A validated LC-MS/MS method for the simultaneous determination of thalidomide and its two metabolites in human plasma: Application to a pharmacokinetic assay.
An accurate and sensitive LC-MS/MS method for determining thalidomide, 5-hydroxy thalidomide and 5'-hydroxy thalidomide in human plasma was developed and validated using umbelliferone as an internal standard. The analytes were extracted from plasma (100 μL) by liquid-liquid extraction with ethyl acetate and then separated on a BETASIL C18 column (4.6 × 150 mm, 5 μm) with mobile phase composed of methanol-water containing 0.1% formic acid (70:30, v/v) in isocratic mode at a flow rate of 0.5 mL/min. The detection was performed using an API triple quadrupole mass spectrometer in atmospheric pressure chemical ionization mode. The precursor-to-product ion transitions m/z 259.1 → 186.1 for thalidomide, m/z 273.2 → 161.3 for 5-hydroxy thalidomide, m/z 273.2 → 146.1 for 5'-hydroxy thalidomide and m/z 163.1 → 107.1 for umbelliferone (internal standard, IS) were used for quantification. The calibration curves were obtained in the concentrations of 10.0-2000.0 ng/mL for thalidomide, 0.2-50.0 ng/mL for 5-hydroxy thalidomide and 1.0-200.0 ng/mL for 5'-hydroxy thalidomide. The method was validated with respect to linear, within- and between-batch precision and accuracy, extraction recovery, matrix effect and stability. Then it was successfully applied to estimate the concentration of thalidomide, 5-hydroxy thalidomide and 5'-hydroxy thalidomide in plasma samples collected from Crohn's disease patients after a single oral administration of thalidomide 100 mg.
['Jiang|Fulin|F|http://orcid.org/0000-0002-0918-805X', 'Peng|Xiang|X|', 'Cai|Dake|D|', 'Wen|Dingsheng|D|', 'Liu|Yao|Y|', 'Zhi|Min|M|', 'Chen|Jiangying|J|', 'Hu|Pinjin|P|', 'Wang|Xueding|X|', 'Gao|Yujie|Y|', 'Huang|Min|M|', 'Gao|Xiang|X|', 'Zhong|Guoping|G|http://orcid.org/0000-0002-5753-4578']
D002853:Chromatography, Liquid D004355:Drug Stability D006801:Humans D016014:Linear Models D008297:Male D015203:Reproducibility of Results D012680:Sensitivity and Specificity D053719:Tandem Mass Spectrometry D013792:Thalidomide
LC-MS/MS thalidomide metabolites pharmacokinetics