摘要
采用ChiralpakAD-H手性柱(250mm×4.6mm,5μm),建立了正相高效液相色谱(NP-HPLC)法直接拆分恩替卡韦与其光学异构体的方法。考察了流动相组成、酸碱性对柱效、分离度、保留时间等参数的影响。经优化,以正己烷-异丙醇-乙醇-三氟乙酸-三乙胺(70∶12∶18∶0.05∶0.05,V/V)为流动相,流速0.5mL/min;检测波长261nm。在此条件下,恩替卡韦与光学异构体分离度>4.2;光学异构体的检出限为0.12mg/L,在0.25～4.0mg/L浓度范围内有良好的线性关系;日内与日间精密度RSD<4.0%;按标准加入法计算,加样回收率在87.0%～100.8%之间;RSD<3.0%;按外标法计算,加样回收率在98.2%～110.4%之间;RSD<3.0%。本方法可作为恩替卡韦原料药中光学异构体杂质限量的控制方法。
The Chiralpak AD-H （ 250 mm × 4.6 mm, 5 μm） column was used to establish an NP-HPLC method for the chiral separation of entecavir and its optical isomer. The composition of mobile phase and its acidity were optimized to reach better column efficiency and resolution. The final chromatographic conditions were established as follows： n-hexane： isopropyl alcohol： ethanol：trifluoroacetic acid： triethylamine （70：12：18： 0. 05：0.05）. The detection wavelength was 261 nm and the flow rate was 0.50 mL/min. The resolution of entecavir and its optical isomer was more than 4.2. The linear range was 0.25 -4 mg/L. The limit of detec- tion （LOD） of optieal isomer was 0.12 mg/L. The relative standard deviations （RSD） of intra- and inter-day precisions were less than 4.0% （n = 3 ）. According to calibration curves by general external standard method, the recoveries of the optical isomer were in the range of 98.2% - 110.4%, RSD 〈 3%. According to calibration curves by standard addition method, the recoveries of the optical isomer were in the range of 87.0% - 100.2%, RSD 〈3%. The limit of the optical isomer was determined using ＂main component self-compare with calibration factor＂ under the chromatographic eonditions established. It was shown that the established HPLC method was reliable, simple and was applicable to the purity determination of entecavir.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2009年第8期1206-1210,共5页
Chinese Journal of Analytical Chemistry
关键词
恩替卡韦
光学异构体
高效液相色谱法
手性拆分
Entecavir, optical isomer, high performance liquid chromatography, ehiral separation
