The Study On Active Ingredients Analysis Of A. Roxburghii (Wall.) Lindl And Quality Standard

In this thesis, Analytical method of the chemical compositions in Anoectocilusroxburghii(A.roxburghii) were investigated. The quality standards and the fingerprint of theA.roxburghii were established. This investgations would provide the basis for drug qualitycontrol. The thesis contain four chapters.Chapter one: Use high performance liquid chromatography–electrospray Ionization/ion trapmass spectrometry (HPLC-ESI-MS)to qualitative and quantitative analysis the nicotinic acidand the Nicotinyl alcohol which extracted from Anoectocilus roxburghii herbs, Thechromatographic separation was achieved on Agilent TC-C18column (5μm,4.6mm×250mm)using methanol:0.05mol/L the ammonium acetate-0.2%acetic acid aqueous solution (5:95,v/v) as the mobile phase at a flow rate of0.6mL/min,254nm as the detection wavelength. Theproposed method is simple, accurate and rapid. To the best of our knowledge, this is firstreported that Nicotinyl alcohol and Nicotinic acid coexisted in A.roxburghii.Chapter two: The important pharmacological active constituents of guanosine and adenosinewere firstly found to be coexisted in A.roxburghii.Guanosine and adenosine were extractedfrom A.roxburghii by ultrasonic-microwave assisted extraction(UMAE). A method ofHPLC-ESI-MS was developed for simultaneous of identification and determination ofguanosine and adenosine. With uridine as internal standard, the chromatographic separationwas achieved on Agilent TC-C18column (5μm,4.6mm×250mm) using acetonitrile-0.1mol/Lammonium acetate (5:95, v/v) as the mobile phase at a flow rate of1.0mL/min,254nm asthe detection wavelength and30℃as the column temperature.The analyte showed good linearregression relationship between relative peak area and concentration in the range of10μg/mL~100μg/mL (r=0.9996). The recovery rate was93.6%and94.3%. measured theaverage content of guanosine and adenosine from Guangxi A.roxburghii were0.1698mg/g and0.3202mg/g.Chapter three: Establishes the quality standard of A.roxburghii, provide the basis for drugquality control. Through the character identification, microscopic identification; Determinationmaterial moisture, ash and the heavy metal content from different origin of A.roxburghii; usingHPLC/ESI-MS to analysis content of guanosine and adenosine from different origin of A.roxburghii. According to "Chinese Pharmacopoeia"(2010), heavy metal lead from six batchof samples from different origin of A.roxburghii did not exceed the standard, but the heavymetal cadmium of different areas are all exceed the standard except Yunnan and Fujian Dehua.The average content of guanosine and adenosine from Guangxi A.roxburghii were0.1698mg/g and0.3202mg/g. The content of guanosine and adenosine from six batch of samples fromdifferent origin of A.roxburghii were0.138mg/g to0.227mg/g,0.261mg/g to0.374mg/g.This study established a reliable standards for quality control of Anoectochilus roxburghii.Chapter four: Establishing HPLC fingerprint to control technics’ and quality’ stability ofA.roxburghii, the method proved practical and feasible;as an indicator of total peak number,relative peak area and similarity to evaluation the quality of6batches of samples. The HPLCfingerprint proved can be used for the quality control of A.roxburghii.