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 C18（100 mm×2.1 mm, 1.9 μm）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1.70~6.80 μg·mL-1（r=0.999 9）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 C18 (100 mm×2.1 mm, 1.9 μm) 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 1.70-6.80 μg·mL-1 (r=0.999 9). 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.
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WS1-XG-056-2011 Nysfungin Tablets[S].2011
