摘要
目的：建立酶促反应中底物胆固醇的高效液相色谱（HPLC）检测方法。方法：HPLC检测胆固醇含量的最佳色谱条件：Dia－monsil C18（250mm×4.6mm，5μm）为固定相，甲醇∶环己烷（90∶10）为流动相，检测波长为215nm，流速为1.0μl·min^-1，柱温为35℃。结果：胆固醇峰呈尖峰，分离较好，理论塔板数为12545；在0.5～10mg·L^-1范围内线性关系良好（A=19.136C-1.2707，r=0.9990）；平均加样回收率为104%，RSD为1.5%（n=6）。结论：该方法专属性强，重复性良好，适合初步用于酶促反应体系中底物胆固醇的含量测定，以胆固醇减少量反映转化率。
Objective：To establish a method for the determination of Cholesterol （CHS） as a substrate in the enzymatic reaction by High Performance Liquid Chromatography （HPLC）. Methods： HPLC method was used to determine the content of CHS, the optimal HPLC conditions were as follows：Diamonsil C18（250mm×4.6mm,5μm）column as the stationary phase, Methanol-Cyclohexane（90：10） as the mobile phase,1.0μl·min^-1 of flow-rate, 35℃ of column temperature, the wavelength of UV detector （215nm）. Results： CHS was well resolved （the number of theoretical plates was 12545）. The calibration curve was in good linearity over the rang of 0.5~10mg·L^-1 for CHS. The equation was （A=19.136C-1.2707,（r=0.9990））. The average recovery of CHS was 104%（RSD=1.5%,n=6）. Conclusion：The method is sensitive and good repeatability,and it could be used for the determination of CHS which depends on its decrement to reflex the conversion rate.
出处
《北方药学》
2014年第12期21-22,共2页
Journal of North Pharmacy
基金
教育部高等学校博士学科点专项科研基金
项目编号20134425110010
