| Objective:To establish a joint mass spectrometry detection in Beagle dog plasma metoprolol concentration in blood by ultra performance liquid chromatography(UPLC-MS)method,and the metoprolol tartaric acid was studied with 12 Beagle dogs in vivo pharmacokinetics by this method.Which is on the Metoprolol Tartrate Sustained-release Tablets release and bioequivalence evaluation.Methods:Seeking 1mL plasma samples with standard solution of 50?L propranolol hydrochloride methanol(4.092?g/ml),vortex mixing for 30s on the solid phase extraction column after activation,and precooling 1mL methanol water solution(3%),collecting methanol elution solution through dry column with 1.5mL methanol with precooled,which was dried under nitrogen at room temperature 20 degrees residue,adding mobile phase acetonitrile water solution(containing 1%formic acid)50?L 3min 13400g 15min scroll,centrifugation,the supernatant of 5?L sample for UPLC-MS analysis,chromatographic conditions were:Agilent SB-C18 RRHD1.8um3*100mm column,acetonitrile(containing 0.1%formic acid)/water(containing 0.1%formic acid)as mobile phase,gradient elution,flow rate was 0.5mL-min-1,sample volume was 5?L,mass spectrometry conditions:pneumatically assisted electrospray ionization;ion polarity:positive ion,spray voltage:3500V,ion detection mode:selected ion monitoring SIM,object detection:[M+H]+m/z268.19,metoprolol,propranolol in standard[M+H]+,m/z260.06.12 Beagle dogs were randomly divided into two groups(6 rats in each group)to take two cycle crossover,and one group to take Metoprolol Tartrate Sustained-release Tablets(the test agent T),while another group received metoprolol tartrate tablets(the reference preparation R),dosage was 100mg,stopping cross medication within the stipulated time of blood sampling in 2 weeks,using ultra performance liquid associated with mass spectrometry(UPLC-MS)method,which was used to detect plasma concentration of metoprolol.Using Kinetica5.5 pharmacokinetic analysis to calculate the pharmacokinetic parameters and relative bioavailability with software and using two one-sided t test and 90%confidence interval method,which was used to evaluate the bioequivalence.Results:in this experiment,the method was specific and impurities in the plasma was without interfere determination.Plasma metoprolol concentration in the range of 15.56-996 ng/mL range,which had a good linear relationship(r=0.9817),the lower limit of quantification(LLOQ)was 15.56ng/ml,the experimental results showed that this method was of high accuracy,good precision,stability of the experimental results of samples showed that samples at-80 degree for cryopreservation of good stability,high recovery rate of absolute.Injecting a single oral dose of metoprolol tartrate sustained-release tablets and conventional tablets of metoprolol in Beagle dogs,plasma Tmax were:2.17± 0.39h and 1 ± Oh respectively,Cmax:223.22 ± 34.03ng/mL and 811.60 ± 119.83ng/mL respectively,t1/2:6.37 ± 2.49h and 2.57 ± 0.90h respectively,AUCO-t:1157.69 ± 189.29ng·h/mL and 1613.56 ± 158.71ng h/mL respectively,AUCO-?:1314.63 ± 213.50ng·h/mL and 1698.34 ± 141.55ng·h/mL respectively,MRT were:8.61 ± 2.13h and 3.22 ± 0.35h respectively.The range of 90%confidence interval didn't fall in tablets of 80%?125%relevant pharmacokinetic parameters,which were analyzed Metoprolol Tartrate Sustained-release Tablets within the range of AUCO-t,90%confidence interval Cmax also didn't fall in the tablets of ordinary 80%?125%,Cmax significantly decreased Tmax was dramticly prolonged.Sustained release tablets compared with ordinary the relative bioavailability was 72.45%.Conclusion:This experiment established by ultra performance liquid chromatography coupled with mass spectrometry(UPLC-MS)method,the tolol method for detecting drug concentration in blood plasma of Beagle dogs was high accuracy,good sensitivity,which can be used for studying clinical medicine generation of dynamics,the measured metoprolol tartrate sustained release tablets and common tablets are bioequivalent with a parentally sustained release effect. |
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