复方盐酸替利定口服液中主要杂质的结构确证与工艺改进

目的：研究复方盐酸替利定口服液主要杂质的结构和产生的原因,提高复方替利定口服液的质量。方法：通过LC-MS、IR和H-NMR检测分析其可能的结构并通过分离、合成等方法对杂质结构进行确认,从而在合成工艺上作出相应改进,减少杂质的含量。结果：按照选定方法,获得复方盐酸替利定口服液三个主要杂质（顺式替利定、反式去甲替利定和顺式去甲替利定）,并与LC-MS、IR、H-NMR结果一一对应。以此依据在合成工艺方面做出了相应改进,减少了盐酸替利定原料杂质的含量,从而提高了复方替利定口服液的质量。结论：本研究提高了盐酸替利定原料的纯度,进一步对提高复方盐酸替利定口服液的质量具有一定价值。     

尼莫地平口服溶液中降解杂质的结构鉴定及分析

目的 对尼莫地平口服溶液中2个未知降解杂质进行结构鉴定和分析测试。方法 通过二维液相色谱-高分辨质谱（2DLC-HRMS）推测未知杂质的化学结构,并采用定向合成的方式获得杂质单体,经核磁共振¹H-NMR、¹³C-NMR对杂质结构进行确证,采用Thermo Syncronis C₁₈（250 mm×4.6 mm,5 μm）色谱柱,以5 mmol·L^-1磷酸二氢钾（pH 2.8）为流动相A,以甲醇-乙腈（50∶50）为流动相B,梯度洗脱;流速为1.0 mL·min^-1;柱温为30℃;检测波长为235 nm。结果 尼莫地平与其降解杂质分离良好,各杂质校正因子均在0.9~1.1。结论 建立的方法专属性好,可有效分离和测定尼莫地平口服溶液中的有关物质。本研究可为尼莫地平口服溶液及其他剂型的杂质控制提供参考。     

正交试验法优化复方熊胆胶囊药材提取工艺

摘 要 目的： 优化复方熊胆胶囊的提取工艺。方法: 分别以得膏率及复方中人参皂苷Rg₁、Re、Rb₁的总含量为评价指标,选择加水量、提取时间、提取次数为考察因素,采用正交试验优选复方熊胆胶囊的水提取工艺。结果: 人参、蜂房加8倍量水煎煮3次,每次1h为最佳水提取工艺参数。将此煎液滤过,合并,减压浓缩汁至相对密度1.20（80～90℃）,干燥,粉碎成细粉。其余五味粉碎成细粉,过五号筛,与上述细粉混匀,加入淀粉适量,混匀,装入胶囊即得。结论:优化的水提取工艺稳定可行,重复性好,为复方熊胆胶囊制剂的工业化生产提供了依据。     

劳拉西泮片的关键质量属性研究

目的 对国产劳拉西泮片的关键质量属性进行分析,为该品种现行质量标准的完善以及质量风险控制提供依据。方法 在了解国内劳拉西泮片整体质量状况的基础上,以问题为导向,围绕安全性和有效性,选择片剂的有关物质、溶出度等质控关键点开展研究。结果 通过对原料杂质谱的分析,确认USP杂质C（ChP杂质Ⅱ）为劳拉西泮最主要的降解杂质,同时确定了该杂质的来源;ChP现行标准有关物质检查方法能够有效分离已知及未知杂质,且杂质的定量方法合理;斑马鱼胚胎发育毒性结果显示杂质C的神经毒性较劳拉西泮强;对现行标准溶出度检查法进行了优化;国产制剂与进口制剂在体外溶出特性方面存在一定差别。结论 国产劳拉西泮片的整体质量状况良好,但与进口制剂间仍存在一定的差距,应提高生产工艺的稳定性。现行标准基本上能够实现对本品关键指标的控制,但溶出度检查项有待优化,以提高对不同质量产品的区分度。     

