摘要
建立一种简便快速的5α-还原酶活性检测方法.采用高效液相色谱法(HPLC)测定酶促反应中底物睾酮(T)量的变化,以此反映酶活性的高低.该方法可用于5α-还原酶抑制剂的高通量体外筛选(HTS).方法的色谱条件为色谱柱:BondapakC18(4.6mm(i.d)×300mm,Waters);流动相中甲醇∶水=70∶30(V V);流速为0.5mL·min-1;紫外检测波长为242nm;柱温32℃;T的线性范围在6-300ng·mL-1,相关系数为0 9992;最小检测限为3ng.
An convenient and quick analytical method has been established for the determination of 5α-reductase activity using high performance liquid chromatography(HPLC). This method is suitable for high throughput screening(HTS) of 5α-reductase inhibitor in vitro. The stainless steel column was 300?mm×4.6?mm (i.d.) packed with totally porous, spherical carbon particles. A solution of methanol and water(70∶30, V/V) was employed as the mobile phase. The flow rate was 0.5?mL·min^(-1). Chromatography was performed with ultra violet detector at 242 nm, column temperature at 32?℃. Linear calibration curve for T was measured within the range of 6-300 ng/?mL with correlation coefficient of 0.999?2. The lowest detection limit was 3?ng.
出处
《福州大学学报（自然科学版）》
CAS
CSCD
2004年第2期216-219,共4页
Journal of Fuzhou University(Natural Science Edition)
基金
福州大学本科生科研实践资助项目(02062)
