Determination of diazepam and its metabolites in human liver microsomal incubation systems by HPLC-MS/MS method
10.13699/j.cnki.1001-6821.2015.12.030
- VernacularTitle:HPLC－MS／MS法同时测定肝微粒体孵育体系中地西泮及其代谢产物的浓度
- Author:
Ya-Fei XU
1
;
Rui-Hua DONG
;
Heng-Yan QU
;
Jing WANG
;
Ze-Yuan LIU
;
Jing-Jie MA
Author Information
1. 军事医学科学院 附属医院 临床药理学研究室
- Keywords:
HPLC-MS/MS;
diazapam;
nordiazepam;
temazepam;
oxazepam
- From:
The Chinese Journal of Clinical Pharmacology
2015;(12):1180-1183
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a HPLC-MS/MS method for simul-taneous determination of diazapam and its three metabolites in human liv-er microsomal.Methods Kromasil C18 column was used at 40 ℃, and the mobile phase was a mixed system consisting of acetonitrile -2 mmol? L-1 mammonium acetate ( 70 ∶30 ) and a flow rate of 0.3 mL? min -1 .The detection of the samples was made using positive ion scan and multiple reaction monitoring mode.The specificity, linearity and limit of quantification, precision and recovery, matrix effect and sta-bility were assessed.Results The sample showed good linearity from 20-1000 ng? mL-1 ( r>0.99 ) and the lower limit of quantitation was 20 ng? mL-1 .The liner calibration curve of nordiazepam, temazepam and oxazepam obtained concentration range of 2 -100 ng? mL-1 (r>0.99) and the low limit of quantitation was 2 ng? mL-1 .The average recovery rates of diazepam, nordiazepam, temazepam and oxaze-pam were 55.47% to 68.76%, 64.84% to 79.11%, 68.32% to 78.27%and 72.66%to 83.82%.The intra-day and inter-day relative standard deviations were both less than 15%.Conclusion The method is simple, rapid and accurate, suitable for measuring the concentration of diazapam and its three metabolites in human liver microsomal.
