Guidance for Industry

Analytical Procedures and Methods Validation
Chemistry, Manufacturing, and Controls Documentation

DRAFT GUIDANCE
This guidance document is being distributed for comment purposes only. Comments and suggestions regarding this draft document should be submitted within 90 days of publication in the Federal Register of the notice announcing the availability of the draft guidance. Submit comments to Dockets Management Branch (HFA-305), Food and Drug Administration, 5630 Fishers Lane, rm. 1061, Rockville, MD 20852. All comments should be identified with the docket number listed in the notice of availability that publishes in the Federal Register. For questions on the contents of this draft document contact (CDER) Radhika Rajagopalan, 301827-5849 or (CBER) Alfred Del Grosso, 301-435-4988.

U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) Center for Biologics Evaluation and Research (CBER) August 2000 CMC #

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Guidance for Industry
Analytical Procedures and Methods Validation
Chemistry, Manufacturing, and Controls Documentation
Additional copies are available from: Office of Training and Communications Division of Communications Management Drug Information Branch, HFD-210 Center for Drug Evaluation and Research (CDER) 5600 Fishers Lane Rockville, Maryland 20857 (Tel) 301-827-4573 (Internet) http://www.fda.gov/cder/guidance/index.htm or Office of Communications Training and Manufacturers Assistance, HFM-40 Center for Biologics Evaluation and Research (CBER) 1401 Rockville Pike Rockville, Maryland 20852-1448 (Fax) 888-CBERFAX or 301-827-3844 (Voice Information) 800-835-4709 or 301-827-1800 (Internet) http://www.fda.gov/cber/guidelines.htm

U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) Center for Biologics Evaluation and Research (CBER) August 2000 CMC #

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Table of Contents
I. II. III. A. B. C. IV. A. B. C. V. VI. A. B. C. D. E. F. G. H. I. J. VII. A. 1. 2. INTRODUCTION...............................................................................................................................................................1 BACKGROUND.................................................................................................................................................................2 TYPES OF ANALYTICAL PROCEDURES .............................................................................................................3 REGULATORY ANALYTICAL PROCEDURE .................................................................................................................. 3 ALTERNATIVE ANALYTICAL PROCEDURE .................................................................................................................. 3 S TABILITY-INDICATING ASSAY.................................................................................................................................... 4 REFERENCE STANDARDS ............................................................................................................................................4 TYPES OF S TANDARDS ................................................................................................................................................... 4 CERTIFICATE OF ANALYSIS ......................................................................................................................................... 4 CHARACTERIZATION OF A REFERENCE S TANDARD .................................................................................................. 4 METHODS VALIDATION FOR INDS ..........................................................................................................................6 CONTENT AND FORMAT OF ANALYTICAL PROCEDURES FOR NDAS, ANDAS, BLAS, AND PLAS ...6 PRINCIPLE....................................................................................................................................................................... 6 S AMPLING ....................................................................................................................................................................... 7 EQUIPMENT AND EQUIPMENT PARAMETERS .............................................................................................................. 7 REAGENTS ...................................................................................................................................................................... 7 S YSTEM S UITABILITY TESTING .................................................................................................................................. 7 PREPARATION OF S TANDARDS ..................................................................................................................................... 7 PREPARATION OF S AMPLES ......................................................................................................................................... 8 PROCEDURE .................................................................................................................................................................... 8 CALCULATIONS .............................................................................................................................................................. 8 REPORTING OF RESULTS.............................................................................................................................................. 8 METHODS VALIDATION FOR NDAS, ANDAS, BLAS, AND PLAS ..............................................................9 NONCOMPENDIAL ANALYTICAL PROCEDURES .......................................................................................................... 9 Validation Characteristics....................................................................................................................................9 Other Methods Validation Information ............................................................................................................10
a. Robustness ............................................................................................................................................................. 11 b. Stress Studies ......................................................................................................................................................... 11 c. Instrument Output/Raw Data ............................................................................................................................... 11

3. B. VIII. A. B. C. IX. X. A. B. C.

Recommended Validation Characteristics for Types of Tests.......................................................................13 COMPENDIAL ANALYTICAL PROCEDURES ............................................................................................................... 15 STATISTICAL ANALYSIS ......................................................................................................................................15 GENERAL ....................................................................................................................................................................... 15 COMPARATIVE S TUDIES ............................................................................................................................................. 16 S TATISTICS ................................................................................................................................................................... 16

REVALIDATION .............................................................................................................................................................16 METHODS VALIDATION PACKAGE: CONTENTS AND PROCESSING........................................................17 METHODS VALIDATION PACKAGE ............................................................................................................................ 17 S ELECTION AND S HIPMENT OF S AMPLES ................................................................................................................ 18 RESPONSIBILITIES OF THE VARIOUS PARTIES ....................................................................................................... 19

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........................ A.................29 REFERENCES ... S PECTROMETRY AND RELATED PHYSICAL METHODOLOGIES .................. F................................................................................................................ ANDA............................ 25 DISSOLUTION ...................................................................................................................... BLA...................................................................... C........28 ATTACHMENT BMETHODS VALIDATION PROBLEMS AND DELAY.............................. 20 GAS CHROMATOGRAPHY (GC) .................................. H......................................................................................................................................................... E........................ 23 OPTICAL ROTATION...... METHODOLOGY................32 J:/guidance/2396dft. 22 S PECTROPHOTOMETRY........ S PECTROSCOPY....................................................................................................................................................................... D................. 26 OTHER INSTRUMENTATION ......... 27 ATTACHMENT ANDA................................................................................................................................. B.............. AND PLA SUBMISSION CONTENTS............................30 GLOSSARY.......................................................................................................................................................................................................doc 08/29/00 ................................................................................................................. G....20 HIGH-PRESSURE LIQUID CHROMATOGRAPHY (HPLC)............................................. 24 METHODOLOGIES RELATING TO PARTICLE S IZE ANALYSIS .........................XI............................................................. 23 CAPILLARY ELECTROPHORESIS (CE) ....................................

The terms drug substance and drug product. and Controls Coordinating Committee (CMC CC) in the Center for Drug Evaluation and Research (CDER) and the Center for Biologics Evaluation and Research (CBER) at the Food and Drug Administration (FDA).doc 08/29/00 1 .gov. and presenting data to support analytical methodologies. as used in this guidance. If a different approach is This guidance has been prepared by the Analytical Methods Technical Committee of the Chemistry. to expedite FDA review of your comments. please: ! ! ! Clearly explain each issue/concern and. If possible. product license applications (PLAs). or both. use the pdf version of the document whenever possible. Identify specific comments by line numbers. when finalized. Sponsors preparing investigational new drug applications (INDs) should also consider the recommendations in this guidance. However. purity.Draft — Not for Implementation 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 Guidance for Industry 1 Analytical Procedures and Methods Validation This draft guidance. abbreviated new drug applications (ANDAs). will represent the Food and Drug Administration=s current thinking on this topic. An alternative approach may be used if such approach satisfies the requirements of the applicable statutes. The recommendations apply to drug substances and drug products covered in new drug applications (NDAs). and potency of drug substances and drug products. Manufacturing. If you plan to submit comments on this draft guidance.2 validation data. submitting samples. regulations. I. the amount and depth of the information that should be submitted to support an IND depends in large part on the phase of the investigation and the specific testing proposed in humans (see section V). e-mail an electronic copy (Word or WordPerfect) of the comments you have submitted to the docket to cunninghamp@cder. and supplements to these applications. biologics license applications (BLAs). quality.4 The principles also apply to drug substances and drug products covered in Type II drug master files (DMFs). strength. 4 J:/guidance/2396dft. when appropriate. include a proposed revision and the rationale and/or justification for the proposed change. INTRODUCTION This guidance provides recommendations to applicants on submitting analytical procedures.3 This guidance is intended to assist applicants in assembling information. It does not create or confer any rights for or on any person and does not operate to bind FDA or the public. and samples to support the documentation of the identity. refer to human drugs and 3 biologics.fda. 2 1 Analytical procedure is interchangeable with method or test procedure.

