| Objective: To establish the qualitative chemical fingerprint of Blumeariparia(BI.) DC. by HPLC. In order to systematic and scientific evaluate theherb and its related product’s efficacy. Moreover, this can be one more step forconfirmation the herb and its related product’s safety for people and also in-crease nationwide people’s welfare.Methods: Series and systematic analyzation by High performance liquidchromatography(HPLC) method with herb samples from difference places dis-tributed in Guangxi province Baise Tianlin and Linyun county of China. Theseherbs are extracted using particular extraction method. Above experiment resultin the proper chromatographic condition. The condition composed of the mostproper mobile phase system, the most proper time period required for recordedeach complete chromatogram, the optimal detecting wavelength, optimal HPLCcolumn, optimum column temperature and proper flow rate. The chromatogra-phic condition mension as followed, Phenomenex synergi4u polar-RP80A(250mm×4.6mm，4μm) column was used under25degree celsius columntemperature,256nm detecting wavelength,20μL HPLC auto-sampler samplesize and recorded60min chromatogram with gradient elution at1ml/min flow rate. The gradient elution was performed by mobile phase system consisted ofmethanol (phase A) and pH2.8solution of acetic acid in water (phase B) thenthe gradient elution program was initially set at10%of methanol (phase A) thengradually increased ratio of phase A up to40%in17min, followed by contin-uously increased ratio of phase A up to60%by37min. After that, method vali-dation and related evaluation were conducted rely on similarity evaluation sys-tem for chromatographic fingerprint of traditional Chinese medicine (version2004A) which was published by Chinese pharmacopoeia com mission. Parame-ters in valuation criteria were within acceptable range then we were establishedthe chemical fingerprint method that not only concise but also practicable andreasonable for Blumea riparia(BI.) DC. and its related products.Results: Fingerprint chromatogram was obtained by12different batchesof glutinous extractions of Blumea riparia(BI.) DC. and11common peaks wereconfirmed in the fingerprint chromatogram. The common peak included peaksof proto-catechuic acid and chlorogenic acid which are two of important watersoluble substances of this herb and these two common peaks had been used asthe index compositions in method development process. The similarity evalua-tion system for chromatographic fingerprint of traditional Chinese medicine (ve-rsion2004A) which was published by Chinese pharmacopoeia commission thatwas elected calculated those chromatogram’s similarity. Calculated chromatogr-aphic condition parameter were acceptable, related parameter mensioned as fol-lowed. Average similarity of precision test and test of reproducibility weregreater than0.99(average similarity of protocatechuic acid and chlorogenic acidin precision test both are equal to1.0000, average similarity in test of reproduci-bility was equal to1.0000too). Parameters of test of stability including overall similarity that was greater than0.98(the Overall similarity within5hours wasequal to1.0000), relative retention time deviation were less than3%(0.04%)and relative peak ratio of significant peak (relative peak area above5%) alsoless than3%.Conclusions: Reasonable and systematic experimental design weregradually practice one’s scientific though and result in novel reliable quanti-tative chemical fingerprint analysis method of Blumea riparia(BI.) DC. Photodiode array detector that able to create3-dimensional chromatogram was alsoused for confirmation the accuracy of the method. This could be a good found-ation for quality assurance at industry level and would be an important step forevaluated safety and efficacy of the herb and it’s related products. For assuredpeople’s health and others further application of Blumea riparia(BI.) DC. |
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