相容性研究中几个关键阈值的简介及转换

目的：通过相容性研究中可提取物与浸出物评估方法研究和评估阈值的转换,将化学物质的毒理学数据转换为化学物质的分析数据,从而实现药品与包装材料相容性研究中对浸出物的控制。方法：文献查阅,汇总美国产品质量研究学会（ PQRI）毒理学家和化学家们确定的几个重要概念、关键阈值,并举例说明。意义：为在药品研发及生产过程中对药品及包装系统的可提取物与浸出物进行研究和控制提供明确的思路和方法。     

化学药品注射剂可提取物和浸出物杂质评估与控制的一般考虑

目的： 从总体思路、研究方法、安全性评估及阈值等方面探讨化学药品注射剂可提取物、浸出物杂质评估和控制的一般考虑,为相关研究人员提供参考。方法：根据我国已发布的相容性研究技术指导原则以及当前国际通行的技术要求,结合技术审评过程中发现和总结的若干典型案例进行分析。结果与结论：注射剂中浸出物杂质所引入的风险是由接触时间、接触条件、制剂和材料特性等各种因素综合决定的。研究者应结合风险评估情况,制定适宜的评估和控制策略。     

无名异炮制乳香的最佳工艺研究

目的探讨无名异制乳香的炮制工艺。方法以水浸提物挥发油含量及醇浸提物量为主要考察指标,采用正交设计筛选工艺。结果用无名异炮制的乳香,加9倍量水,提取6 h,以中火炒制,20%无名异和5%醋的炮制品中挥发油的含量和醇浸出物含量最高。结论火候、无名异和醋的用量是影响无名异制乳香中挥发油含量和醇提取物量的重要因素。该工艺是在生产条件下优选出的,对指导生产有参考价值。     

广东紫珠超临界提取物的GC-MS成分分析及体外抗菌活性

采用超临界CO2萃取法提取广东紫珠低极性成分,应用气相色谱-质谱法分析提取物的化学组成,结合计算机检索技术初步鉴定所分离的化合物,应用色谱峰面积归一化法计算各成分的相对百分含量,并考察各分离釜提取物对金黄色葡萄球菌、大肠杆菌和白色念珠菌的体外抗菌活性。结果从超临界CO2流体萃取分离釜Ⅰ提取物中初步鉴定了27个化合物,占总离子流图峰面积的81.12%;超临界CO2流体萃取分离釜Ⅱ提取物初步鉴定了20个化合物,占总离子流图峰面积的74.60%。广东紫珠超临界CO2萃取分离釜Ⅰ、Ⅱ提取物及二者1︰1混合物对金黄色葡萄球菌、大肠杆菌和白色念珠菌显示出抗菌活性。     

不同采收法对蟾衣水溶性和脂溶性浸出物含量的影响和比较

目的不同采收法对蟾衣水溶性和脂溶性浸出物含量的影响。方法分别检测传统采集方法和经过改进的方法收集到的蟾衣样品之水、95%乙醇、无水乙醚浸出物干重占样品干重百分比。结果改进的方法采集到的蟾衣水浸出物和95%乙醇浸出物干重占样品干重百分比明显高于传统方法 ,而无水乙醚浸出物干重百分比在两种方法之间无明显差异。结论水漂洗的方法可导致蟾衣中极性成分大量流失,但对非极性成分则影响不大。     

HS-SPME-GC-MS联用分析向天果壳、仁中挥发性成分的差异

摘要：目的:通过顶空-固相微萃取-气相-质谱联用技术（HS-SPME-GC-MS）分析向天果果壳、果仁挥发性成分的差异。方法:采用顶空-固相微萃取（HS-SPME）萃取向天果果壳和果仁的挥发物,并结合气相色谱-质谱（GC-MS）联用技术分析挥发性成分的组成,用面积归一化法计算各成分的相对百分含量。结果:向天果果壳中鉴定出48个化合物,占挥发物总量的88.55%;果仁中鉴定出34个化合物,占挥发物总量的62.29%。果壳和果仁中共鉴定出化合物72个,两者共有的成分为10个,其中62个为在向天果成分中首次报道。果壳的挥发物绝对含量较高,是果仁的约2倍。结论:向天果果壳和果仁挥发物的化学成分在种类和含量上差异较大。     

