| It was studied that the process of effective composition from Celastrus aculeatus Merr with the methods of medicinal chemistry of natural products. The results were sumaried as follows:A new method was established for the determination of total flavonoids and total triterpenes from Celastrus aculeatus Merr;the oxidation resistant effect of total flavonoid from Celastrus aculeatus Merr was measured;The extraction technology of total triterpenes from Celastrus aculeatus Merr;Screen the effective composition from different polatity parts of Celastrus aculeatus Merr.1.Extraction of total flavonoids:Weigh 20g powder of Celastrus aculeatus Merr, dissolve the sample in 70% alcohol according to the ratio of powder/liquid as 1:10, extract refluently for 2h, and then filter. The filter residue was extracted twice for 1h each time according to the above method. The mixture of filtrate was settled to a certain volume with 95%alcohol.2. Determination of total flavonoids:The content of total flavonoids in Celastrus aculeatus Merr was determined by ultraviolet spectrophotometry with rutin as standard control. The calibration curve was linear (r=0.9995) in the range of 20-60μg/ml and the curve was y=0.0128x - 0.0266. The method is simple, rapid and accurate. It can be used for determining the content of total flavonoids of Celastrus aculeatus Merr.3. Studies on the oxidation resistant of extraction of total flavonoids from Celastrus aculeatus Merr:The results showed that the general falvone from celastrus aculeatus Merr exhibited a powerful scavenging effect on those three kind of radicals. In addition, the scavenging effects on them had certain dose dependent relationship with flavonoids concentration at the selected concentration. When the concentration of flavonoids reached 18.15μg/ml,11μg/ml,18.15μg/ml,the scavenging rate on DPPH-and ABTS+ and-OH was 87.43%,85.84%,92.56%,respectively, which may be outweigh vitamin C of the same concentration. The general falvone from celastrus aculeatus Merr has strong antioxidative ability.4. Determination of total triterpenes:Total triterpenes was measured by spectrophotometer with 5% vanillin-glacial acetic acid solution and perchloric acid as color-developing agent and detected at 554 nm wavelength with Oleanolic acid as standard control. The experiment showed that the calibration curve was linear (r=0.9988) in the range of 21-74ug/ml and the curve was Y=56.269x-0.0376.The method is simple, rapid and accurate. It can be used for determining the content of total triterpenes of Celastrus aculeatus Merr.5. The extraction and purification technology of total triterpenes from Celastrus aculeatus Merr:The optimum conditions for extraction of total triterpenes from Celastrus aculeatus Merr were:ethanol concentration,95%; solid to liquid ratio,1:1 2; temperature,40℃; time,2h; frequency,3; method, reflux extraction. Under optiml conditions, the extraction ratio of total was 3.8%.According to the adsorption capacity and elution ratio of the resin, three types of macroreticular resin were used to refne the triterpenes from Celastrus aculeatus Merr. The results indicated AB-8 type maeroreticular resin were the best on the adsorption capacity and elution ratio. By the determination, adsorption ratio Was 2 ml/min, pH=6.0, solvent of elution was 40%ethanl. The AB-8 type of macroreticular resin showed better performance for repeated use on being used to refine the triterpenes from Celastrus aculeatus Merr. The result showed that the purity of total triterpenes reached 78.39%, far higher than lavonoid by macroreticular resin refined.6.GSF-A,GSF-B,GSF-C and MTX can significantly inhibit mouse splenic lymphocyte proliferation, and compared with ConA group, their inhibition ratio is significantly different(P<0.05). Among all, GSF-A shows the best inhibition effects. |
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