Determination of ketotifen in human plasma by LC-MS
Feras Q Alali1, Bassam M Tashtoush, Naji M Najib
1Department of Medicinal Chemistry and Pharmacognosy, Faculty of Pharmacy, Jordan University of Science and Technology, PO Box 3030, Irbid 22110, Jordan.falali@just.edu.jo
Abstract:
Analytical validation of a new liquid chromatographic-mass spectrometric (LC-MS) method for determination of total amount of ketotifen (unchanged and conjugated) in human plasma is presented. Pizotifen was used as an internal standard. An enzyme hydrolysis of conjugated ketotifen was conducted with a combination of beta-glucuronidase and aryl sulfatase. After enzyme hydrolysis a liquid-liquid extraction was performed as a cleaning step. The quantitative determination was obtained using selected ion monitoring (SIM) LC-MS. Chromatographic condition was a combination of reverse phase gradient system and a switching column technique. A satisfactory hydrolysis, acceptable accuracy, improved precision in the linear range from 0.5 to 20.0 ng/ml plasma, absolute recovery of 98.04% for ketotifen and 95.13% for pizotifen and stability for 7 months at -20 degrees C have been achieved.