四季感冒片中3味药材超临界CO_2复方萃取物挥发性成分分析

目的:分析四季感冒片中紫苏叶、陈皮、荆芥的超临界CO2复方萃取物挥发性成分。方法:采用超临界CO2萃取技术萃取四季感冒片中的3味药材的挥发性成分,采用气相色谱-质谱法对复方提取物进行成分分析。结果:超临界CO2复方提取法的提取率为2.21%,从复方萃取物中分离到21个化合物,鉴定出16个化合物,主要成分是甜橙黄酮(36.56%)和亚油酸(19.52%)。水蒸气蒸馏法提取率为1.035%,从复方提取物中共分离出51个化合物,鉴定出32种化合物,主要成分是D-柠檬烯(62.40%)和侧柏酮(15.49%)。结论:超临界CO2复方萃取技术可提取紫苏叶、陈皮、荆芥3味药材中的挥发性成分,但与水蒸气蒸馏法所得复方挥发油的成分存在差别。     

仙鹤草提取物镇痛抗炎试验的实验研究

目的:初步探讨中药仙鹤草提取物的镇痛抗炎作用.方法:取仙鹤草的水提取物和乙醇提取物,应用醋酸致小鼠扭体法、小鼠热板法、二甲苯致小鼠耳廓肿胀法和大鼠角叉菜胶足跖肿胀法研究其镇痛抗炎作用.结果:仙鹤草乙醇提取物和水提起物可减少醋酸致小鼠扭体次数,延长小鼠舔足时间,减轻二甲苯致小鼠耳廓肿胀程度,减小角叉菜胶致足跖肿胀程度;乙醇提取物作用强于水提取物.结论:中药仙鹤草乙醇提取物具有明显的镇痛抗炎作用.     

丹参颗粒饮片制备工艺的研究

目的研究丹参颗粒饮片制备工艺条件,筛选最适颗粒粒度和制备方法。方法通过正交实验以浸出物得率和丹酚酸B、丹参酮ⅡA的含量作为评价指标,对提取效率影响因素和干燥灭菌方法对成分影响进行考察。结果提取次数对浸出物的得率和丹酚酸B含量影响显著,颗粒粒度对提取物中丹酚酸B无显著影响。干燥灭菌时粒度对丹酚酸B、丹参酮ⅡA含量有一定影响。结论制备丹参颗粒饮片的粒度应以粗粉为宜。     

神香草水提取物在模拟胃肠道环境中的稳定性

目的:研究神香草水提取物在模拟胃肠道环境下的稳定性。方法:采用HPLC法分别测定不同时间神香草水提取物中迷迭香酸及迷迭香酸单体在不同p H、人工胃液、人工肠液、大鼠肠道内容物及大鼠肠粘膜中的含量。结果:神香草水提取物中迷迭香酸和迷迭香酸单体均随p H升高稳定性降低;在人工胃液中均稳定,在人工肠液中稳定性差;在大鼠小肠内容物、大鼠大肠内容物及大鼠肠粘膜中的稳定性依次为:大鼠肠粘膜>大鼠小肠内容物≈大鼠大肠内容物;在灭菌后的大鼠小肠内容物、大肠内容物及肠粘膜中的稳定性基本一致;两者在灭菌后大鼠肠内容物和肠粘膜中均比灭菌前更加稳定;在上述各条件下,神香草水提取物中的迷迭香酸与迷迭香酸单体相比较更加稳定。结论:该研究比较了单体化合物和其在维药提取物中的稳定性,结果明确,为进一步研究神香草提取物的吸收代谢情况及成药后的剂型设计,奠定良好基础。     

除菌过滤器验证(三):溶出物/析出物验证

溶出物/析出物的评估是制药领域中除菌过滤器的工艺特定验证的非常重要的组成部分。文中针对溶出物/析出物的验证要求,综述了影响溶出物/析出物水平的工艺参数、溶出物/析出物的验证方法、可能的溶出物/析出物种类等。为制药领域过滤器用户提供参考。     

Evaluation of safety and quality impact of extractable and leachable substances in therapeutic biologic protein products: a risk-based perspective