On request from FDA.doc 08/29/00 2 . intermediates. strength.Draft — Not for Implementation 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 chosen. excipients. biotechnological. For questions on appropriate validation approaches for analytical procedures or submission of information not addressed in this guidance. drug substance. will replace the FDA guidance for industry on Submitting Samples and Analytical Data for Methods Validation (February 1987). botanical. FDA investigators inspect the analytical laboratory testing sites to ensure that the analytical procedures used for release and stability J:/guidance/2396dft. when finalized. immunogenicity assessments.50(d)(1) and 314. or other immunoassays that have unique features that should be considered when submitting analytical procedure and methods validation information.94(a)(9)(i)). the specific recommendations in this guidance may not be applicable to certain unique analytical procedures for products such as biological. including bioavailability of the drug product (21 CFR 314. Data must be available to establish that the analytical procedures used in testing meet proper standards of accuracy and reliability (21 CFR 211. and blanks so that the applicant=s drug substance and drug product analytical procedures can be evaluated by FDA laboratories (21 CFR 314. The FDA laboratory analysis demonstrates that the analytical procedures are reproducible by laboratory testing. quality.94(a)(10)). container closure components. For example. BACKGROUND Each NDA and ANDA must include the analytical procedures necessary to ensure the identity. The principles of methods validation described in this guidance apply to all types of analytical procedures. purity. Methods validation is the process of demonstrating that analytical procedures are suitable for their intended use. The review chemists and laboratory analysts determine the suitability of the analytical procedures for regulatory purposes. many bioassays are based on animal challenge models.194(a)(2)).50(e) and 314. specific recommendations for biological and immunochemical tests that may be necessary for characterization and quality control of many drug substances and drug products are beyond the scope of this guidance document. Although this guidance does not specifically address the submission of analytical procedures and validation data for raw materials. validated analytical procedures should be used to analyze these materials. an NDA or ANDA applicant must submit samples of drug product. This guidance. or radiopharmaceutical drugs. the applicant is encouraged to discuss the matter in advance with the center with product jurisdiction to prevent the expenditure of resources on preparing a submission that may later be determined to be unacceptable. Furthermore. and potency of the drug substance and drug product. However. applicants should consult with the appropriate chemistry review staff at FDA. The methods validation process for analytical procedures begins with the planned and systematic collection by the applicant of the validation data to support the analytical procedures.165(e) and 211. The review chemist evaluates the analytical procedures and validation data submitted in the NDA or ANDA. noncompendial reference standards. II. and other materials used in the production of drug substances and drug products.

For BLAs. Drug..2(a) and 601. and Cosmetic Act (the Act) as the regulatory analytical procedures for compendial items. validation J:/guidance/2396dft. TYPES OF ANALYTICAL PROCEDURES A. In general. release. Information on submission of the methods validation package to the NDA or ANDA and samples to the FDA laboratories is provided in section X. PLAs. B. Alternative Analytical Procedure An alternative analytical procedure is an analytical procedure proposed by the applicant for use instead of the regulatory analytical procedure.2(a) and 601. the regulatory analytical procedure is used. validated analytical procedures should be used. Pharmacopeia/National Formulary (USP/NF) are those legally recognized under section 501(b) of the Food. If an alternative analytical procedure is submitted. and their supplements. that demonstrate that the manufactured product meets prescribed standards of safety. purity.194(a)(2)).2(c)(1)(iv)). Regulatory Analytical Procedure A regulatory analytical procedure is the analytical procedure used to evaluate a defined characteristic of the drug substance or drug product. acceptance. A validated alternative analytical procedure should be submitted only if it is shown to perform equal to or better than the regulatory analytical procedure. Representative samples of the product must be submitted and summaries of results of tests performed on the lots represented by the submitted sample must be provided (21 CFR 601. and potency (21 CFR 601. Analytical procedures and validation data are submitted in the sections of the application on analytical procedures and controls.Draft — Not for Implementation 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 testing comply with current good manufacturing practices (CGMPs) (21 CFR part 211) or good laboratory practices (GLPs) (21 CFR part 58). or stability testing. irrespective of whether they are for in-process.g. The review committee chair may request analytical testing by CBER laboratory analysts to evaluate the applicant=s analytical procedures and verify the test results. as appropriate. including analytical procedures. the applicant should provide a rationale for its inclusion and identify its use (e.doc 08/29/00 3 . For purposes of determining compliance with the Act.2(c)(1)(vi)). The analytical procedures in the U. the analytical procedures and their validation are submitted as part of the license application or supplement and are evaluated by the review committee. Each BLA and PLA must include a full description of the manufacturing methods. All analytical procedures are of equal importance from a validation perspective. Data must be available to establish that the analytical procedures used in testing meet proper standards of accuracy and reliability (21 CFR 211. III.S. Recommendations on information to be submitted are included in sections III through IX and XI of this guidance. release. Each quantitative analytical procedure should be designed to minimize assay variation. stability testing).

doc 08/29/00 4 . Assay analytical procedures for stability studies should be stability-indicating. A working standard (i. without interference from degradation products. and potency of the drug substance or the drug product. primary standard) may be obtained from the USP/NF or other official sources (e. including degradation products. strength.20).. 21 CFR 610.g. quality.Draft — Not for Implementation 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 data. the user should ensure the suitability of the reference standard. B... If there are questions on whether a source of a standard would be considered by FDA to be an official source. A reference standard that is not obtained from an official source should be of the highest purity that can be obtained by reasonable effort.e. or other potential impurities. quality. purity. and comparative data to the regulatory analytical procedure. Characterization of a Reference Standard Reference standards from USP/NF and other official sources do not require further characterization. If an applicant submits a non-stability-indicating analytical procedure for release testing. REFERENCE STANDARDS A. For standards from official sources. applicants should contact the appropriate chemistry review staff. excipients. Stability-Indicating Assay A stability-indicating assay is a validated quantitative analytical procedure that can detect the changes with time in the pertinent properties of the drug substance and drug product. in-house or secondary standard) is a standard that is qualified against and used instead of the reference standard. should complement it. strength. C. The standard should be stored correctly and used within the established use interval. and potency. A stability-indicating assay accurately measures the active ingredients. When there is no official source. purity. Analytical procedures used to J:/guidance/2396dft. those used to control the identity. CBER. and it should be thoroughly characterized to ensure its identity. Certificate of Analysis A certificate of analysis (COA) for reference standards from non-official sources should be submitted in the section of the application on analytical procedures and controls.e. process impurities. a reference standard should be of the highest possible purity and be fully characterized. IV. C. unless scientifically justified. and more extensive than. The qualitative and quantitative analytical procedures used to characterize a reference standard are expected to be different from. Types of Standards A reference standard (i. then an analytical procedure capable of qualitatively and quantitatively monitoring the impurities.

additional and/or different tests would be important to assess physicochemical characteristics. if the manufacturing process differs from that of the drug substance. However. peptide map. The data should be obtained by using appropriate tests. such as elemental analysis. amino acid composition.doc 08/29/00 5 . cell cultureJ:/guidance/2396dft. Biological and/or immunochemical activity should be assessed using the same analytical procedures used to determine product potency. and optical rotation. Legible reproductions of the relevant spectra. nuclear magnetic resonance spectroscopy (NMR). appropriate thermometric analytical procedures. structural characteristics. this characterization information should include: ! A brief description of the manufacture of the reference standard. Any additional purification procedures used in the preparation of the reference standard should be described. the recommendations on characterization information above may apply and should be considered. and others as necessary. infrared spectrophotometry (IR). high-pressure liquid chromatography (HPLC). empirical formula. Information to substantiate the proof of structure should include appropriate analytical tests. Physicochemical determinations may include isoform. Appropriate chemical attribute information. Structural characterization may include a determination of amino acid sequence. phase solubility analysis. chromatograms. dissociation constants (pK values). ! ! ! ! ! ! For biotechnological/biological product reference standards. and liquid chromatographic patterns. Data establishing purity. and carbohydrate structure. Detailed interpretation of the test data in support of the claimed structure should be provided. boiling range. as well as spectroscopic profiles. Appropriate physical constants such as melting range. refractive index. A physical description of the material. These can include animal-based. and molecular weight. A detailed description of the analytical procedures used to characterize the reference standard. electrophoretic. and mass spectrometry (MS). such as structural formula. such as TLC. and other appropriate instrumental recordings. thin-layer chromatogram (TLC) photographs or reproductions.Draft — Not for Implementation 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 characterize a reference standard should not rely solely on comparison testing to a previously designated reference standard. including its color and physical form. as well as applicable functional group analysis. biological activity. gas chromatography (GC). and/or immunochemical activity. Generally. ultraviolet spectrophotometry (UV).

