5. VALIDATION
5.驗證
5.1 Specificity/Placebo Interference
5.1專屬性/安慰劑（輔料）幹擾
5.2 Linearity and Range
5.2線性和範圍
5.3 Accuracy/Recovery
5.3準確度/回收率
5.4 Precision
5.4精密度
5.4.1 REPEATABILITY OF ANALYSIS
5.4.1重複性
5.4.2 INTERMEDIATE PRECISION/RUGGEDNESS
5.4.2中間精密度/耐用性
5.4.3 REPRODUCIBILITY
5.4.3重現性
5.5 Robustness
5.5耐用性
5.6 Stability of Standard and Sample Solutions
5.6樣品溶液和標準溶液的穩定性
5.7 Considerations for Automation
5.7自動操作注意事項
6. ACCEPTANCE CRITERIA
6.可接受標準
6.1 Immediate-Release Dosage Forms
6.1速釋劑型
6.2 Delayed-Release Dosage Forms
6.2延遲釋放劑型
6.3 Extended-Release Dosage Forms
6.3延長釋放劑型
6.4 Multiple Dissolution Tests
6.4多個溶解度試驗
6.5 Interpretation of Dissolution Results
6.5溶出結果說明
6.5.1 IMMEDIATE-RELEASE DOSAGE FORMS
6.5.1即時釋放劑型
6.5.2 DELAYED-RELEASE DOSAGE FORMS
6.5.2延遲釋放劑型
6.5.3 EXTENDED-RELEASE DOSAGE FORMS
6.5.3延長釋放劑型
5. VALIDATION5.驗證Thevalidation topics described in this section are typical but not all-inclusiveand can be viewed in the context of Validation of Compendial Procedures <1225>,as well as the International Conference on Harmonization (ICH) document, Validationof Analytical Procedures (18). Validation for both parts of the dissolutionprocedure, the analytical finish and the dissolution step, will bediscussed in this section. The dissolution step is the release of the drug inthe dissolution medium and sampling. The analytical finish is defined insection 3. Analytical Finish. Validation of the analytical finish willevaluate the attributes, linearity and range, precision, specificity,accuracy/recovery, robustness, and stability of the sample and standardsolutions. Validation of the dissolution step will include evaluation ofprecision and robustness of the dissolution sample preparation. Validation ofthe analytical finish is performed either using a standard solution orspiked placebo or by the method of standard addition (spiked drugproduct as described in Accuracy in <1225>), as specified in thesections below. Validation of the dissolution step requires the use of awell-characterized dosage form (e.g., having tight content uniformity anduniform performance). Depending on the parameter of interest, validationof the sample handling and analytical procedure can be performed in situ, e.g.,within the dissolution vessel. The validation parameters addressed andthe extent of the validation may vary, depending on the phase of developmentor the intended use for the data.
本章節所涉及的驗證是一些典型的驗證，但不包括所有的驗證，這些驗證在藥典分析方法驗證<1225>的上下文中和ICH指導原則分析方法驗證。本章節討論的溶出實驗的驗證包括溶出和分析兩部分驗證。溶出步驟是指藥物在溶出介質中的釋放和取樣，分析方法的定義詳見第3章節分析方法。分析方法的驗證包括專屬性、線性和範圍、精密度、準確定/回收率、耐用性、對照品溶液和供試品溶液的穩定性。而溶出步驟的驗證主要是對溶出樣品製備的精密度和耐用性評估。分析方法驗證一般使用標準溶液或者空白輔料溶液或通過下面章節中指定的標準加入法（按照<1225>準確度試驗中描述的加入標準的藥品）。溶出步驟的驗證需要使用具有良好特性的產品（例如：具有良好的含量均勻度和溶出均一性）。根據關注的參數，在原來位置，比如在溶出儀裡進行分析方法和樣品處理的驗證。驗證參數處理和驗證程度會有所不同，這取決於開發階段或數據的使用目的。
The acceptance criteria are presented as guidelinesonly, and may differ for some products. Manufacturers should document theappropriate acceptance criteria for their products in pertinent StandardOperating Procedures (SOPs) or in validation protocols.Other considerations maybe important for special dosage forms. Validation studies should be performedacross the range of profile time points. For products containing more than asingle active ingredient, the dissolution procedure needs to
be validatedfor each active ingredient. It is expected that investigations into filtersuitability and the potential for glass adsorption will have been undertakenalready (see 1.1 Performing Filter Compatibility). Validation of theseassessments may occur during spiked recovery experiments.
