1、（1092）溶出度试验的开发和验证【中英文对照版】INTRODUCTION前言Purpose目的The Dissolution Procedure: Developmentand Validation provides a comprehensive approach covering items to considerfor developing and validating dissolution procedures and the accompanyinganalytical procedures. It addresses the use of automation througho
2、ut the testand provides guidance and criteria for validation. It also addresses thetreatment of the data generated and the interpretation of acceptance criteriafor immediate- and modified-release solid oral dosage forms.溶出实验 :开发和验证（ 1092）指导原则 提供了在溶出度方法开发和验证过程中以及采用相应分析方法时需要考虑的因素。本指导原则贯穿溶出度实验的全部过程，并对方
3、法提供了指导和验证标准。同时它还涉及对普通制剂和缓释制剂所生成的数据和接受标准进行说明。Scope范围Chapter addresses the development andvalidation of dissolution procedures, with a focus on solid oral dosage forms.Many of the concepts presented, however, may be applicable to other dosageforms and routes of administration. General recommendations
4、are given with theunderstanding that modifications of the apparatus and procedures as given in USPgeneral chapters need to be justified.章节讨论了溶出度实验的开发和验证，重点是口服固体制剂。所提出的许多概念也可能适用于其他剂型和给药途径。关于设备和方法的修改部分在 USP 通则中给出了合理的说明。The organization of follows the sequence of actions often performed inthe developme
5、nt and validation of a dissolution test. The sections appear inthe following sequence.在进行溶解度实验的开发和验证时，常遵循指导原则，具体内容如下：1. PRELIMINARY ASSESSMENT (FOR EARLY STAGES OF PRODUCTDEVELOPMENT/DISSOLUTION METHOD DEVELOPMENT)1. 前期评估（对产品开发以及溶出度方法开发的前期研究评估）1.1 Performing Filter Compatibility1.1 滤膜相容性研究1.2 Determ
6、ining Solubility and Stability of DrugSubstance in Various Media1.2 原料药在不同溶出介质中溶解度测定和稳定性研究1.3 Choosing a Medium and Volume1.3 溶出介质和体积选择1.4 Choosing an Apparatus1.4 溶出设备选择（桨法和篮法以及其他方法）2. METHOD DEVELOPMENT2. 方法开发2.1 Deaeration2.1 脱气2.2 Sinkers2.2 沉降篮2.3 Agitation2.3 转速2.4 Study Design2.4 研究设计2.4.1 Ti
7、mePoints2.4.1 取样时间点2.4.2 Observations2.4.2 观察2.4.3 Sampling2.4.3 取样2.4.4 Cleaning2.4.4 清洗2.5 Data Handling2.5 数据处理2.6 Dissolution Procedure Assessment2.6 溶出方法评估3. ANALYTICAL FINISH3.完成分析3.1 Sample Processing3.1 样品处理3.2 Filters3.2 过滤3.3 Centrifugation3.3 离心3.4 Analytical Procedure3.4 分析方法3.5 Spectrop
8、hotometric Analysis3.5 光谱分析3.6 HPLC3.6HPLC 法4. AUTOMATION4.自动化4.1 Medium Preparation4.1 介质的配制4.2 Sample Introduction and Timing4.2 定时进样4.3 Sampling and Filtration4.3 取样和过滤4.4 Cleaning4.4 清洗4.5 Operating Software and Computation of Results4.5 操作软件和计算的结果5. VALIDATION5.验证5.1 Specificity/Placebo Interfe
9、rence5.1 专属性/安慰剂（辅料）干扰5.2 Linearity and Range5.2 线性和范围5.3 Accuracy/Recovery5.3 准确度/回收率5.4 Precision5.4 精密度5.4.1 REPEATABILITY OF ANALYSIS5.4.1 重复性5.4.2 INTERMEDIATE PRECISION/RUGGEDNESS5.4.2 中间精密度/耐用性5.4.3 REPRODUCIBILITY5.4.3 重现性5.5 Robustness5.5 耐用性5.6 Stability of Standard and Sample Solutions5.6
10、 样品溶液和标准溶液的稳定性5.7 Considerations for Automation5.7 自动操作注意事项6. ACCEPTANCE CRITERIA6.可接受标准6.1 Immediate-Release Dosage Forms6.1 速释剂型6.2 Delayed-Release Dosage Forms6.2 延迟释放剂型6.3 Extended-Release Dosage Forms6.3 延长释放剂型6.4 Multiple Dissolution Tests6.4 多个溶解度试验6.5 Interpretation of Dissolution Results6.5
11、 溶出结果说明6.5.1 IMMEDIATE-RELEASE DOSAGE FORMS6.5.1 即时释放剂型6.5.2 DELAYED-RELEASE DOSAGE FORMS6.5.2 延迟释放剂型6.5.3 EXTENDED-RELEASE DOSAGE FORMS6.5.3 延长释放剂型1. PRELIMINARYASSESSMENT (FOR EARLY STAGES OF PRODUCT DEVELOPMENT/DISSOLUTION METHODDEVELOPMENT)1. 前期评估（产品开发/溶出度方法开发的初期阶段）Beforemethod development can b
