Simultaneous determination of five antiviral drug residues and stability studies in honey using a two-step fraction capture coupled to liquid chromatography tandem mass spectrometry.

ABSTRACT An effective sample pretreatment method followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) was first developed for simultaneous determination of five antiviral drug residues including ribavirin, moroxydine, amantadine, rimantadine and memantine in honey. To adsorb analytes with different binding properties and overcome the interference of sugars and uridine as endogenous ribavirin structural analogs in honey, the target drugs were extracted with 1% formic acid and then purified by a phenylboronic acid (PBA) solid phase extraction cartridge using two-step fraction capture prior to LC-MS/MS analysis. This method was validated by analyzing honey samples from nine floral origins including miscellaneous flowers, citrus, vitex, rape, acacia, sunflower, linden, buckwheat and jujube spiking at multiple levels, and the recoveries ranged from 82.46% to 116.34%, with relative standard deviations (RSDs) less than 14.58%. The limits of detection (LODs) and limits of quantitation (LOQs) of moroxydine, ribavirin, amantadine, rimantadine, and memantine were 0.1–2 µg/kg and 0.2–5 µg/kg, respectively. Depletion experiments of five antiviral drugs in honey at different storage and process temperatures demonstrated that moroxydine can potentially be used as a drug to cure sacbrood disease in honeybees.