本章節的驗證標準僅作為指導，對有些產品可能有所不同。生產廠家應該在相關的標準操作規程（SOP）中或在驗證方案中對製劑產品提供合適的可接受標準。對特殊劑型的其他考慮也是重要的。所進行的驗證研究應橫跨溶出曲線時間點範圍。對於複方或者多組分製劑，每一種活性成分的溶出方法均需要進行驗證。過濾器的相容性以及玻璃器的潛在吸收已經進行研究（見1.1濾膜的選擇和相容性），在加標回收率試驗中將對這些評估進行驗證。
5.1 Specificity/PlaceboInterference
5.1專屬性/安慰劑（輔料）幹擾It isnecessary to demonstrate that the results are not unduly affected by placeboconstituents, other active drugs, or degradants. The placebo consists of allthe excipients and coatings, with inks and capsule shells included ifappropriate, without the active ingredient. Placebo interference can beevaluated by using a spiked placebo that is prepared by weighing samples of theplacebo blend, dissolving or dispersing them in dissolution medium atconcentrations that would be encountered during testing, and adding a knownamount of the drug in solution. It may be preferable to perform this experimentat 37°, comparing the solution to a standard solution at the concentrationexpected to be encountered during testing, by using the formula:
Result = (AP/AS) × CS × (V/L) × 100
AP =absorbance of the placebo
AS =absorbance of the standard
CS =concentration of the standard (mg/mL)
V =volume of the medium (mL)
L =label claim (mg)
證明安慰劑（空白輔料）成分、其他活性藥物或降解產物並非影響試驗結果是很有必要的。安慰劑是指除了活性成分以外的所有輔料和包衣材料，在適當的時候還包括油墨和膠囊殼。安慰劑的幹擾可以使用加入標準的安慰劑進行評價，通過稱取安慰劑的混合樣品，將該混合樣品溶解或分散在溶出介質中，製備的濃度為測試過程中使用的濃度，然後向溶液中加入一定量藥物，優選在37℃試驗條件進行這個試驗，使用如下公式比較樣品溶液和標準溶液在測試過程中預計濃度點的吸光度：
結果= (AP/AS)×Cs×(V/L) ×100
AP為安慰劑的吸光度
AS為標準溶液的吸光度
Cs為標準溶液的濃度（mg/ml）
V為溶出介質的體積（ml）
L為標示量（mg）
Theinterference should not exceed 2%. Note that for extended-release products, aplacebo version of the finished dosage form may be more appropriate than blendsbecause this placebo formulation will release the various excipients in amanner more nearly reflecting the product than will a simple blend of theexcipients. In this case, it may be appropriate to evaluate potentialinterference at multiple sampling points in the release profile, withworst-case interference expected at the later sampling points.
幹擾不能超過2%。值得注意的是：緩釋製劑，空白劑型可能比混合物更合適，因為空白劑型比各種輔料簡單的混合物更能反映出不同輔料製劑的釋放方式。在這種情況下，在多個取樣點的釋放曲線更適合評估潛在幹擾，尤其是後面的取樣點預計出現的最壞幹擾。
The blank is the dissolution medium withoutdissolved sample, and it is treated in the same manner as the sample. Theeffect of the absorbance of the blank at the analytical wavelength should beevaluated. In most cases, the absorbance of the dissolution medium blank maynot exceed 1% of the standard solution at the concentration used for analysis.Values >1% should be evaluated on a case-by-case basis.