Including Specified Therapeutic Biotechnology-Derived Products. ANDA.g. strength.194). METHODS VALIDATION FOR INDs For an investigational new drug. All analytical procedures should be fully developed and validation completed when the NDA. A description of analytical procedures from any other published sources should be provided. Aspects of the analytical procedure that require special attention should be described. Therapeutic. a statement indicating the analytical procedure and reference may be provided rather than a description of the method (21 CFR 211. ANDA. Chemistry. While these tests may be needed for complete characterization of certain reference standards. biochemical. or ligand/receptor-binding assays. General guidance regarding analytical procedures and methods validation information to be submitted for phase 2 or phase 3 studies will be provided in the FDA guidance for industry INDs for Phase 2 and 3 Studies of Drugs.23(a)(7)). Principle 6 J:/guidance/2396dft. VI. Including WellCharacterized. Biotechnology-Derived Products (November 1995). specific recommendations for validation of biological and immunochemical tests are not contained in this guidance document.. ANDAs. or PLA is submitted. BLA. when finalized (draft guidance published April 1999). A. The amount of information on analytical procedures and methods validation necessary will vary with the phase of the investigation (21 CFR 312. sponsors should refer to the FDA guidance for industry on Content and Format of Investigational New Drug Applications (INDs) for Phase 1 Studies of Drugs.Draft — Not for Implementation 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 based. BLA. purity. CONTENT AND FORMAT OF ANALYTICAL PROCEDURES FOR NDAs. For general guidance on analytical procedures and methods validation information to be submitted for phase 1 studies. V. If the analytical procedure used is in the current revision of the USP/NF or other FDA recognized standard references (e. or PLA should be described in sufficient detail to allow a competent analyst to reproduce the necessary conditions and obtain results comparable to the applicant=s.doc 08/29/00 . sufficient information is required in each phase of an investigation to ensure proper identification. and/or potency. Manufacturing. because the referenced sources may not be readily accessible to the reviewer. The following is a list of information that should typically be included in a description of an analytical procedure. AOAC International Book Of Methods) and the referenced analytical procedure is not modified. quality. and Controls Content and Format. AND PLAs Any analytical procedure submitted in an NDA. BLAs.

Unstable or potentially hazardous reagents should be identified. System Suitability Testing System suitability test parameters and acceptance criteria are based on the concept that the equipment. A drawing representing the experimental configuration (e. as well as a list of equipment parameters (e. not just those that involve chromatographic techniques.e. Regardless of the type of analytical procedure. USP/NF. System suitability testing ensures that the system is working properly at the time of analysis. D.g. B. For example. electronics.Draft — Not for Implementation 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 A statement of the principle of the analytical procedure should be included. If in-house or modified commercial reagents are used. and samples to be analyzed constitute an integrated system. as individual or composite samples). run time. For example. temperatures. when appropriate. titration analytical procedures should always include the evaluation of a blank (commonly referred to as a blank titration). C. instrument type. Appropriate system suitability criteria should be defined and included in the analytical procedure.. directions for their preparation should be included. separation is based on isocratic reversed phase HPLC with detection by UV. illustrating positions for a spray pattern analytical procedure) should be provided. analytical operations.g.. vials.doc 08/29/00 . American Chemical Society (ACS) Analytical Reagent) should be included. and storage conditions. testing should be used to confirm that the system will function correctly independent of the environmental conditions.g. how they are used (i. Preparation of Standards 7 J:/guidance/2396dft. E. wavelength settings).. Reagents A list of reagents and their grades (e. and the number of replicate analyses per sample should be described. directions for safe use. All chromatographic analytical procedures should include system suitability testing and criteria.. Parameters typically used in system suitability evaluations are defined and discussed in the CDER reviewer guidance on Validation of Chromatographic Methods (November 1994). and usable shelf life for these reagents should be specified. flow rate. F. tablets) selected.g. Equipment and Equipment Parameters A listing of all equipment (e. column type.. System suitability testing is recommended as a component of any analytical procedure. detector. dimensions) should be included. Sampling The number of samples (e.g..

g. The detection limit (DL) or quantitation limit (QL) should be stated. or the use of multiple order regression analyses. parts per million (ppm)) including the specific number of significant figures to be reported should be provided. derivatization).g. Reporting of organic impurities should cover (1) specified identified impurities by name.. and system suitability or start-up parameters.. retention time (RT). Reporting of Results 1. Unusual hazards should be identified. (2) specified unidentified impurities by location/identifier. The description should include. process. and specific instructions for the calculation of degradation products and impurities should be included. weight/volume. percent label claim. General The format used to report results (e. injection sampling sequence. Specific details should be provided for unusual sample preparations (e.g.Draft — Not for Implementation 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 Procedures for the preparation of all standard solutions (e.g. Calculations Representative calculations.. weight/weight. The total organic impurities for the drug product or J:/guidance/2396dft. These may include logarithmic transformations used to obtain a linear relationship from exponential data.g. with a tabulation defining all symbols and numerical factors. (3) any unspecified impurities. relative retention time (RRT)) of impurities and the type of impurity (e.doc 08/29/00 8 . The DL or QL can be set using the drug substance's detection response. 2. stock. H. and (4) total impurities. where appropriate. Procedure A step-by-step description of the procedure should be provided. Any mathematical transformations or formulas used in data analysis should be described in detail. Preparation of Samples Sample preparation for individual tests should be clearly described. G. Impurities Analytical Procedures The name and location/identifier (e.. working standard solutions. excipient degradant) should be included in the analytical procedures for impurities in the drug substance and drug product.. as appropriate. degradant. I. J. solid-phase extraction. equilibration times. internal standards) should be included.

. ANDA. or PLA should contain methods validation information to support the adequacy of the analytical procedures. 1.g. ANDA.Draft — Not for Implementation 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 drug substance is the sum of all impurities equal to or greater than their individual QL. ANDAs.194(a)(2)). For the drug product. data must be submitted to establish that the analytical procedures used in testing meet proper standards of accuracy and reliability (21 CFR 211. drug substance process impurities may be excluded from reporting if an acceptable rationale is provided in the sections on analytical procedures and controls. Analytical procedures outside the scope of the ICH guidances should still be validated. Methods validation is the process of demonstrating that analytical procedures are suitable for their intended use. but any significant process and product-related impurities should be determined and reported. AND PLAs A. biotechnological. botanical. radiopharmaceutical drugs). VII.A. packaging. Drug product impurities from the drug product manufacturing process. METHODS VALIDATION FOR NDAs. See recommendations regarding appropriate QLs in FDA impurities guidances (see references). The above reporting information may not be strictly applicable to all products (e. BLA. biological. BLAs. and labeling should be addressed. Validation Characteristics Applicants should submit information on the validation characteristics of their proposed analytical procedures (see ICH Q2A and ICH Q2B). The International Conference on Harmonisation (ICH) guidance Q2A Text on Validation of Analytical Procedures (March 1995) and Q2B Validation of Analytical Procedures: Methodology (November 1996) provide recommendations on validation of analytical procedures. BLA. Noncompendial Analytical Procedures In an NDA.3). Although not all of the validation characteristics are needed for all types of tests (see section VII. At the time of submission. the NDA. Inorganic impurities and residual solvents should also be addressed.doc 08/29/00 Accuracy Precision (repeatability and intermediate precision) Specificity Detection limit Quantitation limit 9 . typical validation characteristics are: ! ! ! ! ! J:/guidance/2396dft. or PLA.

