Extraction And Separation Of Effective Components From Bergamot And Their Biological Activities Study In Vitro

Bergamot is the dry fruit of citron of rutaceae plants, chemical componentsmainly contain volatile oil, coumarone, flavonoids, polysaccharides and so on, possessversatile biological activities including anticancer, anti-tumor, anti-inflammatory,antifungal, antioxidant and immune regulation effects. At present, the development andutilization of finger citron plants stay mostly in terms of its volatile oil composition, andthe research about its active chemical components and their corresponding biologicalactivity is less. On this basis, this paper reviewed the related research progress aboutbergamot, studied the active chemical composition and biological activity of bergamot，provided the scientific basis for further exploitation and utilization of bergamotresources.The main research works and achievements included the following sections：1. Taken wateras solvent, water extract of bergamot was obtained through heatingreflux extraction and stage extraction. Integrated extraction and separation repeatedof the water extract which was extracted with ethyl acetate and petroleum-etherwere carried out by column chromatography, recrystallization and preparative thinlayer chromatography. The possible structures of the compounds obtained wereinvestigated by NMR, MS and related document, and they were identified aslimettin, scoparone, umbelliferone, bergapten and daucosterol.2. By single factor tests and orthogonal experiment design, it is ensured that theoptimum temperature is100℃, solid-liquid ratio is1:15, extraction time is2hoursand the extraction time number is3.The content of polysaccharides in sample wascalculated on the base of criterion curve and conversion expression, results showthat the content of total polysaccharide in crude polysaccharide was61.51%.Combined with the method of stepwise addition of ethanol, three crudepolysaccharide(FSP-1、FSP-2、FSP-3)were got and the yields of thesewere5.75%,3.57%and4.34%, respectively. In addition, BP-1and BP-2were obtained afterdealing with by DEAE-cellulose column with eluent of water and NaCI solution,dialyzing, concentrating and desiccating. By Thin-layer chromatograms, BP-1andBP-2have the same composition of glucose, rhamnose and arabinose.3. The antitumor activity for limettin, scoparone, umbelliferone, bergapten andbergamot polysaccharide was assayed by cytotoxicity test method using 143Bosteosarcoma cells. In the bioassay, compounds limettin, scoparone andbergapten could lead different extent inhibition to the proliferation of143Bosteosarcoma cells.4. The evaluation system of antioxidant activity test in vitro was established by testingthe capability of scavenging three kinds free radical and by the method ofchemiluminescence. According to the Inhibition degree of the chemiluminescencesystem to evaluate the ability of scavenging free radicals of bergamotpolysaccharide. By optimizing initial parameters of the above evaluation system,the antioxidation effect of three kinds of bergamot polysaccharide (FSP-1, FSP-2and FSP-3)was studied by the flow-injection chemiluminescence. The resultsshowed that three kinds of bergamot polysaccharide (FSP-1, FSP-2and FSP-3)could give significant scavenging effects on OH, H2O2and O2-, OH scavengingability is stronger, H2O2scavenging ability are relatively weak, and the FSP-3wasthe largest of three kinds of bergamot polysaccharide on free radical scavengingeffect. This system can also be used for other antioxidant screening of activeingredients. Bergamot polysaccharide can be used as potential antioxidant activityof natural product.