CAJ | 학술논문

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°C。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。
목적：건립매촉반응중저물담고순적고효액상색보（HPLC）검측방법。방법：HPLC검측담고순함량적최가색보조건：Dia－monsil C18（250mm×4.6mm，5μm）위고정상，갑순∶배기완（90∶10）위류동상，검측파장위215nm，류속위1.0μl·min-1，주온위35°C。결과：담고순봉정첨봉，분리교호，이론탑판수위12545；재0.5～10mg·L-1범위내선성관계량호（A=19.136C-1.2707，r=0.9990）；평균가양회수솔위104%，RSD위1.5%（n=6）。결론：해방법전속성강，중복성량호，괄합초보용우매촉반응체계중저물담고순적함량측정，이담고순감소량반영전화솔。
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°C 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.