Determination of berberine in human plasma by liquid chromatography-electrospray ionization-mass spectrometry

A liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method for the determination of berberine in human plasma using chlorobenzylidine as the internal standard (IS) has been developed and validated. The plasma samples were prepared by LLE and the analytes were chromatographically separated on a Hanbon Lichrospher 5-C18 HPLC column under gradient elution with a mobile phase consisted of acetonitrile and 10mm ammonium acetate buffer containing 0.1% formic acid. Berberine was determined with electrospray ionisation-mass spectrometry (ESI-MS). LC-ESI-MS was performed in the selected-ion monitoring (SIM) mode using target ions at M(+)m/z 336.1 for berberine and M(+)m/z 464.1 for the IS. Calibration curve was linear over the range of 0.020-3.0 ng/ml. The lower limit of quantification (LLOQ) was 0.020 ng/ml. The intra- and inter-run variability values were less than 6.7 and 7.7%, respectively. The method has been successfully applied to determine the plasma concentration of berberine in healthy Chinese volunteers.