Determination of dexamethasone and its metabolite in human liver microsomal incubation systems by HPLC-MS/MS method
10.13699/j.cnki.1001-6821.2015.13.022
- VernacularTitle:HPLC-MS／MS法同时测定肝微粒体孵育体系中地塞米松及其代谢产物的浓度
- Author:
Ya-Fei XU
1
;
Rui-Hua DONG
;
Ze-Yuan LIU
;
Jing-Jie MA
;
Heng-Yan QU
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;
dexamethasone;
6-beta-hydroxy dexamethasone;
in vitro;
liver microsomal
- From:
The Chinese Journal of Clinical Pharmacology
2015;(13):1292-1295
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a HPLC-MS/MS method for simul-taneous determinations of dexamethasone and its metabolite in human liver microsomal incubation system.Methods Kromasil C18 column (2.1 mm ×150 mm, 5 μm) was used at 40 ℃, the mobile phase was a mixed system consisting of acetonitrile and 0.01%formic acid ( 45∶55 ) and the flow rate was 0.35 mL? min -1 .The detection of the samples was made using positive ion scan and multiple reaction monitoring mode .The specificity , standard curve and lower limit of quantification , precision and recovery rate and stability as well as the matrix effect were investiga-ted.Results The liner calibration curve of dexamethasone and 6 -beta-hydroxydexamethasone obtained a concentration range of 20-1000 ng? mL-1 and 1-50 ng? mL-1 ( r =0.998 0 , r=0.998 5 ) and the lower limit of quantification was 20 and 1 ng? mL-1 , the regression equation was y =3.93 ×10 -3 x +9.88 ×10 -3 , y =1.88 ×10 -2 x +2.21 ×10 -4 . The average recovery rates of dexamethasone and 6-beta-hydroxydexamethasone were 73.87% - 92.18% and 86.33%-87.22%.Intra-day and inter-day relative standard devia-tions ( RSD ) of dexamethasone and 6 -beta -hydroxydexamethasone were less than 15%.Conclusion The method is simple , rapid and accurate , suitable for measuring the concentration of dexamethasone and its metabolite in human liver microsomal .