Leachables are chemical entities that migrate spontaneously from the final container closure system, packaging components and/or processing equipment under recommended conditions of product use and storage. Unlike leachables, extractables are generated under exaggerated temperature and time conditions in the presence of an appropriate solvent. Increasing evidence suggests that leachables may pose a safety risk by causing toxicity, carcinogenicity, immunogenicity and/or endocrine dysregulation. These substances may also alter product physico-chemical properties via interaction with the active pharmaceutical ingredient or the excipients in product vehicle, thereby adversely affecting the final product quality. The evaluation of leachable compounds begins with a thorough identification of extractable compounds released from the production and packaging components under exaggerated conditions. The set of observed extractables helps to identify possible targets to be monitored in a subsequent leachables study over extended time periods. Although extractables and leachables also present a challenge for the safe use of device components (e.g., metered dose inhalers, dry powder inhalers, nasal spray devices or various implants), this review focusses on a safety risk assessment for specified therapeutic biological protein products. Regulatory, safety and scientific considerations in evaluating extractables and leachables are discussed, along with strategies for the analytical identification, quantification and monitoring.     

Evaluation of safety and quality impact of extractable and leachable substances in therapeutic biologic protein products: a risk-based perspective

Leachables are chemical entities that migrate spontaneously from the final container closure system, packaging components and/or processing equipment under recommended conditions of product use and storage. Unlike leachables, extractables are generated under exaggerated temperature and time conditions in the presence of an appropriate solvent. Increasing evidence suggests that leachables may pose a safety risk by causing toxicity, carcinogenicity, immunogenicity and/or endocrine dysregulation. These substances may also alter product physico-chemical properties via interaction with the active pharmaceutical ingredient or the excipients in product vehicle, thereby adversely affecting the final product quality. The evaluation of leachable compounds begins with a thorough identification of extractable compounds released from the production and packaging components under exaggerated conditions. The set of observed extractables helps to identify possible targets to be monitored in a subsequent leachables study over extended time periods. Although extractables and leachables also present a challenge for the safe use of device components (e.g., metered dose inhalers, dry powder inhalers, nasal spray devices or various implants), this review focusses on a safety risk assessment for specified therapeutic biological protein products. Regulatory, safety and scientific considerations in evaluating extractables and leachables are discussed, along with strategies for the analytical identification, quantification and monitoring.     

Impact of extractables/leachables from filters on stability of protein formulations

Aqueous extractables/leachables from three sterilizing-grade filter membranes [polyvinylidene fluoride (PVDF), polyethersulfone (PES), and mixed cellulose ester (MCE)] were found to significantly reduce the surface tension of aqueous solutions. To evaluate the effect of these extractables/leachables from filter membranes on stability of protein formulations, model IgG2 formulations (with or without added surfactant) were spiked with different levels of filter extractables from stock solutions as a stress study. The stock solutions of extractables were created by processing the filter membranes through autoclaving and soaking steps. The IgG2 formulations were subsequently subject to agitation and temperature stress. Extractables/leachables from the filters were found to have a significant protective (PVDF, PES) and destabilizing (MCE) impact on both visible and subvisible particulates formation under agitation stress for formulations that did not contain any additional surfactant such as polysorbate 80. The impact of filter extractables/leachables on chemical stability of the antibody formulation displayed a more complicated pattern, but was generally destabilizing, causing increases in aggregation, oxidation, and acidic species. In conclusion, extractables/leachables from filter membranes may have impact on protein formulation stability and caution should be exercised during protein filtration, especially when filtering small volumes and in preformulation or high-throughput screening studies.     

Extractables/leachables from plastic tubing used in product manufacturing

While the ability of packaging systems to contribute leached substances to finished drug products is well established, increasing interest is being focused on the potential contamination of drug substances by plastic materials encountered during their production. The direct contact of such plastic parts (such as tubing, gaskets, filters and temporary storage containers) with the drug substance at some point in its production raises the possibility that plastic-related contaminants (leachables) may be present in the finished drug product. In this study, eight tubing materials potentially encountered in pharmaceutical production facilities, including six silicone materials and two Santoprene materials, were characterized for their extractable substances by static extraction coupled with comprehensive chemical characterization of the resulting extracts. Based on the extractables profiles thus generated, target leachables were identified for each tubing material. The accumulation of these target leachables was studied by subjecting the tubing to dynamic flow, simulated use extractions. The primary organic extractables from the silicone tubing were a homologous series of silicone oligomers, with most of the tubings demonstrating a unique distribution of oligomers. Several of the silicone tubings also possessed extractable dioctyl phthalate and dioctyl adipate. The primary organic extractables from the Santoprene-type tubing included a number of phthalates, a series of alkyl phenols and decomposition products of Irganox-type antioxidants. Inorganic extractables associated with many of the tubings included Ca, Mg, Zn and B. In general, the levels of targeted leachables extracted from the tubing materials under simulated use (flow) conditions was much smaller than the total amount of these leachables in the tubing.     

