Preparation Of The Standard Components And The Study Of The Material Basis Of Phascolosoma Esulenta

As our country’s famous snack, Phascolosoma esculenta belongs to belonging tothe Genus Sipuncula and phylum Sipuncula, also known as mud garlic, soilnailing, sea cordyceps, also is recorded in much China Pharmacopoeia. Phascolosomaesculenta has the pharmacological activity of antioxidant, antifatigue, high-temperature resistance and physiological function regulation. Thispaper adopted HPLC technology to characterize the different constituents ofPhascolosoma esculenta. Established the HPLC fingerprint of every constituentsthrough optimizing the HPLC conditions. We studied the anticoagulant andantioxidant activity of Phascolosoma esculenta, and established its functionalcomponent libraries. It provided a reliable method for the quality evaluation ofPhascolosoma esculenta, provided the scientific basis for Phascolosomaesculenta becoming drug, and also laid a solid foundation for the further study. Thispaper mainly includes three aspects following:We got the crude extract components E1, E2, E3, E4, E5, and W, M, J withultrasonic extraction method, using different polar solvents (from petroleum ether towater) to extract the constituents from esculenta. Adopted multiple (solventdistribution, MCI resin, silica gel) separating methods to separate the the crude extractcomponents further, we obtained8components by solvent distribution,12components separated by MCI resin,17components separated by silica-gel column,we got45components in total. Finally we identified the system separation method,and established the standard components library of Phascolosoma esculenta.We characterized the above components By HPLC and TLC techniques. In thinlayer chromatography, the Chromogenic agent is Vanillin-sulfuric acid reagent, theeluent of all E2components separated by MCI resin is n-butanol-ethyl acetate-water(9:1:10, The upper), and the eluent of E4components separated by silica gel column is petroleum ether-ethyl acetate-methanol(2:3:0.2), we can get the better points andsignificantly expand. In high performance liquid chromatography, we used SymmetryC18column, the flow rate with1mL/min, the water-methanol system eluent, and206nm,208nm and214nm detection wavelength to detect respectively (E1, E2, E3,E4, E5). As can be seen from the HPLC chromatogram, the components got wellseparation. In addition, the PMP derivative method was adopted for monosaccharidecomposition analysis of the polysaccharide component W, J and M. The study shows,there are8monosaccharide varieties at least in Phascolosoma esculenta. Weestablished the P. esculenta HPLC fingerprint by the above method. It Improved theTCM fingerprint database and provided a scientific basis for the pharmacologicalactivity study of P. esculenta.The pharmacological activity of P.esculenta focused on the activity of theanticoagulant and anti-oxidation. Using activated partial thromboplastin time (APTT)assay indicated the anticoagulant activity of P.esculenta. The experimental resultsshowed that the activity of E3is best in all components, but the clotting time onlyprolongs1.08s than the blank group, So all components of Phascolosoma esculentacan be considered no anticoagulant activity almostly. In this paper, we used theclearance of hydroxyl radical and DPPH radical to indicate the antioxidant capacity ofcomponents. The scavenging ability of polysaccharide components is limited, theenzyme component M is strongest, the Clearance rate is up to55.44%with theconcentration of2mg/mL, but the Vc’s Clearance rate is more than99%with thesame concentration. All components have the good ability to scavenge DPPH radicals,the clearance rate of component E1is highest with more than99%. ComponentsW, M, J and E1can promote the self oxidation of Pyrogallol, and componentE3, E4and E5could scavenge superoxide free radicals, but activity is low. Insummary, each component of P.esculenta has no anticoagulant activity almostly, butthere are different levels of antioxidant capacity.