Abstract
A sensitive and reliable liquid chromatography–tandem mass spectrometry (LC–MS/MS) has been developed and validated for the determination of andrographolide sodium bisulphite (ASB) in dog plasma using dehydroandrographolide (DAG) as an internal standard. Chromatographic separation was achieved on a Hypersil Gold C18 column (50mm×2.1mm, 1.9μm) with gradient elution that consisted of methanol and water at a flow rate of 0.2mL/min. Quantification was done using selected reaction monitoring (SRM) mode to monitor precursor–product ion transitions of m/z 413.2→287.2 for ASB and 331.2→303.3 for DAG at negative ionization mode. Good linearity was obtained over the range of 10–1000ng/mL and the correlation coefficient was better than 0.99. The intra- and inter-day accuracies ranged from 97.2% to 107.8% and precisions (RSD) were within 13.9%. ASB was found stable under three freeze–thaw cycles, short-term temperature, post-preparative and long-term temperature conditions. The method was successfully applied to a pharmacokinetic study of ASB intravenously administered to Beagle dogs.
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