Assessing safety of extractables from materials and leachables in pharmaceuticals and biologics – Current challenges and approaches

Leachables from pharmaceutical container closure systems can present potential safety risks to patients. Extractables studies may be performed as a risk mitigation activity to identify potential leachables for dosage forms with a high degree of concern associated with the route of administration. To address safety concerns, approaches to toxicological safety evaluation of extractables and leachables have been developed and applied by pharmaceutical and biologics manufacturers. Details of these approaches may differ depending on the nature of the final drug product. These may include application, the formulation, route of administration and length of use. Current regulatory guidelines and industry standards provide general guidance on compound specific safety assessments but do not provide a comprehensive approach to safety evaluations of leachables and/or extractables. This paper provides a perspective on approaches to safety evaluations by reviewing and applying general concepts and integrating key steps in the toxicological evaluation of individual extractables or leachables. These include application of structure activity relationship studies, development of permitted daily exposure (PDE) values, and use of safety threshold concepts. Case studies are provided. The concepts presented seek to encourage discussion in the scientific community, and are not intended to represent a final opinion or “guidelines.”     

Structural identification of extractables from rubber closures used for pre-filled semisolid drug applicator by chromatography, mass spectrometry, and organic synthesis

Extractables or leachables from rubber closures used for drug products can potentially be detrimental since some of these rubber extractables are known to be toxic and pyrogenic. In addition, these extractables could interfere with analysis or even affect the stability profile of the active ingredients. For these reasons, it became necessary to identify structures of the possible extractables or leachables from rubber closures so that their potential impact on the drug products that are in direct contact with rubber closures could be evaluated. In this report, five species, 4-(1,1-dimethyl-propyl)-phenol (1), sulfur (2), 2,6-di-tert-butyl-[1,4] benzoquinone (3), furan-2-yl-(5-hydroxymethyl-furan-2-yl)-methanol (4), and 2-bromo-4-(1,1-dimethyl-propyl)-phenol (5), were found to be possible extractables from the rubber closures which were used in a selected pre-filled semisolid drug applicator. A strategy combining HPLC, GC, mass spectrometry, and organic synthesis was utilized.     

Evaluation of the chemical compatibility of plastic contact materials and pharmaceutical products; safety considerations related to extractables and leachables

A review is provided on the general topic of the compatibility of plastic materials with pharmaceutical products, with specific emphasis on the safety aspects associated with extractables and leachables related to such plastic materials.     

Generation of the Extractables Profile for an Elastomeric Material and Investigation of the Accumulation Behavior of Targeted Leachables Including Bis(2,2,6,6-tetramethyl-4-piperidyl) Sebacate (Tinuvin 770) and a Related Substance

The organic extractables profile of a synthetic polyisoprene material being considered for use as a closure on a bag-type packaging system has been delineated. The predominant organic extractables associated with the test material were bis(2,2,6,6-tetramethyl-4-piperidyl) sebacate (Tinuvin 770), several Tinuvin-related substances, fatty acids, and antioxidant-related compounds. Based on their potential product safety impact, Tinuvin and one of its related substances were chosen as target leachables. In order to establish the accumulation behavior of these target leachables under conditions that simulate the desired application, monobags (100-mL fill volume) and multichambered bags (1000-mL fill volume) were constructed with injection sites made from the test material, filled with water, and subjected to accelerated aging including multiple sterilization cycles and long-term storage at 40 °C. Even under the worse-case contact conditions, the accumulation levels of the target leachables were much less than their total available pool in the injection sites.     

Linking extractables and leachables in container/closure applications

Establishing a link between extractables and leachables may be necessary to understand, interpret, assess, quantify, or control the interaction between a drug product and its container/closure system. This paper considers the various factors that affect the rigor with which such a linkage is established and justified. Such an assessment considers the origin and/or genesis of the leachable or extractable, enumerates situations in which extractables/leachables linkages are useful, ties such situations to the drug product's lifecycle, defines a hierarchy of linkages based on the rigor with which the linkages are established and justified, and establishes guidelines for how to determine what kind of linkage is appropriate for certain circumstances and situations. Additionally, this paper gives several examples of linkages relevant to flexible plastic drug product containers.