Other impurities (e. ! ! ! ! ! C ! For drug products: J:/guidance/2396dft. Instrument output for placebo. Legible reproductions of representative instrument output or recordings (e. and sample should also be provided (see section VII.g.g.2. C A list of known impurities. with structure if available.b). residual solvents) should be addressed and quantitated..g..g.. Recommendations on submitting information on impurities is provided in various FDA guidances such as the ICH guidance Q3A Impurities in New Drug Substances (January 1996).A.. standard.2. integrated areas). Linearity Range Robustness Other Methods Validation Information Methods validation information should also include: ! Data to demonstrate the stability of all analytical sample preparations through the time required to complete the analysis. Identification and characterization of each organic impurity.A. to show how the impurities in drug substance are calculated..g. chromatograms) and raw data output (e. This information may not be needed for all products (e.Draft — Not for Implementation 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 ! ! ! 2. as appropriate.c).doc 08/29/00 10 . degradants. Impurities labeled with their names and location identifiers (e. botanicals). as appropriate. For drug substances: C A discussion of the possible formation and control of polymorphic and enantiomeric substances. RRT for chromatographic data) for the impurity analytical procedure. Representative calculations using submitted raw data. and possible isomers. inorganics. Information from stress studies (see section VII. including process impurities.

Such testing should be performed during development of the analytical procedure and the data discussed and/or submitted. Instrument output and the raw numerical values (e. In cases where an effect is observed.. representative instrument output (e.g.Draft — Not for Implementation 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 C A degradation pathway for the drug substance in the dosage form. Representative data for residual solvents are generally not needed. Areas that should be provided in more detail are described below.g. is described in ICH Q2A and Q2B. where possible. degradation information obtained from stress studies) should be submitted in the sections on analytical procedures and controls. Data demonstrating recovery from the sample matrix as illustrated by the accuracy studies. Representative instrument output (e. chromatograms) and/or other appropriate data (e. chromatograms) should be submitted.g. products of acid and base hydrolysis..g.. a. The design of the stress studies and the results should be submitted to the stability section of the application.g. Data demonstrating that neither the freshly prepared nor the degraded placebo interferes with the quantitation of the active ingredient.. thermal degradation. Stress studies are described in various FDA guidances relating to the stability of drug products (see references). peak area) J:/guidance/2396dft. Organic Impurities Representative data should be submitted to support an assessment of the organic impurities. C C ICH Q2A and Q2B address almost all of the validation parameters. Robustness Robustness. Stress Studies Degradation information obtained from stress studies (e. Instrument Output/Raw Data i.. The stress studies should demonstrate that impurities and degradants from the active ingredient and drug product excipients do not interfere with the quantitation of the active ingredient. a measure of the analytical procedure's capability to remain unaffected by small but deliberate variations. c.doc 08/29/00 11 . oxidation) for the drug substance and for the active ingredient in the drug product should be provided to demonstrate the specificity of the assay and analytical procedures for impurities. photolysis. b.

Additional information should be provided to confirm that the impurity profile is adequately characterized. in fact..Draft — Not for Implementation 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 J:/guidance/2396dft.g. appropriate information for the batches used in the biobatch or submission batch should be provided. For ANDAs and related submissions. equivalent detector response). peaks) should be labeled. appropriate information for the biobatch or submission batch should be provided. this practice may still be acceptable. such as 205 nm. 12 . integrated peak areas) of representative batches should be submitted in the sections on analytical procedures and controls for the drug substance.g. chromatograms) and raw data (e.. a representative chromatogram using detection at a low wavelength.. and date of analysis should be provided.. degradant.g. The quantitation limit and the type of organic impurity (e. For quantitation purposes. chromatograms) should be included in the stability section of the application. Acceptance criteria and analytical procedures used to estimate identified or unidentified impurities often are based on analytical assumptions (e. Assumptions should be discussed and justified. For ANDAs and related submissions. manufacturing date and site. chromatograms) and raw data (e.g. provided a correction factor is applied or the impurities are.g. Drug Substance Data should be submitted showing the separation and detection of impurities using spiked or stress samples. In cases where the response factors are not close. batch number. The analytical procedure used should be capable of differentiating changes. integrated peak areas) from representative batches under long-term and accelerated stability conditions. The impurity profile should be assessed at the quantitation limit and the instrument output provided. All responses (e.g. References to the raw data (e..doc 08/29/00 with appropriate identification and labeling (e. Drug Product Information such as instrument output (e. and double the proposed total run time could be submitted to support the specificity of the analytical procedure. iii. ii. The analytical procedure number.. the response factor of the drug substance may be used for impurities without a reference standard. being overestimated.g. chemical shift (δ) and coupling constant (J) for NMR) should be provided. process impurity) should be stated.g.. Complete impurity profiles as graphic output (e.. between past and present batches. RT for chromatographic peaks. and stressed samples should be submitted in the sections on analytical procedures and controls of the drug product.g. For example. if any.

if Raman spectroscopy is the methodology selected to quantitate polymorphic forms as impurities. RT. In addition. manufacturing site. The analytical procedures used should be capable of differentiating changes..g. J:/guidance/2396dft. if Raman spectroscopy or chiral HPLC are used for the purpose of identification or as specific tests. 3. batch number of the drug product. manufacturing date. the potential locations/identifier (e.g. and container/closure information should be provided. if any. leachables from packaging) should be reported. For example.. All responses (e. However. Multiple methodologies can be used. the recommended validation characteristics in Table 1 under quantitative testing for impurities would apply. source and batch number of drug substance. The quantitation limit and the type (e. the analytical procedure number.doc 08/29/00 13 . the submission should include instrument output and raw data for release testing and at the latest available time point for the same batch. between past and present batches. The validation information should support the intended purpose of the test. or chiral HPLC for enantiomeric impurities.Draft — Not for Implementation 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 At a minimum. Recommended Validation Characteristics for Types of Tests Table 1 is a summary of the validation characteristics that should be addressed during validation of different types of analytical procedures. date of analysis. degradant. RRT) of these compounds should be described and listed in the analytical procedure. peaks) should be labeled and identified. If process impurities from the drug substance and excipients with their related impurities are not reported in the impurities analytical procedure..g. the recommended validation characteristics listed for those types of tests would apply. The same methodology can be used for several purposes.

chloride test for a counterion) a single test is acceptable. A specific identification test should be included for the active ingredient whenever possible. For other identification tests (e.doc 08/29/00 14 . if justified. In cases where reproducibility has been performed. Recommended Validation Characteristics of the Various Types of Tests. X-ray diffraction (XRD). UV. May be needed in some cases.Draft — Not for Implementation 533 534 535 Table 1. a.g. In cases where a nonspecific identification analytical procedure is proposed for the active ingredient. two independent analytical procedures are generally sufficient. Lack of specificity for an assay for release may be compensated for by impurities testing. differential scanning calorimetry (DSC). Identification Identification analytical procedures may include tests such as IR. This concept of the number of identification tests is J:/guidance/2396dft. and HPLC retention time. Signifies that this characteristic is normally evaluated.. a chiral HPLC retention time as confirmation for the presence of an enantiomer. May not be needed in some cases. intermediate precision is not needed. Lack of specificity for an analytical procedure may be compensated for by the addition of a second analytical procedure. Type of Tests / Characteristics Identification Testing for Impurities Assay Dissolution (Measurement Only). Content/Potency Specific Tests Quantitative Accuracy Precision-Repeatability Precision-Intermediate Precision Specificity Detection Limit Quantitation Limit Linearity Range Robustness +2 + + +1 Limit + + +1 +5 + + + +4 +4 +4 +4 +4 + -3 + + + + + + -3 536 537 538 539 540 541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 NOTE: + 1 2 3 4 5 Signifies that this characteristic is not normally evaluated.

