Journal
BIOMEDICAL CHROMATOGRAPHY
Volume 27, Issue 11, Pages 1532-1539Publisher
WILEY-BLACKWELL
DOI: 10.1002/bmc.2956
Keywords
topotecan; liquid chromatography-tandem mass spectrometry; Ostro(TM) sorbent plate; bioequivalence
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Funding
- National Natural Science Foundation of China [81120108025]
- Department of Education of Guangdong [GDJX1121]
- International Science and Technology Cooperation Base [2010JD035]
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Topotecan (TPT) is an important anti-cancer drug that inhibits topoisomerase I. A sensitive and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method that potentially determines TPT in beagle dog plasma is needed for a bioequivalence study of TPT formulations. We developed and validated LC-MS/MS to evaluate TPT in beagle dog plasma in terms of specificity, linearity, precision, accuracy, stability, extraction recovery and matrix effect. Plasma samples were treated with an Ostro(TM) sorbent plate (a robust and effective tool) to eliminate phospholipids and proteins before analysis. TPT and camptothecin (internal standard) were separated on an Acquity UPLC BEH C-18 column (1.7 mu m, 2.1x50mm) with 0.1% formic acid and methanol as the mobile phase at a flow rate of 0.25mL/min. TPT was analyzed using positive ion electrospray ionization in multiple-reaction monitoring mode. The obtained lower limit of quantitation was 1ng/mL (signal-to-noise ratio>10). The standard calibration curve for TPT was linear (correlation coefficient>0.99) at the concentration range of 1-400ng/mL. The intra-day and inter-day precision, accuracy, stability, extraction recovery and matrix effect of TPT were within the acceptable limits. The validated method was successfully applied in a bioequivalence study of TPT in healthy beagle dogs. Copyright (c) 2013 John Wiley & Sons, Ltd.
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