空白是指不含溶解樣品的溶出介質，按照供試品溶液的處理方法處理空白介質，在分析檢測波長評估空白介質對吸光度的影響。在大多數情況下，空白溶出介質的吸光度不得超過用於分析的標準溶液濃度吸光度的1%。如果超過1%，應根據情況進行評估。
If the placebo interference exceeds 2%, modificationof the method may be necessary. Possible modifications include choosing anotherwavelength, subtracting baseline using a longer wavelength, transformingabsorbance values (e.g., first derivative),and using an alternative analyticaltechnique such as HPLC. Other means for minimizing the placebo interferencewould be acceptable with appropriate justification. When other active drugsubstances or significant levels of degradants are present,it is necessary toshow that these do not significantly affect the results. One procedure fordoing this is to measure the matrix in the presence and absence of the otheractive drug substance or degradant: any interference should not exceed 2%.Similar approaches may be used if other techniques are used for the analyticalfinish.
如果安慰劑幹擾超過2%，有必要對該方法進行改進，可能的改進方法包括選擇另一個檢測波長、通過使用較大波長以減少空白吸收、轉化吸光度值（例如：一階導數）或者使用高選擇性的方法如HPLC法。如果有合適的理由，其他降低安慰劑幹擾的方法也可以使用。當存在其他活性成分或者降解顯著時，證明這些不會對結果產生顯著影響是有必要的。另外解決的方法是不管其他活性成分或者存在存在的降解產物，對主要成分的幹擾均不得超過2%。如果其他的方法用於分析，也可使用類似的方法。
5.2 Linearity and Range
5.2線性和範圍Linearity istypically established by preparing solutions of the drug substance, ranging in concentrationfrom less than the lowest expected concentration to more than the highestconcentration during release. The solutions may be prepared either using eithera standard solution or spiked solution or by the method of standard addition. Aminimum of five concentrations is normally used (see <1225>). Typically,solutions are made from a common stock if possible. The concentration range maynot exceed the linearity limits of the method, including the instrumentation.Organic solvents may be used to enhance drug solubility for the preparation ofthe linearity standard solutions. However, no more than 5% (v/v) of organicsolvent should be present in the final solution unless validated. Linearity istypically calculated by using an appropriate least-squares regression program.Typically, a square of the correlation coefficient (r2 ³0.98) demonstrates linearity. In addition, the y-intercept must not beimportantly different from zero.
線性通常製備一系列原料藥溶液，濃度範圍為低於藥物釋放過程中的最低點濃度至高於藥物釋放過程中最高點的濃度。可以使用標準溶液或加標溶液，或者通過標準加入法製備的溶液。通常至少使用5個濃度點（參見<1225>）。通常情況下，如果可能，溶液有一個共同的線性貯備溶液稀釋製得。濃度範圍不得超過線性方法範圍包括儀器的測量範圍。線性貯備溶液製備過程中為了增加藥物的溶解度，可能會用到有機溶劑，除非經過驗證外，有機溶劑的量均不得超過總體積的5%（v/v）。線性方程一般通過最小二乘法計算，相關係數（r2≥0.98）證明線性較好,此外，y軸截距應接近於0。
The range ofthe procedure is the interval between the upper and lower concentrations of thedrug substance (including these levels) that has been demonstrated to have asuitable level of precision, accuracy, and linearity using the procedure aswritten.
線性範圍內包括原料藥的最低點和最高點在內的所有濃度，均應證明精密度和準確度水平，並且在報告中記錄。
5.3 Accuracy/Recovery
5.3準確度/回收率Accuracy/recoveryis typically established by preparing multiple samples containing the drugsubstance and any other constituents present in the dosage form (e.g.,excipients, coating materials, capsule shell) ranging in concentration fromless than the lowest expected concentration to more than the highestconcentration during release. Accuracy/recovery may be done in conjunction withlinearity determination. The method of standard addition can also be used.Before this activity, it is expected that filter assessment will already havebeen performed, and adsorption of drug onto the glass has also beeninvestigated and ruled out.
