[PDF] METHOD VALIDATION AND QUALITY CONTROL PROCEDURES FOR

美国FDA分析方法验证指南中英文对照Guide to the U.S. FDA method validation Chinese and English目录ContentsI. INTRODUCTION (3)II. BACKGROUND (4)III. TYPES OF ANALYTICAL PROCEDURES (6)IV. REFERENCE STANDARDS (7)V. METHODS VALIDATION FOR INDs (10)VI. CONTENT AND FORMAT OF ANALYTICAL PROCEDURES FOR NDAs, 230ANDAs, BLAs, AND PLAs (11)VII. METHODS VALIDATION FOR NDAs, ANDAs, BLAs, AND PLAs (15)VIII. STATISTICAL ANALYSIS (23)IX. REVALIDATION (24)X. METHODS VALIDATION PACKAGE: CONTENTS AND PROCESSING (25)XI. METHODOLOGY (30)ATTACHMENT ANDA, ANDA, BLA, AND PLA SUBMISSION CONTENTS (40)ATTACHMENT BMETHODS VALIDATION PROBLEMS AND DELAY (41)GLOSSARY (42)美国FDA分析方法验证指南中英文对照I. INTRODUCTIONThis guidance provides recommendations to applicants on submittinganalytical procedures, validation data, and samples to support thedocumentation of the identity, strength, quality, purity, and potencyof drug substances and drug products.1. 绪论本指南旨在为申请者提供建议，以帮助其提交分析方法，方法验证资料和样品用于支持原料药和制剂的认定，剂量，质量，纯度和效力方面的文件。

质量控制现代食品检测中农药残留技术及质量控制措施胡　爽1，李　燕2（1.阳谷县检验检测中心，山东聊城 252300；2.内蒙古华测质检技术服务有限公司，内蒙古呼和浩特010000）摘　要：文章系统阐述了不同类型的样品前处理技术、仪器分析方法和快速检测技术，重点介绍了QuEChERS法、液相色谱-质谱联用技术的基本原理和典型应用。

针对农药残留检测中的关键质控环节，从标准物质的选用、方法验证和不确定度评估、实验室内部质量控制等方面进行了探讨，以确保检测数据质量。

关键词：农药残留；样品前处理；色谱-质谱；质量控制Pesticide Residue Technology and Quality Control Measuresin Modern Food TestingHU Shuang1, LI Yan2(1.Yanggu County Inspection and Testing Center, Liaocheng 252300, China;2.Inner Mongolia Huace Quality Inspection Technology Service Co., Ltd., Hohhot 010000, China)Abstract: The article systematically elaborates on different types of sample pretreatment techniques, instrument analysis methods, and rapid detection techniques, with a focus on the basic principles and typical applications of QuEChERS method and liquid chromatography-mass spectrometry technology. We have explored the key quality control aspects in pesticide residue detection, including the selection of reference materials, method validation and uncertainty assessment, and internal quality control in the laboratory, to ensure the quality of detection data.Keywords: pesticide residue; sample pretreatment; chromatography-mass spectrometry; quality control农药在现代农业生产中发挥着不可或缺的作用，其应用有效地防治了农作物病虫害，提高了农产品产量和质量，但同时农药残留也已成为影响食品安全的重要因素之一。

标准样品英文Standard Sample English。

In the field of scientific research and development, the use of standard samples is crucial for ensuring the accuracy and reliability of experimental results. Standard samples, also known as reference materials, are substances with well-defined properties that are used as a basis for comparison in analytical measurements. They play a vital role in quality control, method validation, and calibration of instruments in various industries such as pharmaceuticals, environmental monitoring, food safety, and material testing.The importance of standard samples lies in theirability to provide a known reference point for measuring the quality and performance of analytical methods and instruments. By using standard samples, researchers and analysts can verify the accuracy of their measurements, identify and correct any errors in their procedures, andensure the consistency and reproducibility of their results. This is particularly important in industries where the reliability of analytical data is critical for publichealth and safety, environmental protection, and product quality.In addition to their role in quality control and method validation, standard samples also serve as a means of ensuring traceability in analytical measurements. Traceability is the ability to demonstrate that a measurement result can be related to a reference through an unbroken chain of calibrations, each contributing to the measurement uncertainty. By using standard samples withwell-established traceability, laboratories and testing facilities can ensure the accuracy and reliability of their measurements, and demonstrate their competence and compliance with international standards and regulations.The selection and use of standard samples requirecareful consideration of their properties, such as purity, stability, homogeneity, and compatibility with theanalytical methods and instruments being used. It isimportant to choose standard samples that are well-characterized and certified by reputable reference material producers, and to handle and store them properly to prevent contamination and degradation. Furthermore, the use of multiple standard samples with different properties and concentrations can help to evaluate the performance and limitations of analytical methods and instruments, and to assess the measurement uncertainty associated with the analysis.In conclusion, standard samples play a critical role in ensuring the accuracy, reliability, and traceability of analytical measurements in various industries. By providing a known reference point for comparison, standard samples enable researchers and analysts to verify the quality and performance of their methods and instruments, and to demonstrate their competence and compliance with international standards and regulations. Therefore, the proper selection, use, and handling of standard samples are essential for achieving high-quality and reliableanalytical results, and for ensuring the safety andintegrity of products and processes.。

