| Objective: Gliquidone is the second generation of sulfonylurea oral hypoglycemic drugs.This paper is to develop a LC-MS/MS method for the determination of gliquidone in plasma samples,and conduct a pharmacokinetics and bioequivalence study of gliquidone tablets and gliquidone dispersible tablets in Chinese healthy volunteers.Methods: The pharmacokinetics and bioequivalence of gliquidone tablets and gliquidone dispersible tablets were studied among 20 healthy male volunteers after taking a single oral dose of 30 mg test and reference formulations according to a randomized two-crossover design.The reference formulation was gliquidone tablet produced by Beijing Manfield DoubleCrane pharmaceutical Co,Ltd.Blood samples were collected at the designed moment before and after administration.Glyburide was selected as internal standard（IS）.The plasma samples were processed by protein-precipitation method and separated on a Capcell PAK MG C18（50×4.6mm I.D.,5μm）analytical column connected with a C18（4×3.0 mm I.D.,5 μm）guard column using acetonitrile-water-formic acid（67: 33:0.1,v/v/v）as the mobile phase at a flow rate of 0.8 ml/min.A tandem quadrupole mass spectrometer equipped with electrospray ionization source was used as the detector and operated in the negative ion mode.Multiple-reaction monitoring（MRM）using the precursor to product ion combinations of m/z 526.2 to m/z 401.2 and m/z492.1to m/z 367.1 were used to quantify gliquidone and IS.Pharmacokinetic parameters were calculated by DAS 2.1.1 sofware.The bioequivalence of two formulations was evaluated by ANOVA,two one-sided t test and[1-2α] confidential interval calculation of logarithmic transformed Cmax and AUC.Tmax was analysized on the base of non-parametric Wilcoxon signedrank test.Results: The linear calibration curves were obtained among the concentration range of 5.00-1000 ng/ml,and the limit of quantification was 5.00ng/ml.For quality control（QC）samples of gliquidone at 10.0,100,and 800ng/ml,the intra-day RSDs（%）were 7.2,3.8 and 3.3,the inter-day RSDs（%）were 10.0,7.0 and 9.5,the accuracies（%）were 97.5,99.9 and 100.4 and extraction recoveries were（73.5±5.0）%,（84.8%±2.3）% and（81.7±4.4）%,respectttively.The extraction recovery of the IS was（93.2±6.0）%.The matrix effect determined at two concentrations（10.0 and 100 ng/ml of gliquidone）were（94.7±7.1）%和 and（103.8±2.9）%,and their relative errors were both within 7.5%.Stability studies revealed that gliquidone was stable in plasma for 6 h at room temperature,at the end of three successive freeze and thaw cycles and for 56 d at-70°C.The main pharmacokinetic parameters of test and reference formulation in 19 healthy male volunteers after taking a dose of30 mg gliquidone tablet and dispersible tablet were as followings: Cmax（773.1±224.1）ng/ml and（668.5±213.6）ng/ml,Tmax（3.3±1.6）h and（3.1±1.0）h,AUC0-t（4853.0±1600.7）ng/ml·h and（4428.8±1335.7）ng/ml·h,AUC0-∞（5501.9±1903.0）ng/ml·h and（5227.6±2473.9）ng/ml·h,t1/2（8.9±7.2）h and（11.9±25.1）h.The ANOVA results indicated that there was no significant difference between gliquidone tablets and gliquidone dispersible tablets（P>0.05）.[1-2α] confidential interval of geometric mean values of AUC0-24 h and Cmax was 88.0%-96.0% and 78.0%-94.4%,respectively.The former met the bioequivalence criteria of 80%-125%,but the latter not.The relative bioavailability of the test compared to reference preparation was（92.5±10.5）% according to AUC0-t.Conclusion: The determination method of gliquidone in human plasma was selective,sensitive,accurate,precise and reliable.It was fully validated on the selectivity,linearity,LLOQ,accuracy,and precision,matrix effect,stability,and was successfully applied in the study of pharmacokinetics and bioequivalence of gliquidone dispersible tablets.According to Chinese Pharmacopoeia（2015 version）bioequivalence criteria,the two formulations ofgliquidone were not bioequivalent. |
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