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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 mm×150 mm, 5μm) with mobile phase composed of methanol / water containing 0.1% formic acid (70/30, v/v) in an isocratic mode at a flow rate of 0.5 ml/min. The detection was performed using an API triple quadrupole mass spectrometer in APCI 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 concentration 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 collecting from Crohn's disease patients after a single oral administration of thalidomide 100mg.
This article was published in the following journal.
Name: Biomedical chromatography : BMC
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Crohn's Disease (CD)
Crohn’s disease (CD) is a long-term condition that causes inflammation of the lining of the digestive system. Inflammation can affect any part of the digestive system, from the mouth to the back passage, but most commonly occurs in the last s...