硫酸氨基葡萄糖标准样品的研制陈伟珠;张怡评;方华;易瑞灶;朱芮琳;洪专【摘要】研制了硫酸氨基葡萄糖标准样品。

以盐酸氨基葡萄糖为原料，制备高纯硫酸氨基葡萄糖，采用红外光谱（IR）、高分辨质谱和核磁共振谱（NMR）进行结构确证。

样品分装成200瓶样品后，采用高效液相色谱–蒸发光散射法进行均匀性检验、稳定性检验和定值分析。

从样品中随机抽取15瓶进行均匀性检验，结果表明在95%的置信区间范围内样品均匀性良好。

按照25℃长期试验稳定性（12个月）进行稳定性考察，结果表明在考察期间内样品稳定性良好。

标准样品经国内8家具有分析资质的实验室进行协同定值，硫酸氨基葡萄糖标准样品定值结果为99.84%，相对扩展不确定度为0.18%（k=1.96）。

该标准样品达到国家标准样品的技术要求，可用于有关硫酸氨基葡萄糖的分析方法校正和质量控制。

%Glucosamine sulfate certified reference material was developed. Glucosamine sulfate was made from the glucosamine hydrochloride. The structure of glucosamine sulfate certified reference material was comfirmed by infrared spectroscopy (IR), high resolution mass spectral (HRMS) and nuclear magnetic resonance (NMR). The sample was divided into 200 bottles, the homogeneity and stability testing and quantitative analysis were carried out by high performance liquid chromatography with evaporative light-scattering detector (HPLC–ELSD). According to analysis procedure of homogeneity, 15 bottles of sample were randomly taken from 200 bottles, and the resultswere validated byF-test statistical methods.The stability inspection was carried on the long-term(12 months), and the results indicated that the period for glucosamine sulfate of storagewas 12 months at 25℃. A cooperative certification was conducted with 8 qualified laboratories. The certified purity value of the reference material of glucosamine sulfate was 99.84% with a relative expaned uncertainty of0.18%(k=1.96). The reference material can conform to the technical requirement of the certified reference material. The material was intended for use in the method validation and quality control regarding glucosamine sulfate.【期刊名称】《化学分析计量》【年(卷),期】2015(000)002【总页数】5页(P1-4,5)【关键词】硫酸氨基葡萄糖;标准样品;均匀性;稳定性;定值;不确定度【作者】陈伟珠;张怡评;方华;易瑞灶;朱芮琳;洪专【作者单位】国家海洋局第三海洋研究所，国家海洋局海洋生物资源综合利用工程技术研究中心，福建厦门 361005; 厦门大学化学化工学院化学系，化学生物学福建省重点实验室，福建厦门 361005;国家海洋局第三海洋研究所，国家海洋局海洋生物资源综合利用工程技术研究中心，福建厦门 361005;国家海洋局第三海洋研究所，国家海洋局海洋生物资源综合利用工程技术研究中心，福建厦门361005;国家海洋局第三海洋研究所，国家海洋局海洋生物资源综合利用工程技术研究中心，福建厦门 361005;国家海洋局第三海洋研究所，国家海洋局海洋生物资源综合利用工程技术研究中心，福建厦门 361005;国家海洋局第三海洋研究所，国家海洋局海洋生物资源综合利用工程技术研究中心，福建厦门 361005【正文语种】中文【中图分类】O653硫酸氨基葡萄糖是由虾、蟹甲壳降解分离得到的一种具有多种功能的高值产品，是一种分子结构明确的天然氨基糖类化合物，是人体合成软骨和关节液基质硫酸软骨素和胶原蛋白所必需的重要成分，可阻止关节软骨退变，延缓病情进展，减轻关节肿痛，促进功能恢复［1-2］。

HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationFLUOXETINE HClC17H18F3NO•HClM.W. = 345.79CAS — 59333-67-4STABILITY INDICATINGA S S A Y V A L I D A T I O NMethod is suitable for:ýIn-process controlþProduct ReleaseþStability indicating analysis (Suitability - US/EU Product) CAUTIONFLUOXETINE HYDROCHLORIDE IS A HAZARDOUS CHEMICAL AND SHOULD BE HANDLED ONLY UNDER CONDITIONS SUITABLE FOR HAZARDOUS WORK.IT IS HIGHLY PRESSURE SENSITIVE AND ADEQUATE PRECAUTIONS SHOULD BE TAKEN TO AVOID ANY MECHANICAL FORCE (SUCH AS GRINDING, CRUSHING, ETC.) ON THE POWDER.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationTABLE OF CONTENTS INTRODUCTION........................................................................................................................ PRECISION............................................................................................................................... System Repeatability ................................................................................................................ Method Repeatability................................................................................................................. Intermediate Precision .............................................................................................................. LINEARITY................................................................................................................................ RANGE...................................................................................................................................... ACCURACY............................................................................................................................... Accuracy of Standard Injections................................................................................................ Accuracy of the Drug Product.................................................................................................... VALIDATION OF FLUOXETINE HCl AT LOW CONCENTRATION........................................... Linearity at Low Concentrations................................................................................................. Accuracy of Fluoxetine HCl at Low Concentration..................................................................... System Repeatability................................................................................................................. Quantitation Limit....................................................................................................................... Detection Limit........................................................................................................................... VALIDATION FOR META-FLUOXETINE HCl (POSSIBLE IMPURITIES).................................. Meta-Fluoxetine HCl linearity at 0.05% - 1.0%........................................................................... Detection Limit for Fluoxetine HCl.............................................................................................. Quantitation Limit for Meta Fluoxetine HCl................................................................................ Accuracy for Meta-Fluoxetine HCl ............................................................................................ Method Repeatability for Meta-Fluoxetine HCl........................................................................... Intermediate Precision for Meta-Fluoxetine HCl......................................................................... SPECIFICITY - STABILITY INDICATING EVALUATION OF THE METHOD............................. FORCED DEGRADATION OF FINISHED PRODUCT AND STANDARD..................................1. Unstressed analysis...............................................................................................................2. Acid Hydrolysis stressed analysis..........................................................................................3. Base hydrolysis stressed analysis.........................................................................................4. Oxidation stressed analysis...................................................................................................5. Sunlight stressed analysis.....................................................................................................6. Heat of solution stressed analysis.........................................................................................7. Heat of powder stressed analysis.......................................................................................... System Suitability stressed analysis.......................................................................................... Placebo...................................................................................................................................... STABILITY OF STANDARD AND SAMPLE SOLUTIONS......................................................... Standard Solution...................................................................................................................... Sample Solutions....................................................................................................................... ROBUSTNESS.......................................................................................................................... Extraction................................................................................................................................... Factorial Design......................................................................................................................... CONCLUSION...........................................................................................................................ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationBACKGROUNDTherapeutically, Fluoxetine hydrochloride is a classified as a selective serotonin-reuptake inhibitor. Effectively used for the treatment of various depressions. Fluoxetine hydrochloride has been shown to have comparable efficacy to tricyclic antidepressants but with fewer anticholinergic side effects. The patent expiry becomes effective in 2001 (US). INTRODUCTIONFluoxetine capsules were prepared in two dosage strengths: 10mg and 20mg dosage strengths with the same capsule weight. The formulas are essentially similar and geometrically equivalent with the same ingredients and proportions. Minor changes in non-active proportions account for the change in active ingredient amounts from the 10 and 20 mg strength.The following validation, for the method SI-IAG-206-02 , includes assay and determination of Meta-Fluoxetine by HPLC, is based on the analytical method validation SI-IAG-209-06. Currently the method is the in-house method performed for Stability Studies. The Validation was performed on the 20mg dosage samples, IAG-21-001 and IAG-21-002.In the forced degradation studies, the two placebo samples were also used. PRECISIONSYSTEM REPEATABILITYFive replicate injections of the standard solution at the concentration of 0.4242mg/mL as described in method SI-IAG-206-02 were made and the relative standard deviation (RSD) of the peak areas was calculated.SAMPLE PEAK AREA#15390#25406#35405#45405#55406Average5402.7SD 6.1% RSD0.1ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::PRECISION - Method RepeatabilityThe full HPLC method as described in SI-IAG-206-02 was carried-out on the finished product IAG-21-001 for the 20mg dosage form. The method repeated six times and the relative standard deviation (RSD) was calculated.SAMPLENumber%ASSAYof labeled amountI 96.9II 97.8III 98.2IV 97.4V 97.7VI 98.5(%) Average97.7SD 0.6(%) RSD0.6PRECISION - Intermediate PrecisionThe full method as described in SI-IAG-206-02 was carried-out on the finished product IAG-21-001 for the 20mg dosage form. The method was repeated six times by a second analyst on a different day using a different HPLC instrument. The average assay and the relative standard deviation (RSD) were calculated.SAMPLENumber% ASSAYof labeled amountI 98.3II 96.3III 94.6IV 96.3V 97.8VI 93.3Average (%)96.1SD 2.0RSD (%)2.1The difference between the average results of method repeatability and the intermediate precision is 1.7%.