The process of vide general information on the verification of compendial assessing the suitability of a compendial analytical test pro-procedures that are being performed for the first time to cedure under the conditions of actual use may or may not yield acceptable results utilizing the personnel, equipment,require actual laboratory performance of each analytical per-and reagents available. This chapter is not intended for ret-formance characteristic.v USP35 The degree and extent of the roactive application to already successfully established labo-verification process may depend on the level of training and ratory procedures. The chapter Validation of Compendial Pro-experience of the user, on the type of procedure and its cedures ?1225? provides general information on

associated equipment or instrumentation, on the specific characteristics that should be considered for various test cat-procedural steps, and on which article(s) are being tested.egories and on the documentation that should accompany v Verification should assess whether the compendial proce-analytical procedures submitted for inclusion in USP–NF. Ver-dure is suitable for the drug substance and/or the drug ification consists of assessing selected analytical performance product matrix, taking into account the drug substance’s characteristics, such as those that are described in chapter synthetic route, the method of manufacture for the drug ?1225?, to generate appropriate, relevant data rather than product, or both, if applicable. Verification should include repeating the validation process.

an assessment of elements such as the effect of the matrix Users of compendial analytical procedures are not re-on the recovery of impurities and drug substances from the quired to validate these procedures when first used in their drug product matrix, as well as the suitability of chromato-laboratories, but documented evidence of suitability should graphic conditions and column, the appropriateness of de-be established under actual conditions of use. In the United tector signal response, etc.v USP35

States, this requirement is established in 21 CFR

As an example, an assessment of specificity is a key pa-211.194(a)(2) of the current Good Manufacturing Practice rameter in verifying that a compendial procedure is suitable regulations, which states that the “suitability of all testing for use in assaying drug substances and drug products. For methods used shall be verified under actual conditions of instance, acceptable specificity for a chromatographic

use.”

method may be verified by conformance with system suita-Verification of microbiological procedures is not covered bility resolution requirements (if specified in the in this chapter because it is covered in USP general test v procedure).v USP35 However, drug substances from different chapters Antimicrobial Effectiveness Testing ?51?, Microbiologi-suppliers may have different impurity profiles that are not cal Examination of Nonsterile Products: Microbial Enumeration addressed by the compendial test procedure. Similarly, the Tests ?61?, Microbiological Examination of Nonsterile Products:excipients in a drug product can vary widely among manu-Tests for Specified Microorganisms ?62?, Sterility Tests ?71?, and facturers and may have the potential to directly interfere in general information chapter Validation of Microbial Recov-with the procedure or cause the formation of impurities that ery from Pharmacopeial Articles ?1227?.

are not addressed by the compendial procedure. In addi-tion, drug products containing different excipients, antioxi-Change to read:

dants, buffers, or container extractives v may affect the re-covery of the drug substance from the matrix.v USP35 In these cases, a more thorough assessment of v the matrix

effects v USP35 may be required to demonstrate suitability of VERIFICATION PROCESS

the v procedure v USP35 for the particular drug substance or product. Other analytical performance characteristics such as v

The verification process for compendial test procedures is an assessment of the limit of detection or quantitation and the assessment of whether the procedure can be used for its precision for impurities procedures may be useful to demon-intended purpose, under the actual conditions of use for a strate the suitability of the compendial v procedure v USP35specified drug substance and/or drug product matrix.v USP35

under actual conditions of use.

Users should have the appropriate experience, knowledge,Verification is not required for basic compendial test pro-and training to understand and be able to perform the cedures that are routinely performed unless there is an indi-compendial procedures as written. Verification should be cation that the compendial procedure is not appropriate for conducted by the user such that the results will provide

the article under test. Examples of basic compendial proce-confidence that the compendial procedure will perform suit-dures include, but are not limited to, loss on drying, residue ably as intended.

on ignition, various wet chemical procedures such as acid If the verification of the compendial procedure is not suc-value, and simple instrumental v determinations v USP35 such as cessful, and assistance from USP staff has not resolved the pH measurements. However, for the application of already problem, it may be concluded that the procedure may not established routine procedures to compendial articles tested be suitable for use with the article being tested in that labo-for the first time, it is recommended that consideration be ratory. It may then be necessary to develop and validate an given to any new or different sample handling or solution alternate procedure as allowed in the General Notices . The preparation requirements.

alternate procedure may be submitted to USP, along with the appropriate data, to support a proposal for inclusion or replacement of the current compendial procedure.

Change to read:

VERIFICATION REQUIREMENTS

Verification requirements should be based on an assess-ment of the complexity of both the procedure and the ma-

USP 35General Information / ?1227? Validation of Microbial Recovery883

culture preparation. The conditions of organism preparation ?1227? VALIDATION OF and storage must be standardized for the neutralizer evalua-

tion and should reflect the conditions of the antimicrobial MICROBIAL RECOVERY FROM assay.

