HPLC柱前衍生法测定兔血浆中海星甾醇(A1998)浓度  

Determination of 3β-Hydroxyandrost -5-en-17-one Succinate (A1998) in Rabbit Plasma with Precolumn Derivatization HPLC

作　　者：张天宇[1] 董善年[1] 章国良[1] 楼雅卿[1] 步秀云[1] 

机构地区：[1]北京大学基础医学院药理系,北京100083

出　　处：《药物分析杂志》2004年第3期304-307,共4页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立高效液相色谱柱前衍生法测定兔血浆中海星甾醇(A1998)浓度。方法:用2,4-二硝基苯肼(2,4-DNPH)将A1998 C-17位上的2个羰基衍生化,形成双腙,以提高检测灵敏度。选用A1998的同系物3β-羟基雄甾-5烯-17酮己二酸酯(C6)作为内标。色谱条件:采用Agilent Zorbax Exend C18反相色谱柱(4.6 mm×150 mm,5μm),流动相为100%甲醇,流速2.0 mL·min-1,检测波长为360 nm。结果:血样中A1998浓度在0.05-10μg·mL-1范围内具有良好的线性关系(r=0.9998),最低检测浓度为0.05μg·mL-1。回收率大于80%,日内及日间精密度RSD分别为3.2%-15%和6.7%-11%。结论:本方法灵敏、准确、实用,能够满足动物药物代谢动力学研究的要求。Objective: To establish a HPLC method with precolumn derivatization for determination of A1998 in rabbit plasma. Methods:Two C -17 carbonyls in A1998 were derivatized by 2,4 - dinitrophenylhydrazine (2,4-DNPH) becoming a double hydrozones, which greatly enhance the detecting sensitivity of A1998. A homolog of A1998(3β- hydroxyandrost -5 - en - 17 - one adipate) (C6) was used as internal standard. The chromatographic condition was listed as follows: mobile phase was 100% methanol; Agilent Zorbax Extend C18 RP - column (4. 6 mm ×150 mm,5 μm) ;a flow -rate was 2. 0 mL·min-1 ;UV detection wavelength was 360 nm. Results: A linearity range of 0. 05 - 10 μg·mL-1 in rabbit plasma was obtained (r =0. 9998). The lowest detection concentration is 0. 05μg·mL -1. The average recovery of A1998 from plasma was more than 80%. The intra - day(n=5) and inter -day(n=5)precision rates were 3.2%-15% and 6.7% - 11% respectively. Conclusion:This method is sensitive, accurate, convenient, and suitable for study of pharmacokinetics of A1998 in animals.

关 键 词：HPLC柱前衍生法 兔 血浆 海星甾醇 A1998 羰基衍生 高效液相色谱 心律失常 

分 类 号：R446.11[医药卫生—诊断学]