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationLINEARITYStandard solutions were prepared at 50% to 200% of the nominal concentration required by the assay procedure. Linear regression analysis demonstrated acceptability of the method for quantitative analysis over the concentration range required. Y-Intercept was found to be insignificant.RANGEDifferent concentrations of the sample (IAG-21-001) for the 20mg dosage form were prepared, covering between 50% - 200% of the nominal weight of the sample.Conc. (%)Conc. (mg/mL)Peak Area% Assayof labeled amount500.20116235096.7700.27935334099.21000.39734463296.61500.64480757797.52000.79448939497.9(%) Average97.6SD 1.0(%) RSD 1.0ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::RANGE (cont.)The results demonstrate linearity as well over the specified range.Correlation coefficient (RSQ)0.99981 Slope11808.3Y -Interceptresponse at 100%* 100 (%) 0.3%ACCURACYACCURACY OF STANDARD INJECTIONSFive (5) replicate injections of the working standard solution at concentration of 0.4242mg/mL, as described in method SI-IAG-206-02 were made.INJECTIONNO.PEAK AREA%ACCURACYI 539299.7II 540599.9III 540499.9IV 5406100.0V 5407100.0Average 5402.899.9%SD 6.10.1RSD, (%)0.10.1The percent deviation from the true value wasdetermined from the linear regression lineHPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::ACCURACY OF THE DRUG PRODUCTAdmixtures of non-actives (placebo, batch IAG-21-001 ) with Fluoxetine HCl were prepared at the same proportion as in a capsule (70%-180% of the nominal concentration).Three preparations were made for each concentration and the recovery was calculated.Conc.(%)Placebo Wt.(mg)Fluoxetine HCl Wt.(mg)Peak Area%Accuracy Average (%)70%7079.477.843465102.27079.687.873427100.77079.618.013465100.0101.0100%10079.6211.25476397.910080.8011.42491799.610079.6011.42485498.398.6130%13079.7214.90640599.413080.3114.75632899.213081.3314.766402100.399.618079.9920.10863699.318079.3820.45879499.418080.0820.32874899.599.4Placebo, Batch Lot IAG-21-001HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::VALIDATION OF FLUOXETINE HClAT LOW CONCENTRATIONLINEARITY AT LOW CONCENTRATIONSStandard solution of Fluoxetine were prepared at approximately 0.02%-1.0% of the working concentration required by the method SI-IAG-206-02. Linear regression analysis demonstrated acceptability of the method for quantitative analysis over this range.ACCURACY OF FLUOXETINE HCl AT LOW CONCENTRATIONThe peak areas of the standard solution at the working concentration were measured and the percent deviation from the true value, as determined from the linear regression was calculated.SAMPLECONC.µg/100mLAREA FOUND%ACCURACYI 470.56258499.7II 470.56359098.1III 470.561585101.3IV 470.561940100.7V 470.56252599.8VI 470.56271599.5(%) AverageSlope = 132.7395299.9SD Y-Intercept = -65.872371.1(%) RSD1.1HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSystem RepeatabilitySix replicate injections of standard solution at 0.02% and 0.05% of working concentration as described in method SI-IAG-206-02 were made and the relative standard deviation was calculated.SAMPLE FLUOXETINE HCl AREA0.02%0.05%I10173623II11503731III10103475IV10623390V10393315VI10953235Average10623462RSD, (%) 5.0 5.4Quantitation Limit - QLThe quantitation limit ( QL) was established by determining the minimum level at which the analyte was quantified. The quantitation limit for Fluoxetine HCl is 0.02% of the working standard concentration with resulting RSD (for six injections) of 5.0%. Detection Limit - DLThe detection limit (DL) was established by determining the minimum level at which the analyte was reliably detected. The detection limit of Fluoxetine HCl is about 0.01% of the working standard concentration.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::VALIDATION FOR META-FLUOXETINE HCl(EVALUATING POSSIBLE IMPURITIES)Meta-Fluoxetine HCl linearity at 0.05% - 1.0%Relative Response Factor (F)Relative response factor for Meta-Fluoxetine HCl was determined as slope of Fluoxetine HCl divided by the slope of Meta-Fluoxetine HCl from the linearity graphs (analysed at the same time).F =132.7395274.859534= 1.8Detection Limit (DL) for Fluoxetine HClThe detection limit (DL) was established by determining the minimum level at which the analyte was reliably detected.Detection limit for Meta Fluoxetine HCl is about 0.02%.Quantitation Limit (QL) for Meta-Fluoxetine HClThe QL is determined by the analysis of samples with known concentration of Meta-Fluoxetine HCl and by establishing the minimum level at which the Meta-Fluoxetine HCl can be quantified with acceptable accuracy and precision.Six individual preparations of standard and placebo spiked with Meta-Fluoxetine HCl solution to give solution with 0.05% of Meta Fluoxetine HCl, were injected into the HPLC and the recovery was calculated.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::META-FLUOXETINE HCl[RECOVERY IN SPIKED SAMPLES].Approx.Conc.(%)Known Conc.(µg/100ml)Area in SpikedSampleFound Conc.(µg/100mL)Recovery (%)0.0521.783326125.735118.10.0521.783326825.821118.50.0521.783292021.55799.00.0521.783324125.490117.00.0521.783287220.96996.30.0521.783328526.030119.5(%) AVERAGE111.4SD The recovery result of 6 samples is between 80%-120%.10.7(%) RSDQL for Meta Fluoxetine HCl is 0.05%.9.6Accuracy for Meta Fluoxetine HClDetermination of Accuracy for Meta-Fluoxetine HCl impurity was assessed using triplicate samples (of the drug product) spiked with known quantities of Meta Fluoxetine HCl impurity at three concentrations levels (namely 80%, 100% and 120% of the specified limit - 0.05%).The results are within specifications:For 0.4% and 0.5% recovery of 85% -115%For 0.6% recovery of 90%-110%HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::META-FLUOXETINE HCl[RECOVERY IN SPIKED SAMPLES]Approx.Conc.(%)Known Conc.(µg/100mL)Area in spikedSample Found Conc.(µg/100mL)Recovery (%)[0.4%]0.4174.2614283182.66104.820.4174.2614606187.11107.370.4174.2614351183.59105.36[0.5%]0.5217.8317344224.85103.220.5217.8316713216.1599.230.5217.8317341224.81103.20[0.6%]0.6261.3918367238.9591.420.6261.3920606269.81103.220.6261.3920237264.73101.28RECOVERY DATA DETERMINED IN SPIKED SAMPLESHPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::REPEATABILITYMethod Repeatability - Meta Fluoxetine HClThe full method (as described in SI-IAG-206-02) was carried out on the finished drug product representing lot number IAG-21-001-(1). The HPLC method repeated serially, six times and the relative standard deviation (RSD) was calculated.IAG-21-001 20mg CAPSULES - FLUOXETINESample% Meta Fluoxetine % Meta-Fluoxetine 1 in Spiked Solution10.0260.09520.0270.08630.0320.07740.0300.07450.0240.09060.0280.063AVERAGE (%)0.0280.081SD 0.0030.012RSD, (%)10.314.51NOTE :All results are less than QL (0.05%) therefore spiked samples with 0.05% Meta Fluoxetine HCl were injected.HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationED. N0: 04Effective Date:APPROVED::Intermediate Precision - Meta-Fluoxetine HClThe full method as described in SI-IAG-206-02 was applied on the finished product IAG-21-001-(1) .It was repeated six times, with a different analyst on a different day using a different HPLC instrument.The difference between the average results obtained by the method repeatability and the intermediate precision was less than 30.0%, (11.4% for Meta-Fluoxetine HCl as is and 28.5% for spiked solution).IAG-21-001 20mg - CAPSULES FLUOXETINESample N o:Percentage Meta-fluoxetine% Meta-fluoxetine 1 in spiked solution10.0260.06920.0270.05730.0120.06140.0210.05850.0360.05560.0270.079(%) AVERAGE0.0250.063SD 0.0080.009(%) RSD31.514.51NOTE:All results obtained were well below the QL (0.05%) thus spiked samples slightly greater than 0.05% Meta-Fluoxetine HCl were injected. The RSD at the QL of the spiked solution was 14.5%HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSPECIFICITY - STABILITY INDICATING EVALUATIONDemonstration of the Stability Indicating parameters of the HPLC assay method [SI-IAG-206-02] for Fluoxetine 10 & 20mg capsules, a