The Effect Of Safflower Injection On The Metabolism Of Carvedilol In Beagles

OBJECTIVEUltra-high performance liquid chromatography tandem mass spectrometry?UPLC-MS/MS?was established to detect the concentration of carvedilol in plasma,and we studied the effect of safflower injection on the metabolism of carvedilol in Beagles.METHODS?1?The effective UPLC–MS/MS separation of the examined compounds was applied on a ZORBAX Eclipse Plus C18?2.1 mm×50 mm,1.7?m?column with a gradient mobile phase system.Agilent 6420 triple quadrupole mass spectrometer equipped with an electrospray ionization?ESI?interface was used for mass spectrometric detection.The MRM transitions of m/z 406.94?99.87?m/z423.10?99.93?m/z 423.03?99.93?m/z 393.03?209.88?m/z 268.10?116.00were used to quantify for CAR?4'-HPC?5'-HPC and IS,respectively.?2?Six healthy beagles were selected,and a single dose of carvedilol tablets?15mg/kg??control group?and 10 consecutive days of safflower injection?0.25 mL/kg?were used.The pharmacokinetic differences of carvedilol tablets?experimental group?were given.Venous blood was collected from the medial cephalic vein of the forelimb or the lateral saphenous vein of the hind limb for 0.25,0.5,0.75,1,1.5,2,4,6,9,12,24 h before and after administration.In the EP tube?heparinized?,mixed and centrifuged at 10000 r·min^-1for 15 min,then took the serum at-20°C for testing.The concentrations of CAR and 4'-HPC,5'-HPC plasma were determined by the validated UPLC-MS/MS method.The pharmacokinetic parameters were calculated by DAS 2.0software,and the two groups of data were calculated by SPSS 20.0 software.RESULTS?1?The CAR?4'-HPC?5'-HPC were eluted at 4.3min?3.0 min?3.0min respectively.Excellent liner relationship was obtained from the range of 1 ng/mL to1000 ng/mL for CAR,0.1 ng/mL to 100 ng/mL for 4'-HPC,0.1 ng/mL to 100 ng/mL for 5'-HPC.The intra-day RSD and inter-day RSD were less than 15%,The recoveries were more than 90%at three concentrations?CAR 2.5?100?500 ng/mL?4'-HPC0.25?10?50 ng/mL?5'-HPC 0.25?10?50 ng/mL?.?2?After the combination of Safflower Injection and Carvedilol,the C_maxax of Carvedilol increased by 39.88%;the C_maxax of 4'-HPC reduced by 71.59%;the 5'-HPC C_maxax was reduced by 79.01%.Other pharmacokinetic parameters also increase or decrease accordingly.CONCLUSIONSThis study successfully established UPLC-MS/MS method for simultaneous detection of carvedilol and its main metabolites?CAR,4'-HPC and 5'-HPC?with high specificity,complete separation and fast detection time;suitable for cardvedilol study in the drug interaction.Safflower injection can inhibit the metabolism of carvedilol in the body.When clinical safflower injection is combined with carvedilol,attention should be paid to the effect of safflower injection on the metabolism of carvedilol and monitoring of therapeutic drugs.