STATISTICAL ANALYSIS A. accuracy. as described in Table 1. B. b. relative standard deviation) of methods J:/guidance/2396dft.doc 08/29/00 15 . Compendial Analytical Procedures The suitability of a compendial analytical procedure must be verified under actual conditions of use (21 CFR 211.194(a)(2)). linear regression analysis. intermediate precision and robustness should be evaluated for molecular size distribution gel permeation chromatography (GPC). droplet distribution. Statistical analysis (e. c. spray pattern. preservative (if used). or drug product can include tests such as particle size analysis. and stability of the sample solution should be included. apply. These additional analytical procedures should be validated (see section VII.g. For compendial items. and this should be considered when developing the specification (see section III. The validation characteristics may differ for the various analytical procedures. Compendial assay analytical procedures may not be stability-indicating. dissolution (excludes measurement). additional analytical procedures. intermediate precision. repeatability. excipient. Assay Assay includes the content of the active ingredient. XRD.Draft — Not for Implementation 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 applicable to both the drug substance and drug product.C). Information on the specificity.A).. and methodologies such as DSC. and measurement of content in dissolution and content uniformity samples. Specific Tests Specific tests to control the drug substance. and Raman spectroscopy. regardless of which methodology is used to quantitate impurities. validation characteristics under limit testing for impurities will apply. optical rotation. such as impurities or osmolality. VIII. For example. Impurities The validation characteristics under quantitative testing for impurities. General Methods validation includes an assessment of the adequacy of the analytical procedure. d. may be requested to support the quality of the drug product or drug substance. If the same analytical procedure is proposed as a limit test. Information to demonstrate that USP/NF analytical procedures are suitable for the drug product or drug substance should be included in the submission.

appropriate literature or texts should be consulted (see references) . specificity) and to demonstrate that the analytical procedure continues to ensure the identity. as appropriate.. The statistical procedures for the analysis of the validation data should be determined prior to the start of any validation study. the new procedure should be validated (see section VII). The degree of revalidation depends on the nature of the change. If during each use an analytical procedure can meet the established system suitability requirements only with repeated adjustments to the operating conditions stated in the analytical procedure. Comparative Studies Comparative studies are performed to evaluate intermediate precision (e. as well as other general information on the interpretation and treatment of analytical data. J:/guidance/2396dft. REVALIDATION When sponsors make changes in the analytical procedure. and revalidated.g.e. reproducibility) when an analytical procedure is used in more than one laboratory or to compare and evaluate the precision and accuracy of two analytical procedures (e. if feasible. the changes may necessitate revalidation of the analytical procedures.. drug substance (e.g.Draft — Not for Implementation 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626 627 628 629 630 631 632 633 634 635 636 637 638 639 640 641 642 validation data is often used to demonstrate the validity of the method. The raw methods validation data and statistical procedures used to analyze the raw data should be provided and discussed in the sections on analytical procedures and controls.doc 08/29/00 16 . or drug product (e. When a different regulatory analytical procedure is substituted (e.g. different equipment.... including the amount of data to collect and the criteria used in determining the acceptability of the analytical procedure. days). homogeneous samples from the same batch should be used. C. quality. Revalidation should be performed to ensure that the analytical procedure maintains its characteristics (e. Statistics For information on statistical techniques used in making comparisons.g. and potency of the drug substance and drug product.g. When comparative studies are performed.. regulatory analytical procedure and an alternative analytical procedure). purity. Comparative studies are also used to evaluate between laboratory variability (i. Comparative results should be statistically analyzed and discussed and any bias explained. HPLC for titration). IX. All statistical procedures used in the analysis of the data should be based on sound principles and be suitable for evaluating the dataset. analysts. route of synthesis). should be specified.g. composition). and the bioavailability of the drug product. B. The procedure followed. amended.. strength. the analytical procedure should be reevaluated.

The methods validation package should include: 1. Individual values as well as summary tables should be provided. X. For BLAs and PLAs. one archival copy and two extra copies of the methods validation package should be submitted. a separate methods validation package need not be submitted. To aid the review chemist. The description should be sufficient to allow the FDA laboratory analysts to perform the analytical procedure (see section VI). J:/guidance/2396dft. Tabular List of All Samples to Be Submitted The list should include the lot number. 2. and quantity of the samples. Analytical Procedures A detailed description of each of the analytical procedures listed in the specifications should be submitted. the archival copy and extra copies of the methods validation packages should be submitted with the application. A. Methods Validation Package The methods validation package will usually include information copied from pertinent sections of the application.doc 08/29/00 17 .A) and samples (see X. one archival copy and three extra copies should be submitted. these copies should retain the original pagination of the application sections. 3. For ANDAs and related supplemental applications. Representative instrument output and raw data and information regarding stress studies should be included (see section VII). date of manufacture. Validation Data Appropriate validation data to support the analytical procedures should be submitted.B) will be needed for this process. A methods validation package (see X. METHODS VALIDATION PACKAGE: CONTENTS AND PROCESSING Part of the methods validation process may include FDA laboratory analysis to demonstrate that an analytical procedure is reproducible by laboratory testing. Information similar to that specified here should be included in the BLA or PLA submission. For ANDA and NDA products. identity (with chemical name and structure where required for clarity).Draft — Not for Implementation 643 644 645 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 681 682 683 684 685 FDA intends to provide guidance in the future on postapproval changes in analytical procedures. package type and size. For NDAs and related supplemental applications.

B. representative samples of the product must be submitted. and summaries of the results of tests performed on the lots represented by the submitted sample must be provided (21 CFR 601.2(c)(1)(vi)). For biological products. Specifications The specifications for the drug substance and the drug product should be included. In the case of toxic or hazardous materials. drug substance. the quantity of samples in each set should be double the amount needed to carry out the testing as performed by the applicant. an NDA or ANDA applicant must submit samples of drug product. MSDSs should be posted on the outside of the package to facilitate safe handling. samples of impurities. For CDER products. standards. Along with the drug substance and the drug product samples. For BLAs and PLAs. In general.2(a) and 601. Composition The components and composition of the drug product should be provided. 7.94(a)(10)). Material Safety Data Sheets The applicant should include material safety data sheets (MSDSs) for all samples. A set of samples will be shipped to each assigned laboratory. COAs could be submitted. Results The results obtained by the applicant for the submitted samples should be provided. As appropriate. CBER should be consulted on the submission of samples and supporting materials. the applicant should submit internal standards. Alternatively. and unusual reagents. the number of sets of samples that should be submitted for methods validation will be identified in the instructions forwarded to the applicant by the FDA laboratory. degradation products. 5. non-USP reference standards. 6.50(e) and 314. noncompendial reference standards. and blanks. so that the suitability of the applicant=s drug substance and drug product analytical procedures can be evaluated by FDA laboratories (21 CFR 314. J:/guidance/2396dft. Selection and Shipment of Samples On request from CDER.doc 08/29/00 18 . and reagents (29 CFR 1910. MSDSs should be provided for other materials used in the analytical procedures listed in the methods validation package.Draft — Not for Implementation 686 687 688 689 690 691 692 693 694 695 696 697 698 699 700 701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 717 718 719 720 721 722 723 724 725 726 727 728 4.1200(g)). The dates of analysis should be stated.

and facsimile number so that samples can be requested.g. preferably samples from an aged batch. telephone number. Applicant In the sections of the application on analytical procedures and controls. drug substances. the contact person and address listed in the NDA. Responsibilities of the Various Parties 1.Draft — Not for Implementation 729 730 731 732 733 734 735 736 737 738 739 740 741 742 743 744 745 746 747 748 749 750 751 752 753 754 755 756 757 758 759 760 761 762 763 764 765 766 767 768 769 770 771 Unless specified differently by the reviewer. Bulk substances (e. an amendment containing a copy of the batch record and certificate of analysis should be provided to the ANDA. a sample of the finished product from a batch being used to support approval of the submission should be used. The drug product should be supplied in its original packaging. samples from several consecutively manufactured batches should be submitted. When an FDA laboratory contacts the applicant for samples. or PLA submission will be used. The methods validation packages should be compiled and submitted with the NDA or ANDA submission. For BLAs and PLAs. If a sample is selected from a batch not described in the application. With the exception of sample delivery arrangements. If this information is not provided. For supplements that do not require submission and review of an exhibit batch record and associated data. For ANDAs and appropriate supplements.g. use of an inert gas blanket) are required to protect sample integrity. C. the samples should be adequately packaged in a sturdy container. J:/guidance/2396dft. or the BLA/PLA committee chair for CBER applications.. To prevent breakage during shipping. address. Samples shipped from outside the United States should contain the appropriate customs forms to reduce delay in delivery. the applicant should provide a name. samples from any batch. may be selected for NDAs and NDA supplemental applications. If special storage precautions (e. excipients) should be stored in opaque nonreactive containers.doc 08/29/00 19 . For biological products. BLA. all communications concerning validation at the FDA laboratories should be made through or with the knowledge of the review chemist for CDER applications. the container should be prominently labeled with an appropriate warning and precautionary handling instructions. the applicant should provide FDA laboratories with the samples within 10 working days. impurities. If a sample is toxic or potentially hazardous. arrangements should be made in advance with the validating laboratory for scheduled direct delivery.. The submitted drug product samples should be from a batch made with the proposed market formulation. ANDA. any commercial batch may be submitted. freezing. a separate methods validation package need not be submitted.

