Study On The Substance Basis Of XiaoErMaTengDingChuanFang

The chemical compositions of traditional chinese medicince (TCM) com-pound prescription are very complex, to reveal the substance basis of traditio-nal chinese medicince compound prescription is the key to modernize the TC-Ms.It is very importance for the development of the TCM science and for constructing scientific quality control standards of TCMs, which would prop-el the internationalization of TCM industry.The "LC-MS/MS" technique is a powerfuil tool for the study of the chemical and efficacy substance of traditio-nal chinese medicince compound prescription.The advantages of the technique involving high efficency, rapidness, low request on sample quantity, and so on, enable it to be well apllied for simultaneous qualitative and quantitative analysis of cmplex TCMs formulae. In the thesis," LC-MS/MS " based method of study on the substance basis of XiaoErMaTengDingChuanFang. The main contents and productions are as followed.PartⅠ:Determination of the active Anti-asthmatic extracts of traditional chinese materials in XiaoErMaTengDingChuanFang1:Determination of the active Anti-asthmatic extracts of Semen Armeniacae Amarum and Identification of the avtive componentsObjective:To study bioactivity of different extracts of Semen Armeniacae Amarum and identify the avtive components.Methods:To evaluate antiasthmatic effect on asthma latency of guinea pigs induced by the mixed solution of acetylcholine and histamine. The "LC-MS/MS" technique is applied for identification of the active components. Results:The butanol extracts significantly prolonged asthma latency induced by the mixed liquor of acetylcholine and histamine. Conclusion:The butanol extracts is the most effective extracts from Semen Armeniacae Amarum on the Anti-asthma.2:Determination of the main active Anti-asthmatic Fractions from Column Chromatography of Extract of EarthwormObjective:To study bioactivity of fractions from column chromatography of extract of earthworm on asthma. Methods:To evaluate antiasthmatic effect on asthma latency of guinea pigs induced by the mixed solution of acetylcholine and histamine. Results:The 5-15% methanol/ethyl acetate eluant from column chromatography significantly prolonged asthma latency induced by the mixed liquor of acetylcholine and histamine. Conclusion:The 5-15% methanol/ethyl acetate eluant from column chromatography is the most effective fraction of extracts of earthworm on the Anti-asthma.3:Determination of the active Anti-asthmatic extracts of Ramulus uncariae cum uncis, Bombyx batryticatus and Radix Et rhizuma nardostachyosObjective:To study bioactivity of their different extracts. Methods:To evaluate antiasthmatic effect on asthma latency of guinea pigs induced by the mixed solution of acetylcholine and histamine. Results:The ethyl acetate extracts significantly prolonged asthma latency induced by the mixed liquor of acetylcholine and histamine. Conclusion:The ethyl acetate extracts is the most effective extracts on the Anti-asthma.Part II:Study on the Water Extraction and Alcohol Precipitation of XiaoErMaTengDingChuanFangObjective:To optimize the water extraction and alcohol precipitation conditions of XiaoErMaTengDingChuanFang. Methods:An extraction procedure was optimized by orthogonal test. The content of Ephedrine Hydro-chloride which was used as the assessment index was determined by HPLC. To evaluate antiasthmatic effect on asthma latency of guinea pigs induced by the mixed solution of acetylcholine and histamine. Results:The optimum water extraction process was as follows:water as extracting solvent,1.5 h each time,10 times solvent to the decoction pieces.The optimum alcohol precipitation process was as follows:60% of precipitation concentration.the alcohol deposit lasted 24 h. The extracts in the optimum process conditions significantly prolonged asthma latency induced by the mixed liquor of acetylcholine and histamine. Conclusion:The optimum process is stable and feasible.PartⅢ:Determination of the main active Anti-asthmatic Fractions from macroporous resin absorption of extracts in the optimum process conditionsObjective:To study bioactivity of fractions from macroporous resin absorption of extracts in the optimum process conditions on asthma. Methods: To evaluate antiasthmatic effect on asthma latency of guinea pigs induced by the mixed solution of acetylcholine and histamine. The content of Ephedrine Hydrochloride in the active Anti-asthmatic Fractions was determined by HPLC. Results:The 70 and 95% ethanol/water eluant from macroporous resin absorption significantly prolonged asthma latency induced by the mixed liquor of acetylcholine and histamine. Conclusion:The 70 and 95% ethanol/ water eluant from macroporous resin absorption is the most effective fraction of extracts of earthworm on the Anti-asthma.Part IV:Identification of chemical compositions in Group of effective compounds from XiaoErMaTengDingChuanFangObjective:To study chemical components in Group of effective compounds.Methods:The "LC-MS/MS" technique is applied for identification of the act-ive components. Results:refering to relevant literature, seventeen compounds were identified.Conclusion:the results reflect the efficacy substance basis of XiaoErMaTengDingChuanFang.Part V:Determination of Ephedrine Hydrochloride in Group of effective compounds by HPLCObjective:To establish the method for detemining the content of ephedrine hydrochloride in Group of effective compounds by HPLC. Methods:Aglient TC-C18 Column was used with acetonitrile-0.1% phosphoric solution(4:96) as the mobile phase, the detection wavelength At 210 nm. and flow rate was 1.0 mL/min. Results:The calibration curve was linear at the range of 0.32-0.74μg for ephedrine hydrochloride and finear equation was Y= 1.2738X+ 0.0539, r=0.9999. The average recovery was 96.4% and RSD was 1.9% (n=6). Conclusion:This method was simple,accurate and proper,with good reproducibility. It can be used for quality control of Group of effective compounds from XiaoErMaTengDingChuanFang.