Analysis of Lycobetaine in Rat Plasma by LC-ESI-MS/MS

In this study, a selective and sensitive liquid chromatography-electrospray ionization-tandem mass spectrometric method was developed and validated for the determination of lycobetaine in rat plasma. Berberine was selected as the internal standard, and rat plasma samples were pretreated via protein precipitation and further separated on a diamonsil octadecyl-silylated silica column using 0.2% (v/v) aqueous formic acid and methanol as the mobile phase. Selected reaction monitoring was performed using the transitions m/z 266.1 → 208.1 and m/z 336.1 → 320.0 to determine the concentrations of lycobetaine and internal standard, respectively. The injection volume was 1 µL, and the calibration curve was linear (R2 = 0.9998), while the validated lower limit of quantification was 25 ng/mL. Precision varied from 3.4% to 9.9%, and accuracy varied from -2.6% to 8.7%. Lycobetaine remained stable under all relevant analytical conditions tested in the study. The method was successfully applied to determine the plasma concentration of lycobetaine in a pharmacokinetic study. After intravenous administration of 10 mg/kg and oral administration of 200 mg/kg lycobetaine in rats, the pharmacokinetic parameters were calculated and the oral bioavailability of lycobetaine was determined as 7.30% ± 1.44%.