| This dissertation focuses on Ziziphi Spinosae Semen and Polygalae Radix with preparation of oligosaccharide esters from Polygalae Radix, preparation of flavonoids and saponins from Ziziphi Spinosae Semen by modern extraction, isolation and purification methods. And the hypnotic effect of effective parts and compatibility of medicines were studied to provide a scientific basis for further development and utilization of Ziziphi Spinosae Semen and Polygalae Radix. The content includes the following parts.1. Study on extraction technology of oligosaccharide esters from Polygalae RadixUsing DISS as the detection index, with the application of cyclic column chromatographic extraction method, extraction technology of oligosaccharide esters from Polygalae Radix was studied. The optimum extraction technological conditions were as follows:20-40 mesh powder of Polygalae Radix was loaded onto glass column and soaked by 70% ethanol (2-fold) for 1 h, then eluted at the speed of 1 BV/h. The first fraction (2-fold) was collected as the final extracts, and the second fraction (2-fold) and third fraction (2-fold) were used to extract the next column. With this method, the extraction rate of DISS was 96.18%, much higher than the rates of ultrasonic extraction (73.58%) and heat reflux extraction (85.46%).2. Study on extraction technology of flavonoids and saponins from Ziziphi Spinosae SemenExtraction technology of flavonoids and saponins from Ziziphi Spinosae Semen was studied applying the cyclic column chromatographic extraction method. Spinosin, jujuboside A, jujuboside B were selected as detection indexes. The optimum extraction technological conditions were as follows:20-40 mesh powder of Ziziphi Spinosae Semen was loaded onto glass column and soaked by 50% ethanol (2.5-fold) for 1 h, then eluted at the speed of 1 BV/h. The first fraction (4-fold) was collected as the final extracts, and the second fraction (2.5-fold) and third fraction (1.5-fold) were used to extract the next column. The extraction rate of spinosin, jujuboside A, jujuboside B were 98.93%,97.06% and 92.71% respectively. Compared with ultrasonic extraction and heat reflux extraction, this method has the advantages of simpler equipment requirements, less solvent usage, higher efficiency and less energy consumption.3. Study on separation and purification technologies of flavonoids and saponins from Ziziphi Spinosae SemenMacroporous resin DM130 was used for the separation and purification of flavonoids and saponins from Ziziphi Spinosae Semen. The optimum technological conditions were as follows:5 BV of solution with crude drug concentration of approximately 1 g/mL was added into resin column at the speed of 2 BV/h. After using 8 BV of water to wash the resin column at the speed of 2 BV/h, apply 40% ethanol to elute 5 BV at the speed of 1 BV/h and then 70% ethanol to elute 4 BV at the speed of 1 BV/h. The 70% ethanol elution would be concentrated and dried to get saponins extract. The 40% ethanol elution would be concentrated up to dryness and dissolved in 0.2-fold water, then liquid-liquid extracted with equivalent water-saturated butanol for five times. And the combined extractions would be decompressed, concentrated and dried to get flavonoids extract. Under these conditions, flavonoids extract yield was 0.43% with flavonoids content of 59.86%, and the saponins extract yield was 0.16% with saponins content of 60.53%.4. Study on hypnotic effect of effective parts and compatibility of medicinesThe hypnotic effect of the following three components (flavonoids of Ziziphi Spinosae Semen, saponins of Ziziphi Spinosae Semen, oligosaccharide esters of Polygalae Radix) and their compatibilities was studied by the test of pentobarbital sodium above threshold dose with active components on hypnotic activities on mice. Uniform design method combined with optimal subset regression was applied. The results indicated that flavonoids of Ziziphi Spinosae Semen, saponins of Ziziphi Spinosae Semen were suitable for compatibility while oligosaccharide esters of Polygalae Radix not. The results of the following verification test were as follows:the sleeping time of flavonoids group, saponins group and compatibility group were 45.86 min,54.57 min and 54.14 min respectively which were significantly different from the blank group (31.00 min). Thus it indicated that all of the three groups had hypnotic effect. While compatibility of flavonoids and saponins of Ziziphi Spinosae Semen had no significant effect on sleeping time. It indicated that flavonoids and saponins of Ziziphi Spinosae Semen had additive effect. |
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