Application of an ultra performance liquid chromatography-tandem mass spectrometry method to the pharmacokinetics study of paeoniflorin-6′-O-benzene sulfonate and its active paeoniflorin metabolite in rat plasma

Ming-yi ZHAO,Jun YU,Jian WANG,Yi-jin WU,Feng XIAO,Chun WANG,Wei WEI

(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Hefei 230032,China)

T2-34

Application of an ultra performance liquid chromatography-tandem mass spectrometry method to the pharmacokinetics study of paeoniflorin-6′-O-benzene sulfonate and its active paeoniflorin metabolite in rat plasma

Ming-yi ZHAO,Jun YU,Jian WANG,Yi-jin WU,Feng XIAO,Chun WANG,Wei WEI

(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory and Immune Medicine,Ministry of Education,Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine,Hefei 230032,China)

OBJECTIVETo establish a rapid,simple,precise and sensitive ultra performance liquid chromatography(UPLC)method for determination of paeoniflorin-6′-O-benzene sulfonate(acylated derivative of Pae,CP-25)and its primary metabolite(Pae,M1)in rat plasma,and to investigate the effects of gender,food and disease status on the pharmacokinetics after oral administration in rats.METHODSPlasma samples and calibrators were extracted with methanol after addition of the internal standard solution.After evaporation of the methanol layer,the residue was reconstituted in mobile phase,and a 2 μL of the sample was injected into a Waters ACQUITY UPLC BEH C18(2.1 mm × 50 mm,1.7 μm)column for separation at a flow rate of 0.5 mL·min-1at 40℃.The mobile phase was composed of 0.1%formic acid in water and methanol(68∶32,V/V).RESULTSThe developed UPLC-MS/MS method was linear in the concentration range of 2-800 mg·L-1.Validation of the method proved that the method′s precision,selectivity and stability were all within the acceptable limits.Pharmacokinetics study showed that CP-25 could have more extensive distribution in females than that in males but no differences in M1.Food intake could also increase the extent of absorption and decrease the rate of clearance of CP-25 and M1 after oral administration in rats.The disease status would decrease the absorption of CP-25 in rats.Comparing single-and multiple-dosing in adjuvant arthritis(AA)model,absorption of CP-25 and M1 was improved with increasing dosage.CONCLUSIONThe developed UPLC-MS/MS method is sensitive,specific and was successfully applied to the pharmacokinetics of CP-25 and M1 in rats.And a significant gender,food intake and disease status differences for CP-25 and M1 were observed in this study.

paeoniflorin-6′-O-benzene sulfonate;paeoniflorin;UPLC-MS/MS;pharmacokinetics

The project supported by National Natural Science Foundation of China(81330081 and 81302845)

Wei WEI,Tel:13605510563,E-mail：wwei@ahmu.edu.cn