美國FDA分析方法驗證指南中英文對照（四）

VII. METHODS VALIDATION FOR NDAs, ANDAs, BLAs, AND PLAs

A. Noncompendial Analytical Procedures

In an NDA, ANDA, BLA, or PLA, data must be submitted to establish that the analytical procedures used in testing meet proper standards of accuracy and reliability (21 CFR 211.194(a)(2)). Methods validation is the process of demonstrating that analytical procedures are suitable for their intended use. At the time of submission, the NDA, ANDA, BLA, or PLA should contain methods validation information to support the adequacy of the analytical procedures.

VII．NDA，ANDA，BLA和PLA中的分析方法驗證

A．非藥典分析方法

在NDA，ANDA，BLA或PLA中，應當要遞交資料以說明檢測中所用的分析方法是滿足適當的準確度和可靠性要求(21 CFR 211.194(a)92))。分析方法驗證是個論述分析方法是適用于其擬定用途的過程。在遞交資料，NDA，ANDA，BLA或PLA中應當要包含分析方法驗證資料以支持分析方法的準確度。

The International Conference on Harmonisation (ICH) guidance Q2A Text on Validation of Analytical Procedures (March 1995) and Q2B Validation of Analytical Procedures: Methodology (November 1996) provide recommendations on validation of analytical procedures. Analytical procedures outside the scope of the ICH guidances should still be validated.

ICH指導原則Q2A：分析方法驗證（1995年3月）和Q2B：分析方法驗證：方法學（1996年11月）給出了分析方法驗證的建議。對于超出ICH指導原則范圍的分析方法也是需要驗證的。

1. Validation Characteristics

Applicants should submit information on the validation characteristics of their proposed analytical procedures (see ICH Q2A and ICH Q2B). Although not all of the validation characteristics are needed for all types of tests (see section VII.A.3), typical validation characteristics are:

1.驗證項目

申請者應當要送交其所擬定分析方法的驗證項目方面的信息（見ICH　Q2A和ICH Q2B）。盡管不是對于所有類型的分析方法都需要進行所有的驗證項目（見第VII.A.3章），但還是有典型的驗證項目，如：

• Accuracy   準確度
• Precision (repeatability and intermediate precision)   精密度（重復性和中間精密度）
• Specificity   專屬性
• Detection limit   檢測限
• Quantitation limit   定量限
• Linearity   線性
• Range   范圍
• Robustnes  耐用性

2. Other Methods Validation Information

Methods validation information should also include:

Data to demonstrate the stability of all analytical sample preparations through the time required to complete the analysis.

2.其它驗證資料

分析方法驗證資料還應當要包括：

說明所有分析制備樣品在完成分析所需的時間內的穩定性的資料。

Legible reproductions of representative instrument output or recordings (e.g., chromatograms) and raw data output (e.g., integrated areas), as appropriate. Instrument output for placebo, standard, and sample should also be provided (see section VII.A.2.c).

清晰可讀的儀器代表性輸出和記錄資料（如色譜圖）和原始資料輸出（積分面積）。安慰劑，對照品和樣品的儀器輸出也都是需要提供的（見第VII.A.2.c章）。

Representative calculations using submitted raw data, to show how the impurities in drug substance are calculated. Information from stress studies (see section VII.A.2.b). Impurities labeled with their names and location identifiers (e.g., RRT for chromatographic data) for the impurity analytical procedure.

代表性計算公式，以表明原料藥中的雜質是如何計算的。(見VII.A.2.b章）。對于雜質分析方法，要標明雜質的名稱和位置標識符（如，色譜中的相對保留時間RRT）。

 
For drug substances:

A discussion of the possible formation and control of polymorphic and enantiomeric substances.

Identification and characterization of each organic impurity, as appropriate. This information may not be needed for all products (e.g., botanicals). Other impurities (e.g., inorganics, residual solvents) should be addressed and quantitated.

對于原料藥：

討論可能會形成的異構體并討論異構體的控制。

對每個有機雜質進行適當的標識和界定。不是所有的產品（如，植物藥）都需要這些資料的。對于其它雜質（如無機雜質，殘留溶劑），應當要進行說明并定量分析。

Recommendations on submitting information on impurities is provided in various FDA guidances such as the ICH guidance Q3A Impurities in New Drug Substances (January 1996).

A list of known impurities, with structure if available, including process impurities, degradants, and possible isomers.