suitable photo-diode array detector was incorporated utilizing a commercial chromatography software managing system2, and applied to analyze a range of stressed samples of the finished drug product.GLOSSARY of PEAK PURITY RESULT NOTATION (as reported2):Purity Angle-is a measure of spectral non-homogeneity across a peak, i.e. the weighed average of all spectral contrast angles calculated by comparing all spectra in the integrated peak against the peak apex spectrum.Purity Threshold-is the sum of noise angle3 and solvent angle4. It is the limit of detection of shape differences between two spectra.Match Angle-is a comparison of the spectrum at the peak apex against a library spectrum.Match Threshold-is the sum of the match noise angle3 and match solvent angle4.3Noise Angle-is a measure of spectral non-homogeneity caused by system noise.4Solvent Angle-is a measure of spectral non-homogeneity caused by solvent composition.OVERVIEWT he assay of the main peak in each stressed solution is calculated according to the assay method SI-IAG-206-02, against the Standard Solution, injected on the same day.I f the Purity Angle is smaller than the Purity Threshold and the Match Angle is smaller than the Match Threshold, no significant differences between spectra can be detected. As a result no spectroscopic evidence for co-elution is evident and the peak is considered to be pure.T he stressed condition study indicated that the Fluoxetine peak is free from any appreciable degradation interference under the stressed conditions tested. Observed degradation products peaks were well separated from the main peak.1® PDA-996 Waters™ ; 2[Millennium 2010]ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationFORCED DEGRADATION OF FINISHED PRODUCT & STANDARD 1.UNSTRESSED SAMPLE1.1.Sample IAG-21-001 (2) (20mg/capsule) was prepared as stated in SI-IAG-206-02 and injected into the HPLC system. The calculated assay is 98.5%.SAMPLE - UNSTRESSEDFluoxetine:Purity Angle:0.075Match Angle:0.407Purity Threshold:0.142Match Threshold:0.4251.2.Standard solution was prepared as stated in method SI-IAG-206-02 and injected into the HPLC system. The calculated assay is 100.0%.Fluoxetine:Purity Angle:0.078Match Angle:0.379Purity Threshold:0.146Match Threshold:0.4272.ACID HYDROLYSIS2.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as in method SI-IAG-206-02 : An amount equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent was added and the solution sonicated for 10 minutes. 1mL of conc. HCl was added to this solution The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with NaOH 10N, made up to volume with Diluent and injected into the HPLC system after filtration.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 98.8%.SAMPLE- ACID HYDROLYSISFluoxetine peak:Purity Angle:0.055Match Angle:0.143Purity Threshold:0.096Match Threshold:0.3712.2.Standard solution was prepared as in method SI-IAG-206-02 : about 22mg Fluoxetine HCl were weighed into a 50mL volumetric flask. 20mL Diluent were added. 2mL of conc. HCl were added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with NaOH 10N, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 97.2%.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSTANDARD - ACID HYDROLYSISFluoxetine peak:Purity Angle:0.060Match Angle:0.060Purity Threshold:0.099Match Threshold:0.3713.BASE HYDROLYSIS3.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as per method SI-IAG-206-02 : An amount equivalent to 20mg Fluoxetine was weight into a 50mL volumetric flask. 20mL Diluent was added and the solution sonicated for 10 minutes. 1mL of 5N NaOH was added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH = 5.5 with 5N HCl, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease. Assay result obtained - 99.3%.SAMPLE - BASE HYDROLYSISFluoxetine peak:Purity Angle:0.063Match Angle:0.065Purity Threshold:0.099Match Threshold:0.3623.2.Standard stock solution was prepared as per method SI-IAG-206-02 : About 22mg Fluoxetine HCl was weighed into a 50mL volumetric flask. 20mL Diluent was added. 2mL of 5N NaOH was added to this solution. The solution was allowed to stand for 18 hours, then adjusted to about pH=5.5 with 5N HCl, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity did NOT decrease - 99.5%.STANDARD - BASE HYDROLYSISFluoxetine peak:Purity Angle:0.081Match Angle:0.096Purity Threshold:0.103Match Threshold:0.3634.OXIDATION4.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as per method SI-IAG-206-02. An equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent added and the solution sonicated for 10 minutes.1.0mL of 30% H2O2 was added to the solution and allowed to stand for 5 hours, then made up to volume with Diluent, filtered and injected into HPLC system.Fluoxetine peak intensity decreased to 95.2%.ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSAMPLE - OXIDATIONFluoxetine peak:Purity Angle:0.090Match Angle:0.400Purity Threshold:0.154Match Threshold:0.4294.2.Standard solution was prepared as in method SI-IAG-206-02 : about 22mg Fluoxetine HCl were weighed into a 50mL volumetric flask and 25mL Diluent were added. 2mL of 30% H2O2 were added to this solution which was standing for 5 hours, made up to volume with Diluent and injected into the HPLC system.Fluoxetine peak intensity decreased to 95.8%.STANDARD - OXIDATIONFluoxetine peak:Purity Angle:0.083Match Angle:0.416Purity Threshold:0.153Match Threshold:0.4295.SUNLIGHT5.1.Sample solution of IAG-21-001 (2) (20mg/capsule) was prepared as in method SI-IAG-206-02 . The solution was exposed to 500w/hr. cell sunlight for 1hour. The BST was set to 35°C and the ACT was 45°C. The vials were placed in a horizontal position (4mm vials, National + Septum were used). A Dark control solution was tested. A 2%w/v quinine solution was used as the reference absorbance solution.Fluoxetine peak decreased to 91.2% and the dark control solution showed assay of 97.0%. The difference in the absorbance in the quinine solution is 0.4227AU.Additional peak was observed at RRT of 1.5 (2.7%).The total percent of Fluoxetine peak with the degradation peak is about 93.9%.SAMPLE - SUNLIGHTFluoxetine peak:Purity Angle:0.093Match Angle:0.583Purity Threshold:0.148Match Threshold:0.825 ED. N0: 04Effective Date:APPROVED::HPLC ASSAY with DETERMINATION OF META-FLUOXETINE HCl.ANALYTICAL METHOD VALIDATION10 and 20mg Fluoxetine Capsules HPLC DeterminationSUNLIGHT (Cont.)5.2.Working standard solution was prepared as in method SI-IAG-206-02 . The solution was exposed to 500w/hr. cell sunlight for 1.5 hour. The BST was set to 35°C and the ACT was 42°C. The vials were placed in a horizontal position (4mm vials, National + Septum were used). A Dark control solution was tested. A 2%w/v quinine solution was used as the reference absorbance solution.Fluoxetine peak was decreased to 95.2% and the dark control solution showed assay of 99.5%.The difference in the absorbance in the quinine solution is 0.4227AU.Additional peak were observed at RRT of 1.5 (2.3).The total percent of Fluoxetine peak with the degradation peak is about 97.5%. STANDARD - SUNLIGHTFluoxetine peak:Purity Angle:0.067Match Angle:0.389Purity Threshold:0.134Match Threshold:0.8196.HEAT OF SOLUTION6.1.Sample solution of IAG-21-001-(2) (20 mg/capsule) was prepared as in method SI-IAG-206-02 . Equivalent to 20mg Fluoxetine was weighed into a 50mL volumetric flask. 20mL Diluent was added and the solution was sonicated for 10 minutes and made up to volume with Diluent. 4mL solution was transferred into a suitable crucible, heated at 105°C in an oven for 2 hours. The sample was cooled to ambient temperature, filtered and injected into the HPLC system.Fluoxetine peak was decreased to 93.3%.SAMPLE - HEAT OF SOLUTION [105o C]Fluoxetine peak:Purity Angle:0.062Match Angle:0.460Purity Threshold:0.131Match Threshold:0.8186.2.Standard Working Solution (WS) was prepared under method SI-IAG-206-02 . 4mL of the working solution was transferred into a suitable crucible, placed in an oven at 105°C for 2 hours, cooled to ambient temperature and injected into the HPLC system.Fluoxetine peak intensity did not decrease - 100.5%.ED. N0: 04Effective Date:APPROVED::。

