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Objective:To explore the macroporous resin adsorption and the membrane separation technologies for the purification of saponins water extract of Anemarrhena asphodeloides. Methods:Ten-fold amount of water was used to extract twice for 120 min each time to extract saponins from Anemarrhena asphodeloides. The macroporous resin adsorption(HP-20,HPD-600,D101,AB-8) and the membrane separation technologies (ceramic microfiltration membranes 0.8 μm and 0.05 μm, and hollow fiber ultrafiltration mem-branes 50,10 and 6 kDa) were adopted to purify the saponins water extract liquid. The physicochemical parameters including electri-cal conductivity,viscosity and turbidity were measured,as well as the contents of total saponins,proteins and polysaccharides were de-termined. Results:The viscosity and turbidity decreased,the value of pH increased and the electrical conductivity of the saponins puri-fication liquid changed irregularly after the membrane filtration. The microfiltration membrane was more advantageous than the ultrafil-tration membrane in removing macromolecular substances. The smaller the pore diameter of microfiltration membrane, the smaller the intercepted molecular weigh,the higher the removal ratio of proteins and the higher the penetration rate of the total saponins,while the polysaccharides content was stable, which was consistent with the results of physicochemical parameters. The ceramic microfiltration membrane could obtain clearer extract,while the ultrafiltration membrane was more suitable for the enrichment of saponins when the in-tercepted molecular weight was 6 kDa. The macroporous resin HPD-600 was the best for the purification of timosaponin water extraction liquid.Conclusion:The selection of membrane for the separation and purification of different substances is particularly important. The change of physicochemical parameters and the content decrease of macromolecular substances have obvious corresponding relationship. Ultrafiltration membrane is better than microfiltration membrane for the purification of timosaponin water extract liquid.
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Objective:To optimize the extraction technology and evaluate antioxidant activity of total flavonoids from Hedyotis dif-fusa. Methods:The content of kaempferol was detected by an HPLC method,and spectrophotometry was used to detect the content of total flavonoids. Results: The optimum extraction conditions were investigated by orthogonal design with the extraction quantities of kaempferol and total flavonoids as the evaluation indices. The antioxidant activities of the extracts were detected based on the clearance of hydroxyl radical and DPPH·model. The optimum extraction conditions were as follows:10-fold amount of 80% methanol and with ultrasonic extraction for 15 min. The clearance rate of 1ml methanol extract for scavenging DPPH· and ·OH was 73.89% and 91.27%,respectively.Conclusion:The optimum technology is stable and feasible for the extraction of Hedyotis diffuse. The extract shows good in vitro antioxidant properties, which provides powerful reference for the future development of relevant antioxidant prod-ucts.
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Objective: To evaluate the cold and hot characteristics of Curcumae longae Rhizoma, Curcumae Radix and Curumae Rhizoma, and study the relationship between the content of curcumin in the three herbs and the hot and cold characteristics.Methods: MTT assay was used to investigate the effect of the three herbs on the growth of SGC-7901 in vitro.Trypan-blue staining was used to analyze the cytotoxicity, and the content of curcumin was detected by HPLC.Results: Curcumae longae Rhizoma and Curumae Rhizoma could promote the proliferation of SGC-7901 at the dose of 5 μg·ml-1 and showed hot characteristics;Curcumae Radix could inhibit the growth and proliferation of SGC-7901 within the concentration range of 5-800 μg·ml-1.Trypan-blue staining showed the three herbs had no cytotoxicity on SGC-7901.The content of curcumin in Curcumae Longae Rhizoma, Curcumae Radix and Curumae Rhizoma was 4.88%, 0.15% and 0.042%, respectively.Conclusion: The content of curcumin in the three herbs affects the hot and cold characteristics, and curcumin may be the important material basis for the hot and cold characteristics.
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Objective:To explore the effect of different separation and purification technology on the physical and chemical proper-ties of water extract of ophiopogonin. Methods:The water extraction process of total ophiopogon saponins was optimized by an orthogo-nal test. The macroporous resin adsorption and membrane separation technology were adopted to purify the ophiopogonin water extract. The physical and chemical parameters, such as electrical conductivity, pH value, viscosity and turbidity, and the contents of total sap-onins, proteins, tannins and polysaccharides were determined. Results:The optimum water extraction technology of total saponins was as follows:6-fold amount of water was added, boiling 3 times with 90 min for each time. The electrical conductivity, pH value and vis-cosity of different purified liquid of total saponins had no significant differences, while the contents of total saponins and the three kinds of macromolecules showed significant differences. Conclusion: The content of macromolecules can be used as the reference index of purification process of total saponins water extract. Compared with macroporous resin separation, membrane separation technology is more suitable for the separation and purification of total saponins water extract.
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Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.
ABSTRACT
Compound composition of several natural products including plants,animals,and minerals.In the different needs of Chinese medicine research and application,different extraction and separation methods have been chosen,however,these technologies lacked of scientific and reasonable evaluation criteria because most of these technologies stem from other disciplines,which is the main reason for limited application of the method in Chinese medicine.So the key scientific issue is to establish the quality evaluation system of extraction and separation technology in line with the modern medicine.This article aims to establish the scientific principles of evaluation system for extraction and isolation of Chinese herbal compound.