| Total saponin of panax japonicus is the main active ingredient of Rhizoma’s Panax japonicus. This study is on the extraction and purification of Total saponin of artificial panax japonicus and its antioxidation function. Extraction by microwave is recognized as the optimal extraction process through the optimization of the process condition among three methords including reflux extraction, ultrasound extraction and microwave extraction.The process condition for purification of Total saponin of panax japonicus by macroporous resin D101 was btained; the panaxsaponin Rgl was seperated by HSCCC;with comparison between the free radicle scavenging ability of Total saponin of panax japonicus before and after purification.1. The optimization of process condition for Total saponin of panax japonicus extracted by refluxThe optimal process condition that extraction time 120min, three times extraction, dosage liquor ration 1:12, ethanol concentration 70% was obtained through orthogonal experiment of four factors and three levels based on single factors including extraction time, extraction times, dosage liquor ration, ethanol concentration. The order for the effect of single factor on the yield of total saponin of panax japonicus is extraction times> extraction time> dosage liquor ration> ethanol concentration.2. Optimization of Extraction Conditions for Total Saponin of Panax Japonicus by UltrasoundThe optimal process condition that dosage liquor ration 1:20, extraction temperature 50℃, ultrasound power 32KHz, three times extraction, extraction time 30min, ethanol concentration 70% were obtained through orthogonal experiment of four factors and three levels based on single factors including extraction time, extraction temperature, dosage liquor ration, ultrasound power. The order for the effect of single factor on the yield of total saponin of panax japonicus is ultrasound power> dosage liquor ration> extraction time> extraction temperature. The effect of ultrasound power on the yield of total saponin of panax japonicus is the most significant.3. The optimization of process condition for Total saponin of panax japonicus extracted by microwave.Response experiment of three factors and three levels (extraction time, extraction temperature, dosage liquor ration) was designed through SAS Data processing software based on single factor using the yield of total saponin of panax japonicus as response value. The optimal process condition for Total saponin of panax japonicus extracted by microwave by response surface analysis is extraction temperature 65℃, dosage liquor ration 1:13, extraction time is 4.2min, three times extraction. The yield of total saponin of panax japonicus is 24.19%.After comparing three extraction processes, extraction by microwave is the optimal process for the extraction of total saponin of panax japonicus, and it has advantage including high yield, lower cost, and high efficiency.4. Study on purification process of total saponin of panax japonicus with macroporous resin D101The process condition for purification of Total saponin of panax japonicus by macroporous resin D101 was obtained by uniform experiment design, which is 50% ethanol for elution, eluting agent dosage 17.11BV, eluting flow 1.25BV/h, sample volume 1.04BV.the expected maximum purity of total saponin of panax japonicus is 91%±7.18%.The optimal expand agents obtained in the experiment of the determination of total saponin of panax japonicus by TLC is chloroform-methanol-water (7:3:0.5 v/v). After comparing the Rf of panaxsaponin with the retention time in HPLC, the seperation effect of trichloromethane-methanol-water system has been verified.5. Separation of ginsengside Rgl from total saponin of panax japonicus by high-speed counter-current chromatographyRgl is seperated from saponin of panax japonicus purified by macroporous resin D101 using HSCCC technique. The retention time for Rgl in trichloromethane-methanol-water system is (170±10) min.Chloroform-methanol-water (7:3:1 v/v) system is the best solvent system comparing to acetidin-n-butanol-water system. After comparing to the traditional silica gel column chromatography, HSCCC is simple, easy to analyse and lower cost for seperation.6. Research the antioxidation function of total saponin of panax japonicusMaking comparison between the free radicle scavenging of the total saponin of panax japonicus before and after purification, saponin of panax japonicus is of certain antioxidation function.As for different free radicle, scavenging ability of saponin of panax japonicus is different. The scavenging effect for superoxide free radical is obvious, and the purified is obviously better.And the scavenging effect on DPPH for the purified increased with the concentration of saponin of panax japonicus, and before purification it decreased with the concentration. The scavenging effect of the total saponin of panax japonicus that on hydroxyl radical before purification is obvious, and it increased with the concentration of the total saponin of panax japonicus, and after purification, the scavenging effect is not obvious. |
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