奥美沙坦酯中基因毒性杂质的检测分离及质量新标准研究

发布时间：2018-09-13 06:27

【摘要】：奥美沙坦酯是一种特异性血管紧张素Ⅱ受体拮抗剂,其工业合成过程中能产生卤代基团的基因毒性杂质。本论文结合原料药工业生产过程,通过高效液相色谱方法,对这类微量的基因毒性杂质进行检测分离,探索了相应的分离条件,并研究了考虑基因毒性杂质控制的原料药奥美沙坦酯质量新标准。具体如下:(1)建立溴代基因毒性杂质的检测分离方法,采用C18柱(250×4.6 mm,5μm);流动相:0.1%冰乙酸水溶液(A):0.1%冰乙酸乙腈溶液(B)=15:85;检测波长:254 nm;流速:1.5 ml·min-1;柱温:25℃;通过对N-(三苯基甲基)-5-(4'-溴甲基联苯-2-基)四氮唑(溴代杂质1)和N-三苯甲基-5-(4',4'-二溴甲基联苯-2-基)四氮唑(溴代杂质2)进行专属性、重复性、定量限和检测限以及回收率等方面的检测分离,溴代杂质1和溴代杂质2的定量限和检测限相对浓度分别可达0.0003%和0.0001%,远远低于其标准限度(0.0037%),杂质1和杂质2均在0.03~0.25μg·mL-1范围内线性良好,平均回收率分别为94.37%和94.43%,相对标准偏差分别为2.38%和2.72%,保证了该方法的灵敏度、可靠性;(2)建立氯代基因毒性杂质的检测分离方法,采用Kromasil Eternity 5-Phenyl Hexyl柱(250×4.6 mm);检测波长:215 nm;流速:1.0 ml/min;柱温:40℃;进样量:10μl。流动相A:乙腈:2.04 g·L-1磷酸二氢钾溶液(pH 3.4)(20:80);流动相B:2.04 g·L-1磷酸二氢钾溶液(pH 3.4):乙腈(20:80);对4-氯甲基-5-甲基-1,3-二氧杂环戊烯-2-酮(氯代杂质1)和4,5二氯甲基-1,3-二氧杂环戊烯-2-酮(氯代杂质2)进行一系列方法验证,结果表明,氯代杂质1和2的检测限的相对浓度分别为0.0003%和0.0004%,低于规定的标准限度(0.0037%),说明该检测方法的灵敏度能达到要求,能有效地监控产品质量;(3)通过三批产品的性状、鉴别、含量、检查等项目的质量研究和检测结果,制定奥美沙坦酯质量新标准,包括有关物质、工艺使用的残留溶剂丙酮、乙酸乙酯、N,N-二甲基乙酰胺等,以及新建立的基因毒性杂质限度标准。最后,综合基因毒性杂质控制,研究了包括基因毒性杂质的奥美沙坦酯质量新标准,更好地保证日常生产的奥美沙坦酯原料药的质量控制。
[Abstract]:Olmesartan ester is a specific angiotensin II receptor antagonist, which can produce halogenated group of genotoxic impurities in the process of industrial synthesis. In this paper, combined with the industrial production process of raw materials, high performance liquid chromatography (HPLC) was used to detect and isolate these trace genotoxic impurities, and the corresponding separation conditions were explored. A new quality standard for olmesartan ester, which considers the control of genotoxic impurities, has been developed. The main contents are as follows: (1) To establish a method for the detection and separation of brominated genotoxic impurities, C18 column (250 *4.6 mm, 5 micron), mobile phase: 0.1% glacial acetic acid aqueous solution (A): 0.1% glacial acetonitrile solution (B) = 15:85, detection wavelength: 254 nm, flow rate: 1.5 ml @ min-1, column temperature 5-(4', 4'-dibromomethylbiphenyl-2-yl) tetrazole (brominated impurity 1) and N-triphenylmethyl-5-(4', 4'-dibromomethylbiphenyl-2-yl) tetrazole (brominated impurity 2) were separated at 25 C. The specificity, repeatability, quantitative limit and detection limit of brominated impurity 1 and brominated impurity 2 were determined. The relative concentrations of impurities 1 and 2 were well linearized in the range of 0.03-0.25 ug.mL-1. The average recoveries were 94.37% and 94.43% respectively. The relative standard deviations were 2.38% and 2.72% respectively, which ensured the sensitivity and reliability of the method. (2) Establishment of chlorinated genotoxicity. The impurities were detected and separated by Kromasil Eternity 5-Phenyl Hexyl column (250 *4.6 mm); detection wavelength: 215 nm; flow rate: 1.0 ml/min; column temperature: 40 sample volume: 10 ml. mobile phase A: acetonitrile: 2.04 g L-1 potassium dihydrogen phosphate solution (pH 3.4) (20:80); mobile phase B: 2.04 g L-1 potassium dihydrogen phosphate solution (pH 3.4): acetonitrile (20:80); p-4-chloromethyl-5-5-phosphate solution (pH 3.4); Methyl-1,3-dioxacyclopentene-2-one (chlorinated impurity 1) and 4,5-dichloromethyl-1,3-dioxacyclopentene-2-one (chlorinated impurity 2) were validated by a series of methods. The results showed that the relative concentrations of chlorinated impurity 1 and 2 were 0.0003% and 0.0004% respectively, which were lower than the prescribed standard limit (0.0037%). To meet the requirements, product quality can be effectively monitored; (3) Through three batches of product properties, identification, content, inspection and other items of quality research and testing results, the development of new quality standards of olmesartan ester, including related substances, residual solvents used in the process acetone, ethyl acetate, N, N-dimethylacetamide, and other newly established genotoxic impurities Finally, the new quality standards of Omexatane Ester including genotoxic impurities were studied to ensure the quality control of Omexatane Ester in daily production.
【学位授予单位】：浙江工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.72;TQ463

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