酒石酸唑吡坦片中2种杂质的合成

摘 要 目的：设计合成酒石酸唑吡坦片中杂质Ⅰ。方法: 以唑吡坦及1 溴代乙酸乙酯为起始原料,经一锅法合成得到酒石酸唑吡坦片中的杂质Ⅰ（4 （二甲胺基） 3 （6 甲基 2 对甲苯基咪唑并[1,2 a]吡啶 3 基） 4 氧代丁酸）,同时杂质Ⅱ（2 羟基 N,N 二甲基 2 （6 甲基 2 对甲苯基咪唑并[1,2 a]吡啶 3 基）乙酰胺）作为副产物被合成得到。结果: 成功获得酒石酸唑吡坦的两个新杂质,纯度分别为95.3%及97.2%,并经质谱和核磁共振(MS,1H NMR)进行结构确证。结论: 首次合成了杂质Ⅰ及杂质Ⅱ,可为酒石酸唑吡坦质量控制和工艺优化提供参考依据。     

基于2D-LC-HR-MS/MS法的头孢丙烯原料药杂质谱分析

目的 基于二维液相色谱-高分辨串联质谱（2D-LC-HR- MS/MS）法分析头孢丙烯原料药杂质谱。方法 一维色谱条件进行样品色谱图采集,确认各杂质的出峰位置,采用 YMC Hydrosphere 色谱柱（250 mm×4.6 mm,5 μm）,以 11.5 g·L^-1磷酸二氢铵（用磷酸调节 pH 为 4.4）为流动相 A,乙腈-流动相 A（50∶50）为流动项 B,梯度洗脱;柱温 40 ℃;体积流量1.0 mL·min-1;检测波长 230 nm;进样量 4 μL。二维液相脱盐后切进高分辨串联质谱进行分析,根据结果推断杂质结构及生成机制,采用 Waters BEH C₁₈色谱柱（50 mm×2.1 mm,1.7 μm）,以 0.01% 甲酸水溶液为流动相 A,乙腈为流动相 B,切峰后开始 A 相由 98% 到 1%,柱温 40 ℃,体积流量 0.3 mL·min-1,质谱采用 Xevo G2-XS QTof MS 系统,离子源为 ESI 源,毛细管电压 3.0 kV,雾化器温度 450 ℃,扫描范围 m/z 100～2 000。结果 头孢丙烯样品中存在 9 个杂质色谱峰,其中 5 个杂质为已知杂质,峰 3 为杂质 B（头孢羟氨苄）、峰 5 为杂质 D、峰 6 为杂质 F、峰 7 为杂质 G、峰 9 为杂质 I;对其中 3 个未知杂质可能的结构式进行了初步推测以及探讨了可能的生成途径,峰 2 分子式为 C₁₈H₁₉N₃O₆S,该化合物比头孢丙烯多一个氧,分析其为头孢丙烯的氧化杂质;峰 4 分子式为 C₁₆H₁₅N₃O₆S,与杂质 B 相比增加了 1 个氧原子,减少了 2 个氢原子,判断其为 7-ACA 内酯与对羟基苯甲甘氨酸甲酯（HPGM）反应产物中的硫原子继续发生了氧化生成的;峰 8 分子式为 C₈H₉NO₂S,该组分为头孢丙烯分子结构的一部分。峰 1 有待进一步研究。结论 该方法有效解决了头孢丙烯流动相中含不挥发性磷酸盐的色谱体系与色谱-质谱快速鉴定杂质不兼容的难题,可以简单、快速地对头孢丙烯有关物质进行定性分析及杂质谱研究。     

