| Allium mongolicum regel is a member of the Liliaceous family, members of which live in desert grassland and arid climates. Its distribution is mainly concentrated in Mongolia, Siberia, and China. Allium mongolicum regel has been shown to have a variety of nutrients for human consumption in addition to having a pleasant taste. According to the Mongolian Pharmacopoeia, Allium mongolicum regel has many important physiological effects, including anti-hypertensive, Hypolipidemic and anti-inflammatory. Thus, development of an effective means to extract and separate the essential components from Allium mongolicum regel has recently been started. Currently, a large number of researchers have focused on the nutritional and biological efficacy of Allium mongolicum regel, as well as its anti-bacterial effects、 anti-oxidant effects%anti-viral and anti-neoplastic effects. Thus, the objective of this study was to use response surface methodology (RSM) to optimize the Allium mongolicum regel flavonoids (AMF) extraction process. We then sought to investigate preliminary anti-oxidant and anti-bacterial abilities of AMF, thereby providing initial data for future studies.(1) The objective of this study was to employ response surface methodology to study the effects of extraction temperature, ultrasonic power, extraction time, solid-liquid ratio, ethanol concentration on total flavonoids yield and to determine the optimum parameters of ultrasonic assisted extract method. And extracting time, extracting temperature, reaction system pH and cellulose enzyme dosage was studied on single factor test, and parameters of cellulase-assisted extraction method was optimized. The best extraction processing parameter of ultrasonic assisted extract method is:15min,40℃,75%alcohol, the ratio of material to liquid is1:30. The projection of the yield of total flavonoid is12.9mg/g. The best process parameters of cellulase-assisted extraction of flavonoids from allium mongolicum regel was: extracting time4hours, extracting temperature39.5℃, reaction system pH4.3, on this condition, extraction efficiency of flavonoids from allium mongolicum regel could achieve to5.5mg/g.(2) The result of the anti-oxidant effect showed that the allium mongolicum regel total flavonoids have good antioxidant effect. Compared to tow extraction method, ultrasonic extraction method AMF antioxidant ability was better than enzymatic method AMF. The result of the antibacterial test was showed that, AMF has good antibacterial effects of salmonella, next was E.coli and staphylococcus aureus, has no significant antibacterial effects to pseudomonas aeruginosa. Compared to tow extraction method, ultrasonic extraction method AMF antioxidant ability is better than enzymatic method AMF. And its immunoregulatory activities test results showed that AMF could increase the rate of lymphocyte proliferation and influence the IL-2,IL-4and INF-ymRNA expression quantity significantly. We could know that different extracting method have significant effects of AMF. Combined with the antioxidant, antibacterial and immunoregulatory test result, they showed that ultrasonic assisted was a relatively novel extracting approach.(3) Used the polyamide column chromatography to separate and purificat the AMF. Used the liquid chromatography tandem mass spectrometry, infrared spectrum detection and ultraviolet spectrum detection to analyze the structure of the different elution group. Analyzes results showed that there were water elution group contained saccharides and naphthenic hydrocarbon.35%elution group contained3’,4’-epoxy group-7-O-5-methoxy flavonols and7-O-5,4’-dimethoxy-3’-hydroxide radical flavone.55%ethanol elution group contained rutin, quercitrin, saccharides and mignonette element-5’-O-glucose-4-hydroxy benzoic acid.75%ethanol elution group has saccharrides and acacia.(4) The result of the anti-oxidant effect showed that the35%ethanol elution group had good antioxidant effect. Compared to other elution groups, it’s total reducing power, clearance of DPPH and OH-was most excellent. The result of the antibacterial test was showed that, all of the different elution group had antibacterial effects of salmonella, next is E.coli and staphylococcus aureus, and has no significant antibacterial effects to pseudomonas aeruginosa. But55%ethanol elution group was best of all. And the immunoregulatory activities test results showed that all group of different elution parts could increase the rate of lymphocyte proliferation and influence the IL-2, IL-4and INF-γmRNA expression quantity, but35%ethanol elution group immunoregulatory activitie was better than other group significantly. Through further study we found that35%ethanol elution group had strong ability of antioxidant and immunoregulatory effect, the reason was that its’mainly composition was3’,4’-epoxy group-7-O-5-methoxy flavonols and7-O-5,4’-dimethoxy-3’-hydroxide radical flavone, they both have antioxidant structure and immunoregulatory structure(C2=C3).While the antibacterial part of allium mongolicum regel total flavonoids is55%ethanol elution group, the reason is in the molecular structure of its, there are so many C4=O, C5-OH and C7-OH structure, but also connect with many kinds of monosaccharide, it could destruct the bacterial cell walls, and inhibit the bacteria growth. |
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