12、egin, it is important to characterize the molecule sothat the filter, medium, volume of medium, and apparatus can be chosen properlyin order to evaluate the performance of the dosage form.在开始溶出方法开发之前，我们对用以评价制剂溶出行为的滤膜、溶出介质、溶出介质体积和溶出设备进行适当的筛选是非常重要的。1.1 Performing Filter Compatibility1.1 滤膜相容性研究Filtrat
13、ionis a key sample-preparation step in achieving accurate test results. Thepurpose of filtration is to remove undissolved drug and excipients from thewithdrawn solution. If not removed from the sample solution, particles of thedrug will continue to dissolve and can bias the results. Therefore, filte
14、ringthe dissolution samples is usually necessary and should be done immediately ifthe filter is not positioned on the cannula.为获得准确试验结果，过滤是样品制备的一个关键步骤。过滤的目的是为了除去溶出液中 未溶解的药物和辅料。如果不把未溶解的药物和辅料从样品溶液中除去，那么未溶解的药物颗粒将会继续溶解使试验结果出现偏差，因此，如果取样管中没有过滤器，应立即对溶出度样品进行过滤。Filtration also removes insolubleexcipients tha
15、t may otherwise interfere with the analytical finish. Selectionof the proper filter material is important and should be accomplished, andexperimentally justified, early in the development of the dissolutionprocedure. Important characteristics to consider when choosing a filtermaterial are type, filt
16、er size, and pore size. The filter that is selectedbased on evaluation during the early stages of dissolution procedure developmentmay need to be reconsidered at a later time point. Requalification has to beconsidered after a change in composition of the drug product or changes in thequality of the
17、ingredients (e.g. particle size of microcrystalline cellulose).过滤也可除去可能会干扰分析测定的不溶性辅料。选择适当的过滤材料是非常重要，应该在早期溶出方法开发的过程中通过实验确定和完成。在选择滤膜时有必要重点考虑滤膜的材料、型号和孔径大小。通常对早期阶段溶出方法开发过程的评价选择过滤器，但在后期试验中如果制剂成分改变或组成成分质量变化可能需要重新考虑过滤器，（例如：微晶纤维素粒径的改变）。Examples of filters used in dissolutiontesting can be cannula filters, f
18、ilter disks or frits, filter tips, or syringefilters. The filter material has to be compatible with the media and the drug.Common pore sizes range from 0.20 to 70 mm, however, filters of other poresizes can be used as needed. If the drug substance particle size is very small(e.g., micronized or nano
19、particles), it can be challenging to find a filterpore size that excludes these small particles.用于溶出试验的过滤器有管路过滤器、过滤盘或玻璃过滤器、滤头或针头式过滤器。过滤材料必须与介质和药物相适合。常见孔径大小范围：0.2070m ，如果需要也可使用其他孔径大小的过滤器。如果原料药的粒度很小（例如，微分化颗粒或纳米颗粒），找到一个合适的过滤器过滤这些小颗粒至今仍具有挑战性。Adsorption of the drug(s) by the filtermay occur and needs to
20、be evaluated. Filter materials will interact withdissolution media to affect the recovery of the individual solutes and must beconsidered on a case-by-case basis. Different filter materials exhibitdifferent drug-binding properties. Percentage of drug loss from the filtratedue to binding may be depen
21、dent on the drug concentration. Therefore theadsorptive interference should be evaluated on sample solutions at differentconcentrations bracketing the expected concentration range. Where the drugadsorption is saturable, discarding an initial volume of filtrate may allow thecollection of a subsequent
22、 solution that approaches the original solutionconcentration. Alternative filter materials that minimize adsorptiveinterference can usually be found. Prewetting of the filter with the medium maybe necessary. In addition, it is important that leachables from the filter donot interfere with the analyt
