Yao T W, Zhou Q, Zeng S
College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, Zhejiang 310006, People's Republic of China.
Biomed Chromatogr. 2000 Nov;14(7):498-501. doi: 10.1002/1099-0801(200011)14:7<498::AID-BMC9>3.0.CO;2-D.
An enantioselective assay for S(+)- and R(-)-propafenone in transgenic Chinese hamster CHL cells expressing human cytochrome P450 was developed. The method involved extraction of propafenone from the S9s incubates, and formation of propafenone diastereomeric derivatives with the chiral reagent 2,3,4, 6-tetra-O-beta-D-glucopranosyl isothiocyanate. Separation and quantitation of diastereomeric propafenone derivatives were carried out in a reverse-phase-HPLC system with UV detection. The assay was linear from 2 to 200 microg/mL for each enantiomer. The analytical method gave average recoveries of 97.5% and 97.0% for S(+)- and R(-)-propafenone, respectively. The limits of detection and quantitation for the method are 0.1 and 2.0 microg/mL for both S(+)- and R(-)-propafenone, respectively. The reproducibility of the assay was good (RSD <10%). The method allowed study of the depletion of S(+)- and R(-)-propafenone in transgenic Chinese hamster CHL cells expressing human cytochrome P450. The stereoselectivity of propafenone phase I metabolism via cDNA-expressed CYP3A4 was observed.
建立了一种对表达人细胞色素P450的转基因中国仓鼠CHL细胞中S(+)-和R(-)-普罗帕酮的对映体选择性分析方法。该方法包括从S9孵育物中提取普罗帕酮，以及用手性试剂2,3,4,6-四-O-β-D-吡喃葡萄糖基异硫氰酸酯形成普罗帕酮非对映体衍生物。非对映体普罗帕酮衍生物的分离和定量在具有紫外检测的反相高效液相色谱系统中进行。该分析方法对每种对映体在2至200μg/mL范围内呈线性。该分析方法对S(+)-和R(-)-普罗帕酮的平均回收率分别为97.5%和97.0%。该方法对S(+)-和R(-)-普罗帕酮的检测限和定量限分别为0.1和2.0μg/mL。该分析方法的重现性良好（相对标准偏差<10%）。该方法可用于研究表达人细胞色素P450的转基因中国仓鼠CHL细胞中S(+)-和R(-)-普罗帕酮的消耗情况。观察到普罗帕酮通过cDNA表达的CYP3A4进行I相代谢的立体选择性。
