Study On HPLC Fingerprint Of Traditional Chinese Medicine Extracts

Establishing the optimal chromatogram conditions was the foundation of establishing HPLC chromatogram fingerprint. First, chromatographic column was considered and C18 reversed phase column was selected commonly. For alkaloids, ion exchange column was selected, and for amylose, gelcolumn was selected. Secondly, type of elution was considered and type of isocratic elution was attempted firstly. When type of isocratic elution was lapsed, type of gradient elution was elected again. There were some definite demands on reproductivity of gradient elution, it could be controlled effectively with some methods e.g. fixing apparatus (low pressure mix or high pressure mix) and chromatographic column, simplifying impregnant system and gradient conditions, fixing temperature. And operating procedure could be standardized with system adaptability experiment and methodology reviewing. Thirdly, probe unit was considered. When chromatogram fingerprint was established with HPLC methods, material methods could be elected according to material substances. These methods consisted of ultraviolet (UV), diode array detector (DAD), scattering of vaporizing light or refraction and so on. Some tasks could be unfolded with DAD, such as optimally selecting measured wavelength, measuring purity of peak and tracing to the source of chemical compositionAfter overall evaluating systematically distilling technics with orthogonal designing (indexes were puerarin and Pueraria Flavonid) and statistical analyzing test results, optimum processing condition was confirmed finally, namely, distilling 3 times with 9 times quantity ethanol (30%) and 1 hour every time. Thus, utilization ratio (extraction rate) of material was increased and flavone was distilled entirely, it was propitious to the research on chromatogram fingerprint. Through the systematical study on various factors of influencing macroporous resin adsorption and desorbing Pueraria Flavonid, the optimal technics of macroporous resin separating was confirmed finally, namely, type of resin was SP70 macroporous resin produced by Mitsubishi Corporation, liquor strength was 0.5g/mL (be equivalent to protosomatic medicine), pH of liquor was 5~6, rate of adsorption was 2BV/h and upper prop quantity was sixtyfold to resin bed volume. The optimal technics of purging was volume 5 times to resin bed volume, concentration of ethanol was 70% and velocity was 2BV/h. The content of Pueraria Flavonid disposed with SP70 could reach 80% upwards. This liquor or extractive material made up could...