藥典分析方法確認與驗證.doc

usp_1225 藥典方法的驗證 2011.02.26驗證: Validation 針對新方法(沒有法定方法, 需要用戶自己建立的) 按照ICH Q2 或USP的方式均可; 主要是需要滿足使用目的. VALIDATION OF COMPENDIAL PROCEDURES藥典規(guī)程的驗證Test procedures for assessment of the quality levels of pharmaceutical articles are subject to various requirements. According to Section 501 of the Federal Food, Drug, and Cosmetic Act, assays and specifications in monographs of the United States Pharmacopeia and the National Formulary constitute legal standards. The Current Good Manufacturing Practice regulations 21 CFR 211.194(a) require that test methods, which are used for assessing compliance of pharmaceutical articles with established specifications, must meet proper standards of accuracy and reliability. Also, according to these regulations 21 CFR 211.194(a)(2), users of analytical methods described in USP-NF are not required to validate the accuracy and reliability of these methods, but merely verify their suitability under actual conditions of use. Recognizing the legal status of USP and NF standards, it is essential, therefore, that proposal for adoption of new or revised compendial analytical procedures be supported by sufficient laboratory data to document their validity. 評估藥品質量水平的實驗方法受到多種要求的影響。要依據(jù)美國食品、藥品、化妝品法案501款以及美國藥典和國家處方集的各論中的含量測定和質量標準來構建法定標準。現(xiàn)行藥品生產(chǎn)質量管理規(guī)范【21 CFR 211.194(a)】要求，用于評價藥物與既有質量標準之間的符合性的分析規(guī)程必須在準確度和可靠性方面達到適當?shù)臉藴?。并且根?jù)【21 CFR 211.194(a)(2)】的這些規(guī)定，不要求USP-NF（美國藥典/國家處方集）中描述的分析方法/規(guī)程的使用者去驗證這些規(guī)程的準確度和可靠性，而僅需確認其在正確實際使用條件下的適用性。認識到USP和NF標準的法定地位，因此，提議采納新的或更改過的藥典分析規(guī)程時，必須有充分的實驗室數(shù)據(jù)作為支持，以記錄其有效性。The text of this information chapter harmonizes, to the extent possible, with the Tripartite International Conference on Harmonization (ICH) documents Validation of Analytical Procedures and the Methodology extension text, which are concerned with analytical procedures included as part of registration applications submitted within the EC, Japan, and the USA. 本信息章節(jié)的內容盡可能地與三方國際協(xié)調會議（ICH）文件分析規(guī)程的驗證和方法學的延伸內容保持一致，那是與包含歐盟、日本和美國注冊申請內的分析方法相關的。SUBMISSIONS TO THE COMPENDIA向藥典提交的文件Submissions to the compendia for new or revised analytical procedures should contain sufficient information to enable members of the USP Council of Experts and its Expert Committees to evaluate the relative merit of proposed procedures. In most cases, evaluations involve assessment of the clarity and completeness of the description of the analytical procedures, determination of the need for the procedures, and documentation that they have been appropriately validated. Information may vary depending upon the type of method involved. However, in most cases a submission will consist of the following sections. 向藥典提交關于新的或更改過的分析規(guī)程的文件應該包括充足的信息，以使USP專家理事會和專家委員會能夠評估擬議規(guī)程的相對優(yōu)越性。在大多數(shù)情況下，這些評估涉及對分析規(guī)程描述的清晰度和完整度的評估，對規(guī)程的需求的確定，以及它們已經(jīng)進行了適當驗證的記錄文件。這些信息可以根據(jù)所涉及規(guī)程的種類而變化。但是，在大多數(shù)情況下，提交的文件將有下面的章節(jié)組成。Rationale- This section should identify the need for the procedure and describe the capability of the specific procedure proposed and why it is preferred over other types of determinations. For revised procedures, a comparison should be provided of limitations of the current compendial procedure and advantages offered by the proposed procedure. 理論依據(jù)- 此部分應該辨明對于該規(guī)程的需求，并描述具體擬議中規(guī)程的能力，以及為什么優(yōu)于其他種類檢測。對于更改的方法/規(guī)程，應該提供對當前藥典規(guī)程之缺陷與擬議中規(guī)程之優(yōu)勢的比較。Proposed Analytical Procedure- This section should contain a complete description of the analytical procedure sufficiently detailed to enable persons “skilled in the art” to replicate it. The write-up should include all important operational parameters and specific instructions such as preparation of reagents, performance of system suitability tests, description of blanks used, precautions, and explicit formulas for calculation of test results.