The specific conditions of the test, including buffers used, PHARMACOPEIAL ARTICLES water, light conditions, and temperature, must be repro-

duced in the validation study. All test conditions also should

be standardized and performed in the validation study ex-

actly as performed in the test.

This chapter provides guidelines for the validation of The conditions of microbial recovery are among the most methods for the estimation of the number of viable micro-crucial in accurately estimating the number of microorgan-organisms, for the detection of indicators or objectionable isms present in a test solution. The first consideration is the microorganisms, for the validation of microbiological meth-recovery medium used to support the growth of survivors. ods used in antimicrobial effectiveness testing, and for the This concern is discussed in detail below. The second con-sterility testing of Pharmacopeial articles. It is generally un-sideration is the incubation conditions. Optimal conditions derstood that if a product possesses antimicrobial properties for growth must be present to ensure complete growth and because of the presence of a specific preservative or because reproducible results.

of its formulation, this antimicrobial property must be neu-

tralized to recover viable microorganisms. This neutralization

may be achieved by the use of a specific neutralizer, by METHODS OF NEUTRALIZING dilution, by a combination of washing and dilution, or by ANTIMICROBIAL PROPERTIES

any combination of these methods.

The tests under Antimicrobial Effectiveness Testing ?51?, Ste-Three common methods are used to neutralize antimicro-rility Tests ?71?, and Microbial Enumeration Tests ?61? and bial properties of a product: (1) chemical inhibition, (2) dilu-Tests for Specified Microorganisms ?62? require the validation tion, and (3) filtration and washing.

of recovery methods. To ensure that the results of the tests

are credible, neutralization of antimicrobial properties of the

test solution is required before estimating the number of Chemical Inhibition

viable microorganisms.

Table 1 shows known neutralizers for a variety of chemical

antimicrobial agents and the reported toxicity of some INFLUENTIAL FACTORS

chemical neutralizers to specific microorganisms. However,

despite potential toxicity, the convenience and quick action Several factors affect the measurement of a test solution’s of chemical inhibitors encourage their use. Chemical inhibi-

antimicrobial activity, and these must be considered in the tion of bactericides is the preferred method for the antimi-validation design. They include the nature of the microor-crobial efficacy test. The potential of chemical inhibitors ganisms used as challenge organisms, the preparation of the should be considered in the membrane filtration and the inoculum of challenge organisms, the specific conditions of direct transfer sterility tests. Antibiotics may not be suscepti-the test, and the conditions of recovery. These factors also ble to neutralization by chemical means, but rather by enzy-affect the validation of recovery methods for aqueous or matic treatment (e.g., penicillinase). These enzymes may be nonaqueous products, irrespective of their antimicrobial used where required.

properties; thus, all test methods should be validated with

these factors in mind.

The nature of the challenge microorganism exerts a Dilution

strong effect upon the response to the antimicrobial agent,

and so upon the neutralization required for recovery. Repre- A second approach to neutralizing antimicrobial proper-sented among these organisms in compendial tests are ties of a product is by dilution, because the concentration of Gram-positive bacteria, Gram-negative bacteria, yeasts, and a chemical bactericide exerts a large effect on its potency. molds. Each organism to be used in the test must be in-The relationship between concentration and antimicrobial cluded in the validation.effect differs among bactericidal agents but is constant for a The preparation of the inoculum of challenge microorgan-particular antimicrobial agent. This relationship is exponen-isms also affects the testing of products having antimicrobial tial in nature, with the general formula:

properties. The growth and preparation of the challenge or-

ganism determines the physiological state of the cell. This Cηt = k

state has a direct influence on the results of any test of

antimicrobial efficacy. Microbial tests do not use individual in which C is the concentration; t is the time required to kill cells; rather, populations of cells are harvested for study. The a standard inoculum; k is a constant; and the concentration data generated from these studies are less variable if the cell exponent, η, is the slope of the plot of log t versus log C. populations are homogeneous. Liquid cultures or confluent Antimicrobial agents with high η values are rapidly neutral-growths on solid medium are best suited for reproducible

Table 1. Some Common Neutralizers for Chemical Biocides

Neutralizer Biocide Class Potential Action of Biocides

Bisulfate Glutaraldehyde, Mercurials Non-Sporing Bacteria

Dilution Phenolics, Alcohol, Aldehydes, Sorbate —

Glycine Aldehydes Growing Cells

Lecithin Quaternary Ammonium Compounds (QACs),Bacteria

Parabens, Bis-biguanides

Mg+2 or Ca+2 ions EDTA—

Polysorbate QACS, Iodine, Parabens—

Thioglycollate Mercurials Staphylococci and Spores

Thiosulfate Mercurials, Halogens, Aldehydes Staphylococci