Process Validation: General Principles and Practices

TABLE OF CONTENTS
I.
INTRODUCTION
1
II.
BACKGROUND
2
III.
STATUTORY AND REGULATORY REQUIREMENTS FOR PROCESS
VALIDATION
4
IV.
RECOMMENDATIONS
6
A.
General Considerations for Process Validation
6
B.
Specific Stages and Activities of Process Validation in the Product Lifecycle
6
1.
Stage 1 – Process Design
7
2.
Stage 2 – Process Qualification
9
3.
Stage 3 – Continued Process Verification
13
V. CONCURRENT RELEASE OF PERFORMANCE QUALIFICATION BATCHES
14
VI.
DOCUMENTATION
15
VII.
ANALYTICAL METHODOLOGY
16
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1 Guidance for Industry1
2
3 Process Validation: General Principles and Practices
4
5
6
This draft guidance, when finalized, will represent the Food and Drug Administration's (FDA's) current
7
thinking on this topic. It does not create or confer any rights for or on any person and does not operate to 8
bind FDA or the public. You can use an alternative approach if the approach satisfies the requirements of 9
the applicable statutes and regulations. If you want to discuss an alternative approach, contact the FDA 10
staff responsible for implementing this guidance. If you cannot identify the appropriate FDA staff, call 11
the appropriate number listed on the title page of this guidance. 12
13 14 15 16 17 18 I. INTRODUCTION 19 20 This guidance outlines the general principles and approaches that FDA considers to be 21 appropriate elements of process validation for the manufacture of human and animal drug and 22 biological products, including active pharmaceutical ingredients (API or drug substance), 23 collectively referred to in this guidance as drugs or products. This guidance incorporates 24 principles and approaches that all manufacturers can use in validating a manufacturing process. 25 26 This guidance aligns process validation activities with the product lifecycle concept and with 27 existing FDA guidance.2 The lifecycle concept links product and process development, 28 qualification of the commercial manufacturing process, and maintenance of the process in a state 29 of control during routine commercial production. This guidance promotes modern 30 manufacturing principles, process improvement, innovation, and sound science. 31
1 This guidance has been prepared by the Division of Manufacturing and Product Quality, Center for Drug Evaluation and Research (CDER), in cooperation with the Center for Biologics Evaluation and Research (CBER) and the Center for Veterinary Medicine (CVM) at the Food and Drug Administration. FDA’s Office of Regulatory Affairs (ORA) also contributed significantly to the development of this guidance.
2 See the FDA/International Conference on Harmonisation (ICH) guidances for industry: Q8 Pharmaceutical Development, Q9 Quality Risk Management, and when finalized, Q10 Pharmaceutical Quality System (a notice of availability for the May 2007 ICH draft guidance, Q10 Pharmaceutical Quality System, published in the Federal Register on July 13, 2007 (72 FR 38604)). We update guidance documents periodically. To make sure you have the most recent version of a guidance, check the CDER guidance page at http://www.fda.gov/cder/guidance/index.htm, the CBER guidance page at http://www.fda.gov/cber/guidelines.htm, or the CVM guidance page at http://www.fda.gov/cvm/Guidance/published.htm.
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32 The following categories of drugs are within the scope of this guidance: 33 34 • Human drugs 35 • Veterinary drugs 36 • Biological and biotechnology products 37 • Finished products and active pharmaceutical ingredients (API or drug substance)3 38 • The drug constituent of a combination (drug and medical device) product 39 40 The following categories of products are not covered by this guidance: 41 42 • Type A medicated articles and medicated feed 43 • Medical devices 44 • Dietary supplements 45 • Human tissues intended for transplantation regulated under section 361 of the Public Health 46 Service Act4 47 48 This guidance does not specify what information should be included as part of a regulatory submission. 49 Interested persons can refer to the appropriate guidance or contact the appropriate Center in determining 50 what information should be included in a submission. 51 52 This guidance also does not specifically discuss the validation of automated process control systems 53 (i.e., computer hardware and software interfaces), which are commonly integrated into modern drug 54 manufacturing equipment. This guidance is relevant, however, to the validation of processes that 55 include automated equipment in processing. 56 57 FDA's guidance documents, including this guidance, do not establish legally enforceable 58 responsibilities. Instead, guidances describe the Agency's current thinking on a topic and should 59 be viewed only as recommendations, unless specific regulatory or statutory requirements are 60 cited. The use of the word should in Agency guidances means that something is suggested or 61 recommended, but not required. 62 63 II. BACKGROUND 64 65 In the Federal Register of May 11, 1987 (52 FR 17638), FDA issued a notice announcing the 66 availability of a guidance entitled Guideline on General Principles of Process Validation (the
3 Separate current good manufacturing practice (CGMP) regulations for drug components such as APIs (drug substances) and intermediates have not published as of the date of this guidance, but these components are subject to the statutory CGMP requirements of section 501(a)(2)(B) of the Federal Food, Drug, and Cosmetic Act (the Act) (21 U.S.C. 351(a)(2)(B)). Process validation for APIs is discussed in the FDA/ICH guidance for industry, Q7A Good Manufacturing Practice Guidance for Active Pharmaceutical Ingredients (ICH Q7A), available on the Internet at http://www.fda.gov/cder/guidance/index.htm. Section XII of ICH Q7A describes in detail the principles to be followed in validating API processes.
4 See the FDA guidance for industry, Validation of Procedures for Processing of Human Tissues Intended for
Transplantation, available on the Internet at http://www.fda.gov/cber/guidelines.htm.
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67 1987 guidance).5 Since then, we have obtained additional experience through our regulatory 68 oversight that allows us to update our recommendations to industry on this topic. This revised 69 guidance conveys FDA’s current thinking on process validation and is consistent with basic 70 principles first introduced in the 1987 guidance. This guidance also provides recommendations 71 that reflect some of the goals of FDA’s initiative entitled “Pharmaceutical CGMPs for the 21st 72 Century – A Risk-Based Approach,” particularly with regard to the use of technological 73 advances in pharmaceutical manufacturing, as well as implementation of modern risk 74 management and quality system tools and concepts. When finalized, this guidance will replace 75 the 1987 guidance. 76 77 FDA has the authority and responsibility to inspect and evaluate process validation performed by 78 manufacturers. The CGMP regulations for validating pharmaceutical (drug) manufacturing 79 require that drug products be produced with a high degree of assurance of meeting all the 80 attributes they are intended to possess (21 CFR 211.100(a) and 211.110(a)). Effective process 81 validation contributes significantly to assuring drug quality. The basic principle of quality 82 assurance is that a drug should be produced that is fit for its intended use; this principle 83 incorporates the understanding that the following conditions exist: 84 85 • Quality, safety, and efficacy are designed or built into the product. 86 87 • Quality cannot be adequately assured merely by in-process and finished-product 88 inspection or testing. 89 90 • Each step of a manufacturing process is controlled to assure that the finished product 91 meets all design characteristics and quality attributes including specifications. 92 93 For purposes of this guidance, process validation is defined as the collection and evaluation of 94 data, from the process design stage throughout production, which establishes scientific 95 evidence that a process is capable of consistently delivering quality products. Process 96 validation involves a series of activities taking place over the lifecycle of the product and 97 process. This guidance describes the process validation activities in three stages. 98 99 • Stage 1 – Process Design: The commercial process is defined during this stage based on 100 knowledge gained through development and scale-up activities. 101 102 • Stage 2 – Process Qualification: During this stage, the process design is confirmed as 103 being capable of reproducible commercial manufacturing. 104 105 • Stage 3 – Continued Process Verification: Ongoing assurance is gained during routine 106 production that the process remains in a state of control.
5 The 1987 guidance was prepared by a working group that included representation from the Center for Devices and Radiological Health (CDRH). Since that time, CDRH elected to publish its own process validation guidance through the Global Harmonization Task Force. The principles and recommendations in that document, Quality Management Systems – Process Validation, edition 2 (available on the Internet at http://www.ghtf.org/sg3/sg3final.html), are also useful to consider for drug manufacturing processes.
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107 108 This guidance describes activities typical in each stage, but in practice, some activities in 109 different stages might overlap. 110 111 Before any batch from the process is commercially distributed for use by consumers, a 112 manufacturer should have gained a high degree of assurance in the performance of the 113 manufacturing process such that it will consistently produce APIs and drug products meeting 114 those attributes relating to identity, strength, quality, purity, and potency. The assurance should 115 be obtained from objective information and data from laboratory-, pilot-, and/or commercial116 scale studies. Information and data should demonstrate that the commercial manufacturing 117 process is capable of consistently producing acceptable quality products within commercial 118 manufacturing conditions, including those conditions that pose a high risk of process failure. 119 120 A successful validation program depends upon information and knowledge from product and 121 process development. This knowledge and understanding is the basis for establishing an 122 approach to control that is appropriate for the manufacturing process. Manufacturers should: 123 124 • understand the sources of variation 125 • detect the presence and degree of variation 126 • understand the impact of variation on the process and ultimately on product attributes 127 • control the variation in a manner commensurate with the risk it represents to the process 128 and product 129 130 Each manufacturer should judge whether it has gained sufficient understanding to provide a high 131 degree of assurance in its manufacturing process to justify commercial distribution of the 132 product. Focusing on qualification efforts without understanding the manufacturing process may 133 not lead to adequate assurance of quality. After establishing and confirming the process, 134 manufacturers must maintain the process in a state of control over the life of the process, even as 135 materials, equipment, production environment, personnel, and manufacturing procedures 136 change.6 137 138 III. STATUTORY AND REGULATORY REQUIREMENTS FOR PROCESS 139 VALIDATION 140 141 Process validation for drugs (finished pharmaceuticals and components) is a legally enforceable 142 requirement under section 501(a)(2)(B) of the Act, which states the following: 143 144 A drug . . . shall be deemed to be adulterated . . . if . . . the methods used in, or the 145 facilities or controls used for, its manufacture, processing, packing, or holding do not 146 conform to or are not operated or administered in conformity with current good 147 manufacturing practice to assure that such drug meets the requirements of this Act as to
6 The statute and regulations described in section III of this guidance explain the requirement that the methods and facilities used for the manufacturing of drugs be operated and maintained under control sufficient to assure that the identity, strength, purity, and quality of a drug are as they purport or are represented to possess.