23、ical procedure. This can be evaluated by analyzingthe filtered dissolution medium and comparing it with the unfiltered medium.过滤时可能会发生药物的吸附，需要进行评估。过滤材料将与溶出介质相互作用，影响每个溶质的回收率应该根据具体问题进行考虑。不同的过滤材料表现出与药物结合的不同特性。由于药物与滤膜结合引起药物从滤液中损失的比例，可能依赖于药物浓度。因此，应采用预期浓度范围内不同浓度的样品溶液来评估滤膜吸附干扰。由于药物吸附是可饱和的，弃去一定体积的初滤液，收集续滤液，
24、以达到接近原来的溶液浓度的样品也是可取的。通常选择适合的过滤材料，最大限度地减少滤膜吸附干扰，润湿滤膜对减少吸附也是必要的。此外，过滤后的溶出物不干扰分析检测也是非常重要的，这可以通过过滤后的溶出介质过滤与未过滤的溶出介质进行比较，评估滤膜是否干扰分析测定。The filter size should be based on thevolume to be withdrawn and the amount of particles to be separated. Use of thecorrect filter dimensions will improve throughput and r
25、ecovery, and also reduceclogging. Use of a large filter for small-volume filtration can lead to loss ofsample through hold-up volume, whereas filtration through small filter sizesneeds higher pressures and longer times, and the filters can clog quickly.根据要过滤样品溶液的体积以及样品溶液中颗粒的量选择滤膜孔径。使用正确的滤膜孔径将提高溶液的通过
26、率和回收率，并减少滤膜堵塞。使用大孔径滤膜过滤小体积溶液，能够导致样品溶液损失量过大而收集不到所用样品量；使用小孔径滤膜过滤，需要更高的压力和较长的时间，并且溶液迅速堵塞滤膜。Filters used for USP Apparatus 4 needspecial attention because they are integrated in the flow-through process.Undissolved particles may deposit on the filters, creating resistance to theflow.USP 仪器 4 中使用的过滤器需要特别
27、注意，因为它们在流动过程中使用。不溶颗粒会沉积在过滤器，产生流动阻力。In the case of automated systems,selection of the filter with regard to material and pore size can be done in asimilar manner to manual filtration. Flow rate through the filter and cloggingmay be critical for filters used in automated systems. Experimentalverificat
28、ion that a filter isappropriate may be accomplished by comparing the responses for filtered andunfiltered standard and sample solutions. This is done by first preparing asuitable standard solution and a sample solution. For example, prepare atypical dissolution sample in a beaker and stir vigorously
29、 with a magneticstirrer to dissolve the drug load completely.For standard solutions, comparethe results for filtered solutions (after discarding the appropriate volume) tothose for the unfiltered solutions. For sample solutions, compare the resultsfor filtered solutions (after discarding the appropr
30、iate volume) to those forcentrifuged, unfiltered solutions.在自动化系统的情况下，关于过滤器滤膜材料和孔径大小可以用类似的方式通过手动过滤进行选择。在自动化系统中通过过滤器的流量和过滤器的堵塞可能是至关重要的。通过试验比较过滤和未过滤的标准溶液和样品溶液的含量差别，验证该过滤器是合适的。首先制备一个合适的标准溶液和样品溶液。例如，在烧杯中制备一个标准溶解样品，用磁力搅拌器搅拌使药物完全溶解。对于标准溶液，比较过滤溶液（弃去的适当体积后）和未过滤溶液的含量测定结果；对于样品溶液，比较过滤（弃去适当体积后）、离心、未过滤样品溶液的含量测定结
31、果。1.2 Determining Solubility and Stability of DrugSubstance in Various Media1.2 原料药在不同溶出介质中的溶解度测定和稳定性研究Physical and chemical characteristics of the drug substance need to be determinedas part of the process of selecting the proper dissolution medium. Whendeciding the composition of the medium for di
32、ssolution testing, it is importantto evaluate the influence of buffers, pH, and if needed, different surfactantson the solubility and stability of the drug substance. Solubility of the drugsubstance is usually evaluated by determining the saturation concentration ofthe drug in different media at 37
33、using the shake-flask solubility method(equilibrium solubility). To level out potential ion effects between the drugand the buffers used in the media, mixtures of hydrochloric acid and sodiumhydroxide are used to perform solubility investigations; this is in addition tothe typical buffer solutions.