準確度/回收率通常是由含有原料藥和製劑中存在的其他成分（如輔料、包衣材料、膠囊殼）製備的多個樣品，濃度範圍的下限為藥物釋放時低於最低預計濃度值，上限高於釋放的最高濃度值。準確度/回收率由線性決定。也可以使用標準加入法。在進行試驗之前，過濾器預計對藥物的吸附要進行評估，同時要考慮並設法排除由於儀器的玻璃材質部分對樣品的吸附而對測試結果造成的影響。
Individual solutions may be directly prepared inthe dissolution medium. Alternatively, to enhance drug solubility it may beappropriate to prepare a stock solution by dissolving the drug substance in asmall amount of organic solvent (typically not exceeding 5% organic solvent inthe final dissolution media) and diluting to the final concentration withdissolution medium.An amount of stock solution equivalent to the targeted labelclaim may be used instead of the drug substance powder. Similarly,for very lowstrengths, it may be more appropriate to prepare a stock solution than toattempt to weigh very small amounts.
直接用溶出介質製備每一個溶液。或者，如果藥物溶解性較差，可以將藥物溶解在少量有機溶劑（一般不超過5%）中製備儲備液，並用溶出介質稀釋到最終濃度。儲備液的量與標示量量相當，可用於代替藥物粉末。同樣地，對於劑量非常小的藥物，製備儲備液比嘗試著稱量非常少量的藥物進行配製更合適。
The measured recovery is typically 95%–105% of theamount added. Bracketing or matrixing of multiple strengths may be useful. Aspecial case for validation is the Acid Stage procedure described in <711>,Delayed-Release Dosage Forms. The limit of NMT 10% needs to be validated.Recovery experiments for drugs that have low solubility in acidic media may bechallenging or impossible to perform and may need to be addressed on a case-by-casebasis. If the compound degrades in acid, the validation experiment must addressthis fact.
回收率測得值通常為加入量的95%~105%。多規格製劑括號法或矩陣化是常用方法。在<711>緩釋劑型中描述了酸性階段的分析驗證的例子。需要對NMT不超過10%這個限度進行驗證。在酸性介質中低溶解度藥物的回收率試驗有挑戰性或者不可能進行，需要根據具體情況進行說明，如果藥物在酸性條件下降解，驗證實驗中須對這一事實進行說明。
5.4 Precision
5.4精密度5.4.1 REPEATABILITY OFANALYSIS
5.4.1重複性
Forthe analytical finish, repeatability is evaluated by obtaining replicatemeasurements of standard and/or spiked placebo/standard addition solutions. Itcan be determined by calculating the RSD of the multiple injections orspectrophotometric readings for each standard solution, or by using theaccuracy or linearity data. ICH guidance, Validation of AnalyticalProcedures: Methodology, recommends that repeatability should beassessed using a minimum of nine determinations covering the specified rangefor the procedure (i.e., three concentrations and three replicates of eachconcentration) or using a minimum of six determinations at 100% of the testconcentration. A typical acceptance criterion is an RSD of <2%. Thedemonstration of the repeatability for the dissolution step is conducted byperforming the dissolution step on separate units of a well-characterized dosageform or equivalent composite.
