摘要
Bergenin, a C-glucoside of 4-O-methyl gallic acid from Bergenia purpurascens, is a naturally antitussive and expectorant agent. A rapid and sensitive liquid chromatography tandem mass spectrometry （LC-MS/MS） method has been developed and validated for the determination of the active component--bergenin, in rat plasma after oral administration of aqueous B. purpurascens extract. The plasma samples were pretreated by protein precipitation with acetonitrile and chromatographic separation was achieved on a Diamonsil C18 column （150mim×4.6mm, 5μm） with isocratic elution using a mobile phase consisting of water- methanol （30：70, v/v） at a flow rate of 0.6 mL/min. The detection was accomplished by a triple- quadrupole tandem mass spectrometer in multiple-reaction monitoring （MRM） scanning via an electrospray ionization （ESI） source operating in the negative mode. The optimized mass transition ion-pairs （m/z） for quantitation were 327.3/192.0 for bergenin, and 431.1/311.1 for IS. The time for each analysis run was only 3.5 min between injections. The calibration curve exhibited good linearity （r2〉 0.99） over a range of 1.00-2000 ng/mL for bergenin. The lower limit of quantitation （LLOQ） was 1.00ng/mL. The intra- and inter-day precisions were no more than 11.8%, and relative errors （RE） were within the range of 0.0-4.4%. The validated method was successfully applied to investigate the pharmacokinetics of bergenin after oral administration of B. purpurascens extract in rats.
Bergenin, a C-glucoside of 4-O-methyl gallic acid from Bergenia purpurascens, is a naturally antitussive and expectorant agent. A rapid and sensitive liquid chromatography tandem mass spectrometry （LC-MS/MS） method has been developed and validated for the determination of the active component--bergenin, in rat plasma after oral administration of aqueous B. purpurascens extract. The plasma samples were pretreated by protein precipitation with acetonitrile and chromatographic separation was achieved on a Diamonsil C18 column （150mim×4.6mm, 5μm） with isocratic elution using a mobile phase consisting of water- methanol （30：70, v/v） at a flow rate of 0.6 mL/min. The detection was accomplished by a triple- quadrupole tandem mass spectrometer in multiple-reaction monitoring （MRM） scanning via an electrospray ionization （ESI） source operating in the negative mode. The optimized mass transition ion-pairs （m/z） for quantitation were 327.3/192.0 for bergenin, and 431.1/311.1 for IS. The time for each analysis run was only 3.5 min between injections. The calibration curve exhibited good linearity （r2〉 0.99） over a range of 1.00-2000 ng/mL for bergenin. The lower limit of quantitation （LLOQ） was 1.00ng/mL. The intra- and inter-day precisions were no more than 11.8%, and relative errors （RE） were within the range of 0.0-4.4%. The validated method was successfully applied to investigate the pharmacokinetics of bergenin after oral administration of B. purpurascens extract in rats.
基金
supported by the Natural Science Foundation of Guangdong Province (Nos. 9151008018000003)