FDA的指導文件，如比ICH指導原則Q3A 新原料藥中的雜質（1996年1月）。

已知雜質列表，包括工藝雜質，降解產物和可能的異構體。如果知道結構的話，也需提供。


For drug products:

A degradation pathway for the drug substance in the dosage form, 419where possible. Data demonstrating recovery from the sample matrix as illustrated by the accuracy studies. Data demonstrating that neither the freshly prepared nor the degraded placebo interferes with the quantitation of the active ingredient. ICH Q2A and Q2B address almost all of the validation parameters. Areas that should be provided in more detail are described below.

對于制劑：

原料藥在制劑中可能的降解途徑。通過準確度實驗論證的樣品回收率資料。要有資料論證無論是新制的安慰劑還是分解了的安慰劑都不會影響活性成分的定量分析。ICH　Q2A和ICH　Q2B幾乎對所有的驗證參數都進行了論述。下面論述的是那些還需要更詳細地進行論述的方面。

a. Robustness

Robustness, a measure of the analytical procedure's capability to remain unaffected by small but deliberate variations, is described in ICH Q2A and Q2B. Such testing should be performed during development of the analytical procedure and the data discussed and/or submitted. In cases where an effect is observed, representative instrument output (e.g., chromatograms) should be submitted.

a.耐用性

ICH　Q2A和ICH　Q2B對耐用性是有論述的，它衡量的是分析方法在細微的變化下不受影響的能力。該實驗應當是在分析方法開發過程中進行的，對實驗結果進行討論和/或遞交。如果觀察到有影響，需提供代表性儀器輸出（如色譜圖）。


b. Stress Studies

Degradation information obtained from stress studies (e.g., products of acid and base hydrolysis, thermal degradation, photolysis, oxidation) for the drug substance and for　the active ingredient in the drug product should be provided to demonstrate the specificity of the assay and analytical procedures for impurities. The stress studies should demonstrate that impurities and degradants from the active ingredient and drug product excipients do not interfere with the quantitation of the active ingredient. Stress studies are described in various FDA guidances relating to the stability of drug products (see references). The design of the stress studies and the results should be submitted to the stability section of the application. Representative instrument output (e.g., chromatograms) and/or other appropriate data (e.g., degradation information obtained from stress studies) should be submitted in the sections on analytical procedures and controls.

b.強降解實驗

FDA很多關于藥品穩定性的指導原則都對強降解實驗進行了論述。

在申請的穩定性一章中應當要提供的資料有：強降解實驗設計和實驗結果。而代表性儀器輸出（如：色譜圖）和/或其它資料（強降解實驗中所得的降解信息）應當要提供在分析方法和控制一章中。


c. Instrument Output/Raw Data

i. Organic Impurities

Representative data should be submitted to support an assessment of the organic impurities. Representative data for residual solvents are generally not needed. Instrument output and the raw numerical values (e.g., peak area) with appropriate identification and labeling (e.g., RT for chromatographic peaks, chemical shift (d) and coupling constant (J) for NMR) should be provided. The impurity profile should be assessed at the quantitation limit and the instrument output provided. Additional information should be provided to confirm that the impurity profile is adequately characterized. For example, a representative chromatogram using detection at a low wavelength, such as 205 nm, and double the proposed total run time could be submitted to support the specificity of the analytical procedure.

c.儀器輸出/原始資料

i. 有機雜質

應當要提供代表性資料以支持有機雜質的評估。一般來說，是不需要殘留溶劑的代表性資料的。儀器輸出和原始數值（比如，峰面積）及合適的標記和標注（比如，色譜峰的保留時間，核磁共振的化學位移和耦合常數）都應當要提供。根據所提供的定量限和儀器輸出對雜質情況進行評估。還應當要提供其它資料以確認雜質情況得到了充分地界定。比如說，比如說，代表性圖譜選用的檢測波長是205nm，則可以將擬定的運行時間延長至兩倍以支持分析方法的專屬性。

For quantitation purposes, the response factor of the drug substance may be
used for impurities without a reference standard. In cases where the response factors are not close, this practice may still be acceptable, provided a correction factor is applied or the impurities are, in fact, being overestimated. Acceptance criteria and analytical procedures used to estimate identified or unidentified impurities often are based on analytical assumptions (e.g., equivalent detector response). Assumptions should be discussed and justified.