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148 safety and has the identity and strength, and meets the quality and purity characteristics, 149 which it purports or is represented to possess. 150 151 FDA regulations describing current good manufacturing practice (CGMP) are provided in 21 152 CFR parts 210 and 211. 153 154 Process validation is required, in both general and specific terms, by the CGMP regulations in 155 parts 210 and 211. The foundation for process validation is provided in § 211.100(a), which 156 states that "[t]here shall be written procedures for production and process control designed to 157 assure that the drug products have the identity, strength, quality, and purity they purport or are 158 represented to possess" (emphasis added). This regulation requires that manufacturers design a 159 process including operations and controls that will result in a product meeting these attributes. 160 Product quality in the context of process validation means that product performance is consistent 161 from batch-to-batch and unit-to-unit. Many products are single-source or involve complicated 162 processes to manufacture. Validation also offers assurance that a process is reasonably 163 safeguarded from sources of variability affecting production output, the loss of which can cause 164 supply problems, thereby negatively affecting public health. 165 166 Other CGMP regulations define the various aspects of validation. Section 211.110(a), Sampling 167 and testing of in-process materials and drug products, requires that control procedures “. . . be 168 established to monitor the output and to validate the performance of those manufacturing 169 processes that may be responsible for causing variability in the characteristics of in-process 170 material and the drug product" (emphasis added). This regulation establishes the requirement 171 that even well-designed processes must include in-process control procedures to assure final 172 product quality. 173 174 CGMP regulations require that batch samples represent the batch under analysis (see, e.g., § 175 211.160(b)(3)) and that the sampling plan result in statistical confidence (§ 211.165(c) and (d)) 176 that the batch meets its predetermined specifications (§ 211.165(a)). Section 211.110(b) 177 provides two principles to follow when establishing in-process specifications. The first principle 178 is that “. . . in-process specifications for such characteristics [of in-process material and the drug 179 product] shall be consistent with drug product final specifications . . . .” Accordingly, in-process 180 material should be controlled to assure that the final drug product will meet its quality 181 requirements. The second principle in this regulation further requires that in-process 182 specifications “. . . shall be derived from previous acceptable process average and process 183 variability estimates where possible and determined by the application of suitable statistical 184 procedures where appropriate.” This requirement, in part, establishes the need for manufacturers 185 to analyze process performance and control batch-to-batch variability.7 186
7 In the Federal Register of September 29, 1978 (43 FR 45013 at 45052), FDA published a final rule on “Current Good Manufacturing Practice in Manufacture, Processing, Packing, or Holding” (available on the Internet at http://www.fda.gov/cder/dmpq/preamble.txt). In the preamble of the final rule, the Agency further explains this principle.
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187 The CGMP regulations also describe and define activities connected with process design, 188 development, and maintenance. Section 211.180(e) requires that information and data about 189 product performance and manufacturing experience be periodically reviewed to determine 190 whether any changes to the established process are warranted. Ongoing feedback about product 191 performance is an essential feature of process maintenance. 192 193 In addition, the CGMP regulations require that facilities in which drugs are manufactured be of 194 suitable size, construction, and location to facilitate proper operations (21 CFR 211.42). 195 Equipment must be of appropriate design, adequate size, and suitably located to facilitate 196 operations for its intended use (21 CFR 211.63). Automated, mechanical, and electronic 197 equipment must be calibrated, inspected, or checked according to a written program designed to 198 assure proper performance (21 CFR 211.68). 199 200 In summary, the CGMP regulations require that manufacturing processes be designed and 201 controlled to assure that in-process materials and the finished product meet predetermined 202 quality requirements and do so consistently and reliably. 203 204 IV. RECOMMENDATIONS 205 206 A. General Considerations for Process Validation 207 208 In all stages of the product lifecycle, good project management and good archiving that capture 209 scientific knowledge will make the process validation program more effective and efficient. 210 These practices should ensure uniform collection and assessment of information about the 211 process, reduce the chance for redundant information gathering and analysis, and enhance the 212 accessibility of such information later in the product lifecycle. 213 214 We recommend an integrated8 team approach to process validation that includes expertise from a 215 variety of disciplines, including process engineering, industrial pharmacy, analytical chemistry, 216 microbiology, statistics, manufacturing, and quality assurance. Project plans, along with the full 217 support of senior management, are essential elements for success. 218 219 Throughout the product lifecycle, various studies can be initiated to discover, observe, correlate, 220 or confirm information about the product and process. All studies should be planned and 221 conducted according to sound scientific principles, appropriately documented, and should be 222 approved in accordance with the established procedure appropriate for the stage of the lifecycle. 223 224 B. Specific Stages and Activities of Process Validation in the Product Lifecycle 225 226 The following subsections describe the recommended stages and specific activities. 227
8 This concept is discussed in more detail in FDA’s guidance for industry, Quality Systems Approach to Pharmaceutical Current Good Manufacturing Practice Regulations, available on the Internet at http://www.fda.gov/cder/guidance/index.htm.
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228 1. Stage 1 – Process Design 229 230 a. Building and Capturing Process Knowledge and Understanding 231 232 Process design is the activity of defining the commercial manufacturing process that will 233 be reflected in the master production and control records. The goal of this stage is to 234 design a process suitable for routine commercial manufacturing that can consistently 235 deliver a product that meets its critical quality attributes. 236 237 Generally, early process design experiments do not need to be performed under CGMP 238 conditions. They should, however, be conducted in accordance with sound scientific 239 methods and principles, including good documentation practices. This recommendation 240 is consistent with ICH guidance for industry, Q10 Pharmaceutical Quality System.9 241 Decisions and justification of the controls should be sufficiently documented and 242 internally reviewed to verify and preserve their value for use later in the lifecycle of the 243 process and product. 244 245 There are exceptions, however. For example, viral and impurity clearance studies have a 246 direct impact on drug safety and should be performed under CGMP conditions, even 247 when performed at small scale. The quality unit should be involved with these studies as 248 is typical during commercial production. 249 250 Product-development activities provide key inputs to the design stage, such as the 251 intended dosage form, the quality attributes, and a general manufacturing pathway. 252 Process information available from the product-development stage can be leveraged in 253 the process-design stage. However, the full spectrum of input variability typical of 254 commercial production is not generally known at this stage. The functionality and 255 limitations of commercial manufacturing equipment should be considered, as well as the 256 contributions of variability by different component lots, production operators, 257 environmental conditions, and measurement systems in the production setting. 258 Laboratory or pilot-scale models designed to be representative of the commercial process 259 can be used to estimate variability. However, it is not a regulatory expectation that the 260 process be developed and tested until it fails, but rather that a process be controlled 261 within commercial manufacturing conditions, including those combinations of conditions 262 posing a high risk of process failure. 263 264 Designing an efficient process with an effective process control approach is dependent on 265 the process knowledge and understanding obtained. Design of Experiment (DOE) 266 studies can help develop process knowledge by revealing relationships, including 267 multifactorial interactions, between the variable inputs (e.g., component10 characteristics
9 A notice of availability for this draft ICH guidance published in the Federal Register on July 13, 2007 (72 FR 38604). When finalized, this guidance will represent FDA’s current thinking on this topic.
10 “Component means any ingredient [raw material] intended for use in the manufacture of a drug product, including those that may not appear in such drug product” (21 CFR 210.3(b)(3)).
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268 or processing parameters) and the resulting outputs (e.g., in-process material, 269 intermediates, or the final product). Risk analysis tools can be used to screen potential 270 variables for DOE studies to minimize the total number of experiments conducted while 271 maximizing knowledge gained. The results of DOE studies can provide justification for 272 establishing ranges of incoming component quality, equipment parameters, and in273 process material quality attributes. 274 275 Other activities, such as experiments or demonstrations at laboratory or pilot scale, allow 276 evaluation of certain conditions and prediction of performance of the commercial 277 process. These activities also provide information that can be used to model or simulate 278 the commercial process. Computer-based or virtual simulations of certain unit operations 279 or dynamics can provide process understanding and avoid problems at commercial scale. 280 It is important to understand the degree to which models represent the commercial 281 process, including any differences that might exist, as this may have an impact on the 282 relevance of information derived from the studies. 283 284 It is essential that activities and studies resulting in product understanding be 285 documented. Documentation should reflect the basis for decisions made about the 286 process. For example, manufacturers should document the variables studied for a unit 287 operation and the rationale for those variables identified as significant. This information 288 is useful during the process qualification and continued process verification stages, 289 including when the design is revised or the strategy for control is refined or changed. 290 291 b. Establishing a Strategy for Process Control 292 293 Process knowledge and understanding is the basis for establishing an approach to process 294 control for each unit operation and the process overall. Strategies for process control can 295 be designed to reduce input variation, adjust for input variation during manufacturing 296 (and so reduce its impact on the output), or combine both approaches. 297 298 Process controls address variability to assure quality of the product. Controls can consist 299 of material analysis and equipment monitoring at significant processing points designed 300 to assure that the operation remains on target and in control with respect to output quality. 301 Special attention to control of the process through operational limits and in-process 302 monitoring is essential (1) where the product attribute is not readily measurable due to 303 limitations of sampling or detectability (e.g., viral clearance or microbial contamination), 304 or (2) when intermediates and products cannot be highly characterized and well-defined 305 quality attributes cannot be identified. These controls are included in the master 306 production and control records (see 21 CFR 211.186(a) and (b)(9)). 307 308 More advanced strategies, such as process analytical technology (PAT), use timely 309 analysis and control loops to adjust the processing conditions so that the output remains 310 constant. Manufacturing systems of this type can provide a higher degree of process 311 control. In the case of PAT strategy, the approach to process qualification will be 312 different from that for other process designs. Further information on PAT processes can
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313 be found in FDA’s guidance for industry on PAT – A Framework for Innovative 314 Pharmaceutical Development, Manufacturing, and Quality Assurance (available on the 315 Internet at http://www.fda.gov/cder/guidance/index.htm). 316 317 The planned commercial production and control records, which contain the operational 318 limits and overall strategy for process control, should be carried forward to the next stage 319 for confirmation. 320 321 2. Stage 2 – Process Qualification 322 323 During the process qualification stage of process validation, the process design is 324 confirmed as being capable of reproducible commercial manufacture. This stage has two 325 elements: (1) design of the facility and qualification of the equipment and utilities, and 326 (2) performance qualification (PQ). During this stage, CGMP-compliant procedures 327 must be followed and successful completion of this stage is necessary before commercial 328 distribution.11 Products manufactured during this stage, if acceptable, can be released. 329 330 a. Design of a Facility and Qualification of Utilities and Equipment 331 332 Proper design of a manufacturing facility is required under 21 CFR part 211, subpart C, 333 of the CGMP regulations on Buildings and Facilities. It is essential that activities 334 performed to assure proper facility design and commissioning precede PQ. Activities 335 undertaken to demonstrate that utilities and pieces of equipment are suitable for their 336 intended use and perform properly is referred to in this guidance as qualification. These 337 activities necessarily precede manufacturing products at the commercial scale. 338 339 Qualification of utilities and equipment generally includes the following activities: 340 341 • Selecting utilities and equipment construction materials, operating principles, and 342 performance characteristics based on whether they are appropriate for their specific 343 use. 344 345 • Verifying that utility systems and equipment are built and installed in compliance 346 with the design specifications (e.g., built as designed with proper materials, capacity, 347 and functions, and properly connected and calibrated). 348 349 • Verifying that the utility system and equipment operate in accordance with the 350 process requirements in all anticipated operating ranges. This should include 351 challenging the equipment or system functions while under load comparable to that 352 expected during routine production. It should also include the performance of 353 interventions, stoppage, and start-up as is expected during routine production.
11 As discussed in section III of this guidance, process validation (including process qualification) is legally enforceable under section 501(a)(2)(B) of the Act. FDA regulations require that process validation procedures be established and followed (21 CFR 211.100) before a batch can be distributed (21 CFR 211.22 and 211.165).
