HPLC法测定氟康唑的有关物质及最大未知杂质结构鉴定

目的：建立HPLC 法测定氟康唑的有关物质，并对最大未知杂质进行结构鉴定。方法：采用CERIL-column ODS（250 mm×4.6 mm，5 μm）色谱柱，以0.01 mol·L^-1 醋酸钠缓冲液（pH 5.0）- 甲醇（95:5）为流动相A，乙腈- 甲醇（60:40）为流动相B，梯度洗脱，流速1.0 mL·min^-1，检测波长261 nm，柱温30 ℃。用LC-MS 分析最大未知杂质，再用半制备液相色谱分离纯化，通过NMR 确定结构。结果：建立的HPLC有关物质分析方法具有良好的方法学验证结果，与中国药典方法比较，杂质检出能力更强；最大未知杂质为2-（2，4-二氟苯基）-1，3- 二（1H-1，2，4- 三唑-1- 基）- 丁-2- 醇。结论：建立的方法简便、灵敏，专属性好，杂质检出能力强，能有效地反应药品质量。

Objective:To establish an HPLC method for determination of related substances in fluconazole and to identify the structure of the largest unknown impurity.Methods:A CERI L-column ODS(250 mm×4.6 mm, 5 μm)was adopted for the study with gradient elution.The mobile phase A was 0.01 mol·L^-1 sodium acetate buffer solution(pH 5.0)-methanol(95:5),and the mobile phase B was acetonitrile-methanol(60:40).The flow rate was 1.0 mL·min^-1,the detection wavelength was 261 nm,and the column temperature was 3℃0 .The largest unknown impurity was analyzed by LC-MS,and was then separated and purified by semi-preparative HPLC,and its structure was confirmed by NMR.Results:The established and validated HPLC method showed stronger capacity for fluconazole related substances control compared to the method recorded in Chinese pharmacopoeia 2010.The largest unknown impurity was 2-(2,4-difluorophenyl)-1,3-bis(1H-1,2,4-triazol-1-yl)-butan-2-ol.Conclusion:The established HPLC method is simple,specific,sensitive and good at impurity inspection,which can be applied in the quality control and comparison of the marketed products from different manufacturers.

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