Studies On Preparation And Biological Activity Of Ophiurasaponin From Ophiomastix Annulosa

In this paper, Ophiomastix annulosa were used as the experimental material. Thenutrient constituents of Ophiomastix annulosa (whole) from KongDong island inYantai were analyzed comprehensively. The optimum extraction technology ofsaponins from Ophiomastix annulosa, the physical and chemical properties ofsaponins from Ophiomastix annulosa, the separation and purification of Ophiomastixannulosa saponins were determined, and its antioxidant activity, antibacterial activity,hemolysis activity and antitumor activity were also studied.The content of water, ash, protein, total accharide and rude fat are2.55%,45%,19.04%,1.72%and2.20%, respectively. Particularly, the content of eicosapentaenoicacid (EPA) and arachidonic acid （AA）are higher, accounting for17.87%of the totalfatty acids. Essential amino acids accounted for25.39%of total amino acids. Thecontent of inorganic elements Ca is the highest. In conclusion, Ophiomastixannulosla (overall) has a good development and utilization of value with its richnutrient.The solvent extraction was applied to obtain the crude ophiurasaponin. Thenused water,20%ethanol,40%ethanol,60%ethanol,80%ethanol through the AB-8macroporous rensin to remove polysaccharides and inorganic salts. Every part elutionwas collected separately.80%ethanol Elution composition was eluted though silicagel column chromatography, TLC (thin layer chromatography) spot plate to get singlecomponent.Through physical and chemical methods to identify the property of Ophiomastixannulosa saponins, the results showed that saponins is steroidal saponins, comparedwith standard saponins, it was scanned in the range from200to800nm UVwavelength, with maximum absorption peak at216nm was obtained. Infrared scan inthe range from400-4000cm-1showed that the saponins from Ophiomastix annulosa is25R type screw sterane alcohol.Hemolysis activity was determinated, suggested that ophiurasaponin withhemolysis activity, the nature of the ophiurasaponin in eliminating blood clots, prevent cardiovascular disease is of great significance, indicates the potentialapplication value and prospect.Using circular filter paper method, the antimicrobial activity of ophiurasaponinon four kinds of microorgnisms were determined in this paper. Ophiurasaponin has acertain inhibitory effect of the selected bacteria. The results show that Escherichiacoli performanced the most sensitive to ophiurasaponin, Staphyloccocus aureusRosenbach and Saccharomyces cerevisiae Hansen characterized by moderatesensitivity, Aspergillus flavus characterized by low sensitivity. Ophiurasaponin onfour kinds of antimicrobial, with the value of inhibitory concentrations (MIC) is0.5mg/mL,1.0mg/mL,4.0mg/mL and2.0mg/mL, respectively.MTT method was used to determine the antitumor activity of the saponins fromOphiomastix annulosa, ophiurasaponin had significant inhibitory effect on tumor cellHCT-116, the MCF-7, LnCAP, HCT-15. The RM-1、Acan had inhibitory effect,but the effect was not obvious.The experimental results showed that four kinds oftumor cells HCT-116, the MCF-7, LnCAP, HCT-15had a very strong sensitivityon ophiurasaponin samples. Therefore, ophiurasaponin had antitumor effect on fourkinds of tumor cell HCT-116, the MCF-7, LnCAP, HCT-15, the result is toprovide theoretical basis for the study of ophiurasaponin from marine animal onantitumor.This paper was mainly to determined antioxidant activity: the total reducing power,hydroxyl radicals, superoxide anion. it was concluded that the ophiurasaponin hadsignificant antioxidant activity. Total reducing power of ophiurasaponin was measured,OD value was1.31. The result was that ophiurasaponin had antioxidant capacity.According to the scavenging effect on oxygen free radical, the results concluded that theophiurasaponin have the scavenging effect on superoxide anion and hydroxyl freeradicals.