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354 Operating ranges should be shown capable of being held as long as would be 355 necessary during routine production. 356 357 Qualification of utilities and equipment can be covered under individual plans or as part 358 of an overall project plan. The plan should consider the requirements of use and can 359 incorporate risk management to prioritize certain activities and to identify a level of effort 360 in both the performance and documentation of qualification activities. The plan should 361 identify (1) the studies or tests to use, (2) the criteria appropriate to assess outcomes, (3) 362 the timing of qualification activities, (4) responsibilities, and (5) the procedures for 363 documenting and approving the qualification. It should also include the firm’s 364 requirements for the evaluation of changes. Qualification activities should be 365 documented and summarized in a report with conclusions that address criteria in the plan. 366 The quality control unit must review and approve the qualification plan and report (21 367 CFR 211.22). 368 369 b. Performance Qualification Approach 370 371 The PQ is the second element of stage 2, process qualification. The PQ combines the 372 actual facility, utilities, equipment (each now qualified), and the trained personnel with 373 the commercial manufacturing process, control procedures, and components to produce 374 commercial batches. A successful PQ will confirm the process design and demonstrate 375 that the commercial manufacturing process performs as expected. 376 377 Success at this stage signals an important milestone in the product lifecycle and needs to 378 be completed before a manufacturer commences commercial distribution of the drug 379 product.12 The decision to begin commercial distribution should be supported by data 380 from commercial batches. Data from laboratory and pilot studies can provide additional 381 assurance. 382 383 The approach to PQ should be based on sound science and the manufacturer’s overall 384 level of product and process understanding. The cumulative data from all relevant 385 studies (e.g., designed experiments; laboratory, pilot, and commercial batches) should be 386 used to establish the manufacturing conditions in the PQ. For example, to have sufficient 387 understanding of the commercial process, the manufacturer will need to consider the 388 effects of scale; however, it is not typically necessary to explore the entire operating 389 range at commercial scale if assurance can be provided by other data. Previous credible 390 experience with sufficiently similar products and processes can also be considered. In 391 addition, we strongly recommend firms employ objective measures (e.g., statistical 392 metrics), wherever feasible and meaningful to achieve adequate assurance. 393 394 In most cases, PQ will have a higher level of sampling, additional testing, and greater 395 scrutiny of process performance. The level of monitoring and testing should be sufficient 396 to confirm uniform product quality throughout the batch during processing. This greater
12 See section III of this guidance, Statutory and Regulatory Requirements for Process Validation.
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397 scrutiny accompanied by a higher level of sampling should continue through the process 398 verification stage, as appropriate. 399 400 The extent to which some materials, such as column resins or molecular filtration media, 401 can be re-used without adversely affecting product quality can be assessed in relevant 402 laboratory studies, and their usable lifetime should be confirmed by an ongoing PQ 403 protocol during commercial manufacture. 404 405 A manufacturing process that uses PAT may warrant a different PQ approach. Such a 406 process is one that is designed to measure in real time the attributes of an in-process 407 material and then adjust the process in a timely control loop so the process maintains the 408 desired quality of the output material. The process design stage and the process 409 qualification stage should have as a focus the measurement system and control loop. 410 Regardless, the goal remains the same: establishing scientific evidence that the process is 411 reproducible and will consistently deliver quality products. 412 413 c. Performance Qualification Protocol 414 415 A written protocol that specifies the manufacturing conditions, controls, testing, and 416 expected outcomes is essential for this stage of process validation. We recommend that 417 the protocol discuss: 418 419 • The manufacturing conditions including operating parameters, processing limits, and 420 component (raw material) inputs. 421 422 • The data to be collected and when and how it will be evaluated. 423 424 • Tests to be performed (in-process, release, characterization) and acceptance criteria 425 for each significant processing step. 426 427 • The sampling plan including sampling points, number of samples, and the frequency 428 of sampling for each unit operation and attribute. The number of samples should be 429 adequate to provide sufficient statistical confidence of quality both within a batch and 430 between batches. The confidence level selected can be based on risk analysis as it 431 relates to the particular attribute under examination. Sampling during this stage 432 should be more extensive than is typical during routine production. 433 434 • Criteria that provide for a rational conclusion of whether the process consistently 435 produces quality products. The criteria should include: 436 437 o A description of the statistical methods to be used in analyzing all collected 438 data (e.g., statistical metrics defining both intra-batch and inter-batch 439 variability). 440
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441 o Provision for addressing deviations from expected conditions and handling of 442 nonconforming data. Data should not be excluded from further consideration 443 in terms of PQ without a documented, science-based justification. 444 445 • Design of facilities and the qualification of utilities and equipment, personnel training 446 and qualification, and verification of material sources (components and 447 container/closures), if not previously accomplished. 448 449 • Status of the validation of analytical methods used in measuring the process, in450 process materials, and the product. 451 452 • Review and approval by appropriate departments and the quality unit. 453 454 d. Protocol Execution and Report 455 456 Protocol execution should not begin until the protocol has been reviewed and approved 457 by all appropriate departments, including the quality unit. Departure from the established 458 protocol must be made according to established procedure or provisions in the protocol. 459 Such departures must be justified and approved by all appropriate departments and the 460 quality unit before implementation (§ 211.100). 461 462 The commercial manufacturing process and routine procedures must be followed (§§ 463 211.100(b) and 211.110(a)). The PQ lots should be manufactured under normal 464 conditions by personnel expected to routinely perform each step of each unit operation in 465 the process. Normal operating conditions should cover the utility systems (e.g., air 466 handling and water purification), material, personnel, environment, and manufacturing 467 procedures. 468 469 A report documenting and assessing adherence to the written protocol should be prepared 470 in a timely manner after the completion of the protocol. This report should: 471 472 • Discuss and cross-reference all aspects of the protocol. 473 474 • Summarize data collected and analyze the data, as specified by the protocol. 475 476 • Evaluate any unexpected observations and additional data not specified in the 477 protocol. 478 479 • Summarize and discuss all manufacturing nonconformances such as deviations, 480 aberrant test results, or other information that has bearing on the validity of process. 481 482 • Describe in sufficient detail any corrective actions or changes that should be made to 483 existing procedures and controls. 484
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485 • State a clear conclusion as to whether the data indicates the process met the 486 conditions established in the protocol and whether the process is considered to be in a 487 sufficient state of control. If not, the report should state what should be accomplished 488 before such a conclusion can be reached. This conclusion should be based on a 489 documented justification for the approval of the process, and release of lots produced 490 by it to the market in consideration of the entire compilation of knowledge and 491 information gained from the design stage through the process qualification stage. 492 493 • Include all appropriate department and quality unit review and approvals. 494 495 3. Stage 3 – Continued Process Verification 496 497 The goal of the third validation stage is to continually assure that the process remains in a 498 state of control (the validated state) during commercial manufacture. A system or 499 systems for detecting unplanned departures from the process as designed is essential to 500 accomplish this goal. Adherence to the CGMP requirements, specifically including the 501 collection and evaluation of information and data about the performance of the process 502 (see below), will allow detection of process drift. The evaluation should determine 503 whether action must be taken to prevent the process from drifting out of control (§ 504 211.180(e)). 505 506 An ongoing program to collect and analyze product and process data that relate to 507 product quality must be established (§ 211.180(e)). The data collected should include 508 relevant process trends and quality of incoming materials or components, in-process 509 material, and finished products. The data should be statistically trended and reviewed by 510 trained personnel. The information collected should verify that the critical quality 511 attributes are being controlled throughout the process. 512 513 We recommend that a statistician or person with adequate training in statistical process 514 control techniques develop the data collection plan and statistical methods and 515 procedures used in measuring and evaluating process stability and process capability. 516 Procedures should describe how trending and calculations are to be performed. 517 Procedures should guard against overreaction to individual events as well as against 518 failure to detect process drift. Production data should be collected to evaluate process 519 stability and capability. The quality unit should review this information. If done 520 properly, these efforts can identify variability in the process and/or product; this 521 information can be used to alert the manufacturer that the process should be improved. 522 523 Good process design and development should anticipate significant sources of variability 524 and establish appropriate detection, control, and/or mitigation strategies, as well as 525 appropriate alert and action limits. However, a process is likely to encounter sources of 526 variation that were not previously detected or to which the process was not previously 527 exposed. Many tools and techniques, some statistical and others more qualitative, can be 528 used to detect variation, characterize it, and determine the root cause. We recommend 529 that the manufacturer use quantitative, statistical methods whenever feasible. We also
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530 recommend that it scrutinize intra-batch as well as inter-batch variation as part of a 531 comprehensive continued process verification program. 532 533 We recommend continued monitoring and/or sampling at the level established during the 534 process qualification stage until sufficient data is available to generate significant 535 variability estimates. Once the variability is known, sampling and/or monitoring should 536 be adjusted to a statistically appropriate and representative level. Process variability 537 should be periodically assessed and sampling and/or monitoring adjusted accordingly. 538 539 Variation can also be detected by the timely assessment of defect complaints, out-of540 specification findings, process deviation reports, process yield variations, batch records, 541 incoming raw material records, and adverse event reports. Production line operators and 542 quality unit staff should be encouraged to provide feedback on process performance. 543 Operator errors should also be tracked to measure the quality of the training program; to 544 identify operator performance issues; and to look for potential batch record, procedural, 545 and/or process improvements that could help to reduce operator error. We recommend 546 that the quality unit meet periodically with production staff to evaluate data, discuss 547 possible trends or drifts in the process, and coordinate any correction or follow-up actions 548 by production. 549 550 Data gathered during this stage might suggest ways to improve and/or optimize the 551 process by altering some aspect of the process or product such as the operating conditions 552 (ranges and set-points), process controls, component, or in-process material 553 characteristics. A description of the planned change, a well-justified rationale for the 554 change, an implementation plan, and quality unit approval before implementation must 555 be documented (21 CFR 211.100). Depending on the significance to product quality, 556 modifications may warrant performing additional process design and process 557 qualification activities.13 558 559 Maintenance of the facility, utilities, and equipment is another important aspect of 560 ensuring that a process remains in control. Once established, qualification status must be 561 maintained through routine monitoring, maintenance, and calibration procedures and 562 schedules (21 CFR part 211, subparts C and D). The data should be assessed periodically 563 to determine whether re-qualification should be performed and the extent of that re564 qualification. Maintenance and calibration frequency should be adjusted based on 565 feedback from these activities. 566 567 V. CONCURRENT RELEASE OF PERFORMANCE QUALIFICATION BATCHES 568 569 In most cases, the PQ protocol needs to be completed before the commercial distribution of a 570 product. In special situations, the PQ protocol can be designed to release a PQ batch for
13 Certain manufacturing changes may call for a formal notification to the Agency before implementation, as directed by existing regulations and filing guidance (i.e., documents that describe procedures for filing information to an application).
