测定体外人肝微粒体中安定代谢物(去甲安定和羟安定)含量的方法学研究  

IN VITRO DETERMINATION OF DIAZEPAM METABOLITES(N-DEMETHYLDIAZEPAM AND TEMAZEPAM)IN HUMAN LIVER MICROSOMES

作　　者：匡唐永[1] 楼雅卿[1] 

机构地区：[1]北京医科大学药理教研室

出　　处：《中国临床药理学杂志》1993年第1期30-34,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：应用高效液相色谱法,建立了测定体外人肝微粒体孵育安定的代谢物—去甲安定和羟安定含量的方法。结果表明,去甲安定、羟安定和内标物各峰分离良好,其保留时间分别为8.4min,10.2min和7.4min;方法的灵敏度较高,能检测出5μg/l的样品;标准曲线线性关系良好(去甲安定:r=0.9981±0.0016,n=8;羟安定:r=0.9981±0.0015,n=8);回收率均在95%以上,其日内、日间变异系数均在10%以内;其它多种药物对样品色谱峰均无干扰。因而为国内开展安定氧化代谢,多态性及与其它药物代谢相关性的研究提供了快速可靠的定量分析方法。A HPLC method was developed for simultaneous in Vitro determination of N-demethyldiazepam (NDZ) and Temazepam (TMZ) in human liver microsomes. Each tube with NDE and TMZ contained 2 mg microsomal protein, and 75ng Clonazepam as internal standard was added. The final volume was adjusted to 1ml with 150mmol/L Tris-HCL buffer, pH7. 4, and 2ml ethyl ether added. The mixture was shaken for 10min, and then centrifuged. The organic phase was transferred to another tube and blown to dryness under N_2.The residues were dissolved in 50μl of the mobile phases for HPLC and 25μl injected into the HPLC system. The column was Ultrasphere ODS (shandon 3μm×4. 6mm×10cm) and the eluent contained 28% acetonitrile in phosphorate buffer (0. 1M; pH2. 8) with a flow rate of 1. 5ml/min. A UV detector operated at 254nm were used. The retention time of I. S, NDZ and TMZ was 7. 4, 8. 4 and 10. 2min respectively, with a detection limit of 5μg/l. Linear and reproducible standard curves were obtained over the concentration range of 10μg/l to 130μg/l (NDZ:r = 0. 9981 ± 0. 0016, n = 8; TMZ: r=0. 9981±0. 0015, n=8,p<0. 001). The recoveries of NDZ and TMZ were all above 95%. The within day and between days coefficients of variation were all less than 10%.

关 键 词：高效液相色谱 苯甲二氮Zhuo 代谢 

分 类 号：R971.3[医药卫生—药品]