Column The following characteristics are useful for defining a particular column and. The laboratory will test the samples according to the submitted analytical procedures to determine whether the analytical procedures are acceptable for quality control and suitable for regulatory purposes. will be forwarded by the review chemist to the applicant on completion of the studies by the laboratories. XI. purity. 3. Investigator The investigator inspects the analytical laboratory testing sites where the release and stability testing are performed to ensure that the analytical procedures are performed in compliance with CGMP/GLP. A. Many of the following points may also apply to other chromatographic analytical procedures. will decide which analytical procedures are to be validated. Any changes in the methods resulting from the review of the application may require resubmission of the methods validation package. 4.Draft — Not for Implementation 772 773 774 775 776 777 778 779 780 781 782 783 784 785 786 787 788 789 790 791 792 793 794 795 796 797 798 799 800 801 802 803 804 805 806 807 808 809 810 811 812 813 814 2. in coordination with the appropriate FDA laboratories. FDA Laboratory An FDA laboratory will contact applicants with instructions on the submission of samples and the addresses to which samples should be mailed.doc 08/29/00 20 . if any. METHODOLOGY Sections II through IX provide general information on the submission of analytical procedures and methods validation information. Comments from the FDA laboratories. Review Chemist The review chemist will review the application to determine that the analytical procedures are adequate to ensure the identity. quality. Results and comments will be forwarded to the review chemist on completion of the studies. including validation characteristics. strength. 1. should be included in the analytical procedure description. if known. Additional information on certain methodologies is provided below. High-Pressure Liquid Chromatography (HPLC) The widespread use of HPLC analytical procedures and the multitude of commercial sources of columns and packings frequently have created problems in assessing comparability. and potency of the drug substance and/or drug product. The review chemist. If method development has J:/guidance/2396dft.

pore diameter Surface modification (e. additional silylation) Recommended pH range for column use System Suitability Testing Each analytical procedure submitted should include an appropriate number of system suitability tests defining the critical characteristics of that system. bonded surface type. the reported average may be taken from injections at the beginning and end of the run. a listing of columns found to be equivalent should be included. ! ! ! 2. plastic Dimensions: length. surface coverage. Criteria for all system suitability testing should be provided. ! ! ! ! ! b. shape.doc 08/29/00 21 . percent carbon. If the analytical procedure is used to control the level of impurities. If more than one column is suitable. stainless steel. or the two peaks J:/guidance/2396dft.Draft — Not for Implementation 815 816 817 818 819 820 821 822 823 824 825 826 827 828 829 830 831 832 833 834 835 836 837 838 839 840 841 842 843 844 845 846 847 848 849 850 851 852 853 854 855 856 857 indicated that columns from only one commercial source are suitable. If an internal standard is used. The system suitability tests listed below are defined in CDER=s reviewer guidance on Validation of Chromatographic Methods (November 1994). Column Parameters Material: glass. this information should be included as part of the analytical procedure. middle. ! ! ! ! ! ! Tailing factor Relative retention Resolution Relative standard deviation (RSD) Capacity factor Number of theoretical plates The RSD is normally performed at the beginning of the run. inner diameter Frit size Filter type Precolumn and/or guard column type. if used Packing Material Particle type: size. and end of the run. a. or at the beginning. However.g. the minimum resolution between the active ingredient and the closest eluting impurity. for assays with lengthy run times or as otherwise justified by the applicant. the minimum acceptable resolution between the internal standard and one or more active ingredients should be specified..

If more than one column is suitable.. including the order of addition of the reagents and the methods of degassing and filtration. If method development has indicated that columns from only one commercial source are suitable.doc 08/29/00 22 .. system suitability standards. such as the use of inert tubing or injection valves. ! ! ! ! 2. should be given. Operating Parameters The sequence of injection of blanks. 3. splitless. on-column) Detector Typical retention time and total run time System Suitability Testing Appropriate system suitability criteria should be defined and included in all analytical procedures. silica.g. The rationale for the use of precolumns and/or guard columns should be provided and justified. temperatures.Draft — Not for Implementation 858 859 860 861 862 863 864 865 866 867 868 869 870 871 872 873 874 875 876 877 878 879 880 881 882 883 884 885 886 887 888 889 890 891 892 893 894 895 896 897 898 899 900 eluting closest to each other.g. Flow rates. B. external diameter Stationary phase Column material (e. J:/guidance/2396dft. The effect of adjustments in mobile phase composition on retention times should be included in the analytical procedure. pressure Temperatures: column. if used) Injection (e. the following parameters should be included in the description of a GC procedure. glass. 1. and samples should be defined. other standards. Additional parameters should be specified if required by the analytical procedure. and gradients should be described. ! ! ! ! ! 3. stainless steel) Column conditioning procedure Operating Parameters Gases: purity. Any special requirements. should be specified. Gas Chromatography (GC) At a minimum. internal diameter. this information should be included as part of the analytical procedure. injector. flow rate. a listing of columns found to be equivalent should be included. Column Column dimensions: length. Complete details should be provided for the preparation of the mobile phase. split. detector (including temperature program.

atomic emission and atomic absorption. If the analytical procedure is used to control the level of impurities. Spectrometric analytical procedures may not be stability-indicating. linearity. Additional parameters may be included as required by the procedure. repeatability. this information should be included as part of the analytical procedure. If method development has indicated that capillaries from only one commercial source are suitable. Capillary Electrophoresis (CE) At a minimum. 1. UV/visible spectrophotometry (UV/Vis). or the two peaks eluting closest to each other. The RSD is normally performed at the beginning of the run. The bias of the analytical procedure should be evaluated by comparing it with a chromatographic procedure. IR spectrophotometry. near IR spectrophotometry (NIR). middle. a listing of capillaries found to be equivalent should be included. and robustness. C. external diameter Capillary material Capillary internal coating (if any) Operating Parameters 23 . the minimum resolution between the active ingredient and the closest eluting impurity. Validation criteria should include specificity (demonstrating no interference of placebo).doc 08/29/00 Capillary Capillary dimensions: length. Spectrophotometry. Appropriate system suitability and/or calibration testing is recommended. the parameters listed below should be specified for a capillary electrophoretic analytical procedure.Draft — Not for Implementation 901 902 903 904 905 906 907 908 909 910 911 912 913 914 915 916 917 918 919 920 921 922 923 924 925 926 927 928 929 930 931 932 933 934 935 936 937 938 939 940 941 942 943 If an internal standard is used. internal diameter. intermediate precision. MS. and XRD. the reported average may be taken from injections at the beginning and end of the run. Spectroscopy. but are not limited to. for assays with lengthy run times or as otherwise justified by the applicant. However. ! ! ! 2. where appropriate. Raman spectroscopy. and end of the run. the minimum acceptable resolution between the internal standard and one or more active ingredient should be specified. Spectrometry and Related Physical Methodologies These analytical procedures include. or beginning. should be given. When manually operated equipment is used. J:/guidance/2396dft. If more than one capillary is suitable. the description of the analytical procedure should include an acceptance criterion for the amount of time that may elapse between sampling and reading. D. NMR. length to detector.