34、In certain cases, it may be necessary to evaluatethe solubility of the drug at temperatures other than 37 (i.e., 25). The pHof the clear supernatant should be checked to determine whether the pH changesduring the solubility test. Alternative approaches for solubility determinationmay also be used.在选
35、择合适溶出介质的过程中，需要确定原料药的物理化学特性。当需要确定溶出度试验中溶出介质的组成时，有必要评估缓冲液、pH 值、以及不同的表面活性剂（如果需要）对药物的溶解度和稳定性的影响。在 37温度条件下，采用摇瓶溶解法（平衡溶解度）测定原料药在不同介质中的饱和浓度，来评估药物的溶解性。为了消除溶出介质中药物和缓冲液之间离子的潜在影响，使用盐酸和氢氧化钠的混合物对溶解度进行研究，这是一种典型的缓冲溶液。在某些情况下，评估药物在 37以外条件下（即，25）的溶解度可能也是必要的。在溶解度试验过程中应检查上清溶液的 pH 值，以确定在溶解过程中 pH 值是否改变。也可使用其他可供选择的方法进行溶解度
36、测定。Typical media for dissolution mayinclude the following (not listed in order of preference): diluted hydrochloricacid, buffers (phosphate or acetate) in the physiologic pH range of 1.27.5, simulatedgastric or intestinal fluid (with or without enzymes),and water. For somedrugs, incompatibility of t
37、he drug with certain buffers or salts may influencethe choice of buffer. The molarity of the buffers and acids used can influencethe solubilizing effect, and this factor may be evaluated.溶出的典型介质包括（未按照优先顺序列出）：稀盐酸、在生理 pH 值范围为 1.2-7.5 缓冲溶液（磷酸盐或者醋酸盐）、模拟胃液或肠液（含有或不含有酶）和水。对于一些药物，与药物不相容的特定缓冲液或盐可能会影响缓冲剂的选择。所
38、使用的缓冲液和酸的体积摩尔浓度能够改变药物的增溶作用，这个因素也需要评估。Aqueous solutions (acidic or buffersolutions) may contain a percentage of a surfactant e.g., sodium dodecylsulfate (SDS),polysorbate, or lauryldimethylamine oxide to enhance thesolubility of the drug. The surfactants selected for the solubilityinvestigations shou
39、ld cover all common surfactant types, i.e., anionic,nonionic, and cationic. When a suitable surfactant has been identified,different concentrations of that surfactant should be investigated to identifythe lowest concentration neededto achieve sink conditions. Typically,the surfactant concentration i
40、s above its critical micellar concentration(CMC). Table 1 shows a list of some of the surfactants used indissolution media. Approximate CMC values are provided with referenceswhenavailable. The list is not comprehensive and is not intended to exclude surfactantsthat are not listed. Other substances,
41、 such ashydroxypropyl b -cyclodextrin,have been used as dissolution media additives to enhance dissolution of poorlysoluble compounds.The U.S. Food and Drug Administration (FDA) maintains adatabase of dissolution methods, including information on dissolution mediathat have been used (1). Typically,