推薦的分析方法/規(guī)程- 此部分包含對該分析規(guī)程的完整描述，應足夠具體以便能讓業(yè)內技術熟練的人重復它。文章應該包括所有重要的操作參數(shù)和具體的指令，例如：試劑制備、系統(tǒng)適用性測試表現(xiàn)、所使用空白對照的描述、預防措施、用于計算測試結果的明確公式。Data Element- This section should provide thorough and complete documentation of the validation of the analytical procedure. It should include summaries of experimental data and calculations substantiating each of the applicable analytical performance characteristics. These characteristics are described in the following section. 數(shù)據(jù)要素- 此部分應該提供完全徹底的分析規(guī)程驗證記錄文件。其應該包括對于證明每一個實用功能特性的實驗數(shù)據(jù)和計算的概況、總結。這些特性在下面的部分描述。VALIDATION驗證Validation of an analytical procedure is the process by which it is established, by laboratory studies, that the performance characteristics of the procedure meet the requirements for the intended analytical applications. Typical analytical performance characteristics that should be considered in the validation of the types of procedures described in this document are listed in Table 1. Because opinions may differ with respect to terminology and use, each of the performance characteristics is defined in the next section of this chapter, along with a delineation of a typical method or methods by which it may be measured. 分析規(guī)程的驗證是，通過實驗室研究，確定該規(guī)程的工作特性達到了預定分析用途要求的過程。在此文件中所描述的規(guī)程種類的驗證中，應當考慮的常見分析工作特性在表1中列出。因為對于術語和使用的意見可能不同，在此通則的下個部分定義了每個工作性能，以及可以對其進行測量的常用的一個或幾個方法的描繪。Table 1. Typical Analytical Characteristics Used in Method Validation表1. 在方法驗證中使用的常用分析特性Accuracy準確度Precision精密度Specificity專屬性Detection Limit檢測限度Quantitation Limit定量限度Linearity線性Range范圍Robustness耐用性In the case of compendial procedures, revalidation may be necessary in the following cases: a submission to the USP of a revised analytical procedure; or the use of an established general procedure with a new product or raw material (see below in Data Elements Required for Validation).在藥典方法的情況下，在下面的情況有必要進行再驗證：向USP提交修改的分析規(guī)程；或將已確立的通用規(guī)程用于新產(chǎn)品或原料（見下面驗證必需的數(shù)據(jù)要素）The ICH documents give guidance on the necessity for revalidation in the following circumstances: changes in the synthesis of the drug substance; changes in the composition of the drug product; and changes in the analytical procedure.ICH文件對于下列情況下再驗證的必要性做出了指導：藥物合成過程有變更；藥品組成的有變更；以及分析規(guī)程中有變更。Analytical Performance Characteristics分析性能特征ACCURACY準確度Definition- The accuracy of an analytical procedure is the closeness of test results obtained by that procedure to the true value. The accuracy of an analytical procedure should be established across its range. 定義- 分析規(guī)程的準確度是由該規(guī)程得到的測試結果與真實值的接近程度。分析規(guī)程的準確度應通過它的適用范圍來建立。Determination- In the case of the assay of a drug substance, accuracy may be determined by application of the analytical procedure to an analyte to known purity (e.g., a Reference Standard) or by comparison of the results of the procedure with those of a second, well-characterized procedure, the accuracy of which has been stated or defined. 測定- 當對一種藥物進行分析時，準確度可以通過該分析規(guī)程來分析一個已知純度的物質（例如，某個標準物質）來進行測定，或者通過比較運用這個分析方法所得的結果與另一個已經(jīng)鑒定的、其準確度已被說明或被解釋過的方法所得的結果來進行測定。In the case of the assay of a drug in a formulated product, accuracy may be determined by application of the analytical procedure to synthetic mixtures of the drug product components to which known amounts of analyte have been added within the range of the procedure. If it is not possible to obtain samples of all drug product components, it may be acceptable either to add known quantities of the analyte to the drug product (i.e., “to spike”) or to compare results with those of a second, well-characterized procedure, the accuracy of which has been stated or defined. 