Pharmacokinetics Of Five Major Active Ingredients In Beagle Dogs Based On Liquid Chromatography - Mass Spectrometry

Majie Pingchuan Babu agent in Qing Dynasty "Zhang Yee Tong" contained in semen Sinapis Albae, mainly for treatment of acute and chronic asthma by acupoint application method, clinical curative effect, the research group on the basis of semen Sinapis Albae on, re composed of Majie Pingchuan decoction, the ephedra, bitter almond, white mustard seed composition of ginger, rhizoma corydalis and five kinds of traditional Chinese medicine, and to optimize the formulation, development of Majie Pingchuan Babu agent, and. This study was carried out in two aspects in the previous basis, first established for simultaneous determination of five kinds of main chemical components of Majie Pingchuan Babu agent: ephedrine, pseudoephedrine, fluid of sinapine thiocyanate, tetrahydropalmatine, amygdalin in Beagle dog plasma content of spectrometry (LC-MS/MS); the second five, the main chemical components of in vivo drug metabolism beagle, objective material foundation and the in vivo process so as to reveal Babu agent, clinical efficacy, and provide scientific basis for its clinical application and further development of new drugs.Objective:To establish a simultaneous determination in Beagle dog plasma five main components (ephedrine. pseudoephedrine, sinapine thiocyanate, bitter almond gluc oside, corydalis b) concentration by liquid chromatography mass spectrometry (LC-MS MS); and for the observation of percutaneous drug administration in beagle dogs in dynamic changes and pharmacokinetic characteristics.Methods:Majie Pingchuan cataplasm agent in beagle dogs is transdermal drug delivery, and in the following 13 at the time of blood sampling:0min, 10min,30min,60min,90min and 120min,150min,3h,6h,9h,12h,24h,48h, the blood sampling from the experience of the plasma samples by methanol extraction method the plasma treatment to be measured. Choice of chromatographic and mass spectrometric conditions:column chromatography:Zorbax rrhd eclipse plus C18 (3.0 humorous 100 mm,1.8 μm)/40 degree, flow phase:A:B:water acetonitrile gradient program:0.0 min.3% B; 10.0 min,6% B; 12.0 min,40% B; 13.0 min, 98%B; 14.0 min,98% B; 14.1 min,3% B. Sample size:5 g l, flow rate of 0.4 ml/min, post column split, enter the mass spectrometer detector and ion polarity is positive ion, atomizing air pressure:N2@3.5 psi, drying air velocity and temperature:N2@15.0 L@ 200 ℃, sheath gas flow rate and temperature:N2@ 11.0 L@ at 350 ℃, capillary voltage of 4KV, nozzle voltage:0 V. For analysis:ephedrine, pseudoephedrine, sinapine thiocyanate, tetrahydropalmatine.:@@@,10%B.,98%B,98%B,40%B,10%B; gradient of acetonitrile water (negative ion mode:the chromatographic column was Zorbax rrhd eclipse plus C183.0 humorous 100 mm.1.8 μ m)/40 degrees, the mobile phase was a:B:Program:0.0 min:3.0 min:5.0 min:6.0 min:6.1 min sample volume of 5 g 1. flow rate of 0.4 ml/min. post column split. enter the mass spectrometer detector and ion polarity is negative ions. atomization gas pressure:N2@ 3.5 psi, drying air velocity and temperature:N215.0 L@ 200 ℃,the sheath gas flow rate and temperature, N211.0 L/min at 350 ℃, capillary voltage poll, spray nozzle voltage 1.5kV. For analysis:bitter almond glycosides. The concentrations of 5 major components in plasma were treated by EXCEL and KINETIC, and the curves and pharmacokinetic parameters were obtained respectively.Results Established method for the simultaneous determination of five compounds by liquid chromatography-mass spectrometry detection methods, the Beagle dogs by percutaneous paste deposited Majie Pingchuan cataplasm agent and ephedrine in 0.01-10ng/ml concentration range of linear good (r= 09989), the lower limit of quantification was 0.01 ng/ml. the major drug dynamic parameters results:Cmax:mountain+0.026ng/ml, Tmax:630+220.454, AUClast:409.393+60.458 ng/ml*min, MRT:3293.275+517.841. Pseudoephedrine good linearity in the concentration range of 0.01-10ng/ml (r=0.9987), the lower limit of quantification was 0.01 ng/ml. The main pharmacokinetic parameters were: Cmax; 0.127+0.020ng/ml, Tmax:450+295.771min, AUClast:175.461+43.273 ng/ml*min, MRT:1763.037+501.097min. The concentration of bitter almond 0.01-50ng/ml was linear within the range of good (r=0.9964), the quantitative limit of the concentration was 0.01 ng/ml. After transdermal delivery, the peak value of the chromatographic peak was below the quantitative limit, and could not make it quantitative.. Tetrahydropalmatine in the concentration range of 0.01-10ng/ml with good linearity (r=0.9994). the limit of quantitation was 0.01 ng/ml. After transdermal delivery, tetrahydropalmatine was not detected. Sinapine thiocyanate with good linearity in the concentration range of 0.01-1 Ong/ml (r=0.9994). the lower limit of quantification was 0.01 ng/ml. The peak of chromatographic peak was the lower limit, and could not make it quantitatively.Conclusion:Liquid mass spectrometry (LC-MS MS) good selectivity, high sensitivity for determination in Beagle dog plasma sample composition content; Majie Pingchuan cataplasm agent of ephedrine and pseudoephedrine percutaneous after administration have bimodal phenomenon and has sustained long-term pharmacokinetic characteristics, reached the pre period can be a long time maintained stable and effective drug concentration in blood, improve drug biological utilization degree, reduce the frequency of taking, reduce the toxic and side effects are designed to, was not detected in the Corydalis B, sinapine thiocyanate. amygdalin, there is no denying that the role of the relieving cough and asthma.