對於分析方法，通過獲得標準和/或加入安慰劑/標準加入溶液的重複測定結果對重複性進行評估，通過多次進樣或者每個標準溶液分光光度計讀數或者使用精密度或者線性數據來計算RSD值， ICH指導原則，分析方法的驗證：方法，推薦重複性測定用覆蓋特定分析範圍的九個確定濃度點（三個濃度點，每個濃度點重複製備三份樣品）或在100%測試濃度點至少製備6份樣品溶液進行測試，通常可接受的標準：RSD<2%。通過採用質量好的製劑或與製劑相等組成（原料+輔料）進行溶出步驟的獨立單元的重複性證明。
5.4.2 INTERMEDIATEPRECISION/RUGGEDNESS
5.4.2中間精密度/耐用性
Assuming thatthe major contributor to the variance is from the dissolution step,intermediate precision may be evaluated to determine the effects of randomevents on the precision of the dissolution procedure. This evaluation istypically done later in the development of the drug product and is required forfull method validation. For many analytical procedures intermediate precisionis typically assessed by determination of contributions to variance and,possibly, by a comparison of means. The use of an experimental matrix design isencouraged for evaluation of intermediate precision because interaction effectsmay be observed more clearly relative to a single variable experiment. Indissolution testing, a ruggedness approach that compares means alone is oftentaken to investigate the factors that contribute to intermediate precision. Theruggedness can be evaluated across the range of product strengths. Typicalvariations to be studied include different days, analysts, and equipment. If possible,ruggedness can be evaluated using a drug product lot if well characterized, forexample, by having tight content uniformity and uniform performance, but ifthis type of lot is not available, a premeasured placebo with activeingredients may be used to investigate the intermediate precision. The use ofsuch a spiked placebo would additionally support the assessment of thecontribution of the analytical finish to the observed variability of results.
假設溶出步驟是產生偏差的主要因素，可以用中間精密度評估，以確定隨機事件對溶出精密度的影響。這種評估通常在製劑開發後完成，需要對方法學進行充分驗證。對於很多分析方法，中間精密度通常通過確定偏差來源進行評估，通過比較分析的方式。鼓勵使用實驗室矩陣設計對中間精密度進行評估，因為與單因素試驗相關的相互作用會更清楚地觀察到。在溶出度測定時，耐用性試驗方法可以採取單獨比較的方法來研究中間精密度的影響因素。這些影響因素可能是由中間精密度引起的。耐用性試驗可以評估製劑的濃度範圍。研究過程中的典型的變化，包括不同天、不同操作人員和設備。如果可能，耐用性試驗可以用較好質量特徵的製劑批次進行評估，例如：較好的含量均勻度。但如果這種批次的不可獲得測，可用活性成分加安慰劑進行中間精密度研究。使用加入標準的安慰劑（空白輔料）將支持觀察到的變化結果對分析方法誤差的影響。
Thedissolution procedure on the same lot of well-characterized dosage form may berun by at least two different analysts from the same laboratory, with eachanalyst preparing the standard solutions and the medium and following thedefined extraction/quantification procedure. Typically, the analysts usedifferent dissolution baths, spectrophotometers or HPLC equipment (includingcolumns), and autosamplers, and they perform the test on different days. Fullprofiles are assessed where relevant to the product. This procedure may not benecessary at each strength; instead, bracketing with high and low strengths maybe acceptable.
同一批次質量特徵較好的製劑的溶出試驗可以由同一實驗室至少兩個不同的分析人員進行，每個分析人員製備標準溶液和溶出介質和依據明確的提取和定量步驟進行。通常情況下，分析人員用不同的溶出液、分光光度計或HPLC（包括色譜柱）和自動進樣器，在不同天進行試驗。與製劑相關的曲線進行全面的評估，這個分析操作對每個濃度可能不是必須的，而是使用高濃度和低濃度進行分析是可以接受的。
Acceptancecriteria for intermediate precision or for ruggedness are predetermined. Atypical acceptance criterion for ruggedness is that the difference in the meanvalue for dissolution results between any two conditions, using the samestrength,does not exceed an absolute 10% at time points with <85% dissolvedand does not exceed 5% for time points >85%. Acceptance criteria may beproduct specific, and other statistical tests and limits may be used.
中間精密度的可接受標準或耐用性是預先確定的。通常耐用性的可接受標準是使用相同濃度，在任何兩個條件之間溶出結果平均值的差，在溶出度小於85%的時間點，絕對誤差不能超過10%；大於85%的時間點，絕對差值不能超過5%，可接受標準可以用於特定產品、也可以使用其他統計方法和範圍。
5.4.3 REPRODUCIBILITY
5.4.3重現性
Reproducibilityfollows the general concepts of intermediate precision, but is performed by twodifferent analysts at different labs.