當分析一種成品藥時，準確度可以通過分析規(guī)程來分析成藥各組分的合成物質來測定，而添加進去的分析物質是在方法的范圍之內的、是已知數(shù)量的。如果不能得到成藥的所有組分，也可以將已知數(shù)量的待分析物加入到該成藥中（例如，“增敏”），或者將結果與用第二種、成熟的、已知準確度的規(guī)程得到的結果進行比較，In the case of quantitative analysis of impurities, accuracy should be assessed on samples (of drug substance or drug product) spiked with known amount of impurities. Where it is not possible to obtain samples of certain impurities or degradation products, results should be compared with those obtained by an independent procedure. In the absence of other information, it may necessary to calculate the amount of an impurity based on comparison of its response to that of the drug substance; the ratio of the response of equal amounts of the impurity and the drug substance (relative response factor) should be used if known. 對于雜質的定量分析，應使用以已知數(shù)量雜質增敏的樣品來評估準確度。當不能獲得特定雜質或降解產(chǎn)物的樣品時，應將結果與另一獨立方法獲得的結果進行比較。在沒有其他信息的情況下，可能必需通過將某種雜質的響應值與藥物的響應值進行比較來計算雜質的數(shù)量；同等數(shù)量的雜質與藥物的響應值的比值（相對響應因子），如果已知，則應使用。Accuracy is calculated as the percentage of recovery by the assay of the known added amount of analyte in the sample, or as the difference between the mean and the accepted true value, together with confidence interval. 通過測定被加入到樣品中的已知數(shù)量的被分析物來計算準確度，得到回收百分比，或得到平均值與可接受的真實值之間的差異，同置信區(qū)間一起。The ICH documents recommended that accuracy should be assessed using a minimum of nine determinations over a minimum of three concentration levels, covering the specified range (i.e., three concentrations and three replicates of each concentration).ICH文件建議精密度的評估應當使用覆蓋規(guī)定范圍的至少三個濃度水平進行至少九次測試（例如，三個濃度并且每個濃度三次重復進樣）。Assessment of accuracy can be accomplished in a variety of ways, including evaluating the recovery of the analyte (percent recovery) across the range of the assay, or evaluating the linearity of the relationship between estimated and actual concentrations. The statistically preferred criterion is that the confidence interval for the slope be contained in an interval around 1.0, or alternatively, that the slope be close to 1.0. In either case, the interval or the definition of closeness should be specified in the validation protocol. The acceptance criterion will depend on the assay and its variability and on the product. Setting an acceptance criterion based on the lack of statistical significance of the test of the null hypothesis that the slop is 1.0 is not an acceptable approach. 準確度的評估可以通過多種不同的方式完成，包括評價在含量測定的整個范圍內被分析物的回收率，或評價估計濃度與實際濃度之間關系的線性。具統(tǒng)計學意義的標準有二，一是斜率的置信區(qū)間被限定在約1.0的區(qū)間，二是此斜率接近1.0。在任意一種情況下，此區(qū)間或接近程度的定義應該在驗證方案中明確規(guī)定。接受標準將取決于含量和其差異性，以及取決于該產(chǎn)品。通過測試證明該斜率為1.0的零假設沒有統(tǒng)計學意義，這樣的方法不能用于設定接受標準。PRECISION精密度Definition- The precision of an analytical procedure is the degree of agreement among individual test results when the procedure is applied separately to multiple samplings of a homogeneous sample. The precision of an analytical procedure is usually expressed as the standard deviation or relative standard deviation (coefficient of variation) of a series of measurements. Precision may be a measure of either the degree of reproducibility or of repeatability of the analytical procedure under normal operating conditions. In this context, reproducibility refers to the use of the analytical procedure in different laboratories, as in a collaborative study. Intermediate precision (as known as ruggedness) express within-laboratory variation, as on different days, or with different analysts or equipment within the same laboratory. Repeatability refers to the use of the analytical procedure within a laboratory over a short period of time using the same analyst with the same equipment. 