Studies On Determination Of Multi-contents Of Oligosaccharide Eesters And Preparation Process Of Polygalae Radix Effective Fractions

Polygalae Radix the dried root of Polygala tenuifolia Willd. or Polygala sibirica L., is used as expectorant, tonic, sedative and preventing dementia agents. Polygalae Radix is enriched with triterpenoid sapionins, oligosaccharide esters, xanthones, ect. Sucrose esters has been reported to be significant in anti-dementia and antidepressant.This dissertation focused on sucrose esters, determination method of multi-contents of oligosaccharide esters was establish, extraction and purification parameters were studied systematically. The dissertation consists the following5parts.1. Establish HPLC analysis method of six sucrose esters in Polygalae RadixEstablished HPLC analysis method of Sibiricose A5, Sibiricose A6, Tenuifoliside B, DISS, Tenuifoliside A, Tenuifoliside C, Methodological study showed the method was specificity, sensitivity, had good precision and accuracy.18batches Polygalae Radix of different areas were determined. It was confirmed that the contents of Sibiricose A5, Sibiricose A6, Tenuifoliside B, DISS, Tenuifoliside A, Tenuifoliside C were in the range of0.08%-0.30%,0.03%-0.21%,0.02%-0.27%,0.18%-1.16%,0.06%-0.60%,0.04%-0.20%.2. Simultaneous determination of six oligosaccharide esters from Polygalae Radix by QAMSDISS was used as the internal reference substance, relative correlationfactors (RCF) of Sibiricose A5, Sibiricose A6, Tenuifoliside B, Tenuifoliside A, Tenuifoliside C to DISS were1.23,1.16,1.56,1.79,0.92. In different experimental conditions RCF had good reproducibility. The method was evaluated by comparison of external standard method and QAMS, there were no significant differences. 3. Studies on extraction parameters of preparation technology of effective fractionsSingle-factor experiment and L9(34) orthogonal test were performed to optimize the extracting process. Using DISS as the index, the relation between the factors and diversion ratio was:extraction times> ethanol concentration> extraction time> ethanol quantity, The optimal extraction process was extracted the drug by70%(V/V) ethanol twice,1.5hours each time, and the quantity of solvent was8times. Optimum conditions were stable and feasible.4. Studies on purification parameters of the preparation technology of effective fractionsTooking the content of Tenuifoliside B, DISS,Tenuifoliside A,Tenuifoliside C as the index of evaluation, ethanol extracting-water precipitating, macroporous resin purification, solvent extraction technology parameters were studied completely and systematically. The optimum method was as followe:ethanol extracts should be concentrated to4times of the raw materials, refrigerated24hours at4℃, centrifugal in the speed of3000r·min-1and collect the supernatant, the solution was added5BV (BV, bed volume) into resin column in the speed of3BV/h. Then5BV of water was added to wash away impurities in the speed of4BV/h.20%ethanol eluted5BV in2BV/h,35%ethanol eluted5BV in2BV/h, the35%fraction was evaporated in vacuum to6mg·mL-1, extracting with LE06, phase ratio is1:3, extraction three times, collect the layer of LE06, evaporated to dry in vacuum, get the effective fractions. The optimized. purification process was proved stable and feasible, can be used to production.5. Establish HPLC analysis method of six sucrose esters in effective fractionsEstablished HPLC analysis method of Tenuifoliside B, DISS, Tenuifoliside A, Tenuifoliside C, Methodological study showed that the method was specificity, sensitivity.3batches of effective fractions were determined, the average content of oligosaccharide esters was64.84%, meet the requirements of effective parts.