| Objective: To proof Chuanxiong Rhizoma(CR) and Salviae miltiorrhizae Radix et Rhizoma(SRR) of in compatibility can increase the clinical efficacy,in order to provide theoretical basis of modern medicine science for reasonable combination of tcm doctors prescribing medicine.Method: 1.To optimize the extraction technology and purification craft of CR and SRR by the orthogonal test and macroporous resin adsorption method. To study the effects of solvent quantity,extract times,extract temperature and extract time for the extraction technology of SRR and CR.To study the effects of the different kinds of resins,sample quantity,cleaning solvent and elution for the edulcoration of SRR and CR. 2. Chromatographic column is Agilent TC-C18 BDS(250x4.6mm,5um), mobile phase is acetonitrile and 0.5% acetic acid aqueous solution, gradient elution, flow rate is 1.0 m L / min; the detection wavelength was 321 nm; the column temperature is 35℃, the indexes is FA and Sal B, which was used to determinating the extraction rate of effective components in the decoctions of SRR and CR at different ratio. 3.Rats were stochastic divided into five groups, respectively, injected FA, FA with the SRR purified, the CR purified, the CR purified with the SRR purified, the SRR purified which is passed through the tail vein,draw blood from the orbital veniplex in different time. To Purify the blood samples by liquid-liquid extraction and to detect concentration of FA and Sal B in rat plasma,at last using Win Nonlin 6.2 to calculate the parameters of pharmacokinetic. 4. Rats were divided into third groups randomly, respectively, injected the purified of CR, the purified of CR with the purified of SRR, the purified of SRR which is passed through the tail vein, gatherred sanguins from the abdominal aorta and get out of heart, livers, spleen, lungs, kidney, brain, muscle.To Purify the blood samples by liquid- liquid extraction and the tissue samples by protein precipitation and liquid-liquid extraction.Determinating the concentration of Sal B, LA, RA and FA in the biological samples of rats by UPLC-MRM.Making the concentration time curve and strip distribution chart of tissue.Result:1.The optimal technical condition were extracted third times with 70% ethanol on the ratio of 8kg:1L for ferulic acid. The purification of ferulic acid with HPD300 macroporous resin was adsorbed effectively.The best proportion of the crude drug with drying macroporous resin was 5:1.Eluted the impurities with water,then washed with 50% alcohol.Collected the 50% alcohol,in which the content of FA was 9.71%.The optimal technical condition were extracted twice times with water on the ratio of 8kg:1L for ferulic acid. The purification of ferulic acid with HPD300 macroporous resin was adsorbed effectively.The best proportion of the crude drug with drying macroporous resin was 1:2. Eluted the impurities by water washed by 40% alcohol then collected the eluent, in which the level of Sal B was 73.6%. 2.The harvest of Sal B increased by the augment of SC. The ratio of SRR with CR is 1:2<3:4<1:0<2:1<3:2<1:1. 3.The methodology including linear equation, method of rate, precision, accuracy, stability and recovery was tested which show the method of UPLC was stable.The parameters of pharmacokinetic like T1/2 and AUC both had significant differences. 4.The methodology including linear equation, method of rate, precision, accuracy, stability and recovery was tested which show the method of UPLC-MRM was stable.Datas displaied the drug content in the tissues was lower than in plasma. The concentration of Sal B in rat’s tissues is kidney﹥lung﹥heart﹥liver﹥muscle﹥spleen ﹥ brain.The concentration of lithospermic acid in rat tissues from high to low is: heart﹥kidney﹥ lung﹥liver﹥muscle﹥spleen﹥ brain;The concentration of rosmarinic acid in rat tissues from high to low is: kidney﹥lung﹥ heart﹥brain﹥muscle﹥ spleen ﹥ liver; The concentration of ferulic acid in rat tissues from high to low is: kidney﹥lung﹥liver﹥heart﹥spleen﹥brain﹥ muscle.Conclusion: SRR and CR used together can significantly affect both the efficacy component remove and the distribution in the organs of rats. |
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