定義- 分析規(guī)程的精密度是當該分析規(guī)程單獨分析均質樣品的多個樣本時，若干檢驗結果的一致程度。分析規(guī)程的精密度通常以一系列測量數(shù)值的標準差或相對標準差（變異系數(shù)）來表示。精密度可以是分析規(guī)程在普通操作條件下可重現(xiàn)性或可重復性程度的度量單位。在這樣的背景下，重現(xiàn)性指的是該分析規(guī)程在不同實驗室的應用，例如在一個協(xié)作實驗室里進行研究。中間精密度（也稱為“耐用性”）體現(xiàn)了在實驗室內的差異，如在相同的實驗室，但在不同的日期，或使用不同的分析員或設備?？芍貜托灾傅氖窃谕粋€實驗室內，一段較短的時間內，相同的分析員使用相同的設備、同一個分析方法的應用情況。Determination- The precision of an analytical procedure is determined by assaying a sufficient number of aliquots of a homogeneous sample to be able to calculate statistically valid estimates of standard deviation or relative standard deviation (coefficient of variation). Assays in this context are independent analyses of samples that have been carried through the complete analytical procedure from sample preparation to final test result. 測定-一個分析方法的精密度是通過對足夠的同一樣品的分析來計算有統(tǒng)計學意義的標準偏差或相對標準偏差來測定的。在這個背景下的分析是從樣品準備到最終實驗結果的完整分析程序的對樣品的獨立分析。The ICH documents recommended that repeatability should be assessed using a minimum of nine determinations covering the specified range for the procedure (i.e., three concentration and three replicates of each concentration or using a minimum of six determinations at 100% of the test concentration).ICH文件建議可重復性的評估應該使用最少九次檢測，覆蓋該分析規(guī)程所規(guī)定的范圍（例如，三個濃度和每個濃度三次重復進樣，或在100%測試濃度上進行最少六次測定）。SPECIFITY專屬性Definition- The ICH documents define specificity as the ability to assess unequivocally the analyt in the presence of components that may be expected to be present, such as impurities, degradation products, and matrix components. Lack of specificity of an individual analytical procedure may be compensated by other supporting analytical procedures. NOTE- Other reputable international authorities (IUPAC, AOAC-I) have preferred the term “selectivity”, reserving “specificity” for those procedures that are completely selective. For the tests discussed below, the above definition has the following implications. 定義- ICH文件將專屬性定義是當待分析物含有預期會有的其他組分（例如，雜質、降解產(chǎn)物、矩陣組分）時，準確可靠地評估待分析物的能力。某個分析規(guī)程缺乏專屬性可以通過其他輔助性分析規(guī)程進行補償。【注意：其他聲譽卓著的國際權威機構（IUPAC、AOAC-I）已經(jīng)提出術語“選擇性”，而將“專屬性”留給已經(jīng)具有完全選擇性的規(guī)程。】對于下面討論的測試，上述定義具有以下的含義。Identification Tests: ensure the identity of the analyte.鑒別檢測：確保待分析物的鑒別。Purity Tests: ensure that all the analytical procedures performed allow an accurate statement of the content of impurities of an analyte (e.g., related substances test, heavy metals limit, organic volatile impurities). 純度檢測：確保執(zhí)行的所有分析規(guī)程能夠令對于待分析物各雜質含量的準確陳述得以做出。Assays: provide an exact result, which allow an accurate statement on the content or potency of the analyte in a sample. 含量檢測：提供準確的結果，令對樣品中待分析物的含量或效力的準確陳述得以做出。Determination- In the case of qualitative analyses (identification tests), the ability to select between compounds of closely related structure that are likely to be present should be demonstrated. This should be confirmed by obtaining positive results (perhaps by comparison to a known reference material) from samples containing the analyte, coupled with negative results from samples that do not contain the analyte and by confirming that a positive response is not obtained from materials structurally similar to or closely related to the analyte. 測定- 對于定性分析（鑒別檢驗），應當論證其在可能存在的、結構密切相關的物質中進行選擇的能力。從含有待分析物的樣品中得到陽性結果（可能通過與已知標準物質的比較），而從不含待分析物的樣品得到陰性結果，以對其選擇能力加以確認，并還要確認陽性響應不是來自與待分析物結構相似或密切相關的物質。