Contains Nonbinding Recommendations
Draft — Not for Implementation
571 distribution before completion of the protocol. The conclusions about the manufacturing 572 process should be made when the protocol is completed and the data is fully evaluated. 573 574 FDA expects that concurrent release will be used rarely. Concurrent release might be 575 appropriate for processes used infrequently because of limited demand for the product (e.g., 576 orphan drugs), processes with necessarily low production volume per batch (e.g., 577 radiopharmaceuticals, including positron emission tomography drugs), and processes 578 manufacturing medically necessary drugs to alleviate a short supply, which should be 579 coordinated with the Agency. 580 581 When warranted and used, concurrent release should be accompanied by a system for careful 582 oversight of the distributed batch to facilitate rapid customer feedback. For example, customer 583 complaints and defect reports should be rapidly assessed to determine root cause and whether the 584 process should be improved or changed. We recommend that each batch in a concurrent release 585 program also undergo stability testing and that this test data be promptly evaluated to ensure 586 rapid detection and correction of any problems. 587 588 VI. DOCUMENTATION 589 590 Documentation at each stage of the process validation lifecycle is essential for effective 591 communication in complex, lengthy, and multidisciplinary projects. Documentation is important 592 so that knowledge gained about a product and process is accessible and comprehensible to others 593 involved in each stage of the lifecycle. In addition to being a fundamental tenet of following the 594 scientific method, information transparency and accessibility are essential so that organizational 595 units responsible and accountable for the process can make informed, science-based decisions 596 that ultimately support the release of a product to commerce. 597 598 The degree and type of documentation required by CGMP is greatest during stage 2, process 599 qualification, and stage 3, continued process verification. Studies during these stages must 600 conform to CGMPs and must be approved by the quality unit in accordance with the regulations 601 (see 21 CFR 211.22 and 211.100). Viral and impurity clearance studies, even when performed at 602 small scale, also require full quality unit oversight as is necessary during routine commercial 603 production. 604 605 CGMP documents for commercial manufacturing (i.e., the initial commercial master batch 606 production and control record (21 CFR 211.186) and supporting procedures) are key outputs of 607 stage 1, process design. We recommend that firms diagram the process flow for the full-scale 608 process. Process flow diagrams should describe each unit operation, its placement in the overall 609 process, monitoring and control points, and the component, as well as other processing material 610 inputs (e.g., processing aids) and expected outputs (i.e., in-process materials and finished 611 product). It is also useful to generate and preserve process flow diagrams of the various scales as 612 the process design progresses to facilitate comparison and decision making about their 613 comparability. 614
Contains Nonbinding Recommendations
Draft — Not for Implementation
615 VII. ANALYTICAL METHODOLOGY 616 617 Process knowledge is dependent on accurate and precise measuring techniques that are used to 618 test and examine the quality of drug components, in-process materials, and finished products. 619 For data to have value in predicting process outcomes, it is essential that the analytical tests be 620 scientifically sound (as required under 21 CFR 211.160). While validated analytical methods are 621 not required during product- and process-development activities, methods should be 622 scientifically sound (e.g., specific, sensitive, and accurate), suitable, and reliable for the specified 623 purpose. There should be assurance of proper equipment function for laboratory experiments. 624 Procedures for analytical method and equipment maintenance, documentation practices, and 625 calibration practices supporting process-development efforts should be documented or described. 626 Analytical methods supporting clinical supply production, particularly stage 2 and 3 studies, 627 must follow appropriate CGMPs in parts 210 and 211.
Contains Nonbinding Recommendations
Draft — Not for Implementation
628 REFERENCES 629 630 FDA, 1987 (CDER, CBER, and Center for Devices and Radiological Health (CDRH)), 631 Guideline on General Principles of Process Validation, guidance for industry, May 1987. 632 633 FDA, 2002 (CBER), Validation of Procedures for Processing of Human Tissues Intended for 634 Transplantation, guidance for industry, May 2002. 635 636 FDA, 2004 (CDER, CVM, and ORA), PAT — A Framework for Innovative Pharmaceutical 637 Development, Manufacturing, and Quality Assurance, guidance for industry, September 638 2004. 639 640 FDA, 2006 (CDER, CBER, CVM, and ORA), Quality Systems Approach to Pharmaceutical 641 Current Good Manufacturing Practice Regulations, guidance for industry, September 642 2006. 643 644 FDA/Global Harmonization Task Force (GHTF; medical devices), 2004, Quality Management 645 Systems – Process Validation, edition 2, guidance, January 2004. 646 647 FDA/ICH, 2001 (CDER and CBER), Q7A Good Manufacturing Practice, Guidance for Active 648 Pharmaceutical Ingredients, ICH guidance for industry, August 2001. 649 650 FDA/ICH, 2006 (CDER and CBER), Q8A Pharmaceutical Development, ICH guidance for 651 industry, May 2006. 652 653 FDA/ICH, 2006 (CDER and CBER), Q9A Quality Risk Management, ICH guidance for industry, 654 June 2006. 655 656 FDA/ICH (CDER and CBER) Q10 Quality Systems, ICH draft guidance for industry, May 2007 657 (when finalized, this guidance will convey FDA’s current thinking on this topic).

Validation of Analytical Procedures for Type C Medicated Feeds

TABLE OF CONTENTS
Page No.
Introduction 4
Discussion 5
1. Specificity 6
2. Linearity 7
3. Range 7
4. Accuracy 8
5. Precision 8
5.1. Repeatability 8
5.2. Intermediate Precision 8
5.3. Reproducibility 9
5.4. Proof of Performance 9
6. Limit of Detection 9
6.1. Based on Visual Evaluation 9
6.2. Based on Signal-to-Noise 9
6.3. Based on the Standard Deviation of the Response and the Slope 10
6.3.1. Based on the Standard Deviation of the Blank 10
6.3.2. Based on the Calibration Curve 10
7. Limit of Quantitation 10
7.1. Based on Visual Evaluation 10
7.2. Based on Signal-to-Noise Approach 11
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7.3. Based on the Standard Deviation of the Response and the Slope 11
7.3.1. Based on the Standard Deviation of the Blank 11
7.3.2. Based on the Calibration Curve 11
8. Robustness / Ruggedness 11
9. System Suitability Testing 12
10. Recommended Data 12
Glossary 15
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Guidance for Industry
Validation of Analytical Procedures for Type C
Medicated Feeds1
INTRODUCTION
The purpose of this guidance is to provide recommendations on how to consider the
various validation characteristics for each analytical procedure used in medicated
feed assays. This guidance is written primarily for chromatographic methods;
however, the guidance does not limit the analytical technique to chromatographic
procedures, as other techniques may be appropriate. In some cases (for example,
demonstration of specificity), the overall capabilities of a number of analytical
procedures in combination may be investigated in order to ensure the quality of the
medicated feed.
Section 512(b) of the Federal Food, Drug, and Cosmetic Act (21 U.S.C. § 360b)
establishes the requirements for new animal drug approval. 21 C.F.R. § 514.1
specifies the information required to be submitted as part of the application and the
proper form for the submission. Section 514.1(b)(5)(vii) requires an applicant to
describe analytical procedures that should be capable of determining the active
component(s) within a reasonable degree of accuracy and of assuring the identity of
such components. Section 514.1(b)(5)(vii)(a) states that a description of practicable
1 This guidance has been prepared by the Office of New Animal Drug Evaluation in the Center for
Veterinary Medicine at the Food and Drug Administration.
This guidance represents the agency’s current thinking on this topic. It does not
create or confer any rights for or on any person and does not operate to bind
FDA or the public. You can use an alternative approach if the approach satisfies
the requirements of the applicable statute(s) and regulation(s). If you want to
discuss an alternative approach, contact the FDA staff responsible for
implementing this guidance. If you cannot identify the appropriate FDA staff,
call the appropriate number listed on the title page of this guidance.
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methods of analysis of adequate sensitivity to determine the amount of the new
animal drug in the final dosage form should be included.
FDA’s guidance documents, including this guidance, do not establish legally
enforceable responsibilities. Instead, guidances describe the Agency’s current
thinking on a topic and should be viewed only as recommendations, unless
specific regulatory or statutory requirements are cited. The use of the word
“should” in Agency guidances means that something is suggested or
recommended, but not required.
DISCUSSION
The objective of the analytical procedure should be clearly understood since this
will govern the validation characteristics that are evaluated. Typical validation
characteristics that may be considered are listed below:
- Specificity
- Linearity
- Range
- Accuracy
- Precision
- Limit of Detection
- Limit of Quantitation
- Robustness
Each of these validation characteristics is defined in the attached Glossary.
Approaches other than those set forth in this guidance may be acceptable. It is the
responsibility of the applicant to choose the validation procedure and protocol most
suitable for the product. However, it is important to remember that the main
objective of validation of an analytical procedure is to demonstrate that the
procedure is suitable for its intended purpose.
It is recommended that a well-characterized reference standard, with documented
purity, be used throughout the validation study. The degree of purity necessary
depends on the intended use.
For the sake of clarity, this document considers the various validation
characteristics in distinct sections. The arrangement of these sections reflects the
process by which an analytical procedure may be developed and evaluated.
In practice, it is recommended to design the experimental work such that the
appropriate validation characteristics can be considered simultaneously to provide a
sound, overall knowledge of the capabilities of the analytical procedure.
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Appropriate validation characteristics may include: specificity, linearity, range,
accuracy, and precision.
1. SPECIFICITY
It is recommended that an investigation of specificity be conducted during the
validation of the medicated feed assay. The procedures used to demonstrate
specificity will depend on the intended objective of the analytical procedure.
Identification of the analyte may be made by means of retention time of the
standard.
For chromatographic procedures, it is recommended that representative
chromatograms be used to demonstrate specificity, and individual feed components
and drug products be appropriately labeled. The chromatographic profile using
peak shape and tailing criteria may be used to indicate either co-eluting peaks or
sample matrix effects. The peak parameters should be in agreement between the
standard and analyte peaks. In addition, to ensure that the peaks are single
components, a diode array detector may be used to obtain peak purity information
for the analyte peaks in a variety of feed matrices. Similar considerations may be
given to other separation techniques.
For the assay, it is recommended that there be a demonstration of a lack of
interference by feed ingredients or other drug products that may be in the feed.
This may be done by demonstrating that the responses of a blank placebo made
from the feed ingredients and/or drug products, either separately or in combination,
are either different from the absorbance (for Ultraviolet (UV) methods) or retention
time (for Gas Chromatography (GC) and High Performance Liquid
Chromatography (HPLC) methods) of the analyte of interest or not significant (i.e.,
that the signal measured as a percent concentration is not greater than 10%). It is
recommended that additional information be provided showing that common feed
ingredients do not interfere with the detection system. If potential interference is
observed, it is recommended that the ingredient be evaluated by the complete
method. Some examples of interfering ingredients are clay agents (for flowability)
and pellet binding, molasses, grass meals (e.g., alfalfa), high mineral content, corn
cob meal, cottonseed by product meal, meat and bone meal, and fish meal. Many
methods developed to give good recovery in a simple corn-soy feed do not work
well when the analyte is added to high mineral feeds, and are not recommended.
Typically, 2-3 feed mixtures, based on the species that will be medicated,
geographical location where the feed may be prepared, and life cycle of the species
(e.g., starter, finisher), should be tested. For drug products, it is recommended that
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applicants consider the most common products that may typically be present within
the feed.
Applicable literature references demonstrating non-interference may be supplied in
lieu of actual testing.
2. LINEARITY
It is recommended that a linear relationship be evaluated across the range (see
section 3) of the analytical procedure. It may be demonstrated directly on the drug
substance by separate weighings (two separate weighings preferred) and/or dilution
of a standard stock solution, using the proposed procedure.
