Study On The Determination Of Bergenin In Human And Pharmacokinetics

AIM: To establish the rapid and sensitive assays for determination of bergenin in human plasma by liquid chromatography-mass spectrometry (LC-MS/MS). Then the method was used to study the relative bioavailability of bergenin in healthy male volunteers, to assess the bioequivalence and to provide scientific evidence for reasonable guideline for clinical application.METHOD: The LC-MS/MS methods with high sensitive and high precision, were developed and validated for Bergenin assay in plasma sample. Bergenin and the internal standard, 5-BrU, were precipitated from the matrix by acetonitrile, then washed with dichloromethane and separated on a Zorbax SB-C18 column employing a 10 mM ammonium acetate/ acetonitrile gradient system. Detection was carried out by multiple reaction monitoring on a LC-MS/MS system. The analytic methodology was evaluated by detecting the limit of quantization for Bergenin in plasma, drawn recovery rate, stability, RSD of inter-precision and intra-precision, and selectivity and proved whether it could be used in studying the bioequivalence of Bergenin.According to a two-period, two-sequence, randomized crossover design, the pharmacokinetics of the 20 healthy male volunteers were studied after each was administrated by a single oral dose of bergenin dripping pills (test drug) and bergenin tablet (reference drug), respectively. The concentrations of bergenin in plasma samples at different time points were determined by LC-MS/MS and the plasma concentrations-time profiles were obtained. The other pharmacokinetic parameters were calculated by pharmacokinetic statistical software BAPP 2.0. Analysis of Variance (ANOVA), two-one side Student's test and (1-2α)% confidential internal were adopted to statistically evaluate the equivalences.RESULTS: After oral administration with single dose of test drug (T) or reference drug (R) at 250 mg bergenin. The Tmax values of bergenin were 1.9±0.8 (mean±SD) and 1.9±0.9 h, respectively. The Cmax values of tranilast were 66.58±22.1 and 82.1± 37.36 ng/mL, respectively. The t1/2 values of tranilast were 5.67±3.76 and 4.8±2.86 h, respectively. The AUC0-t values were 282.33±92.53 and 314.91±140.83 ng?h/mL and the AUC0-∞values were 348.51±116.56 and 355.47±161.59 ng?h/mL, respectively. According to AUC0-t, the relative bioavailability of test drug was 100.6±38.8 %.CONCLUSION: The pharmacokinetics of bergenin test drug and reference drug were similar. Analysis of Variance (ANOVA), two-one side Student's test and (1-2α)% confidential internal were adopted to statistically evaluate the equivalences. Based on the pharmacokinetic and statistical results, we could conclude that the test drug is bioequivalent to the reference drug.