Analysis On Active Substances And Evaluation On Quality Of A. Roxburghii (Wall.) Lindl

Anoectocilus roxburghii(A.roxburghii) is a precious rare materia medica inChina.The medicine value of A.roxburghii is pretty significant. In this thesis, thecontents of gastrodine and4-hydroxybenzyl alcohol in A. roxburghii were determinedby high performance liquid chromatography-diode array detector(HPLC-DAD). Highperformance liquid chromatography-electrospray ionization-mass spectrometry(HPLC/ESI/MS) method was developed for identification and determinationnucleosides in A. roxburghii. Based on the systemic chemical research of A.roxburghii,the HPLC fingerprint of it was established. Through researching thecommonly used pesticides in the whole cultivation process of A. roxburghii, themethod for determination of common pesticide residues in A.roxburghii wasdeveloped. This investigations would provide the material base for pharmacologicalactivities of A.roxburghii and reliable basis for drug quality control. The thesiscontains four chapters.Chapter one: Determination of gastrodine and4-hydroxybenzyl alcohol inA.roxburghii by HPLC-DAD.Ultrasound-microwave was applied to the extraction of A. roxburghii and theextraction conditions were optimized in this study. A method of HPLC-DAD wasdeveloped for simultaneous of identification and determination of gastrodine and4-hydroxybenzyl alcohol. The chromatographic conditions were achieved on AgilentTC-C18column (5μm,4.6mm×250mm) using gradient elution with mobile phase of0.1%acetic acid and methanol. The assay was carried out at a flow rate of1.0mL/min, at30℃,with the diode-array detection at270nm. The content of gastrodineand4-hydroxybenzyl alcohol from A.roxburghii were1.430mg/g and0.562mg/g. Theproposed method is simple, accurate, rapid and reproducible.Chapter two: Analysis on the nucleosides substances of A.roxburghii byHPLC-ESI-MS.A.roxburghii powder extracted using ultrasonic-microwave assisted extraction,High performance liquid chromatography–electrospray Ionization/ion trap massspectrometry (HPLC-ESI-MS) was developed for analysis nucleosides substances in A. roxburghii with thymine as internal standard,the chromatographic separation wasperformed on Agilent TC-C18column (5μm,4.6mm×250mm), using mobile phase of0.02mol/L ammonium acetate aqueous solution including0.05%triethylamine andmethanol (5:95,v/v). The assay was carried out at a flow rate of0.6mL/min,columntemperature was at30℃,detection wavelength:260nm. To the best of our knowledge,the important pharmacological active constituents of cytidine, hypoxanthine, guanineand adenine were firstly found to be coexisted in A.roxburghii.Chapter three: Study on HPLC fingerprint of A. roxburghii.Based on the systemic chemical research of A. roxburghii, the HPLC fingerprint ofit was established. The best chromatographic conditions were optimized including theextraction condition, the column, the mobile phase systems, detection wavelength, etc.The HPLC fingerprint of A. roxburghii was established preliminarily with goodspecificity, characteristic, stability and reproducibility.Chapter four: Determination of pesticide residues in A. roxburghii by HPLC.Through researching the commonly used pesticides in the whole cultivation processof A.roxburghii, the method for determination of common pesticide residuesincluding carbendazol, topsin-M and streptomycin mixed with oxytetracycline inA.roxburghii were developed. According to the physical and chemical properties ofcarbendazol, topsin-M and streptomycin mixed with oxytetracycline, the method fordetermination of them of pesticide residues in A.roxburghii were established byHPLC.The optimization of the extraction system, purification system andchromatographic conditions were the focus of the experiment.