42、the amount of surfactant added issufficient to achieve sink conditions in the desired volume of dissolutionmedium.有时候水溶性介质中（酸性水溶液或缓冲溶液）可能添加一定比例的表面活性剂（如十二烷基硫酸钠（SDS），聚山梨醇酯，或十二烷基二甲基氧化胺）以提高药物的溶解度。选择用于溶解度研究的表面活性剂时应涵盖所有常用种类的表面活性剂，比如阴离子、非离子型和阳离子，当已经确定一个合适的表面活性剂时，应对表面活性剂的不同浓度进行研究，以确定达到漏槽条件所需的最低浓度。一般情况下，表面活性
43、剂的浓度高于它的临界胶束浓度（CMC）。表 1 列出了溶出介质中常用的表面活性剂，表中提供了 CMC 的近似临界值，以便我们参考，此外，表中所列表面活性剂并不全面，不能排除未列出的表面活性剂。其他表面活性剂，如羟丙基 -环糊精，已被用来作为溶出介质添加剂提高难溶性化合物的溶解度，美国食品药品管理局（FDA）溶出度数据库中，已经收载含有羟丙基 -环糊精的溶出介质（ 1）。通常情况下，表面活性剂的加入量以满足达到漏槽条件所需的溶出介质体积。It is important to control thegrade and purity of surfactants because use of dif
44、ferent grades could affectthe solubility of the drug. For example, SDS is available in both a technicalgrade and a high-purity grade. Obtaining polysorbate 80 from different sourcescan affect its suitability when performing high-performance liquidchromatography (HPLC) analysis.由于使用不同级别的表面活性剂会影响药物的溶解
45、度，因此要控制表面活性剂的级别和纯度。例如，SDS 只有在工业级和高纯度级才可以使用。在使用HPLC 方法进行分析时，不同来源的聚山梨酯（吐温）80 会影响它的适用性。There may be effects of counter-ions orpH on the solubility or solution stability of the surfactant solutions. Forexample, a precipitate forms when the potassium salt for the phosphate bufferis used at a concentratio
46、n of 0.5 M in combination with SDS. This can beavoided by using the sodium phosphate salt when preparing media with SDS.反离子或 pH 值可能会影响表面活性剂溶液的溶解性或稳定性。例如，当含有 SDS 的磷酸盐缓冲液中钾盐浓度为 0.5mol/L 时，就形成了沉淀析出，但是使用磷酸钠制备含有 SDS 的介质时，可以避免这种现象发生。Table 1. Commonly Used Surfactants with Critical Micelle Concentrations表
47、1 常见表面活性剂的临界胶束浓度Routinely, the dissolution medium is buffered; however, the useof purified water as the dissolution medium is suitable for products with adissolution behavior independent of the pH of the medium. There are severalreasons why purified water may not be preferred. The water quality can
48、varydepending on its source, and the pH of the water is not as strictly controlledas the pH of buffer solutions. Additionally, the pH can vary from day to dayand can also change during the run, depending on the drug substance andexcipients. Use of an aqueousorganic solvent mixture as a dissolution m
49、ediumis discouraged; however,with proper justification this type of medium may beacceptable.通常，溶出介质为缓冲盐溶液，但是，对于非 pH 值依赖性的制剂可以使用纯化水作为溶出介质。不推荐使用纯化水作为溶出介质的原因：水的质量变化取决于它的来源，而水的 pH 值不像缓冲溶液能够严格控制；此外，若药物和辅料的溶出对 pH 值敏感时需要考虑使用缓冲液。另外使用水 -有机溶剂混合物作为溶出介质也是不推荐的，但是，特殊情况下（有充分适当的理由），也是可以接受的。Investigations of the stability of thedrug substance should be carried out, when needed, in the selected dissolutionmedium with excipients present, at 37. This elevated temperature has thepotential to decrease solution stability (degradation). Stability should allowfor sufficient time to complete or repeat