It is recommended that linearity be evaluated by visual inspection of a plot of
signals as a function of analyte concentration or content. If there is a linear
relationship, it is recommended that test results be evaluated by appropriate
statistical methods, for example, by calculation of a regression line by the method
of least squares. Data from the regression line itself may be helpful to provide
mathematical estimates of the degree of linearity. It is recommended that the
correlation coefficient (R) be at least 0.995. The regression line intercept should
not differ from zero if a single point calibration technique is used. This may be
demonstrated if the confidence limits of the intercept include zero or if the intercept
value is a small percentage of the target level. If the intercept is significantly
different from zero, then a single point calibration technique is not recommended.
For the establishment of linearity, a minimum of 5 concentrations, covering the
intended dosing range with one concentration 50% of the lowest dose, is
recommended. It is recommended that the sponsor contact CVM if other
approaches are used.
3. RANGE
The specified range should be derived from linearity studies and depends on the
intended application of the procedure. It may be established by confirming that the
analytical procedure provides an acceptable degree of linearity, accuracy, and
precision when applied to samples containing amounts of analyte within or at the
extremes of the specified range of the analytical procedure.
For the assay of a drug in a medicated feed, the range should be from 50 to 150
percent of the labeled concentration.
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4. ACCURACY
It is recommended that accuracy be established across the specified range of the
analytical procedure used for medicated feed assays.
It is recommended that two (2) typical feed matrices with known quantities of the
drug added be analyzed.
It is recommended that accuracy be assessed using a minimum of 15 - 20
determinations over the concentration levels covering the specified range for each
feed matrix tested (e.g. 3 - 4 concentrations (depending on the dose range) / 5
replicates each of the total analytical procedure). Recovery from fortified blank
matrix samples should be between 80 - 110%.
5. PRECISION
Validation of tests for assay of medicated feeds should include an investigation of
precision.
5.1. Repeatability
For fortified medicated feed samples, it is recommended that repeatability be
assessed using a minimum of 15 determinations covering the specified range for
the procedure (e.g., 3-4 concentrations / 5 replicates each).
For drugs incorporated into medicated feeds at greater than 10 ppm, the withinlaboratory
variation coefficient should be less than 5.0%. For drugs
incorporated into medicated feeds at less than 10 ppm, the within-laboratory
variation coefficient should be less than 7.5%.
5.2. Intermediate Precision
The extent to which intermediate precision should be established depends on the
circumstances under which the procedure is intended to be used. It is
recommended that the applicant establish the effects of random events on the
precision of the analytical procedure. It is recommended that variations to be
studied include days, analysts, equipment, etc. It is not recommended to study
these effects individually. Instead, the use of a statistical experimental design
(matrix) is encouraged (see Statistical Manual of the AOAC by W.J. Youden
and E.H. Steiner, 1975, page 33 for more information on statistical design of
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experiments). The performance of the method by a second independent
laboratory is encouraged.
5.3. Reproducibility
It is recommended that reproducibility be assessed by means of an interlaboratory
trial. It is recommended that reproducibility be considered in the
case of standardization of an analytical procedure.
5.4. Proof of Performance
It is recommended that proof of performance of the assay be demonstrated by
testing two (2) batches of the proposed medicated feed manufactured in mixing
equipment of the appropriate size and under conditions representative of typical
commercial processing. When feasible, batches should be manufactured using
different configurations of mixers.
It is recommended that a minimum of 10 determinations covering the specified
range for the procedure be made (e.g., 2 concentrations (high and low) / 5
replicates each). If the feed is pelletized, it is recommended that the mash and
the pelletized feed be tested separately. Results should be reported in both
concentration and percent label claim.
6. LIMIT OF DETECTION
There are several approaches for determining the limit of detection (LOD),
depending on whether the procedure is non-instrumental or instrumental.
Approaches other than those listed below may be used.
6.1. Based on Visual Evaluation
Visual evaluation may be used for non-instrumental or instrumental methods.
The detection limit may be determined by the analysis of samples with known
concentrations of analyte and by establishing the minimum level at which the
analyte can be reliably detected.
6.2. Based on Signal-to-Noise
It is recommended that this approach be applied only to analytical procedures
that exhibit baseline noise.
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Determination of the signal-to-noise ratio may be performed by comparing
measured signals from samples with known low concentrations of analyte with
those of blank samples, and establishing the minimum concentration at which
the analyte can be reliably detected. A signal-to-noise ratio of between 3 or 2:1
is generally recommended for estimating the detection limit.
6.3. Based on the Standard Deviation of the Response and the Slope
It is recommended that the LOD be expressed as:
LOD = 3.3 δ / S
where δ = the standard deviation of the responses and S = the slope of the
calibration curve. The slope S may be estimated from the calibration curve of
the analyte. The estimate of δ may be carried out in a variety of ways, for
example:
6.3.1. Based on the Standard Deviation of the Blank
It is recommended that the measurement of the magnitude of analytical
background response be performed by analyzing an appropriate number of
blank samples and calculating the standard deviation of these responses.
6.3.2. Based on the Calibration Curve
It is recommended that a specific calibration curve be studied using samples
containing an analyte in the range of the LOD. The residual standard deviation
of a regression line or the standard deviation of y-intercepts of regression lines
may be used as the standard deviation.
7. LIMIT OF QUANTITATION
Several approaches for determining the limit of quantitation (LOQ) are possible,
depending on whether the procedure is non-instrumental or instrumental.
Approaches other than those listed below may be used.
7.1. Based on Visual Evaluation
Visual evaluation may be used for non-instrumental or instrumental methods.
The LOQ may be determined by the analysis of samples with known
concentrations of analyte, and by establishing the minimum level at which the
analyte can be quantified with acceptable accuracy and precision.
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7.2. Based on Signal-to-Noise Approach
It is recommended that this approach be applied only to analytical procedures
that exhibit baseline noise. Determination of the signal-to-noise ratio may be
performed by comparing measured signals from samples with known low
concentrations of analyte with those of blank samples, and establishing the
minimum concentration at which the analyte can be reliably quantified. A
signal-to-noise ratio of 10:1 is recommended.
7.3. Based on the Standard Deviation of the Response and the Slope
The LOQ may be expressed as:
LOQ = 10 δ / S
where δ = the standard deviation of the responses and S = the slope of the
calibration curve. The slope S may be estimated from the calibration curve of
the analyte. The estimate of δ may be carried out in a variety of ways, for
example:
7.3.1. Based on the Standard Deviation of the Blank
Measurement of the magnitude of analytical background response may be
performed by analyzing an appropriate number of blank samples and calculating
the standard deviation of these responses.
7.3.2. Based on the Calibration Curve
It is recommended that a specific calibration curve be studied using samples
containing an analyte in the range of the LOQ. The residual standard deviation
of a regression line or the standard deviation of y-intercepts of regression lines
may be used as the standard deviation.
8. ROBUSTNESS / RUGGEDNESS
It is recommended that the evaluation of robustness be considered during the
development phase and demonstrated during the analytical validation phase.
Robustness depends on the type of procedure under study. It should show the
reliability of an analysis with respect to deliberate variations in method parameters.
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If measurements are susceptible to variations in analytical conditions, it is
recommended that the analytical conditions be suitably controlled or a
precautionary statement be included in the procedure. One consequence of the
evaluation of robustness should be that a series of system suitability parameters
(e.g., resolution test) are established to ensure that the validity of the analytical
procedure is maintained whenever used.
Examples of typical variations are:
-stability of analytical solutions and feed extracts; and
-extraction time.
(Note: it is recommended that results of the stability studies of the
analytical solutions and feed extracts be included in the procedure)
In the case of high performance liquid chromatography (HPLC), examples of
typical variations are:
-influence of variations of pH in a mobile phase;
-influence of variations in mobile phase composition;
-different columns (different lots and/or suppliers); and
-temperature-flow rate.
In the case of gas chromatography (GC), examples of typical variations are:
-different columns (different lots and/or suppliers); and
-temperature-flow rate.
9. SYSTEM SUITABILITY TESTING
System suitability testing is an integral part of many analytical procedures. The
tests are based on the concept that the equipment, electronics, analytical operations,
and samples to be analyzed constitute an integral system that can be evaluated as
such. System suitability test parameters to be established for a particular procedure
depend on the type of procedure being validated and may include, for example, data
acceptability testing for feed controls. It is recommended that system suitability
tests and criteria for the HPLC or GC detection system be evaluated. Performance
specifications for critical reagents and steps, such as solid phase extraction, should
be included when appropriate. If specific tests and criteria are used, then the
recommended actions taken if performance does not meet the criteria should be
determined. Additional information is available in Pharmacopoeias.
10. RECOMMENDED DATA
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It is recommended that data collected during validation and formulae used for
calculating validation characteristics be submitted for each feed type and discussed
as outlined below:
Specificity:
It is recommended that representative sample sets of chromatograms be
provided so that recalculation can be performed, including:
Baseline / mobile phase
Extraction solvent
Feed ingredient placebo that cause interference
Other drug product placebo that causes interference
Standards
Samples (high and low concentration, different feed mixtures)
Retention times and a comparison of relative retention times should be
provided.
Tabular listing of feed mixture ingredients and other drug products tested
should be provided.
Linearity & Range:
It is recommended that the correlation coefficient, y-intercept, slope of the
regression line, and residual sum of squares be submitted. A plot of the data
should be included. In addition, an analysis of the deviation of the actual data
points from the regression line may also be helpful for evaluating linearity.
Accuracy:
It is recommended that accuracy be reported as percent recovery by the assay of
known added amount of analyte in the sample or as the difference between the
mean and the accepted true value together with the confidence intervals.
Tabular listing of feed mixture ingredients used should be provided.
For each feed matrix studied, it is recommended that the complete set of data
including weighings, sample and standard preparation, chromatography,
calculations, and results be provided. A representative set of chromatograms
should be provided and, for the concentration(s) in between, a table of relevant
parameters should be provided. All individual area or height measurements for
controls, standards, and samples and all other information such as sample
weights, standard concentrations, and dilutions should also be provided.
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Precision:
It is recommended that the standard deviation, relative standard deviation
(coefficient of variation), and confidence interval be reported for each type of
precision investigated.
It is recommended that the complete set of data including weighings, sample
and standard preparation, chromatography, calculations, and results be
provided. A representative set of data should be provided and, for the
concentration(s) in between, a table of relevant parameters should be provided
for each type of precision investigated.
Limit of Detection:
It is recommended that the limit of detection and the method used for
determining the detection limit be presented. If the LOD is determined based
on visual evaluation or based on signal-to-noise ratio, the presentation of the
relevant chromatograms may be considered acceptable for justification.
In cases where an estimated value for the LOD is obtained by calculation or
extrapolation, this estimate may subsequently be validated by the independent
analysis of a suitable number of samples known to be near, or prepared at, the
LOD.
Limit of Quantitation:
It is recommended that the limit of quantitation and the method used for
determining the LOQ be presented. The limit should be subsequently
confirmed by the analysis of a suitable number of samples known to be near, or
prepared at, the LOQ.
Robustness/Ruggedness:
It is recommended that tabular representation including conditions tested,
retention times, tailing factors, effects on resolution, and potency be presented.
