Simultaneous determination of itraconazole and hydroxyitraconazole in human plasma by liquid chromatography-isotope dilution tandem mass spectrometry method.
A rapid and sensitive liquid chromatography-isotope dilution tandem mass spectrometry method was developed and validated for quantification of itraconazole (ITZ) and its active metabolite hydroxyitraconazole (OH-ITZ ) in human plasma. The plasma samples were extracted with tert-butyl methyl ether and two isotope-labeled internal standards (D5-itraconazole and D5-hydroxyitraconazole) were used. The chromatographic separation was performed on a Capcell Pak C(18) MG III (100 x 2 mm, 5 microm, Shiseido). The protonated ions of analytes were detected in positive ionization in multiple reaction monitoring mode. The plasma method has a lower limit of quantification of 1 ng/mL with a linearity range of 1-500 ng/mL for ITZ and OH-ITZ using 100 microL of plasma. The recoveries of the method were found to be 69.47-71.98% for ITZ and 75.68-82.52% for OH-ITZ. The intra- and inter-batch precision was less than 11% for all quality control samples at concentrations of 2.5, 200 and 400 ng/mL. These results indicate that the method was efficient with a short run time (4.5 min) and acceptable accuracy, precision and sensitivity.The validated method was successfully applied to analysis of human plasma samples in pharmacokinetics study.
Jia
Jing-Ying
JY
Central Laboratory, Shanghai Xuhui Central Hospital,966 Huaihai Middle Road, Shanghai 200031, China.
Lu
Chuan
C
Liu
Gang-Yi
GY
Zhang
Meng-Qi
MQ
Liu
Yan-Mei
YM
Wang
Wei
W
Weng
Li-Ping
LP
Li
Shui-Jun
SJ
Yu
Chen
C
D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007553:Isotope Labeling; D017964:Itraconazole / Q000031:analogs & derivatives* / Q000097:blood* / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods*
C084794:hydroxyitraconazole; D017964:Itraconazole