方法验证指南The process of method validation is an essential part of ensuring the accuracy and reliability of analytical results in various fields such as pharmaceuticals, clinical testing, and environmental monitoring. 方法验证的过程是确保在制药、临床测试和环境监测等各个领域的分析结果准确性和可靠性的重要环节。

As a crucial step in the development and implementation of a new analytical method, validation helps to demonstrate that the method is suitable for its intended use and produces consistent and reproducible results. 作为一种新分析方法开发和实施过程中的关键步骤，验证有助于证明该方法适合其预期用途，并能产生一致和可重复的结果。

It involves a series of experiments and evaluations that assess the performance characteristics of the method, including accuracy, precision, specificity, detection limits, quantitation limits, linearity, range, and robustness. 它涉及一系列实验和评估，评估方法的性能特征，包括准确性、精密度、特异性、检测限、定量限、线性、范围和稳健性。

Comments concerning revised textspublished in Supplement 9.2The following information details the technical modifications that have been made to revised texts adopted by the European Pharmacopoeia Commission at the June 2016 session and published in Supplement 9.2.When a text has been technically revised, this is indicated by horizontal or vertical lines inthe margin of the supplement. The details given below complete this information, but are not necessarily exhaustive.The following details can also be consulted in the Knowledge database under View history.GENERAL CHAPTERS2.2.1. Clarity and degree of opalescence of liquidsGeneral revision to restructure the text and eliminate unnecessary repetition. The requirements for accuracy and repeatability of the instrument have been changed.2.6.27. Microbiological examination of cell-based preparationsThe revised chapter redefines the scope to exclude preparations covered by the EU Directives for human blood or blood components.The approach to microbiological examination of cell-based preparations that was previously described in the general chapter took particular account of constraints linked to preparations of limited volume and restricted shelf-life. Provisions applicable to other types of cell-based preparations are now included:- preparations for which the volume available for testing is not a limiting factor;- preparations for which administration to the patient does not have to take place before results of the microbiological examination are available; and/or- preparations whose production process makes them more prone to environmental contamination.The main changes to the automated growth-based method include greater flexibility for the incubation temperature(s) and examples of temperature settings where the test volume allows 2 incubation conditions. In addition, in the list of micro-organisms used for method validation, Yersinia enterocolitica is replaced by Micrococcus sp., because it is more appropriate asan example of a common contaminant of cell-based preparations. Information about the sensitivity to be achieved during validation has also been included.The revision has also been an opportunity to refer to general chapter 2.6.1. Sterility, which may be applied, and to introduce alternative rapid test methods, to be used with or without a pre-incubation step, by referring to general chapter 5.1.6. Alternative methods for control of microbiological quality.An introduction has been added with a rationale for method selection according to the characteristics and constraints inherent to the cell-based preparation to be tested. The revised general chapter also includes considerations and recommendations concerning sampling, the sample composition, and ‘negative-to-date’ results.2.6.30. Monocyte-activation testAs a result of a survey distributed by the EDQM in 2013 to users of the Ph.Eur., the following improvements have been made.Introduction: the wording ‘very steep dose-response curves’ has been changed to ‘very steep or non-linear dose-response curves’ as the latter better characterises the responses when non-endotoxin contaminants are present.Definitions: clarification added that calculation of the maximum valid dilution (MVD) is based on the endotoxin reference standard; the possibility of using an estimated limit of detection (LOD) based on historical data when calculating MVD has been introduced.Cell sources and qualification- Additional cross-references to sections relating to the qualification of cells according to their origin, preparation and/or intended use (i.e. for the detection of endotoxin and/or non-endotoxin contaminants) have been included in sections 5-1, 5-2, 5-4, 5-5 and 5-6.- Section 5-4. Qualification of cells pooled from a number of donors: a caution statement has been added regarding the need to consider the averaging effect when cells are pooled.- Section 5-5. Qualification of cryo-preserved cells: the repetitive description regarding the preparation of cell pools has been deleted.- Section 5-6. Monocytic continuous cell lines: a statement regarding the limited use of monocytic cell lines for the detection of non-endotoxin pyrogens has been introduced. Preparatory testing- Section 6-1. Assurance criteria for the endotoxin standard curve: numerical example provided to illustrate the term ‘as low as possible’, which defines the blank.- Section 6-2. Test for interfering factors: text revised so that the concentration of added endotoxin in the preparation or the diluent is to be justified and can be estimated before starting the test. In Method C, it is stated that the type of analysis used to compare the test and reference lots must be justified and validated for each preparation, that assay validity criteria are to be included and that the dilutions tested depend on the type of analysis used. In addition, more information is given on how to test preparations with an inherently high pyrogen content.- Section 6-3. Method validation for non-endotoxin monocyte-activating contaminants: text revised to note that during preparatory testing, at least 2 non-endotoxin ligands for toll-like receptors must be used to validate the test system, 1 of which is also used to spike the test preparation, and that the choice of non-endotoxin pyrogens used should reflect the most likely contaminant(s) of the test preparation. In addition, more information is given regarding the available ligands that can be used.Methods- Section 7-1-1. Method A, Test procedure: regarding the qualification procedure applied to monocytes of different origin, the term ‘qualified cells’ has now been introduced throughout the text. Changes to Table 2.6.30.-1 have been made so that all 3 test solutions (A, B and C)are to be spiked and not just the highest concentration. Solution D has therefore been deleted and replaced by solutions AS, BS and CS (i.e. spiked solutions A, B and C).- Section 7-1-2. Calculation and interpretation: text reworded to reflect the changes in Table 2.6.30.-1. In addition, the text now states that dilutions with an invalid spike recovery are deleted from further analyses and that at least 1 valid dilution is required for a valid test.- Section 7-1-3. Pass/fail criteria of the preparation: conditions for the use of monocytic cell lines have been deleted.- Section 7-2-1/2. Method B, Table 2.6.30.-2: text updated accordingly, as above for Method A. - Section 7-3. Method C. Reference lot comparison test: although there is flexibility on the type of analysis used, the analysis must be justified and validated for each product and is to include assay validity criteria; the text has been changed to reflect this. A statement has also been included to emphasise that the description of the test method includes just an example of a type of analysis which could be used.- Section 7-3-2. Calculation and interpretation: numerical example provided to show a possible acceptance value.Guidance notes- Section 2-1. Information regarding the choice of methods: further clarification is given on the inappropriateness of Method A if the dose-response curve for the preparation to be examined is not parallel to that of the standard endotoxin curve. In addition, a notice has been added regarding the product specific validation and capacity of the chosen method to identify non-responders along with low and high responders to a particular product/contaminant(s) combination(s).- Section 2-5. Cross-validation has been added. Regarding the presence of non-endotoxin pyrogens in the product, a recommendation to perform cross-validation of the monocyte-activation test together with the bacterial endotoxins test has been introduced. In the context of the 3Rs, the rabbit pyrogen test can be performed for cross-validation purposes where the monocyte-activation test cannot be validated.- A new entry has been included in Table 2.6.30.-4 for ‘Parenteral formulations administered per square metre of body surface’, in accordance with the recently revised general chapter 5.1.10. Guidelines for using the test for bacterial endotoxins.