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GLOSSARY
1. ANALYTICAL PROCEDURE
The analytical procedure refers to the way an analysis is performed. It describes in
detail the steps that should be followed to perform each analytical test. This may
include, but is not limited to, the sample, the reference standard and the reagents
preparations, use of the apparatus, generation of the calibration curve, and use of
the formulae for the calculation.
2. SPECIFICITY
Specificity is the ability to assess unequivocally the analyte in the presence of
components that may be expected to be present. Typically, these might include
impurities, degradation products, matrix, other approved drugs, etc.
Lack of specificity of an individual analytical procedure may be compensated for
by other supporting analytical procedure(s).
This definition includes the following:
Identification: to ensure the identity of an analyte.
Assay (content or potency): to provide an exact result which allows an
accurate statement on the content or potency of the analyte in a sample.
3. LINEARITY
The linearity of an analytical procedure is its ability (within a given range) to obtain
test results that are directly proportional to the concentration (amount) of analyte in
the sample.
4. RANGE
The range of an analytical procedure is the interval between the upper and lower
concentration (amounts) of analyte in the sample (including these concentrations)
for which it has been demonstrated that the analytical procedure has a suitable level
of precision, accuracy, and linearity.
5. ACCURACY
The accuracy of an analytical procedure refers to the closeness of agreement
between the value that is accepted either as a conventional true value or an accepted
reference value, and the value found.
This is sometimes termed trueness.
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6. PRECISION
The precision of an analytical procedure expresses the closeness of agreement
(degree of scatter) between a series of measurements obtained from multiple
sampling of the same homogenous sample under the prescribed conditions.
Precision may be considered at three levels: repeatability, intermediate precision,
and reproducibility.
Precision is investigated using homogenous, authentic samples. However, if it is
not possible to obtain a homogenous sample, it may be investigated using
artificially prepared samples or a sample solution (although extraction variability
will not be measured).
The precision of an analytical procedure is usually expressed as the variance,
standard deviation, or coefficient of variation of a series of measurements.
6.1. Repeatability: Repeatability expresses the precision under the same
operating conditions over a short interval of time. Repeatability is also termed
intra-assay precision.
6.2. Intermediate precision: Intermediate precision expresses within-laboratories
variations: different days, different analysts, different equipment, etc.
6.3. Reproducibility: Reproducibility expresses the precision between
laboratories (collaborative or transfer studies, usually applied to standardization
of methodology).
7. LIMIT OF DETECTION
The limit of detection of an individual analytical procedure is the lowest amount of
analyte in a sample that can be detected, but not necessarily quantitated as an exact
value.
8. LIMIT OF QUANTITATION
The limit of quantitation 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 quantitation limit is a parameter of quantitative assays for low
levels of compounds in sample matrices and is used particularly for the
determination of impurities and/or degradation products.
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9. ROBUSTNESS / RUGGEDNESS
The robustness of an analytical procedure is a measure of its capacity to remain
unaffected by small, but deliberate variations in method parameters. Robustness
provides an indication of its reliability during normal usage.
10. SYSTEM SUITABILITY
A procedure run prior to the individual analytical analysis to demonstrate that the
instrument, column, mobile phase, etc., parameters are within defined criteria.
Adequate system suitability is demonstrated before proceeding with the analysis.

Sterilization Process Validation in Applications for Human and Veterinary Drug Products

TABLE OF CONTENTS
I. INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
A. Purpose . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
B. Documenting Sterilization Process Validation . . . . . . . . . . . . . . . . . . . 2
C. Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
II. INFORMATION FOR TERMINAL MOIST HEAT STERILIZATION
PROCESSES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
A. Description of the Process and Product . . . . . . . . . . . . . . . . . . . . . . . . 3
1. The Drug Product and Container-Closure System . . . . . . . . . . 3
2. The Sterilization Process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. The Autoclave Process and Performance Specifications . . . . . 4
4. Autoclave Loading Patterns . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
5. Methods and Controls to Monitor Production Cycles . . . . . . . . . 4
6. Requalification of Production Autoclaves . . . . . . . . . . . . . . . . . . 4
7. Reprocessing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
B. Thermal Qualification of the Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
1. Heat Distribution and Penetration Studies . . . . . . . . . . . . . . . . . 4
2. Thermal Monitors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3. The Effects of Loading on Thermal Input . . . . . . . . . . . . . . . . . . 5
4. Information Included in the Batch Record . . . . . . . . . . . . . . . . . 5
C. Microbiological Efficacy of the Cycle . . . . . . . . . . . . . . . . . . . . . . . . . . 5
1. Identification and Characterization of Bioburden Organisms . . . 6
2. Specifications for Bioburden . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3. Identification, Resistance, and Stability of Biological
Indicators . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
4. The Resistance of the Biological Indicator Relative to That
of Bioburden . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
5. Microbiological Challenge Studies . . . . . . . . . . . . . . . . . . . . . . . 7
D. Microbiological Monitoring of the Environment . . . . . . . . . . . . . . . . . . . 7
E. Container-Closure and Package Integrity . . . . . . . . . . . . . . . . . . . . . . . . 7
1. Simulation of the Stresses From Processing . . . . . . . . . . . . . . . . 7
2. Demonstrate Integrity Following the Maximum Exposure . . . . . . 8
3. Multiple Barriers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
4. The Sensitivity of the Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
5. Integrity Over the Product Shelf Life . . . . . . . . . . . . . . . . . . . . . . 8
F. Bacterial Endotoxins Test and Method . . . . . . . . . . . . . . . . . . . . . . . . . . 8
G. Sterility Testing Methods and Release Criteria . . . . . . . . . . . . . . . . . . . . 8
H. Evidence of Formal, Written Procedures . . . . . . . . . . . . . . . . . . . . . . . . 9
III. OTHER TERMINAL STERILIZATION PROCESSES . . . . . . . . . . . . . . . . . . . . 9
A. Ethylene Oxide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
1. Description of the Sterilizer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
2. Cycle Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
3. Microbiological Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
4. Stability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
B. Radiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
1. The Facility and the Process . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2. The Packaging of the Product . . . . . . . . . . . . . . . . . . . . . . . . . . 10
3. Multiple-Dose Mapping Studies . . . . . . . . . . . . . . . . . . . . . . . . . 10
4. Microbiological Methods and Controls . . . . . . . . . . . . . . . . . . . 11
5. Monitoring Stability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
IV. INFORMATION FOR ASEPTIC FILL MANUFACTURING PROCESSES
WHICH SHOULD BE INCLUDED IN DRUG APPLICATIONS . . . . . . . . . . . 11
A. Buildings and Facilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1. Floor Plan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
2. Location of equipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
B. Overall Manufacturing Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
1. Drug Product Solution Filtration . . . . . . . . . . . . . . . . . . . . . . . . . 12
2. Specifications Concerning Holding Periods . . . . . . . . . . . . . . . . 12
3. Critical Operations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
C. Sterilization and Depyrogenation of Containers, Closures,
Equipment, and Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
1. Bulk Drug Solution Components That are Sterilized
Separately . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
2. Sterilization Information in the Batch Records . . . . . . . . . . . . . . 13
D. Procedures and Specifications for Media Fills . . . . . . . . . . . . . . . . . . . 13
E. Actions Concerning Product When Media Fills Fail . . . . . . . . . . . . . . . 14
F. Microbiological monitoring of the environment . . . . . . . . . . . . . . . . . . . 15
1. Microbiological Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2. Yeasts, Molds, and Anaerobic Microorganisms . . . . . . . . . . . . . 15
3. Exceeded Limits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
G. Container-Closure and Package Integrity . . . . . . . . . . . . . . . . . . . . . . . 15
H. Sterility Testing Methods and Release Criteria . . . . . . . . . . . . . . . . . . . 16
I. Bacterial Endotoxins Test and Method . . . . . . . . . . . . . . . . . . . . . . . . . 16
J. Evidence of Formal Written Procedures . . . . . . . . . . . . . . . . . . . . . . . . 16
V. MAINTENANCE OF MICROBIOLOGICAL CONTROL AND QUALITY:
STABILITY CONSIDERATIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
A. Container-Closure Integrity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
B. Preservative Effectiveness . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
C. Pyrogen or Endotoxin Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
VI. ADDITIONAL INFORMATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
1This guidance has been prepared by the Sterility Technical Committee of the
Chemistry Manufacturing Controls Coordinating Committee of the Center for Drug
Evaluation and Research (CDER), and the Center for Veterinary Medicine (CVM), at the
Food and Drug Administration. Although this guidance does not create or confer any
rights for or on any person and does not operate to bind FDA or the industry, it does
represent the agency’s current thinking on sterilization process validation documentation.
For additional copies of this guidance, contact the Division of Communications
Management, HFD-210, CDER, FDA, 5600 Fishers Lane, Rockville, MD 20857 (Phone:
301-594-1012) Send one self-addressed adhesive label to assist the office in processing
your request. An electronic version of this guidance is also available via Internet via World
Wide Web (WWW) (connect to the FDA Home Page at WWW.FDA.GOV/CDER and go
to the “Regulatory Guidance” section).
GUIDANCE FOR INDUSTRY1
FOR THE SUBMISSION OF
DOCUMENTATION FOR STERILIZATION PROCESS
VALIDATION IN APPLICATIONS FOR HUMAN AND
VETERINARY DRUG PRODUCTS
I. INTRODUCTION
A. Purpose
This document is intended to provide guidance for the submission of
information and data in support of the efficacy of sterilization processes in
drug applications for both human and veterinary drugs. The
recommendations in the guidance apply to applications for sterile drug
products (new drug applications, new animal drug applications,
abbreviated new drug applications, abbreviated antibiotic applications,
and abbreviated new animal drug applications). These recommendations
also apply to previously approved applications when supplements
associated with the sterile processing of approved drugs are submitted.
Information and data in support of sterility assurance may also be
necessary in investigational new drug and investigational new animal
drug applications.
In the FEDERAL REGISTER of October 11, 1991 (56 FR 51354), the
agency published a proposed rule entitled "Use of Aseptic Processing
and Terminal Sterilization in the Preparation of Sterile Pharmaceuticals
for Human and Veterinary Use." This guidance is not a substitution for or
a supplement to that proposed rule. Regardless of whether the applicant
uses terminal sterilization or aseptic processing to manufacture a drug
product that is purported to be sterile, certain information about the
validation of that process should be submitted for both of those types of
sterilization.
B. Documenting Sterilization Process Validation
The efficacy of a given sterilization process for a specific drug product is
evaluated on the basis of a series of protocols and scientific experiments
designed to demonstrate that the sterilization process and associated
control procedures can reproducibly deliver a sterile product. Data
derived from experiments and control procedures allow conclusions to be
drawn about the probability of nonsterile product units (sterility assurance
level). Based on the scientific validity of the protocols and methods, as
well as on the scientific validity of the results and conclusions, the agency
concludes that the efficacy of the sterilization process is validated.
Whether a drug product is sterilized by a terminal sterilization process or
by an aseptic filling process, the efficacy of the sterilization process may
be validated without the manufacture of three production batches.
Sterilization process validation data, however, should be generated using
procedures and conditions that are fully representative and descriptive of
the procedures and conditions proposed for manufacture of the product in
the application.