在定量分析時，原料藥的響應因子可用于沒有相應對照品的雜質。如果響應因子不接近的話，只要應用了較正因子或或者雜質實際上是被高估的話，這樣做也是可行的。用于評估指定雜質或未指定雜質的合格標準和分析方法經常都是基于分析假設的（比如，相當的檢測器響應）。應當要對這些假設進行討論和合理性說明。

 
ii. Drug Substance

Data should be submitted showing the separation and detection of impurities using spiked or stress samples. Complete impurity profiles as graphic output (e.g., chromatograms) and raw data (e.g, integrated peak areas) of representative batches should be submitted in the sections on analytical procedures and controls for the drug substance. For ANDAs and related submissions, appropriate information for the batches used in the biobatch or submission batch should be provided. All responses (e.g., peaks) should be labeled. The analytical procedure used should be capable of differentiating changes, if any, between past and present batches. The quantitation limit and the type of organic impurity (e.g., degradant, process impurity) should be stated. The analytical procedure number, batch number, manufacturing date and site, and date of analysis should be provided.

ii.原料藥

ANDA和相關的遞交，用于生物利用度研究的批次（biobatch）或者提交批次（submission batch）的相關資料應當要提供。所有的響應（比如，色譜峰）都應當要進行標注。所用的分析方法應當要有能力區分先前批次和當前批次之間的變化，如果有這些變化的話。應當要說明定量限和有機雜質的類型（比如：降解物，工藝雜質）。還需提供分析方法編號，批號，生產日期和生產地點及分析日期。


iii. Drug Product

Information such as instrument output (e.g., chromatograms) and raw data (e.g., integrated peak areas) from representative batches under long-term and accelerated stability conditions, and stressed samples should be submitted in the sections on analytical procedures and controls of the drug product. For ANDAs and related submissions, appropriate information for the biobatch or submission batch should be provided. References to the raw data (e.g., chromatograms) should be included in the stability section of the application.

iii.制劑

在制劑的分析方法和控制一章中，應要當提供的資料有：代表性批號在長期和加速穩定性實驗條件下及強降解實驗條件下的儀器輸出（如，圖譜）和原始資料（如峰面積）。對于ANDA和相關遞交，應當要提供生物利用度實驗批次或遞交批次的適當資料。在申請的穩定性章節中應當要引用原始資料（比如：圖譜）。

At a minimum, the submission should include instrument output and raw data for release testing and at the latest available time point for the same batch. All responses (e.g., peaks) should be labeled and identified. In addition, the analytical procedure number, batch number of the drug product, manufacturing date, date of analysis, source and batch number of drug substance, manufacturing site, and container/closure information should be provided. The analytical procedures used should be capable of differentiating changes, if any, between past and present batches. The quantitation limit and the type (e.g., degradant, leachables from packaging) should be reported. Multiple methodologies can be used.

至少，遞交材料中應當要包括放行檢測(Release testing)的儀器輸出和原始資料. 需標注所有的響應信號（如色譜峰）。此外，還要提供，分析方法編號，制劑的批號，生產日期，分析日期，原料藥的來源和批號，生產地點及容器/密閉系統信息。如在過往批次和現批次之間存在差異的話，所用的分析方法應當要有能力區分出來。應當要報告檢測限和類型（如，降解物，包裝時的漏出物）。


3. Recommended Validation Characteristics for Types of Tests

Table 1 is a summary of the validation characteristics that should be addressed during validation of different types of analytical procedures. The same methodology can be used for several purposes. The validation information should support the intended purpose of the test. For example, if Raman spectroscopy is the methodology selected to quantitate polymorphic forms as impurities, or chiral HPLC for enantiomeric impurities, the recommended validation characteristics in Table 1 under quantitative testing for impurities would apply. However, if Raman spectroscopy or chiral HPLC are used for the purpose of identification or as specific tests, the recommended validation characteristics listed for those types of tests would apply.

3.各類檢測的推薦驗證項目

表1概述了在不同分析方法的驗證過程中所需要的驗證項目。同一分析方法可用于多個用途。驗證資料需要能支持該分析方法的擬定用途。比如說，如果拉曼光譜用于定量分析多晶型雜質，或手性HPLC用于分析異構體雜質，則要應用表1中雜質定量分析　中所推薦的驗證項目。然而，如果拉曼光譜或手性HPLC被用于鑒定或特征實驗的話，則要應用表1中所推薦的這些類型的驗證項目。


