Studies On Using Substitute Reference Substance In Quality Control Of Rhubarb And Rhubarb Preparations

This thesis includes five major parts, the identification and quality evaluation ofrhubarb species by HPLC fingerprint and chemometrics; the identification ofchromatography peaks and purity determination of reference substance; preparation ofrhubarb reference extract; methodology validation of determination method ofmulti-target compounds in rhubarb crude drugs, preparation and extract; developmentand application of substitute reference substance.Part Ⅰ is identification and quality evaluation of rhubarb species by HPLCfingerprint and chemometrics. After collecting6different species of rhubarb crude drugsand decoction pieces, HPLC fingerprint of rhubarb had been established. Compared theHPLC fingerprints of rhubarb and its decoction pieces at254nm and430nm werecompared and analyzed with CHROMAP software. By using chemometrics analysis,official and unofficial rhubarb species can be identified. It had been proved that thisHPLC fingerprint and chemometrics analysis can be used to distinguish official andunofficial rhubarb. This provide a good reference for quality evaluation of rhubarb andrevision of the quality control standards.Part Ⅱ is the identification of chromatography peaks and purity determination ofreference substance. By using the modern chromatographic techniques, the maincomponents of the chromatographic peaks were analysed, and then purchased chemicalreference standard to verify the results and got accurate identification results at last. Allstructures of the chemical reference standard have been confirmed by NMR; as far as thepurity analysis, did determination of purity by HPLC. These all made good preparationsfor establishing next step of assay methods.Part Ⅲ is rhubarb reference extract preparation. By using macroporous adsorptiveresin, rhubarb extract was purified, which contains combined anthraquinones and4-(4′-hydroxyphenyl)-2-butanone-4′-O-β-D-(2′′-O-galloyl-6″-O-cinnamoyl)-glucopyran-oside, then got free anthraquinones by acidolysis. rhubarb extract and acidolysis productswas mixed and rhubarb reference extract was gotten at last.Part Ⅳ is methodology validation of determination method of multi-compounds inrhubarb crude drugs、preparation and extract as well as standardization of rhubarbreference extract. Validated the established method of content determination and thefeasibility of application in Rhubarb crude drugs、preparation and extract. Rhubarbreference extract had been standardized finally. Part ⅴ is development and application of substitute reference substance. By usingrhubarb reference extract, we established the assay method of rhubarb, validatedruggedness of the method and apply this method into five collected crude drugs to provethe feasibility of this method. Established the method of two reference substances forsimultaneous determination of multiple components, relative correction factor is used tocalculate the content of8compounds in rhubarb. Through the retention time ofrhein-8-O-glucopyranoside and aloe-emodin, the retention time of the other compoundscan be predicted. We also validated ruggedness of this method and apply it into crudedrugs to prove the feasibility of this method.