E.. but has expanded to virtually any wavelength. parameters (e.. the pH to which the solution should be adjusted. In addition to the provisions of USP <781>. Optical Rotation Optical rotation is used for the measurement of stereochemical purity.e. before each run (e. time) Detector Typical migration time and total run time Model of CE equipment used Voltage (if constant voltage) Current (if constant current) Polarity (e. Other parameters may be included at the discretion of the applicant. concentration. If the analytical procedure is used to control the level of impurities. before the first run of the day. pressure. or the two peaks eluting closest to each other.g. an explanation for selecting the wavelength should be given. along with a comparison of the specific rotation at sodium D and the wavelength to be used. hydrodynamic). using the following formula as an example: J:/guidance/2396dft.Draft — Not for Implementation 944 945 946 947 948 949 950 951 952 953 954 955 956 957 958 959 960 961 962 963 964 965 966 967 968 969 970 971 972 973 974 975 976 977 978 979 980 981 982 983 984 985 986 ! ! ! ! ! ! ! ! ! 3. and. should be given. The enantiomeric purity can be expressed as enantiomeric excess (e.g. polarity of electrode by detector) System Suitability Testing Each analytical procedure should include the appropriate system suitability tests defining the critical characteristics of that system. If an internal standard is used.doc 08/29/00 24 ... If measurements are to be made at a wavelength other than sodium D. The conditions and equipment should be shown to be suitable to confirm the stereochemical identity of a racemate or an enantiomer. Circular dichroism (CD) spectra may suffice for this purpose. for aqueous solutions. electrokinetic.g. procedures for measurement of specific rotation should include the solvent. the minimum acceptable resolution between the internal standard and one or more active ingredient should be specified.g. the minimum resolution between the active ingredient and the closest eluting impurity. including a detailed preparation procedure with the order of addition of the components Injection: mode (e. flush with 100 millimolar sodium hydroxide. Visual polarimeters rely on a monochromatic source. voltage. which traditionally was sodium D. flush with running buffer) Running buffer: composition.). Capillary preparation procedure: procedure to be followed before the first use.

Draft — Not for Implementation 987 988 989 990 991 992 993 994 995 996 997 998 999 1000 1001 1002 1003 1004 1005 1006 1007 1008 1009 1010 1011 1012 1013 1014 1015 1016 1017 1018 1019 1020 1021 1022 1023 1024 1025 1026 1027 1028 1029 e.doc 08/29/00 25 . and intermediate precision. but are not limited to: a. Validation criteria should include specificity. 1. F. for example. J:/guidance/2396dft.[m]}/{[M] + [m]} where [M] and [m] are the concentrations of the major and minor enantiomers. morphology.e. respectively. = 100% * {{M} . Particle size evaluation can include characteristics of size. of 94 percent. However.e. The normal concepts of validation may differ for particle size methodologies as compared to other analytical methodologies such as HPLC. An intermediate concentration gives intermediate values. Appropriate system suitability and/or calibration testing is recommended. photon correlation. laser diffraction) Electrozone sensing Photozone sensing Fractionation methods that use physical techniques to separate particles on the basis of size Sieving Cascade impactor Sedimentation Size exclusion chromatography Calibration and Validation Characteristics ! ! ! ! 2. 97:3 would give an e. and population of particles. This yields values of zero for a racemate and 100 percent for a pure enantiomer. surface. Nonfractionation methods that evaluate an entire population of particles Microscopy (optical. Methodologies Relating to Particle Size Analysis Particle size analysis is an important element for quality control and regulatory evaluation of certain drug substances and drug products. ! ! ! ! b. a standard mixture may be used for calibration. Particle Size Methods Types of particle size methods include. electron) Light scattering (dynamic. The following parameters are useful for describing particle size analysis for characterization of drug substances and drug products.

Blank and standard solution spectra or chromatograms should be included.g. J:/guidance/2396dft.g. the system should be calibrated according to the manufacturer's and/or the laboratory's specification. Data should be submitted to support the stability of the dissolution sample during the procedure. Assurance should be provided that the data generated are reproducible and control the product's quality. handling of samples. Validation Characteristics Both the dissolution procedure and the method of analysis should be validated. G. as appropriate.. Procedure A dissolution test consists of a dissolution procedure and method of analysis (automated on-line analysis or manual sampling followed by HPLC analysis). filtration. HPLC) Sampling procedure (e.Draft — Not for Implementation 1030 1031 1032 1033 1034 1035 1036 1037 1038 1039 1040 1041 1042 1043 1044 1045 1046 1047 1048 1049 1050 1051 1052 1053 1054 1055 1056 1057 1058 1059 1060 1061 1062 1063 1064 1065 1066 1067 1068 1069 1070 1071 1072 To ensure proper instrument operation. The time needed for the completion of the sample analysis should be stated in the procedure. dilutions) Calculations Acceptance criteria Regardless of the method of analysis. system suitability criteria should be described. 3. See additional information in sections V and VII. The methods validation usually involves evaluation of intermediate precision and robustness. The written procedure should cover the following items: ! ! ! ! ! ! ! Apparatus Preparation of standard Preparation of sample Method of analysis (e. 1.. If filters are used on-line or during sample preparation. Dissolution The equipment used for dissolution is covered by USP <711> or USP <724>. 2. The dissolution procedure description and validation should include the following.doc 08/29/00 26 . UV. Dissolution Medium A brief discussion of the reasons for selecting the medium. intervals. appropriate recovery studies should be performed and documented and any bias should be addressed.

complete performance assessment procedures should be provided. the applicant's cooperation with the FDA laboratories in helping facilitate duplication of the analytical procedure is important. the instrumentation should be capable of being constructed from commercially available components at a reasonable cost. 2. Other Instrumentation 1. When noncommercial instrumentation is used. the use of rare or exotic systems not only places an undue burden on the regulatory laboratory. However.Draft — Not for Implementation 1073 1074 1075 1076 1077 1078 1079 1080 1081 1082 1083 1084 1085 1086 1087 1088 1089 1090 1091 1092 1093 1094 1095 H. Noncommercial Instrumentation FDA encourages the development and use of the most appropriate instrumentation. To avoid this delay.doc 08/29/00 27 . may lead to delay in regulatory methods validation because FDA laboratories have to assemble and validate the system before running samples. In addition to design and equipment specifications. if possible. but also may delay the validation process. J:/guidance/2396dft. For unique methodologies or instrumentation requiring contract fabrication. applicants should demonstrate the equivalence of a manual procedure to the automated procedure based on the same principle whenever possible. Such systems may be found suitable for regulatory use. although desirable for control testing. Automated Analytical Procedures The use of automated analytical procedures.

a separate methods validation package need not be submitted. and PLAs is identified below with a cross-reference to the section of this guidance that provides recommendations and/or discussion on the topics.c Representative instrument output and raw data for initial and oldest sample of a batch Section VII.Draft — Not for Implementation 1096 1097 1098 1099 1100 1101 1102 1103 1104 1105 1106 1107 1108 1109 1110 1111 1112 1113 1114 1115 1116 1117 1118 1119 1120 1121 1122 1123 1124 1125 ATTACHMENT A NDA.A.A. 5 J:/guidance/2396.2.2. ANDA.b Information that should be included in the stability section ! ! Stress study designs and results Reference (volume and page number of submission) to instrument output and raw data submitted to the section dedicated to analytical procedures and controls Section VII.2.b Section VIII Section IX • • • • • • Information that should be included in the methods validation package5 • • ! Contents of the MV Package Section XI Representative instrument output/data for stress studies Section VII. BLA.A 2.dft.A.2. BLAs. as needed Section IV Section III. VI Section VII Section VII. Information that should be included in the analytical procedures and controls sections ! Reference standard information Analytical procedures Validation data Stress studies Instrument output/raw data for impurities Statistical analysis Revalidation.b Section VII.c Section VII. Information similar to what is listed here should be included in the BLA or PLA submission. ANDAs.2.A.A.wpd 08/29/00 28 . AND PLA SUBMISSION CONTENTS The information relating to analytical procedures and methods validation that should be submitted in NDAs.c For BLAs and PLAs.

