Development and validation of a sensitive liquid chromatography-tandem mass spectrometry method for the determination of naringin and its metabolite, naringenin, in human plasma.

A sensitive and specific method was developed for the simultaneous determination of naringin and its metabolite, naringenin, in human plasma by liquid chromatography-tandem mass spectrometry. Hesperidin was used as the internal standard, plasma samples were extracted with ethyl acetate and the analytes were chromatographically separated by using acetonitrile-0.1% formic acid (gradient elution) as the mobile phase. Detection was performed by electrospray ionization mass spectrometry in negative ion mode with multiple reaction monitoring. The lower limit of quantification was 0.5 ng/mL for naringin and naringenin and the linear calibration curves ranged from 0.5 to 200 ng/mL. The intra-run and inter-run precision values were within 8.6 and 7.7% for naringin and between 13.1 and 10.3% for naringenin. The accuracy ranged from 91.3 to 98.2% for naringin and from 90.2 to 97.6 % for naringenin. The validated method was successfully applied to determine concentrations of naringin and naringenin in clinical patients.