- It is now specified that MAT is considered as a replacement for the rabbit pyrogen test.3.1.3. PolyolefinsProduction: information about suitable types of silica introduced.IR identification: absorption maxima deleted since the chapter covers a variety of materials which are further specified under chapters 3.1.4, 3.1.5, 3.1.6 and 3.1.7. Possibility of recording spectra directly on granules or hot pressed films introduced, since this is the technique most often used in current practice.Solution S1: water for injections R replaced by water R.Substances soluble in hexane: test deleted for technical reasons as it can be difficult to perform on certain materials due to the formation of gel which impairs the filtering step and therefore compromises the performance of the test.Phenolic antioxidants: procedure C deleted; additives 11 and 12 are now determined using procedure B; quantitative expression of acceptance criteria introduced.Non-phenolic antioxidants; Amides and stearates: TLC plate replaced.3.1.4. Polyethylene without additives for containers for parenteral preparations and for ophthalmic preparationsIR identification: absorption maxima revised and tolerance added; possibility of recording spectra directly on granules or hot pressed films introduced, since this is the technique most often used in current practice.Solution S1: water for injections R replaced by water R.Substances soluble in hexane: test deleted for technical reasons as it can be difficult to perform on certain materials due to the formation of gel which impairs the filtering step and therefore compromises the performance of the test.3.1.5. Polyethylene with additives for containers for parenteral preparations and for ophthalmic preparationsProduction: information about suitable types of silica introduced.IR identification: absorption maxima revised and tolerance added; possibility of recording spectra directly on granules or hot pressed films introduced, since this is the technique most often used in current practice.Solution S1: water for injections R replaced by water R.Substances soluble in hexane: test deleted for technical reasons as it can be difficult to perform on certain materials due to the formation of gel which impairs the filtering step and therefore compromises the performance of the test.Phenolic antioxidants: procedure C deleted; additives 11 and 12 are now determined using procedure B; quantitative expression of acceptance criteria introduced.Non-phenolic antioxidants; Amides and stearates: TLC plate replaced.3.1.6. Polypropylene for containers and closures for parenteral preparations and ophthalmic preparationsProduction: information about suitable types of silica introduced.IR identification: absorption maxima revised and tolerance added; possibility of recording spectra directly on granules or hot pressed films introduced, since this is the technique most often used in current practice.Solution S1: water for injections R replaced by water R.Substances soluble in hexane: test deleted for technical reasons as it can be difficult to perform on certain materials due to the formation of gel which impairs the filtering step and therefore compromises the performance of the test.Phenolic antioxidants: procedure C deleted; additives 11 and 12 are now determined using procedure B; quantitative expression of acceptance criteria introduced.Non-phenolic antioxidants; Amides and stearates: TLC plate replaced.3.1.7. Poly(ethylene - vinyl acetate) for containers and tubing for total parenteral nutrition preparationsIR identification: absorption maxima revised and tolerance added; possibility of recording spectra directly on granules or hot pressed films introduced, since this is the technique most often used in current practice.Solution S2: water for injections R replaced by water R.Amides and stearates: concentration of reference solutions (b) and (c) corrected for consistency with limits set in the Production section; TLC plate replaced.Substances soluble in hexane: test deleted for technical reasons as it can be difficult to perform on certain materials due to the formation of gel which impairs the filtering step and therefore compromises the performance of the test.5.1.1. Methods of preparation of sterile productsThis text has undergone a general revision and has been completely rewritten. The sections on the different sterilisation processes, where appropriate, now have the same format: principle, equipment, sterilisation cycle, cycle effectiveness and routine control; where required, specific information has been added.Sterility assurance level: the reference to exponential inactivation has been removed as membrane filtration is not a first-order process.Steam sterilisation: modern concepts for validation have been added.Dry heat sterilisation: a wider description of the suitable equipment has been provided. Ionising radiation sterilisation: the reference to European Notes for Guidance has been removed.Gas sterilisation: 2 types of agents are defined: alkylating agents and oxidising agents; the establishment of the cycle effectiveness has been described in more detail.Membrane filtration: the description of the microbial challenge test has been moved to general chapter 5.1.2. Biological indicators and related microbial preparations used in the manufacture of sterile products, published in the same supplement.Aseptic assembly: freeze-drying under aseptic conditions is added.5.1.2. Biological indicators and related microbial preparations used in the manufacture of sterile productsThe general chapter has undergone significant revision as listed below.Title: it has been adapted to take into account microbial preparations used for sterilisation grade filtration.Introduction: describes when biological indicators (BIs) are intended to be used and what is outside the scope of the general chapter, including that BIs are in most cases only to be used for development of the sterilisation process and are not to be employed for routine monitoring unless otherwise stated in this general chapter. A definition of BIs is given and the processes in which they can be used are described. Importantly, the Introduction section introduces the concept of the use of reduced sterilisation process conditions in order to ensure the validity of the sterilisation process. It is also made clear that there should be no surviving micro-organisms when the biological indicator is subject to a full sterilisation process.BIs for sterilisation processes: this section gives guidance on how BIs are selected and how they are used to characterise sterilisation processes.A description is provided of 4 types of BIs for sterilisation processes: inoculated carriers, self-contained BIs, characterised spore suspensions and custom-made BIs.Information regarding the quality requirements for BIs and user requirement specifications have been introduced.BIs for heat sterilisation: the parameters of BIs for heat sterilisation are described and how a validation cycle is established. Further information on biological validation with reduced sterilisation cycles has been included.BIs for moist heat sterilisation: it is recognised that Geobacillus stearothermophilus maynot be suitable for sterilisation processes delivering an F0 between 8 and 15, therefore adifferent test micro-organism may be used.BIs for dry heat sterilisation: description of the reference conditions and an example of how survivor rates of typical BIs are affected by temperature variations are given.BIs for gas sterilisation: this section sets out that the use of gas sterilisation for disinfection is outside the scope of the general chapter. There are a number of different types of gas sterilisation processes and no reference cycles, therefore no criteria to which the BIs shall comply have been defined. Suitable micro-organisms for ethylene oxide sterilisation are given. It is, however, the responsibility of the user to define the cycle and the suitability of any BI used.BIs for ionising radiation sterilisation: it is recognised that BIs are not considered to be necessary for defining the suitability of the radiation sterilising dose, but their use may be required for the development and validation of ionising radiation sterilisation in specific cases. Information on test micro-organisms is given.Microbial preparations for sterilisation grade filtration: information on test micro-organisms is now given for the validation of retention of micro-organisms using a membrane.Indicators for depyrogenation processes: this section has been removed from this general chapter and will be published elsewhere in the Ph. Eur.The following article was published in Pharmeuropa Bio & Scientific Notes and can be consulted on http://pharmeuropa.edqm.eu/PharmeuropaBioSN/ (registration required) for further information:K. Haberer, H. van Doorne. Biological indicators, tools to verify the effect of sterilisation processes - position paper prepared on behalf of Group 1 (biological methods and statistical analysis). Pharmeur Bio Sci Notes 2011(2):26-39.5.1.6. Alternative methods for control of microbiological qualityThe chapter has been completely revised and rewritten to take account of technological developments in alternative microbiological methods.The introduction and the sections concerning the 3 major types of determinations specific to microbiological tests have been reworded and expanded. In addition, information on the use of alternative methods for process analytical technology (PAT) has also been given.Under Identification tests, addition of:- paragraph dealing with databases and their validation;- some requirements on micro-organism culture for identification purposes;- remark on a potential disadvantage of traditional biochemical and phenotypic techniques versus genotypic methods.Under General principles of alternative methods:- some methods have been removed, namely microcalorimetry and phage-based methods, and the former section on media development has been replaced by a section on growth detection using selective and/or indicative media;- in the section on direct measurement, a new autofluorescence method has been added;- the section on biochemical assays based on physiological reactions (2-3-1-5) has been expanded to include alternatives to the traditional Gram staining method;- the section on genotypic techniques (2-3-2) has been extensively revised to reflect improvements in this field, including current DNA or RNA-based detection methods; there has also been revision of the critical aspects and potential uses to make them more relevant for users.The section on validation of alternative microbiological methods has been restructured and now gives details of the validation process (both primary validation (3-2-3) and validationfor the intended use (3-2-4)) as well as details on the validation of the different types of microbiological tests (3-3). As microbiological tests have 3 basic applications (qualitative, quantitative and identification), 3 separate sets of validation criteria are now included.The validation example section has been removed from the chapter