The Center for Drug Evaluation and Research's (CDER's) and the Center
for Veterinary Medicine's (CVM's) review of the validation of the
sterilization process consists of a scientific evaluation of the studies
submitted in the applications. This review is conducted by FDA's review
staff, and is part of a cooperative effort between the review staff,
compliance staff, and field investigators to ensure the overall state of
control of the sterile processing of human and veterinary drug products.
Information and data in support of sterility assurance may be provided
directly to the application or by specific reference to a drug master file
(DMF), a veterinary master file (VMF), or another application. Letters of
authorization to refer to the referenced files should be included.
C. Remarks
This guidance is intended to provide recommendations for the types of
information applicants should include in human and animal drug
applications. Regulatory requirements for the submission of information
and data in various applications are specified in the sections listed below:
1. Human Drugs:
Investigational new drug applications 21 CFR 312.23(a)(7)
New drug applications 21 CFR 314.50
Abbreviated new drug and abbreviated
antibiotic drug applications 21 CFR 314.94 and 314.50
Supplements to NDA's and ANDA's 21 CFR 314.70
2. Animal Drugs:
Investigational new animal drug applications 21 CFR Part 511
New animal drug applications 21 CFR 514.1
Supplements to NADA's 21 CFR 514.8
II. INFORMATION FOR TERMINAL MOIST HEAT STERILIZATION
PROCESSES
The following types of information should be submitted in support of sterility
assurance for products produced using terminal moist heat sterilization.
Although the following outline directly addresses moist heat processes, the
same types of information would generally pertain to other terminal sterilization
processes (e.g., ethylene oxide or radiation). (See section III of this guidance.)
The following information should be submitted for each facility to be used in the
manufacture of the proposed drug product:
A. Description of the Process and Product
1. The Drug Product and Container-Closure System
Descriptions of the drug product and the container-closure
system(s) to be sterilized (e.g., size(s), fill volume, or secondary
packaging).
2. The Sterilization Process
A description of the sterilization process used to sterilize the drug
product in its final container-closure system, as well as a
description of any other sterilization process(es) used to sterilize
delivery sets, components, packaging, bulk drug substance or bulk
product, and related items. Information and data in support of the
efficacy of these processes should also be submitted. (See also
sections II.B. and II.C. of this guidance.)
3. The Autoclave Process and Performance Specifications
A description of the autoclave process, including pertinent
information such as cycle type (e.g., saturated steam, water
immersion, and water spray), cycle parameters and performance
specifications including temperature, pressure, time, and minimum
and maximum Fo. Identify the autoclave(s) to be used for
production sterilization, including manufacturer and model.
4. Autoclave Loading Patterns
A description of representative autoclave loading patterns should
be provided.
5. Methods and Controls to Monitor Production Cycles
Methods and controls used to monitor routine production cycles
(e.g., thermocouples, pilot bottles, and biological indicators) should
be described, including the number and location of each as well as
acceptance and rejection specifications.
6. Requalification of Production Autoclaves
A description of the program for routine and unscheduled
requalification of production autoclaves, including frequency,
should be provided.
7. Reprocessing
A description and validation summary of any program that provides
for reprocessing (e.g., additional thermal processing) of product
should be provided. Please note that the stability program is also
affected by additional thermal processing. For further information
concerning the stability program, reference is made to the Center
for Drug Evaluation and Research "Guideline for Submitting
Documentation for the Stability of Human Drugs and Biologics"
and to the Center for Veterinary Medicine "Drug Stability
Guideline."
B. Thermal Qualification of the Cycle
1. Heat Distribution and Penetration Studies
Heat distribution and penetration study protocols and data
summaries that demonstrate the uniformity, reproducibility, and
conformance to specifications of the production sterilization cycle
should be provided. Results from a minimum of three consecutive,
successful cycles should be provided to ensure that the results are
consistent and meaningful.
2. Thermal Monitors
The number of thermal monitors used and their location in the
chamber should be described. A diagram is helpful.
3. The Effects of Loading on Thermal Input
Data should be generated with minimum and maximum load to
demonstrate the effects of loading on thermal input to product.
Additional studies may be necessary if different fill volumes are
used in the same container line. Data summaries are acceptable
for these purposes. A summary should consist of, for example,
high and low temperatures (range), average temperature during
the dwell period, minimum and maximum F0 values, dwell time, run
date and time, and identification of the autoclave(s) used. These
data should have been generated from studies carried out in
production autoclave(s) that will be used for sterilization of the
product that is the subject of the application.
4. Information Included in the Batch Record
The batch record supplied with the chemistry, manufacturing, and
controls section of the application should identify the validated
processes to be used for sterilization and for depyrogenation of
any container-closure components. This information can be
included in the batch record by reference to the validation protocol
or standard operating procedure (SOP). Validation information
should be provided as described above.
C. Microbiological Efficacy of the Cycle
Validation studies that demonstrate the efficacy (lethality) of the
production cycle should be provided. A sterility assurance of 10-6 or
better should be demonstrated for any terminal sterilization process. This
level of sterility assurance should be demonstrated for all parts of the
drug product (including the container and closure, if applicable), which
are claimed to be sterile. The specific type of study and the methods
used to carry out the study (or studies) are product and process specific
and may vary from manufacturer to manufacturer. In general, the
following types of information and data should be provided.
1. Identification and Characterization of Bioburden Organisms
Describe the methods and results from studies used to identify and
characterize bioburden organisms. The amount and type of
information supplied may be dependent on the validation strategy
chosen. For example, more information may be needed for
bioburden-based autoclave processes than for overkill processes.
Information concerning the number, type, and resistance of
bioburden organisms may be necessary, including those
organisms associated with the product solution and the container
and closure. It may be necessary to identify the most heatresistant
bioburden organisms.
2. Specifications for Bioburden
Specifications (alert and action levels) for bioburden should be
provided. A description should be included of the program for
routinely monitoring bioburden to ensure that validated and
established limits are not exceeded (e.g., frequency of analysis
and methods used in bioburden screening). The methods
provided should be specific.
3. Identification, Resistance, and Stability of Biological Indicators
Information and data concerning the identification, resistance (D
and Z values), and stability of biological indicators used in the
biological validation of the cycle should be provided. If biological
indicators are purchased from a commercial source, it may be
necessary to corroborate the microbial count and resistance, and
provide performance specifications.
4. The Resistance of the Biological Indicator Relative to That of
Bioburden
Studies characterizing the resistance of the biological indicator
relative to that of bioburden may be necessary. Resistance in or
on the product (i.e., in the product solution, or on the surface of
container or closure parts or interfaces) should be determined as
necessary. If spore carriers are used (e.g., spore strips), the
resistance of spores on the carrier relative to that of directly
inoculated product should be determined, if necessary.
5. Microbiological Challenge Studies
Microbiological validation studies should be submitted that
demonstrate the efficacy of the minimum cycle to provide a sterility
assurance of 10-6 or better to the product under the most difficult to
sterilize conditions (e.g., the most difficult to sterilize load with
biological indicators at microbiological master sites or in master
product or both). Use of a microbiological master product or site
should be supported by scientific data. Microbiological master
sites or solutions are those sites or solutions in which it is most
difficult to kill the biological indicator under sterilization cycles that
simulate production conditions.
D. Microbiological Monitoring of the Environment
Section 211.160 of the Code of Federal Regulations requires, in part, the
establishment of scientifically sound and appropriate specifications,
standards, sampling plans, and test procedures designed to ensure that
components, drug product containers, closures, in-process materials, and
drug products conform to appropriate quality standards. Therefore, a
microbiological monitoring program for production areas along with a
bioburden monitoring program for product components and process water
should be established. Process water includes autoclave cooling water.
Applicants should provide information concerning this program.
Frequency, methods used, action levels, and data summaries should be
included. A description of the actions taken when specifications are
exceeded should be provided.
E. Container-Closure and Package Integrity
An applicant should provide scientific validation studies (and data) in
support of the microbial integrity of the drug packaging components. The
following types of information should be included:
1. Simulation of the Stresses from Processing
Experimental designs should simulate the stresses of the
sterilization process, handling, and storage of the drug and their
effects on the container-closure system. Physical, chemical, and
microbiological challenge studies may be necessary.
2. Demonstrate Integrity Following the Maximum Exposure
Container-closure integrity should be demonstrated on product
units that have been exposed to the maximum sterilization
cycle(s). If a product is exposed to more than one process, then
exposure to the maximum cycle of all processes should be
incorporated into the study design.
3. Multiple Barriers
Each barrier that separates areas of the drug product claimed to
be sterile should be separately evaluated and validated.
4. The Sensitivity of the Test
The sensitivity of the experimental method used for containerclosure
integrity testing should be specified and provided.
5. Integrity Over the Product Shelf Life
Microbial integrity of the container-closure system should be
demonstrated over the shelf life of the product. (See section V.A.
of this guidance.)
F. Bacterial Endotoxins Test and Method
The bacterial endotoxins test used for the product should be described.
The description should include qualification of the laboratory, inhibition
and enhancement testing and results, determination of noninhibitory
concentration and maximum valid dilution. For further information see the
agency guidance entitled "Guideline on Validation of the Limulus
Amebocyte Lysate Test As An End-Product Endotoxin Test for Human
And Animal Parenteral Drugs, Biological Products, and Medical Devices."
G. Sterility Testing Methods and Release Criteria
Sterility test methods should be described and should include the protocol
for the selection of representative units during production. When test
methods differ significantly from compendial test methods, a
demonstration of the equivalency to the compendial method should be
provided. Testing performed within barrier systems should be described,
and information concerning validation of the barrier system may be
necessary.
H. Evidence of Formal, Written Procedures
Section 211.113(b) of the Code of Federal Regulations requires that
written procedures, designed to prevent microbiological contamination of
drug products purporting to be sterile, be established and followed. Such
procedures should include validation of any sterilization process.
Therefore, evidence should be provided that there are formal, written
procedures describing the elements listed above and that these
procedures are followed. Such evidence may consist of SOP's, listing of
SOP's, and protocols submitted as part of these elements.
III. OTHER TERMINAL STERILIZATION PROCESSES
Although the information above (sections I.A. through I.G. of this guidance)
directly addresses moist heat processes, the same type of information would
pertain to other terminal sterilization processes used singly or in combination to
sterilize a drug product. The types of information outlined are, in general, also
applicable to ethylene oxide and radiation (gamma and electron beam). These
other processes should be addressed as each applies to the drug product,
sterile packaging and in-process sterilization of components. Examples of such
information might include: descriptions of loading configurations; qualification
and validation of master load configurations; determination and validation of the
efficacy of the minimum cycle to provide sterility assurance at the product
master sites; requalification of the cycle; provisions for resterilization;
specifications and monitoring program for product bioburden; and containerclosure
integrity. Specific examples are provided below to demonstrate the
application of these concepts to other sterilization processes.
Additional information relating to the effects of the sterilization process on the
chemical and physical attributes of the drug substance or drug product may be
applicable, and should be supplied to the chemistry, manufacturing, and controls
section of the application.
A. Ethylene Oxide
1. Description of the Sterilizer
The sterilizer(s) and controlled site(s) for prehumidification and
aeration of the product load should be described.
