Analysis Of Chemical Constituents Of Three Tibetan Medicine

Tibetan medicine has thousands of years history, clinical application extensively, is one of the most important and indivisible part of traditional Chinese medicine and commonly reconised by the majority of patients. The research of the active components from Tibetan medicine is relatively lagged behind for historical and geographical reasons. The research explore the material basis that the combined treatment is better than unilateral treatment, according to the principle of Chronopharmacology in clinical application.The three classic Tibetan medicine prescription are:Ershiwuwei shanhu pill,Wilwishfui-treaure pill and 20-Herb Chenxiang; Ershiwuwei shanhu pilll was collected in China Pharmacopeia (2015); Wilwishfui-treaure pill and 20-Herb Chenxiang were collected in the the first volume of Tibetan medicine standard by Ministry of health. In order to control the accuracy and stability of the three Tibetan medicine’s chemical composition in vivo, the study is going to control the three Tibetan medicine’s quality and explore the prototype components and metabolites in the blood circulation, so that we can further clarify the rationality of Chronopharmacology and the material basis of three priscription in curing Cerebral ischemia.The study contains two parts:The first part is to identify the effective constitue-nts of Classical Tibetan medicine and establish the determination method; The secon-d part is to track the effective component in blood. Part-1:identify the effective constituents of Classical Tibetan medicine and establish the determination method 1. To establish a method of the Gallic acid determination of three Tibetan medicine by HPLC:Global Chromatography C18 (4.6mm×250mm,5μm) column with methanol-0.2% formic acid (2:98) as the mobile phase.The detection wavelength was set at 280nm, The linear was 0.896g-4.48g;To established the determination method of Hydroxy Safflo wer Yellow A in Wilwishfui-treaurepill and 20-Herb Chenxiang:Global Chromatography Ci 8 chromatographic column with methanol acetonitrile-0.5% phosphoric acid (29:2:69) as the mobile phase.The detection wavelength was set at 403nm.The linear was 0.1286mg-0.6 432mg; To established establish a determination method of Hydroxy Safflower YellowA,Crocin I,and CrocinⅡ in Eshiwuwei shanhu pill.It was carried out Global Chromatograp y C18 chromatographic column, mobile phase A-methanol, B-acetonitrile C-0.2% for mic acid, gradient elution:0-80min,2-20%A,80-160min,20-38%A,160-200min,38-68%A,200-220min,68-75%A. The detection wavelength was set at 400nm and 440nm. 2. To establish the characteristic spectrum of three Tibetan medicine. It was carried out Global chromatography C18 chromatographic column(4.6mmx250mm,5μm). The mobile phase A was acetonitrile and B was 0.2% formic acid water.gradient elution program for 0-80min,2-20%A,80-160min,20-38%A,160-200min,38-68%A,200-220min, 68-75%A; DAD detector scanning range for 190-440nm, real-time detection wavele ngth 250nm and 400nm, the column temperature was 35℃ and and the flow rate was about 1.0 mL/min. The results the similarity of 10 batches of the three Tibetan samples average values were above 0.9, indicating that the samples of the respectie its main chemical composition similar, and sample quality and stability are in good agreement.Part-2:tracking the effective component in blood.The study is to track the effective components in blood by comparing the the characteristic spectrum of those three Tibetan medicine in vivo and vitro. The result indicates that there were 12、14、15 effective components in blood in Ershiwuwei shanhu pill; Wilwishfui-treaurepill; 20-Herb Chenxiang respectively. Ershiwuwei shanhu pill has 12 components in blood, but only 8 components are prototype, Wilwishfui-treaurep ill has 8 prototype constituents absorbed into blood,20-Herb Chenxiang has five proto type components in blood.