Study On Pharmacokinetics Of The Chemical Composition In XuanMai GanJu Particles

Xuan Mai Gan Ju Particles is a variety of Chinese medicine Loading in Chinese Pharmacopoeia(2015)derived from Ancient Xuan Mai Gan Ju Decoction,which Constituted by Four herbs od the Scrophulariaceae,Radix,licorice,and Campanulaceae,has the effect of expectorant pharynx,Yin and Lung Nourishing to treat pharyngitis,cough and other respiratory diseases caused by Yin deficiency and Huo exuberance.The clinical use of the medicine is wild,and the effect of it is clear.Currently,domestic scholars' research mostly focused on determination and quality control of the medicine,however,there is no report about the research on active component in pharmacokinetics of this medicine.The experiment was established in UPLC-MS / MS method of liquiritin and harpagoside in rat plasma after oral administration.The method is sensitive,operable to obtain the pharmacokinetic parameters of main active component in Xuan Mai Gan Ju Particles.Fill the blank of vivo studies of the medicine and provide evidence for clinical application of the medicine.?Part One Content Determination of Two kinds of chemical composition in Xuan Mai Gan Ju ParticlesObjective: To measure and establish content analysis of the two active components(harpagoside,liquiritin)of Xuan Mai Gan Ju Particles.Method: Accurately weighed Xuan Mai Gan Ju granules 0.1 g,added 50% ethanol 30 m L,said that given quality,sonicated 0.5 h,cooled to room temperature,used a mass of 50% ethanol and to complement loss of,shake,14 000 r/min centrifuged 5min,and took supernatant through 0.22 ?m microporous membrane,the filtrate as the test solution.mobile phase acetonitrile(7)eluteUsed UPLC-MS/MS method,with Phenomenex KenetixC18 column(50 mm × 3 mm,2.6 ?m),gradient eluting the mobile phase gradient of aqueous solution of acetonitrile containing 0.1% formic acid,volume flow 0.3 mL/ min;and used MRM scan mode of ESI to analyze with injection volume5 ?L.Results: liquiritin and harpagoside shows a good linear relationship respectively in 2.1~420.0 ng/mL,1.6~320.0 ng/mL,r?0.9945.average recoveries of liquiritin and harpagoside are greater than 95%,RSD less than 4.6%,which indicated good recovery rate.Conclusion: This method is feasible,rapid,sensitive,specific and can be well used for the component content analysis of blood component of liquiritin and harpagoside in Xuan Mai Gan Ju Particles.Part Two Pharmacokinetic study on Two kinds of chemical composition of Xuan Mai Gan Ju ParticlesObjective: Establish a UPLC-MS/MS method for the determination to research pharmacokinetics of the active ingredient in Xuan Mai Gan Ju Particles in rats' vivo.Methods: Collected plasma samples after oral administration Xuan Mai Gan Ju Particles in rats.Analyze samples after quantitative aspirating the sample plasma proteins precipitated with acetonitrile.Rutin as an internal standard,acetonitrile-0.1 % aqueous solution as a mobile phase to do a gradient elution,which the flow rate was 0.3 mL/min,the column was Phenomenex KenetixC18 column(50 mm×30 mm,2.6 ?m),column temperature 20 ?.Used ESI source,MRM and negative ion mode to monitor.Injection volume: 10 ?L.And pharmacokinetic parameters were calculated by DAS 3.0 software.Results: The liquiritin and harpagoside shows a good linear relationship respectively 0.78-50.00 ng · mL-1(r = 0.9979),1.66-106.10 ng · m L-1(r = 0.9958)by this method.Precision,accuracy,detection limit and quantitation limit meet the requirements.Stability of two kinds of analyzed plasma samples is good,and RE are less than ± 15%.Recoveries of liquiritin in low middle high concentrations of are(61.4%,67.7%,74.3%);Low middle high concentrations of matrix effects are respectively(93.2%,93.57%,95.6%);Recoveries of harpagoside in low middle high concentrations of are(71.5%,70.6%,82.3%);Low middle high concentrations of matrix effects are respectively(89.5%?87.6%?90.2%);which are all within the allowable range.After rats' intragastric administration with Xuan Mai Gan Ju Particles,Cmax of liquiritin is 27.55±21.12 ng/mL?Tmax is 0.31±0.19 h?T1/2 is 3.46±2.08 h?AUC0?t is 48.82±11.35 ng·h/mL?AUC0-? is 54.17±16.31 ng·h/mL;Cmax of Harpagoside is 16.28±14.53 ng/mL?Tmax is 0.33±0.18 h?T1/2 is 3.28±1.17 h?AUC0?t is 64.85±46.70 ng·h/mL?AUC0-? is 69.82±48.2 ng·h/mL.Conclusion: The method is sensitive,simple,rapid and can be used in pharmacokinetic studies of two-active component plasma in rats after intragastric administration with Xuan Mai Gan Ju Particles.The results of pharmacokinetic studies can provide reference for clinical treatment.