制霉素中未知成分的结构鉴定及UHPLC法含量测定  

Identification and determination of unknown component in nysfungin by UHPLC

作　　者：孙胜男 冷霄云 李怡然 孙忠浩 李晓茜 孟祥燕 解云英[1] 山广志[1] SUN Sheng-nan;LENG Xiao-yun;LI Yi-ran;SUN Zhong-hao;LI Xiao-qian;MENG Xiang-yan;XIE Yun-ying;SHAN Guang-zhi(Institute of Medicinal Biotechnology,Chinese Academy of Medical Science and Peking Union Medical Colege,Beijing 100050,China;Zhejiang Zhenyuan Pharmaceutical Research Institute,Zhejiang Zhenyuan Pharmaceutical Co.,Ltd.,Shaoxing 312071,China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所,北京100050 [2]浙江震元制药有限公司浙江震元制药研究院,绍兴312071

出　　处：《药物分析杂志》2021年第8期1375-1380,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：十三五新药创制重大专项(2017ZX09101001)。

摘　　要：目的:鉴定制霉素中未知成分(RT6)的结构,并建立该成分的UHPLC含量测定方法。方法:从原料药中分离纯化未知成分,利用核磁共振波谱和高分辨质谱对未知成分(RT6)进行结构鉴定和解析。采用Thermo Hypersil GOLD C_(18)(100 mm×2.1 mm,1.9μm)色谱柱,甲醇-乙腈-28 mmol·L^(-1)醋酸铵溶液(0.8%乙酸)为流动相,梯度洗脱,流速0.2 mL·min^(-1),柱温30℃,检测波长305 nm。结果:经NMR和HRMS鉴定,制霉素中未知成分为白诺氏菌素。在建立的色谱条件下,该成分与其他成分分离良好,质量浓度在1.70~6.80μg·mL^(-1)(r=0.9999)范围内与峰面积呈良好线性关系,低、中、高3个水平下平均回收率分别为103.6%、99.4%和100.1%,RSD分别为0.33%、1.6%和0.90%。3批供成品中白诺氏菌素的含量检测结果分别为3.97%、3.41%、3.72%。结论:制霉素原料中存在的未知成分为白诺氏菌素。经方法学验证,建立的UHPLC方法可用于制霉素原料药中白诺氏菌素的测定与控制。Objective:To identify the structure of unknown component(RT6)in nysfungin and establish UHPLC method for content determination.Methods:The unknown component(RT6)was identified by NMR and HRMS.And the determination was performed by Thermo Hypersil GOLD C_(18)(100 mm×2.1 mm,1.9μm)column,with the mobile phase consisted of methanol,acetonitrile and 28 mmol·L^(-1) ammonium acetate(0.8%acetic acid)by gradient elution.The flow rate was 0.2 mL·min^(-1),the column temperature was 30℃,and the detection wavelength was 305 nm.Results:The unknown component(RT6)in nysfungin was identified as albonoursin.RT6 was well separated from other component and showed a good linearity in the concentration range of 1.70-6.80μg·mL^(-1)(r=0.9999).The average recoveries at low,medium and high levels were 103.6%,99.4%and 100.1%,respectively,and the RSDs were 0.33%,1.6%and 0.90%,respectively.The result showed that the contents of albonoursin in three batches of samples were 3.97%,3.41%and 3.72%.Conclusion:The unknown component in nysfungin is albonoursin.The established UHPLC method can be used for the determination and control of albonoursin in nysfungin.

关 键 词：制霉素 白诺氏菌素 结构鉴定 液相色谱-质谱 核磁共振 超高效液相色谱 含量测定 

分 类 号：R917[医药卫生—药物分析学]