and will be added tothe knowledge database at a later date. This will allow for the examples to evolve in a more flexible way and eliminate any misinterpretation of the purpose of the examples.5.8. Pharmacopoeial harmonisationInformation modified for several excipients.5.15. Functionality-related characteristics of excipientsChapter completely reviewed and numerous modifications introduced to align better with ICH guideline Q8 Pharmaceutical Development.5.22. Names of herbal drugs used in traditional Chinese medicineTable updated to include a new monograph published in Supplement 9.2.GENERAL MONOGRAPHSHerbal drugs (1433)Definition: section slightly modified to take account of the fact that algae, fungi and lichen do not belong to the plant kingdom.Dried herbal drugs: when used for the production of essential oils, an exemption has been introduced to evaluate on a case-by-case basis if a particular test needs to be performed or not.Fresh herbal drugs: new section introduced as not all tests required for dried herbal drugs are considered equally suitable for fresh herbal drugs.DOSAGE FORMSGlossary (1502)Basis: the definition has been reworded, including the notion of single-phase and multiphase systems.Dispersion: the terms ‘Colloidal dispersion’, ‘Emulsion’ and ‘Suspension’ have been placed under the new entry ‘Dispersion’ with revised definitions.Solution: some additional information on the state of the dissolved substances has been included.Standard term: a more detailed definition has been elaborated.VACCINES FOR VETERINARY USEFurunculosis vaccine (inactivated, oil-adjuvanted, injectable) for salmonids (1521)Batch potency test (section 2-3-1) : revised to clarify that alternative methods are not limited to serological methods.Vibriosis (cold-water) vaccine (inactivated) for salmonids (1580)Batch potency test (section 2-3-1) : revised to clarify that alternative methods are not limited to serological methods.Vibriosis vaccine (inactivated) for salmonids (1581)Batch potency test (section 2-3-1) : revised to clarify that alternative methods are not limited to serological methods.Yersiniosis vaccine (inactivated) for salmonids (1950)Batch potency test (section 2-3-1) : revised to clarify that alternative methods are not limited to serological methods.RADIOPHARMACEUTICAL PREPARATIONS AND STARTING MATERIALS FOR RADIOPHARMACEUTICALPREPARATIONSCopper tetramibi tetrafluoroborate for radiopharmaceutical preparations (2547) The test for bacterial endotoxins (2.6.14) has been deleted from the monograph. The requirements for bacterial endotoxins (2.6.14) as outlined in the general monograph Chemical Precursors for Radiopharmaceutical Preparations (2902) are applicable.Iobenguane sulfate for radiopharmaceutical preparations (2351)Identification test A: the use of an iobenguane sulfate CRS has been introduced replacing the reference spectra.The test for bacterial endotoxins (2.6.14) and the labelling section have been deleted from the monograph. The requirements for bacterial endotoxins (2.6.14) and labelling as outlined in the general monograph Chemical Precursors for Radiopharmaceutical Preparations (2902) are applicable.Medronic acid for radiopharmaceutical preparations (2350)The test for bacterial endotoxins (2.6.14) and the labelling section have been deleted from the monograph. The requirements for bacterial endotoxins (2.6.14) and labelling as outlined in the general monograph Chemical Precursors for Radiopharmaceutical Preparations (2902) are applicable.Sodium iodohippurate dihydrate for radiopharmaceutical preparations (2352) The test for bacterial endotoxins (2.6.14) and the labelling section have been deleted from the monograph. The requirements for bacterial endotoxins (2.6.14) and labelling as outlined in the general monograph Chemical Precursors for Radiopharmaceutical Preparations (2902) are applicable.Technetium (99m Tc) bicisate injection (2123)Radiochemical purity: inclusion of a further group of impurities determined by the TLC test. Limit for the sum of all impurities widened.Technetium (99m Tc) mebrofenin injection (2393)Impurity A: modification of the preparation of reference solution (b) to achieve full dissolution of the CRS.Tetra-O-acetyl-mannose triflate for radiopharmaceutical preparations (2294)The test for bacterial endotoxins (2.6.14) and the labelling section have been deleted from the monograph. The requirements for bacterial endotoxins (2.6.14) and labelling as outlined in the general monograph Chemical Precursors for Radiopharmaceutical Preparations (2902) are applicable.HERBAL DRUGS AND HERBAL DRUG PREPARATIONSAcanthopanax bark (2432)Definition: currently accepted botanical name introduced.Identification B: illustration of powdered herbal drug introduced and its legend integrated into text.Aniseed (0262)Water: limit for water content increased.Astragalus mongholicus root (2435)Definition: currently accepted botanical name introduced.Identification B: illustration of powdered herbal drug introduced and its legend integrated into text of Identification B.Assay: extraction procedure modified to improve efficiency (soxhlet extraction replaced by sonication, and solid phase extraction step deleted).Aucklandia root (1797)Identification: more detailed description provided for Identification A; illustration of powdered herbal drug introduced and its legend integrated into text of Identification B.Coix seed (2454)Identification B: illustration of powdered herbal drug introduced and its legend integrated into text.Drynaria rhizome (2563)Identification B: illustration of powdered herbal drug introduced and its legend integrated into text of Identification B.Eclipta herb (2564)Identification B: illustration of powdered herbal drug introduced and its legend integrated into text.Eucommia bark (2412)Identification B: illustration of powdered herbal drug introduced and its legend integrated into text.Myrrh (1349)Commiphora mukul: TLC replaced by HPTLC allowing distinction between different resins. Matter insoluble in ethanol: limit increased based on batch data.Myrrh tincture (1877)Identification: TLC replaced by same HPTLC used for Myrrh (1349), allowing distinction between different resins.Peppermint leaf (0406)Identification C: TLC replaced by HPTLC to allow differentiation between different Mentha species.Peppermint leaf dry extract (2382)Identification: HPTLC method updated to allow differentiation between different Mentha species.MONOGRAPHSAmiloride hydrochloride dihydrate (0651)Related substances: impurity C is now a specified impurity; the limit for total impurities has been increased accordingly.Benzylpenicillin potassium (0113)Definition: means of production amended and lower content limit adjusted based on revised limits for total impurities.Characters: appearance and solubility updated.Identification: TLC test updated in line with current Style guide.Appearance of solution: test introduced as substance can be for parenteral use.Specific optical rotation, Absorbance: tests removed as deemed no longer required based on improved LC for related substances.Related substances: improved LC introduced allowing for identification of 2 additional impurities; impurity limits updated to reflect current batches on market.Bacterial endotoxins: test removed according to Ph. Eur. policy.Impurities: impurities G and H added.Benzylpenicillin sodium (0114)Definition: means of production amended and lower content limit adjusted based on revised limits for total impurities.Characters: appearance and solubility updated.Identification: TLC test updated in line with current Style guide.Appearance of solution: test introduced as substance can be for parenteral use.Specific optical rotation, Absorbance: tests removed as deemed no longer required based on improved LC for related substances.Related substances: improved LC introduced allowing for identification of 2 additional impurities; impurity limits updated to reflect current batches on market.Bacterial endotoxins: test removed, according to Ph. Eur. policy.Impurities: impurities G and H added.Cellulose acetate (0887)Identification (IR): solvent modified to ensure adequate solubility.Cholesterol for parenteral use (2397)Benzoyl ureas: ‘rotary evaporator’ replaced by ‘evaporate by suitable means’.Colistimethate sodium (0319)Chemical formula: parent structure introduced to show disubstitution at N4 in 2 to 5 of the DAB residues.Definition: updated to reflect the current understanding of the substance’s molecular structure.Production: section introduced to control the composition and purity of colistin starting material.Identification: former tests A, B and C replaced by test for composition.Specific optical rotation: test removed as the substance is adequately controlled by the tests for composition and related substances.Composition, Related substances: LC method introduced which is capable of separating and quantifying the polymyxin components and impurities; limits for the components CMSE1ASM8, CMS E1ASM6, CMS E1ASM4, CMS E2ASM8, CMS E2ASM6, CMS E2ASM4 and also limits for any other impurity and sum of impurities introduced based on available batch data.Total sulfite: test removed.Ethylcellulose (0822)Definition: possibility of adding antioxidants is now stated.Identification A: sample preparation added.Assay: chromatographic conditions modified to involve the use of a wide-bore capillary GC column instead of a packed column.Functionality-related characteristics (FRCs): section added for ethylcellulose used as binder and film former.Glycerol monostearate 40-55 (0495)Functionality-related characteristics: this section has been added. Glycerol monostearate 40-55 is used as matrix former in prolonged-release oral solid dosage forms and as consistency agent in dosage forms for cutaneous application. For use as matrix former, the tests for composition of fatty acids, powder flow and particle-size distribution are cross-referenced. For use as consistency agent, the test for composition of fatty acids is cross-。