重現性遵照中間精密度的一般概念，但在不同實驗室由兩位不同的分析人員進行試驗。
5.5 Robustness
5.5耐用性Evaluation ofrobustness, which assesses the effect of making small, deliberate changes tothe dissolution conditions, typically is done later in development of the drugproduct and is a requirement for full method validation. It is performed usinga well-characterized lot of drug product, for example having tight contentuniformity and uniform performance. The number of
replicates(typically 3 or 6) is dependent on the intermediate precision. All profilepoints should be evaluated.
評估溶出條件做一下小的、故意改變溶出條件對耐用性的影響，通常是在製劑開發出來以後進行，需要進行充分的方法學驗證。用具有良好表徵的製劑批次進行，例如，具有較好的含量均勻度。重複性數量（通常3或6）取決於中間精密度，所有的曲線點都應進行評估.
Selection ofparameters to be varied depends on the dissolution procedure and analysis type.The parameters may include medium composition (e.g., buffer or surfactantconcentration, pH, deaeration), volume, agitation rate, sampling time, and temperature.Statistical analysis of the data generated will help determine the extent towhich the parameters must be controlled in the method. The robustnessassessment is well suited to Design of Experiments (DoE) methodologies toefficiently investigate the impact of the individual parameters and/or theirinteraction
參數選擇是多種多樣的取決於溶出過程和分析類型，參數包括溶出介質組合物（例如，緩衝液、表面活性劑濃度、pH、脫氣），體積、轉速、取樣時間和溫度。得到的數據進行統計分析將有助於確定參數在方法中控制的程度，試驗設計方法（DOE）非常適合耐用性評估,以有效研究各個參數和他們相互作用的影響。
Robustness of analytical finish is referenced in <1225>.HPLC analysis parameters may include mobile phase composition (percentageorganic, buffer concentration, pH), flow rate, wavelength, column temperature,and multiple columns (of the same type). For spectrophotometric analysis, thewavelength may be varied.
分析方法的耐用性參考<1225>，HPLC分析參數包括流動相成分（有機相的百分比、緩衝液濃度、pH）、流速、波長、柱溫、同一類型的多根色譜柱、對於分光光度計方法，波長可能不同。
5.6 Stability of Standard andSample Solutions
5.6樣品溶液和標準溶液的穩定性The standardsolution is stored under conditions that ensure stability. The stability of thestandard solution is analyzed over a specified period of time (for at least thetime of the entire dissolution procedure), using a freshly prepared standardsolution at each time interval for comparison. The acceptable range forstandard solution stability is influenced by the concentration and is typicallybetween 98% and 102% at the expected final concentration.
標準溶液貯藏在能保證其穩定性的條件下，應在指定的時間內（完全溶出的最少時間）分析標準溶液的穩定性，在每個時間間隔使用新配製的標準溶液進行比較，標準溶液穩定性可接受的範圍受濃度的影響，通常是預期最終濃度的98%-102%。
The sample solution is typically stored at roomtemperature. The sample is analyzed over a specified period of time, using theoriginal sample solution response for comparison. The typical acceptable rangefor sample solution stability may be between 98% and 102%, compared with theinitial analysis of the sample solutions. If the solution is not stable,aspects to consider include temperature (refrigeration may be needed), lightprotection, and container material (plastic or glass).
樣品溶液一般在室溫下貯存，樣品應在指定時間段進行分析，與最初分析的樣品溶液進行比較，樣品溶液穩定性通常的可接受範圍98%-102%，，如果溶液不穩定，需要考慮溫度（需要冷藏）、避光、以及容器材料（塑料或玻璃）。
The proceduremay state that the standards and samples need to be analyzed within a timeperiod demonstrating acceptable standard and sample solution stability.
該過程表明，需要分析對照品和樣品在一段時間內的可接受標準和樣品溶液穩定性。
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