摘要
目的:考察不同溶剂对发酵类脂溶性药物中残留DNA测定准确性的影响并确定最佳溶剂。方法:以脂溶性药物-环孢素为例,通过研究DNA检测全过程中各步骤溶解因素对检测结果的影响,从供试品溶液制备、供试品溶液与DNA试剂盒试剂相溶性及DNA溶液稳定性3个方面入手,对不同种类的溶剂进行考察和筛选,并根据荧光染色法的原理,采用Qubit■DNA检测试剂盒对环孢素供试品溶液中残留DNA进行测定,探讨溶剂选择的适用性和合理性。此外,应用筛选的溶剂对另一种脂溶性药物——两性霉素B中残留DNA进行了测定,进一步验证可靠性。结果:0.1 mol·L^-1盐酸-N,N-二甲基甲酰胺(1∶4)、0.1 mol·L^-1醋酸-N,N-二甲基甲酰胺(1∶4)和0.1 mol·L^-1柠檬酸-N,N-二甲基甲酰胺(1∶4)3种混合溶剂对脂溶性药物均有较好溶解性并能与DNA试剂较好相溶;进一步的DNA稳定性研究结果显示,0.1 mol·L^-1柠檬酸-N,N-二甲基甲酰胺(1∶4)对DNA结构与性质不产生影响,稳定性最好。结论:采用0.1 mol·L^-1柠檬酸-N,N-二甲基甲酰胺(1∶4)作溶剂,不仅对环孢素和两性霉素B具有较好的溶解性,而且对DNA不产生影响,稳定性较高,可实现终产品中残留DNA的准确测定。本文研究结果为脂溶性药物中残留DNA检测溶剂的选择提供了一定科学参考。
Objective:To investigate the influence of solvents on the accuracy of measurement of residual DNA in lipid-soluble drugs and then determine the suitable solvents.Methods:In this paper,we took a case study on-cyclosporine,a liposoluble drug,investigating various kinds of solvents with focus on test sample solution,inter-compatibility of test sample solution with Qubit■working solution in DNA kit,and DNA stability in solvents as well.Residual DNA in cyclosporine samples was measured using Qubit■DNA kit based on fluorescence staining method.The applicability and rationality of solvent selection was also discussed.In addition,the selected solvent was also used for the determination of residual DNA in amphotericin B for the purpose of reliability verification.Results:0.1 mol·L^-1 hydrochloric acid-DMF(1∶4),0.1 mol·L^-1 acetic acid-DMF(1∶4)and 0.1 mol·L^-1 citric acid-DMF(1∶4)had better solubility for cyclosporine and better inter-compatibility with Qubit■working solution.However,further investigation showed that it had no effect on the structure and properties of DNA and presented the best stability when it took 0.1 mol·L^-1 citric acid-DMF(1∶4)as the solvent.Conclusion:Not only does 0.1 mol·L^-1 citric acid-DMF(1∶4)show good solubility for lipid-soluble drugs,but it also has high stability for DNA,which is an appropriate solvent for more accurate determination of residual DNA in fermented lipidsoluble drugs.
作者
王琰
张培培
崔生辉
姚尚辰
胡昌勤
许明哲
张彦民
WANG Yan;ZHANG Pei-pei;CUI Sheng-hui;YAO Shang-chen;HU Chang-qin;XU Ming-zhe;ZHANG Yan-min(National Institutes for Food and Drug Control,Beijing 102629,China;Shool of Pharmacy,Xi’an Jiaotong University,Xi’an 710061,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2020年第4期651-658,共8页
Chinese Journal of Pharmaceutical Analysis
基金
国家“重大新药创制”科技重大专项资助项目(2017ZX09101001-007)。
关键词
发酵类脂溶性药物
环孢素
两性霉素B
残留DNA
测定影响因素
溶剂筛选
质量控制
fermented lipid-soluble drugs
cyclosporine
amphotericin B
residual DNA
determine influencing factors
solvents screening
quality control
