采用固相萃取-高效液相色谱法同时测定大鼠尿液中扑米酮及其三种主要代谢物。

Simultaneous determination of primidone and its three major metabolites in rat urine by high-performance liquid chromatography using solid-phase extraction.

作者信息

Ferranti V, Chabenat C, Ménager S, Lafont O

机构信息

Laboratoire de Pharmacochimie, Faculté de Médecine et de Pharmacie de Rouen, Saint-Etienne-du-Rouvray, France.

出版信息

J Chromatogr B Biomed Sci Appl. 1998 Oct 23;718(1):199-204. doi: 10.1016/s0378-4347(98)00356-9.

DOI:10.1016/s0378-4347(98)00356-9

PMID:9832377

Abstract

A new high-performance liquid chromatographic method for simultaneous determination of primidone (PRM) and of its three major metabolites, phenobarbital (PB), p-hydroxyphenobarbital (p-HO-PB) and phenylethylmalonamide (PEMA), in rat urine, was developed. After acid hydrolysis, these compounds were extracted from urine by means of a Bond Elut Certify LRC column with good clean-up. The extracts were chromatographed on a C18 reversed-phase column using isocratic elution at 40 degrees C, with UV detection at 227 nm. The limit of detection was 0.5 mg/ml for the four compounds. Good linearity (r2>0.99) was observed within the calibration ranges studied: 37.4-299.3 microg/ml for PRM, 26.4-211.2 microg/ml for PB, 12.5-100.2 microg/ml for p-HO-PB and 12.1-97.0 microg/ml for PEMA. Repeatability was in the range 3.1-6.8%. This method constitutes a useful tool for studies on the influence of various parameters on primidone metabolism.

摘要

建立了一种高效液相色谱法，用于同时测定大鼠尿液中的扑米酮（PRM）及其三种主要代谢物苯巴比妥（PB）、对羟基苯巴比妥（p-HO-PB）和苯乙基丙二酰胺（PEMA）。酸水解后，这些化合物通过Bond Elut Certify LRC柱从尿液中提取，净化效果良好。提取物在C18反相柱上进行色谱分离，于40℃采用等度洗脱，在227nm处进行紫外检测。四种化合物的检测限均为0.5mg/ml。在所研究的校准范围内观察到良好的线性关系（r2>0.99）：PRM为37.4 - 299.3μg/ml，PB为26.4 - 211.2μg/ml，p-HO-PB为12.5 - 100.2μg/ml，PEMA为12.1 - 97.0μg/ml。重复性在3.1% - 6.8%范围内。该方法为研究各种参数对扑米酮代谢的影响提供了一种有用的工具。