甾体激素ESI-MS分析中酮基衍生方法研究及替比培南相关杂质的分析

发布时间：2018-10-14 08:55

【摘要】：甾体激素是一类具有环戊烷多氢菲母核结构的四环脂肪烃化合物,广泛存在于人和动物体内,对人体内新陈代谢、内环境的稳定、生长发育以及生殖行为有重要的调节作用。一般情况下,甾体激素在人体内维持正常范围,过高或过低容易造成内分泌功能紊乱,引发各种疾病。因此,内源性甾体激素的测定对疾病的预防、诊断和治疗有重要意义。药品作为一种特殊商品,其质量与人们的身体健康及生命安全息息相关,因此,对其质量的控制必须贯穿药品研发、生产、流通及储存各个阶段。药品杂质的研究是药品质量控制的一项重要内容。通过对药品相关杂质的分析检测对药品的生产工艺的优化及质量控制提供依据。本文综述部分首先介绍了甾体激素的种类和作用,展开叙述了国内外甾体激素的分析检测方法。其次对药品杂质的分类及来源做了简要介绍,重点阐述了药品杂质的检测方法以及高效液相色谱-质谱联用技术(HPLC-MS)在杂质分析中的应用。实验部分分为两部分,第一部分选择一种甾体激素的衍生试剂,优化了衍生反应条件及质谱条件;第二部分分析替比培南复杂中间体侧链和母核(MAP)的相关杂质,优化了分析条件,通过质谱数据、二级碎片信息推测了几种可能杂质。具体研究内容及结果包括以下两个方面:1.首先选择本校药学院合成的10种肼类化合物与睾酮进行衍生反应,通过ESI-MS检测,发现各个肼类化合物衍生效果不理想,然后选择市售的相对分子质量较小的7种肼类化合物与睾酮反应,结果显示水杨酰肼衍生效果最好。以水杨酰肼作为衍生试剂,与孕酮、孕烯醇酮、雌酮、脱氢表雄酮和双氢睾酮五种甾体激素标准品进行衍生反应,优化了反应溶剂、反应温度、反应时间、反应pH和水杨酰肼的用量等衍生反应的条件。该方法简单快速、灵敏度高,水杨酰肼与甾体激素反应条件简单、高效,可以作为甾体激素的衍生试剂来检测甾体激素的含量。2.对合成替比培南的复杂中间体侧链和母核进行杂质分析,确立了HPLC-MS条件,优化了流动相和洗脱程序,实现了20 min内对药品杂质的分析检测。根据药品的合成工艺信息、液相色谱保留时间、液相色谱图对应的质谱图以及二级裂解碎片信息,推测了杂质的可能结构,并分析推测了杂质的裂解途径,为替比培南新药研发及质量控制提供参考依据。
[Abstract]:Steroid hormones are a class of tetracyclic fatty hydrocarbons with the nuclear structure of cyclopentane polyhydrophenanthrene which widely exist in human and animal bodies and play an important role in regulating metabolism stability of internal environment growth and development as well as reproductive behavior. In general, steroid hormones in the human body maintain a normal range, too high or too low prone to endocrine disorders, leading to various diseases. Therefore, the determination of endogenous steroid hormones is of great significance in the prevention, diagnosis and treatment of diseases. As a special commodity, the quality of drugs is closely related to people's health and life safety. Therefore, the quality control must run through all stages of drug development, production, circulation and storage. The study of drug impurity is an important content of drug quality control. The analysis and detection of drug related impurities provide basis for the optimization of drug production process and quality control. In the part of review, the types and functions of steroid hormones are introduced, and the analytical methods of steroid hormones at home and abroad are described. Secondly, the classification and source of drug impurity are briefly introduced, and the detection method of drug impurity and the application of high performance liquid chromatography-mass spectrometry (HPLC-MS) in impurity analysis are emphasized. The experiment is divided into two parts. In the first part, a steroid hormone derivative reagent is selected to optimize the derivatization reaction conditions and mass spectrometry conditions, and the second part is to analyze the related impurities in the side chain and mother nuclear (MAP) of the complex intermediate of tibipenem. The analysis conditions were optimized and several possible impurities were inferred by mass spectrometry data and secondary fragment information. The specific research contents and results include the following two aspects: 1. Firstly, 10 kinds of hydrazine compounds synthesized in our school of pharmacy were selected for derivatization reaction with testosterone. By ESI-MS detection, it was found that the derivative effect of each hydrazine compound was not satisfactory. Seven hydrazine compounds with relatively small molecular weight were selected to react with testosterone. The results showed that salicylic hydrazide derivatives were the best. Salicylic hydrazide was used as derivatization reagent. The reaction solvent, reaction temperature and reaction time were optimized by derivatization reaction with five steroidal hormone standard compounds, progesterone, pregnenolone, estradione, dehydroepiandrosterone and dihydrotestosterone. The reaction conditions such as the amount of pH and salicylic hydrazide were obtained. The method is simple, rapid and sensitive. The reaction condition of salicylhydrazide with steroid hormone is simple and efficient. It can be used as the derivative reagent of steroid hormone to detect the content of steroid hormone. 2. The side chain and mother nucleus of the complex intermediate of tibipenem were analyzed, the HPLC-MS conditions were established, the mobile phase and elution procedure were optimized, and the analysis and detection of drug impurities within 20 min were realized. According to the synthetic process information of drugs, retention time of liquid chromatography, mass spectrogram corresponding to liquid chromatogram and information of secondary pyrolysis fragment, the possible structure of impurity was deduced, and the pathway of impurity cracking was analyzed. To provide a reference for the R & D and quality control of new drugs of tibipenem.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R927.1

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