[UPLC-MS/MS method for the determination of icaritin in rat plasma]
Shuangqing Zhang1, Zhenwu Huang
1National Institute for Nutrition and Food Safety, China Center for Disease Control and Prevention, Beijing 100050, China. zhangshq@hotmail.com
Objective:
To establish an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the quantification of icairitin (ICT), and investigate pharmacokinetics of ICT in rats following a single intravenous administration.
Methods:
ICT and an internal standard coumestrol (CMT) were extracted from rat plasma using acetonitrile and separated on a BEH C18 column (2.1 mm x 50 mm, 1.7 μm) using a gradient mobile phase of acetonitrile, water, ammonium formate and formic acid at a flow rate of 0.3 ml/min. At negative electrospray ionization mode, multiple reaction monitoring of the precursor-product ion transitions of m/z 367.1-->297.1 for ICT, 267.0 --> 211. 1 for CMT was used for the quantification. Plasma was collected after rats were intravenously administered with ICT at a single dose of 10 mg/kg.
Results:
The linear calibration curve was obtained in a concentration range of 0.5 - 20 ng/ml with a lower limit of quantification of 0. 5 ng/ml. The value of intra- and inter-day precision was less than 8.1% and accuracy fell in the ranges of 95.3% - 99.3%. The recovery ranged from 90.1% to 92.1% and the matrix effects from 97.5% to 101.2%. After intravenous administration of ICT to rats, t½ was(1.13 ± 0.32) h, V was(4.54 ± 0.27) L/kg, and CL was (2.91 ± 0.68) L/(h x kg).
Conclusion:
The method was rapid, specific and accurate, suitable for preclinical pharmacokinetics of ICT.