 NOTE注：

- Signifies that this characteristic is not normally evaluated. 表示通常不需要驗證的項目。

+ Signifies that this characteristic is normally evaluated. 表示通常需驗證的項目。

1 In cases where reproducibility has been performed, intermediate precision is not needed. 假如已經論證了重現性，可不需要再論證中間精密度。

2 Lack of specificity for an analytical procedure may be compensated for by the addition of a second analytical procedure. 如果一分析方法缺少專屬性的話，則需要另一分析方法進行補償。

3 May be needed in some cases. 有些情況下是需要的。

4 May not be needed in some cases.  有些情況下是不需要的。

5 Lack of specificity for an assay for release may be compensated for by impurities testing.  缺少專屬性數據時可用雜質測試的數據來補償。


a:Identification

Identification analytical procedures may include tests such as IR, differential scanning calorimetry (DSC), X-ray diffraction (XRD), UV, and HPLC retention time. A specific identification test should be included for the active ingredient whenever possible. In cases where a nonspecific identification analytical procedure is proposed for the active ingredient, two independent analytical procedures are generally sufficient, if justified. For other identification tests (e.g., a chiral HPLC retention time as confirmation for the presence of an enantiomer, chloride test for a counterion) a single test is acceptable. This concept of the number of identification tests is applicable to both the drug substance and drug product.

a. 鑒別

定性分析方法有：紅外(IR)，差示熱分析(DSC)，X-射線衍射(XRD)和HPLC保留時間。對于原料藥，盡可能要有專屬性強的鑒別實驗，如果用的是專屬性不強的定性分析方法，則通常來說運用兩個獨立的分析方法應當是足夠的。對于其它的一些定性分析方法(如，手性HPLC的保留時間用于確認是否存在異構體，平衡離子的氯化物實驗)，就一個檢測也是可可行的。定性實驗數目的概念既適用于原料藥，也適用于制劑。


b. Impurities

The validation characteristics under quantitative testing for impurities, as described in Table 1, apply, regardless of which methodology is used to quantitate impurities. If the same analytical procedure is proposed as a limit test, validation characteristics under limit testing for impurities will apply.

b. 雜質

表1中雜質定量分析 項下所說的相應驗證項目。如果同樣的方法還用做定性檢測的分析方法的話，則要對表1中雜質定性分析 項下的驗證項目進行驗證。


c. Assay

Assay includes the content of the active ingredient, preservative (if used), and measurement of content in dissolution and content uniformity samples.

c. 含量

含量分析包括活性成分的含量，所用到的防腐劑的含量，及溶出度和含量均勻度檢測樣品的含量測定。


d. Specific Tests

Specific tests to control the drug substance, excipient, or drug product can include tests such as particle size analysis, droplet distribution, spray pattern, dissolution (excludes measurement), optical rotation, and methodologies such as DSC, XRD, and Raman spectroscopy. The validation characteristics may differ for the various analytical procedures. For example, accuracy, repeatability, intermediate precision and robustness should be evaluated for molecular size distribution gel permeation chromatography (GPC).

d.特定實驗

用于原料藥，賦形劑或制劑控制的特定檢測包括粒徑分析，霧化狀態，溶出度，旋光度和比如DSC，XRD及拉曼光譜等方法。不同的分析方法所需的驗證項目是不盡相同的。比如說，對于凝膠滲透色譜(GPC)，需要考察其準確度，重復性，中間精密度和耐用性。



B. Compendial Analytical Procedures

The suitability of a compendial analytical procedure must be verified under actual conditions of use (21 CFR 211.194(a)(2)). Information to demonstrate that USP/NF analytical procedures are suitable for the drug product or drug substance should be included in the submission. Information on the specificity, intermediate precision, and stability of the sample solution should be included. Compendial assay analytical procedures may not be stability-indicating, and this should be considered when developing the specification (see section III.C). For compendial items, additional analytical procedures, such as impurities or osmolality, may be requested to support the quality of the drug product or drug substance. These additional analytical procedures should be validated (see section VII.A).

B．藥典分析方法(21CFR 211.194(a)(2))。

在申請中應當要提供藥典分析方法適用于該藥品或原料藥分析的論證資料。還應該要包括專屬性，中間精密度，和樣品溶液穩定性方面的資料。藥典含量分析方法可能是沒有穩定性指示能力的，因此在開發質量標準的過程中因當要考慮這個(見第III.C.章)。對于藥典項目，可能需要補充方法，比如雜質或滲透度，來控制制劑或原料藥的質量。這些補充方法也是需要驗證的(見第VII.A.章)。