2. Cycle Parameters
The parameters and limits for all phases of the cycle, e.g.,
prehumidification, gas concentration, vacuum and gas pressure
cycles, exposure time and temperature, humidity, degassing,
aeration, and determination of residuals should be specified.
Specific procedures used to monitor and control routine production
cycles to assure that performance is within validated limits should
be provided.
3. Microbiological Methods
The microbiological methods (growth medium, incubation
temperature, and time interval) for cultivating spores from
inoculated samples during validation experiments should be
described as well as the microbiological methods used as part of
routine production cycles.
4. Stability
The program for monitoring the stability of packaging and the
integrity of the container-closure system barrier over the claimed
shelf life should be described.
B. Radiation
1. The Facility and the Process
The radiation facility should be identified. The radiation source,
method of exposure (i.e., movement through the irradiator), and
the type and location of dosimeters used to monitor routine
production loads should be described. If the low dose site is not
used for routine monitoring, data that show the dose relationship
between the two sites should be provided.
2. The Packaging of the Product
The packaging of the drug product within the shipping carton and
within the carrier should be described.
3. Multiple-Dose Mapping Studies
Multiple-dose mapping studies for identification of low and high
dose sites and demonstration of uniformity and reproducibility of
the process should be described.
4. Microbiological Methods and Controls
The microbiological methods and controls used to establish,
validate, and audit the efficacy of the cycle should be described.
5. Monitoring Stability
The program for monitoring the stability of packaging and the
integrity of the container-closure system barrier over the claimed
shelf life should be described.
IV. INFORMATION FOR ASEPTIC FILL MANUFACTURING PROCESSES
WHICH SHOULD BE INCLUDED IN DRUG APPLICATIONS
The following types of information should be submitted in support of sterility
assurance for products manufactured by aseptic processing.
A. Buildings and Facilities
A brief description of the manufacturing building and facilities should be
provided. The following information should be included:
1. Floor Plan
A floor plan of the areas holding the aseptic filling facilities
including preparation and holding areas, filtering and filling areas,
and gowning rooms should be included. The air cleanliness class
of each area should be identified (e.g., Class 100, Class 10,000,
Class 100,000). Isolators or barrier systems should be identified.
2. Location of Equipment
The placement of all critical equipment, including, but not limited to,
laminar flow hoods, autoclaves, lyophilizers, and filling heads,
should be identified. Equipment within barrier or isolation systems
should be noted.
B. Overall Manufacturing Operation
The overall manufacturing operation including, for example, material flow,
filling, capping, and aseptic assembly, should be described. The normal
flow (movement) of product and components from formulation to finished
dosage form should be identified and indicated on the floor plan
described above. The following information should be considered when
describing the overall manufacturing operation:
1. Drug Product Solution Filtration
The specific bulk drug product solution filtration processes,
including tandem filter units, prefilters, and bacterial retentive
filters, should be described. A summary should be provided
containing information and data concerning the validation of the
retention of microbes and compatibility of the filter used for the
specific product. Any effects of the filter on the product formulation
should be described (e.g., adsorption of preservatives or active
drug substance, or extractables).
2. Specifications Concerning Holding Periods
Section 211.111 of the Code of Federal Regulations requires, in
part, when appropriate, the establishment of time limits for
completing each phase of production to ensure the quality of the
drug product. Therefore, specifications concerning any holding
periods between the compounding of the bulk drug product and its
filling into final containers should be provided. These
specifications should include, for example, holding tanks, times,
temperatures, and conditions of storage. Procedures used to
protect microbiological quality of the bulk drug during these holding
periods should be indicated. Maintenance of the microbiological
quality during holding periods may need verification.
3. Critical Operations
The critical operations that expose product or product contact
surfaces to the environment (such as transfer of sterilized
containers or closures to the aseptic filling areas) should be
described. Any barrier or isolation systems should be described.
C. Sterilization and Depyrogenation of Containers, Closures,
Equipment, and Components
The sterilization and depyrogenation processes used for containers,
closures, equipment, components, and barrier systems should be
described. A description of the validation of these processes should be
provided including, where applicable, heat distribution and penetration
summaries, biological challenge studies (microbiological indicators and
endotoxin) and routine monitoring procedures. Validation information for
sterilization processes other than moist heat should also be included.
Methods and data (including controls) demonstrating distribution and
penetration of the sterilant and microbiological efficacy of each process
should be submitted. The section of this guidance concerning terminal
sterilization contains information that may be of further assistance.
1. Bulk Drug Solution Components That are Sterilized Separately
If the bulk drug solution is aseptically formulated from components
that are sterilized separately, information and data concerning the
validation of each of these separate sterilization processes should
be provided.
2. Sterilization Information in the Batch Records
The completed batch record supplied with the chemistry,
manufacturing, and controls section of the application should
identify the validated processes to be used for sterilization and
depyrogenation of any container-closure components. This
information may be included in the batch record by reference to the
validation protocol or SOP.
D. Procedures and Specifications for Media Fills
The procedures and specifications used for media fills, and summaries of
results for validation using the same container- closure system and filling
process that is to be used for the product should be described. The
microbiological testing method(s) used should be described. Any
procedural differences between the media fill and the production process
should be indicated. A summary of recent media fill results, including
failures, should be provided. These data should be obtained using the
same filling line(s) that are to be used for the drug product. The following
are recommended to be included with the data summary for each media
fill run described:
1. The filling room
Identify the aseptic filling area used and relate this to the floor plan
provided in section IV.A.1 of this guidance.
2. Container-closure type and size
3. Volume of medium used in each container
4. Type of medium used
5. Number of units filled
6. Number of units incubated
7. Number of units positive
8. Incubation parameters
The incubation time and temperature for each group of units
incubated and specifications for any group of units subjected to
two (or more) different temperatures should be specified.
9. Date of each media fill
10. Simulations
The procedures used to simulate any steps of a normal production
fill should be described. This might include, for example, slower
line speed, personnel shift changes, equipment failure and repair,
mock lyophilization and substitution of vial headspace gas.
11. Microbiological monitoring
The microbiological monitoring data obtained during the media fill
runs should be provided (see section IV.F. of this guidance).
12. Process parameters
The parameters used for production filling and for media fills (e.g.,
line speed, fill volume, number of containers filled, or duration of
fill) should be compared.
E. Actions Concerning Product When Media Fills Fail
The disposition of product made before and after a failed media fill should
be described. The description should include details of investigations,
reviews, and how decisions are made to reject or release product.
F. Microbiological Monitoring of the Environment
The microbiological monitoring program used during routine production
and media fills should be described. The frequency of monitoring, type of
monitoring, sites monitored, alert and action level specifications, and
precise descriptions of the actions taken when specifications are
exceeded should be included.
1. Microbiological Methods
The microbiological materials and methods used in the
environmental monitoring program should be described. Methods
may include sample collection, transport, neutralization of
sanitizers, incubation, and calculation of results. The following are
sources of microbial contamination and their monitoring that should
be addressed, including specifications:
a. Airborne microorganisms
b. Microorganisms on inanimate surfaces
c. Microorganisms on personnel
d. Water systems
e. Product component bioburden
2. Yeasts, Molds, and Anaerobic Microorganisms
A description of periodic or routine monitoring methods used for
yeasts, molds, and anaerobes should be provided.
3. Exceeded Limits
A description of the actions taken when specifications are
exceeded should be provided.
G. Container-Closure and Package Integrity
The methods and results demonstrating the integrity of the
microbiological barrier of the container-closure system should be
summarized. This should include testing for initial validation. The
procedures used for the stability protocol also should be described. For
initial validation of microbiological integrity of container-closure systems,
product sterility testing is not normally considered sufficient. The
sensitivity of the experimental method used for container-closure integrity
testing should be specified and provided.
H. Sterility Testing Methods and Release Criteria
Sterility test methods should be described and should include the protocol
for the selection of representative units during production. For a drug
product represented to be a drug recognized in an official compendium,
when test methods differ significantly from official compendial test
methods, a demonstration of the equivalency to the official compendial
method should be provided. Testing performed within barrier systems
should be discussed, and information concerning validation of the barrier
system may be necessary.
I. Bacterial Endotoxins Test and Method
The bacterial endotoxins test used for the product should be described, if
applicable. This description should include qualification of the laboratory,
inhibition and enhancement testing and results, determination of
noninhibitory concentration and maximum valid dilution. For further
information see the agency guidance entitled "Guidance on Validation of
the Limulus Amebocyte Lysate Test As An End-Product Endotoxin Test
for Human And Animal Parenteral Drugs, Biological Products, and
Medical Devices."
J. Evidence of Formal Written Procedures
Evidence should be provided that there are formal, written procedures
describing the above elements and that these procedures are followed.
Such evidence may consist of SOP's or a listing of SOP's or protocols
submitted as part of the elements listed above.
V. MAINTENANCE OF MICROBIOLOGICAL CONTROL AND QUALITY:
STABILITY CONSIDERATIONS
A. Container-Closure Integrity
The ability of the container-closure system to maintain the integrity of its
microbial barrier, and, hence, the sterility of a drug product throughout its
shelf life, should be demonstrated. Reference is made to sections II.E.
and IV.G. of this guidance. As previously stated, sterility testing at the
initial time point is not considered sufficient to demonstrate the microbial
integrity of a container-closure system. Documentation of the sensitivity
of the container-closure integrity test should be provided.
B. Preservative Effectiveness
The efficacy of preservative systems to control bacteria and fungi
inadvertently introduced during drug product use should be demonstrated
at the minimum concentration specified for drug product release or at the
minimum concentration specified for the end of the expiration dating
period, whichever is less. Since the efficacy of preservative systems is
judged by their effect on microorganisms, microbial challenge assays
should be performed. The United States Pharmacopeia (USP) provides
a microbial challenge assay under the title "Antimicrobial Preservatives-
Effectiveness." For purposes of the stability protocol, the first three
production lots should be tested with a microbial challenge assay at the
beginning and end of the stability period. Chemical assays to monitor the
concentration of preservatives should be performed at all test intervals.
For subsequent lots placed on stability, chemical assays may be
adequate to demonstrate the presence of specified concentrations of
preservatives, and such testing should be carried out according to the
approved stability study protocol.
C. Pyrogen or Endotoxin Testing
For drug products purporting to be pyrogen free, it is recommended that
pyrogen or endotoxin tests be carried out at the beginning and end of the
stability period as part of the approved stability study protocol.
VI. ADDITIONAL INFORMATION
Further information concerning content and format of drug applications is
available in the form of guidances and other publications. The following
documents contain information related to the topics discussed in this guidance:
"Guideline for Submitting Documentation for the Stability of Human Drugs and
Biologics" (CDER).
"Guideline on Validation of the Limulus Amebocyte Lysate Test as an End-
Product Endotoxin Test for Human and Animal Parenteral Drugs, Biological
Products, and Medical Devices" (CDER, CVM, CBER, CDRH).
"Guideline on Sterile Drug Products Produced by Aseptic Processing" (CDER).
"Drug Stability Guideline" (CVM).
Submitted by:
_____________________________
Peter H. Cooney, PhD
Center for Drug Evaluation and Research
______________________________
William G. Marnane
Center for Veterinary Medicine
Approved by CMC CC:
_______________________________ ______________________________
Charles Kumkumian, Ph.D. Roger L. Williams, M.D.