. For example: C C C Use of arbitrary arithmetic corrections Failure to provide system suitability tests Differing content uniformity and assay analytical procedures without showing equivalence factors for defining corrections as required by the current USP chapter <905> . For example: C C Failure to label instrument output to indicate sample identity Failure to label the axes ! Inappropriate shipping procedures. or novel reagent Failure to submit well-characterized reference standards for noncompendial drugs Failure to provide sufficient detail or use of unacceptable analytical procedures. For example: C C C Failure to properly label samples Failure to package samples in accordance with product storage conditions Inadequate shipping forms (e.Draft — Not for Implementation 1126 1127 1128 1129 1130 1131 1132 1133 1134 1135 1136 1137 1138 1139 1140 1141 1142 1143 1144 1145 1146 1147 1148 1149 1150 1151 1152 1153 1154 1155 1156 1157 1158 ATTACHMENT B METHODS VALIDATION PROBLEMS AND DELAY Listed below are examples of common problems that can delay successful validation. internal standard. missing customs form for samples from outside the United States) ! Failure to describe proper storage conditions on shipping containers J:/guidance/2396.g.wpd 08/29/00 29 . ! Failure to provide a sample of a critical impurity.dft. degradation product.Uniformity of Dosage Units ! ! ! Failure to submit complete or legible data.

6 J:/guidance/2396. September 1998). June 1998). December 1998). Guidance for Industry: ANDAs: Impurities in Drug Substances (February 2000). December 1997). Biotechnologyderived Products (February 1995).1 Requesting Methods Validation for ANDAs (November 1998). and Controls Information for Synthetic Peptide Substances (November 1994). Manufacturing. As draft documents. Guidance for Industry: Submitting Documentation for the Stability of Human Drugs and Biologics (February 1987).wpd 08/29/00 30 . Reviewer Guidance: Validation of Chromatographic Methods (November 1994). Guidance for Industry: Investigating Out of Specification (OOS) Test Results for Pharmaceutical Production (Draft. Guidance for Industry: Submission of Chemistry.Draft — Not for Implementation 1159 1160 1161 1162 1163 1164 1165 1166 1167 1168 1169 1170 1171 1172 1173 1174 1175 1176 1177 1178 1179 1180 1181 1182 1183 1184 1185 1186 1187 1188 1189 1190 1191 1192 1193 1194 1195 1196 1197 REFERENCES FDA Documents 6 Guidance for Industry: ANDAs: Impurities in Drug Products (Draft. Guidance for Industry: Stability Testing of Drug Substances and Drug Products (Draft. Guidance for Industry: CMC Content and Format of INDs for Phase 2 and 3 Studies of Drugs. Including Well-Characterized. Therapeutic. Including Specified Therapeutic Biotechnology-Derived Products (Draft.dft. International Conference on Harmonization Guidances ICH Q1A: Stability Testing of New Drug Substances and Products (November 1994) ICH Q1B: Photostability Testing of New Drug Substances and Products (November 1996) ICH Q1C: Stability Testing for New Dosage Forms (May 1997) Draft guidances have been included for completeness only. Guidance for Industry: Content and Format of Investigational New Drug Applications (INDs) for Phase 1 Studies of Drugs. they are not intended to be implemented until published in final form. FDA CDER MAPP 5221.

Inc. United States Pharmacopeial Convention. United States Pharmacopeial Convention. US Pharmacopeia 23.dft. 3rd edition. Volume 24.C. United States Pharmacopeial Convention.. Rockville MD: 1994 Interpretation and Treatment of Analytical Data. US Pharmacopea 23. MD: 1994 Chapter <1225> Validation of Compendial Methods. USP Pharmacopeial Forum.. and E.. J:/guidance/2396.Draft — Not for Implementation 1198 1199 1200 1201 1202 1203 1204 1205 1206 1207 1208 1209 1210 1211 1212 1213 1214 1215 1216 1217 1218 1219 1220 1221 1222 1223 1224 1225 1226 1227 1228 1229 1230 1231 1232 1233 1234 1235 1236 1237 ICH Q2A: Text on Validation of Analytical Procedures (March 1995) ICH Q2B: Validation of Analytical Procedures: Methodology (May 1997) ICH Q3A: Impurities in New Drug Substances (January 1996) ICH Q3B: Impurities in New Drug Products (May 1997) ICH Q3C: Impurities: Residual Solvents (December 1997) ICH Q5C: Quality of Biotechnological Products: Stability Testing of Biotechnological/Biological Products (July 1996) ICH Q6A: Specifications: Test Procedures and Acceptance Criteria for New Drug Substances and New Drug Products: Chemical Substances (Draft (Step 2) November 1997) ICH Q6B: Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products (March 1999) U. Pharmacopeia/National Formulary Chapter <621> Chromatography. B. 7051 . Basic and Clinical Biostatistics. Prentice Hall. Number 5. Rockville MD: 1994.wpd 08/29/00 31 .. Trapp. Miller.D..N. Inc. 1994. Inc. Saunders. J. 2nd edition. J.S. Statistics for Analytical Chemistry. 1993. Horwood. Appleton and Lange. US Pharmacopeia 23.7056 Other Miller. Rockville MD: 1994 Chapter <781> Optical Rotation. pp.G. United States Pharmacopeial Convention. and R. Rockville.. Inc.

and other components including container closure systems. Reagent: For analytical procedures.108(a)). Specification: The quality standards (i. responsible for the physiological or pharmacological action of the drug substance (21 CFR 314. tests. measuring. The term includes those components that may undergo chemical change in the manufacture of the drug product and be present in the drug product in a modified form intended to furnish the specified activity or effect (21 CFR 210. Quantitation Limit: The quantitation limit of an individual analytical procedure is the lowest amount of analyte in a sample that can be quantitatively determined with suitable precision and accuracy. The active ingredient does not include intermediates used in the synthesis of such ingredient. treatment.dft. mitigation. chelate. but not necessarily quantitated as an exact value. any substance used in a reaction for the purpose of detecting. Placebo (or Blank): A dosage form that is identical to the drug product except that the drug substance is absent or replaced by an inert ingredient or a mixture of the drug product excipients quantitatively equivalent to those found in the drug product dosage form. cure. and in-process materials. analytical procedures.3(b)).e. capsule. but not necessarily. and is used particularly for the determination of impurities and/or degradation products. or clathrate) of the molecule. excluding those appended portions of the molecule that cause the drug to be an ester. not the active site. Detection Limit: The detection limit of an individual analytical procedure is the lowest amount of analyte in a sample that can be detected..3(b)(7) and 314.Draft — Not for Implementation 1238 1239 1240 1241 1242 1243 1244 1245 1246 1247 1248 1249 1250 1251 1252 1253 1254 1255 1256 1257 1258 1259 1260 1261 1262 1263 1264 1265 1266 1267 1268 1269 1270 1271 1272 1273 1274 1275 1276 1277 1278 1279 1280 GLOSSARY Acceptance Criteria: Numerical limits. or solution that contains a drug substance. J:/guidance/2396. salt (including a salt with hydrogen or coordination bonds). or other noncovalent derivative (such as a complex.3(b)). The active moiety is the entire molecule or ion. raw materials. intermediates. ranges. The quantitation limit is a parameter of quantitative assays for low levels of compounds in sample matrices. generally. drug products. reagents. in association with one or more other ingredients (21 CFR 314. and acceptance criteria) provided in an approved application to confirm the quality of the drug substances. examining. for example. Drug Product: A finished dosage form. Active moiety: The molecule or ion. Drug Substance/Active Ingredient: An active ingredient that is intended to furnish pharmacological activity or other direct effect in the diagnosis. or other suitable measures for acceptance of the results of analytical procedures. or analyzing other substances. a tablet.wpd 08/29/00 32 . or prevention of disease or to affect the structure or any function of the human body.

process impurities.dft. typically for the purpose of confirming the performance of an analytical procedure or the calibration of an instrument. Stability-Indicating Assay: A validated quantitative analytical procedure that can detect the changes with time in the pertinent properties (e.wpd 08/29/00 33 . sample. Working Standard: A standard that is qualified against and used instead of the reference standard (also known as in-house or secondary standard). active ingredient.g. or placebo.. excipients. A stability-indicating assay accurately measures the active ingredients without interference from degradation products. preservative level) of the drug substance and drug product.Draft — Not for Implementation 1281 1282 1283 1284 1285 1286 1287 1288 1289 1290 1291 1292 1293 1294 1295 1296 1297 Spiking: The addition of a small known amount of a known compound to a standard. or other potential impurities. J:/guidance/2396.

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