左乙拉西坦缓释片的制备及其Beagle犬体内药动学研究

目的 制备左乙拉西坦缓释片,进行体外释放特性以及Beagle犬体内药动学研究。方法 按照筛选处方制备左乙拉西坦缓释片,以市售左乙拉西坦缓释片（Keppra-XR）为参比制剂,采用相似因子（f₂）法进行体外释放行为相似度评价;并将自制左乙拉西坦缓释片与市售普通片进行Beagle犬体内药动学参数比较。结果 自制左乙拉西坦缓释片与市售左乙拉西坦缓释片体外释放行为相似。市售左乙拉西坦普通片和自制左乙拉西坦缓释片的药动学参数T_max分别为（1.75±0.50）和（5.50±1.00）h,C_max分别为（30.33±2.00）和（17.29±3.56）mg·L^-1,AUC_0-t分别为（186.88±8.83）和（202.50±34.20）mg·L^-1·h。结论 自制的左乙拉西坦缓释片具有缓释效果。     

色瑞替尼有关物质遗传毒性（Q） SAR评价及Ames试验研究

目的 对色瑞替尼合成工艺中具有警示结构的潜在有关物质进行遗传毒性（致突变性）评价研究，为色瑞替尼有关物质的分类分级控制提供指导和依据。方法 分别采用基于专家规则和统计学原理的2种互补的（定量）构效关系[（Q） SAR]评价系统（Derek和Sarah）对色瑞替尼有关物质的遗传毒性进行初步预测和分类；对（Q） SAR预测结果为阳性的有关物质，开展Ames试验进一步验证。结果 杂质6 （CAS：1032903-50-6）、杂质7 （CAS：1032903-62-0）和杂质13 （CAS：1032903-63-1）的Derek预测结果均为阳性，依据ICH M7指导原则的分类为第3类。在Ames试验中，在加代谢活化系统S9和不加S9的情况下，3个杂质（每皿22~1 800 μg）回复突变的菌落数小于溶剂对照组的2倍，由此判定Ames试验提示的致突变性为阴性。结论 杂质6 、杂质7 和杂质13 的致突变性均为阴性，可推翻基于结构的疑虑，依据ICH M7中的第5类即非突变性杂质进行控制。     

2D-LC-IT-TOF-MS鉴定他克莫司原料有关物质

目的 应用在线脱盐-飞行时间-液质联用法，对他克莫司原料中检出的一杂质峰进行结构定性。方法 采用Thermo Hypersil GOLD C₁₈色谱柱（4.6 mm×150 mm，3 μm），乙腈-叔丁基甲醚-磷酸溶液为流动相，梯度洗脱；然后采用InertSustain C₁₈色谱柱（2.1 mm×50 mm，2 μm），0.1%甲酸溶液-乙腈（30∶70）为流动相，等度洗脱；电喷雾电离离子源，正离子检测模式，雾化气流速度1.5 L·min^-1，干燥气流速度10 L·min^-1，脱溶剂管温度200℃。根据多级图谱信息推定特定杂质峰结构。结果 推定出他克莫司原料中特定保留时间处色谱峰的物质结构，该检出峰不是美国药典给出的子囊霉素杂质而是有药理活性的他克莫司异构体Ⅰ，应参与含量计算。结论 建立的方法可用于他克莫司原料杂质的鉴定，为更加精准控制产品质量和工艺优化研究提供可靠的技术保证。     

复方岩白菜素片有关物质HPLC方法的建立和研究

目的 建立复方岩白菜素片有关物质检查的HPLC方法。方法 色谱柱为SHISEIDO CAPCELL PAK C₈（4.6×250 mm,5 µm）,流动相为甲醇-水（16︰84）,流速为1.0 mL?min^-1,检测波长为215 nm,柱温为30 ℃,进样量为20 μL;以岩白菜对照品峰面积按外标法计算有关物质含量。结果 复方岩白菜素片主成分峰及各降解产物峰间能较好分离,12批次样品中最大杂质含量为0.08%～0.28%,总杂质为0.09%～0.29%,并对总杂质含量进行了控制。结论 建立的有关物质方法专属性强、灵敏高,能满足各峰的良好分离,可用于复方岩白菜素片的有关物质检查。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.