头孢拉定原料及制剂的聚合物杂质分析
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摘要
目的建立头孢拉定原料及制剂中聚合物杂质的分析方法。方法采用碱降解法制备头孢拉定强制降解溶液;采用高效凝胶色谱法(TSK G2000 SWxl)和柱切换-LC/MS法对头孢拉定强制降解溶液中的聚合物杂质进行分离和结构鉴定;采用Agilent ZORBAX SB-C_(18)型色谱柱,以磷酸盐缓冲液-甲醇为流动相,进行梯度洗脱,建立头孢拉定聚合物的RP-HPLC分析方法,采用二维液相色谱法和柱切换-LC/MSn法对该方法的专属性进行分析;进行方法学验证。结果在头孢拉定强制降解物中鉴定出头孢拉定二聚体、三聚体、四聚体;高效凝胶色谱法分离头孢拉定聚合物杂质时,易受到小分子杂质的干扰,同时分离的聚合物杂质色谱峰拖尾严重,存在肩峰,定量准确性差;RP-HPLC法分析头孢拉定聚合物杂质时,在20～22min范围内检出头孢拉定二聚体、三聚体、四聚体;方法定量限为500μg,最低检测限为150μg。结论高效凝胶色谱法不能对头孢拉定中的聚合物杂质进行有效质控,建立的反相色谱法分析头孢拉定聚合物杂质时专属性良好、灵敏度高、方法耐用性好,可用于头孢拉定原料及制剂的聚合物杂质质控;头孢拉定强制降解溶液可作为头孢拉定聚合物分析的系统适用性溶液。
Objective To establish a method for the determination of polymer impurities in cefradine raw materials and preparations. Methods Using the base degradation method to prepare cefradine stress degradation solution, the high performance size exclusion chromatography method(HPSEC, using TSK-gel G2000 SWxl column)and the Column Switching-LC/MSn method were applied to separate and identify the polymer impurities in cefradine stress degradation solution. A new RP-HPLC method for cefradine polymer analysis was established with an agilent ZORBAX SB-C_(18) column, using a mobile phase gradient elution by phosphate buffer and methanol. The specification of RP-HPLC method was assessed by the dimensional chromatography method and the Column Switching-LC/MSn method as well as other method validation experiments. Results A series of polymer impurities in cefradine stress degradation solution were identified including cefradine dimers, trimers, and tetramers. Polymer impurities were liable to co-elute with small molecular impurities severe tailing peaks and shoulder peaks were found, which made a poor quantification accuracy. Cefradine dimers, trimers, and tetramers were detected with the retention time from20.0 to 22.0 min when the RP-HPLC method was used to separate cefradine stress degradation solution. The limit of quantification(LOQ) was 500μg and the limit of qualification was 150μg. Conclusion The HPSEC method is not suitable for the quality control of polymer impurities in cefradine raw materials and productions, while the new established RP-HPLC method is specific, sensitive, and robust, which can effectively control the polymer impurities.Cefradine stress degradation solution can be used to identify polymer peaks as the system suitability solution.
Objective To establish a method for the determination of polymer impurities in cefradine raw materials and preparations. Methods Using the base degradation method to prepare cefradine stress degradation solution, the high performance size exclusion chromatography method(HPSEC, using TSK-gel G2000 SWxl column)and the Column Switching-LC/MSn method were applied to separate and identify the polymer impurities in cefradine stress degradation solution. A new RP-HPLC method for cefradine polymer analysis was established with an agilent ZORBAX SB-C_(18) column, using a mobile phase gradient elution by phosphate buffer and methanol. The specification of RP-HPLC method was assessed by the dimensional chromatography method and the Column Switching-LC/MSn method as well as other method validation experiments. Results A series of polymer impurities in cefradine stress degradation solution were identified including cefradine dimers, trimers, and tetramers. Polymer impurities were liable to co-elute with small molecular impurities severe tailing peaks and shoulder peaks were found, which made a poor quantification accuracy. Cefradine dimers, trimers, and tetramers were detected with the retention time from20.0 to 22.0 min when the RP-HPLC method was used to separate cefradine stress degradation solution. The limit of quantification(LOQ) was 500μg and the limit of qualification was 150μg. Conclusion The HPSEC method is not suitable for the quality control of polymer impurities in cefradine raw materials and productions, while the new established RP-HPLC method is specific, sensitive, and robust, which can effectively control the polymer impurities.Cefradine stress degradation solution can be used to identify polymer peaks as the system suitability solution.
引文
[1]金少鸿.头孢菌素过敏反应的研究进展[J].国外医药抗生素分册,1984,5(3):191-197.
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[7]胡昌勤.β-内酰胺抗生素聚合物分析技术的展望[J].中国新药杂志,2008,17(24):2098-2102.
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[11]李进,张培培,崇小萌,等.阿莫西林克拉维酸钾复方制剂中聚合物杂质的分析[J].药物分析杂志,2017,37(8):1430-1440.
[12]杨美琴,金少鸿,胡昌勤.HPLC-柱切换法归属青霉素钠有关物质中的聚合物分析[J].药物分析杂志,2009,29(10):1615-1620.
[13]国家药典委员会.中华人民共和国药典[S].(2015年版二部).北京:化学工业出版社,2015:249.
[2]金少鸿.抗生素的过敏反应--Ⅰ.β-内酰胺类抗生素的过敏反应[J].中国临床药理学杂志,1986,2(4):251-257.
[3]金少鸿,胡昌勤.β-内酰胺类抗生素过敏反应的研究[J].医学研究杂志,2002,31(4):22-23.
[4]金少鸿.β-内酰胺类抗生素的质量和过敏反应关系的评述[J].中国抗生素杂志,1988,13(1):65-71.
[5]胡昌勤,金少鸿.β-内酰胺抗生素中的高分子杂质及其分析技术研究进展[J].国外医药抗生素分册,1996,17(5):336-343.
[6]王勇跃,陈悦,洪丽娅.β-内酰胺类抗生素中高分子杂质的分离与检测方法进展[J].中国药事,2015,29(6):608-612.
[7]胡昌勤.β-内酰胺抗生素聚合物分析技术的展望[J].中国新药杂志,2008,17(24):2098-2102.
[8]国家药典委员会.中华人民共和国药典[S].(2015年版二部).北京:化学工业出版社,2015:267.
[9]The United States Pharmacopeia 41 edi[S].Roehville MD:the United States Pharmacopeial Convention Inc,2018.
[10]Europeon Pharmacopeia 9.0[S].Strasbourg:European directorate for the quality of medicines,2017:VII 1994.
[11]李进,张培培,崇小萌,等.阿莫西林克拉维酸钾复方制剂中聚合物杂质的分析[J].药物分析杂志,2017,37(8):1430-1440.
[12]杨美琴,金少鸿,胡昌勤.HPLC-柱切换法归属青霉素钠有关物质中的聚合物分析[J].药物分析杂志,2009,29(10):1615-1620.
[13]国家药典委员会.中华人民共和国药典[S].(2015年版二部).北京:化学工业出版社,2015:249.