In the case of analytical procedures for impurities, specificity may be established by spiking the drug substance or product with appropriate levels of impurities and demonstrating that these impurities are determined with appropriate accuracy and precision. 對于檢測雜質的分析規(guī)程，專屬性可以通過以適當水平的雜質將原料藥或成藥增敏，并論證這些雜質的測定達到了適當?shù)臏蚀_度和精密度。In the case of the assay, demonstration of specificity requires that it can be shown that the procedure is unaffected by the presence of impurities or excipients. In practice, this can be done by spiking the drug substance or product with appropriate levels of impurities or excipients and demonstrating that the assay result is unaffected by the presence of these extraneous materials. 對于含量檢測，對專屬性的論證要求能夠顯示出該分析規(guī)程不受各雜質或輔料的影響。在實際操作中，通過以適當水平的雜質或輔料將原料藥或成藥增敏，并證明含量檢驗結果不受這些外來物質的影響，來完成論證。If impurity or degradation product standards are unavailable, specificity may be demonstrated by comparing the test results of samples containing impurities or degradation products to a second well-characterized procedure (e.g., Pharmacopeial or other validated procedure). These comparisons should include samples stored under relevant stress conditions (e.g., light, heat, humidity, acid/base hydrolysis, oxidation). In the case of assay, the results should be compared; in the case of chromatographic impurity test, the impurity profiles should be compared. 如果沒有雜質或降解產(chǎn)物的標準品，可以通過將含有雜質或降解產(chǎn)物的樣品的測試結果與第二種、成熟規(guī)程（例如，藥典或其他驗證過的規(guī)程）的結果進行比較，來論證專屬性。這些比較應該包括在相關破壞性條件下（例如，光、熱、濕度、酸/堿水解、氧化作用）存儲的樣品。對于含量測定，應比較其結果；對于色譜法雜質檢測，應比較雜質概況。The ICH documents state that when chromatographic procedures are used, representative chromatograms should be presented to demonstrate the degree of selectivity, and peaks should be appropriately labeled. Peak purity tests (e.g., using diode array or mass spectrometry) may be useful to show that the analyte chromatographic peak is not attributable to more than one component. ICH文件聲明，當使用色譜分析規(guī)程時，應提交具代表性的色譜圖，以論證選擇性的程度，而且應對色譜峰作適當?shù)臉俗R。也可以使用色譜峰純度測試（例如，使用二極管陣列或質譜儀），來顯示待分析物的色譜峰僅產(chǎn)生于一個組分。DETECTION LIMIT檢測限度Definition- The detection limit is a characteristic of limit tests. It is the lowest amount of analyte in a sample that can be detected, but not necessarily quantitated, under the stated experimental conditions. Thus, limit tests merely substantiate that the amount of analyte is above or below a certain level. The detection limit is usually expressed as the concentration of analyte (e.g., percentage, parts per billion) in the sample. 定義- 檢測限度是限度檢測的特性。它是指在規(guī)定的試驗條件下，樣品中可被檢測到的待分析物的最小數(shù)量，但是無需定量。因此，限度檢測僅僅說明了待分析物的數(shù)量高于或低于某個特定水平。檢測限度通常以在樣品中的待分析物濃度（例如，百分比、十億分率）表示。Determination- For non-instrumental procedures, the detection limit is generally 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. 測定- 對于非儀器分析規(guī)程，檢測限度的測定方法通常為，對含有已知濃度待分析物的樣品進行分析，并確立能夠可靠地被檢測出來的待分析物的最低水平。For instrumental procedures, the same approach may be used as for non-instrumental procedures. In the case of procedures submitted for consideration as official compendial procedures, it is almost never necessary to determine the actual detection limit. Rather, the detection limit is shown to be sufficiently low by the analysis of samples with known concentrations of analyte above and below the required detection level. For example, if it is required to detect an impurity at the level of 0.1%, it should be demonstrated that the procedure will reliably detect the impurity at that level.對于儀器分析規(guī)程，可以使用與非儀器分析規(guī)程相同的方法。對于提交用于官方藥典規(guī)程的備選方法，其幾乎從來不需要確定實際的檢測限度。而是，通過分析含有高于和低于必需的檢測水平的、已知待分析物濃度的樣品，以顯示檢測限度足夠低。例如，如果必需檢測濃度在0.1%的雜質，則應當證明該分析規(guī)程將可靠地檢測在這個水平的雜質。In the case of instrumental analytical procedures that exhibit background noise, the ICH documents describe a common approach, whic