Research On Citrus Bergamot's Mineral Ingredient, Optimum Extraction Of Bioactive Substances And Its Antioxidation

With the rising of living standard,there is a gradually increasing awareness of health care among people,who are paying more and more attention on the nutrient health food.Bergamot,with a certain function in nutrition and health care,is gradually arousing people's concern.The antioxidant property of bergamot is one of the most important effects,while bergamot may contain some mineral compositions,which are the reason for its function in nutrition and health care.In this study,citrus bergamot's fruit and leaf are taken as materials for element analysis by ICP-MS method, while by the method of response surface analysis,the best extraction way of flavonoids,polyphenols and polysaccharides of citrus bergamot's fruit and leaf are studied,and by UV spectrophotometer method,the content of flavonoids,polyphenols and polysaccharides,and its antioxidant capacity are determined as well.The main findings are as follows:1.By ICP-MS method,mineral ingredient in the fruit and leaf of Citrus Bergamot are determined and analyzed.Major elements K,Ca,Mg Na,trace elements Zn,Fe,Cr,Cu,Co,Ni,Mo, Mn,Se,and heavy metals elements Pb,Al,Cd are respectively determined.The result shows that: the content of Na in leaf is less than that of the fruit,while K,Ca and Mg are more than that of the fruit;the content of determined trace elements in leaf are more than that of the fruit;Exceeded phenomenon of the harmful elements content does not existed.2.Through Single factor and multi-factor experiments,and response surface experimental design method,the best extraction process of polyphenols,flavonoids and polysaccharides of the Citrus Bergamot is studied.The result shows that:for polyphenols,the optimal extraction conditions is 63%of ethanol,180 minutes of extracting time,80℃of extracting temperature;for flavonoids, optimal extraction conditions is 85%of ethanol,171 minutes of extracting time,81℃of extracting temperature;for polysaccharides,the optimal extraction conditions is 247 minutes of extracting time, 91℃of extracting temperature,and the ratio of sample to extracting reagent with 1:64.Through reliable verified experiment,the optimized condition of the above extraction method is finally indicated to be reliable.3.By UV spectrophotometer method,the content of flavonoids,polyphenols and polysaccharides of Citrus Bergamot's fruit and leaf are determined.One gram of fruit,leaf powder of Citrus Bergamot polyphenols yield 1.084 and 1.302 grams,respectively,per 100 grams. Flavonoids yield 0.404 and 1.292 grams,respectively,per 100 grams.Polysaccharides yield 4.542 and 1.776 grams,respectively,per 100 grams.4.By DPPH method,the total antioxidant capacity of flavonoids and polyphenols of Citrus Bergamot's fruit and leaf are determined.Fruit flavonoids and leaf flavonoids are in good ffect on scavenging DPPH,and the IC₅₀ are respectively 8.25 and 9.03 mg/l,with less scavenging capacity than Vc(IC₅₀=5.99mg/l) and rutin(IC₅₀=5.90mg/l),and fruit flavonoids scavenging capacity is a little stronger than leaf flavonoids.Fruit polyphenols and leaf polyphenols' IC₅₀ are respectively 8.22 and 8.74,with less scavenging capacity than Vc and rutin,in addition,fruit polyphenols is a little stronger than leaf polyphenols.5.Capacity of scavenging superoxide anion free radical:fruit flavonoids and leaf flavonoids' IC₅₀ are respectively 17.74 and 20.77mg/l,stronger than rutin(IC₅₀=25.01mg/l),but less than Vc (IC₅₀=5.32mg/l) for scavenging capacity.With the concentration of fruit and leaf flavonoids in the reaction solution increasing,uperoxide anion free radical scavenging rate is also on the rise.Fruit and leaf polyphenols' IC₅₀ are respectively 26.79 and 28.16mg/l,with less scavenging capacity than Vc and rutin.With the concentration of fruit and leaf polyphenols in the reaction solution increasing,uperoxide anion free radical scavenging rate is also on the rise.Fruit and leaf polysaccharides' IC₅₀ are respectively 11.83 and 12.26mg/l,a little stronger than rutin in scavenging capacity,but less than Vc.With the concentration of fruit and leaf polyphenols in the reaction solution increasing,the rate of uperoxide anion free radical clearance upward trend is almost in a straight line.When the concentration reaches 20mg/l,the scavenging rate is highest for 66.47%and 64.26%respectively.Further increase of the concentration will cause the scavenging rate to tend to be slow.Experimental results of variance analysis show that among the flavonoids,polyphenols and polysaccharides of fruit and leaf,there is a significantly difference of concentration scavenging rate.6.Scavenge hydroxyl radical capacity:With the concentration fruit and leaf flavonoids in the reaction solution increasing,hydroxyl radical scavenging rate is also on the rise.Their IC₅₀ are respectively 17.06 and 15.86mg/l,stronger than Vc(IC₅₀=24.30mg/l),and as much as rutin (IC₅₀=16.17mg/l) in scavenging capacity.When the concentration of rutin and flavonoids of the fruit and leaf are more than 18mg/l,the scavenging capacity of hydroxyl radical will decline,on the contrary,Vc is always on the rise.Therefore,for fruit and leaf flavonoids to the role of hydroxyl radical system,the best dose for scavenging hydroxyl radical of fruit and leaf flavonoids is 18mg/l.Fruit and leaf polyphenols' IC₅₀ are respectively 24.72 and 25.67mg/l,less than rutin in scavenging capacity,and as much as Vc.With their concentration in the reaction solution increasing, hydroxyl radical scavenging rate is also on the rise.Fruit and leaf polysaccharides' IC₅₀ are respectively 11.09 and 12.22mg/l,stronger than rutin and Vc in scavenging capacity.When the concentration reaches 20mg/l,the scavenging rates are highest for 61.35%and 59.58%respectively.Further increase of the concentration will cause the scavenging rate to tend to be slow.Experimental results of variance analysis show that among the flavonoids,polyphenols andpoly saccharides of fruit and leaf,there is a significantly difference of concentration scavenging rate.To sum up,the present research results has provided a scientific theory base for citrus bergamot development and utilization,especially for the development of bergamot's leaf.