Studies On The Extraction And Purification And Ingredient Analysis Of Polyphenols In Pomegranate Peel And Its Activities

Pomegranate peel polyphenols are all the sum of polyphenol hydroxyl compounds in pomegranate peel, It contains gallic tannins,ellagic tannins,ellagic acid,chlorogenic acid,gallic acid,catechin,epicatechin,anthocyanin,ferulic acid and quercetin etc. It content about 10%-20% of it dry weight。Many research show that pomegranate peel polyphenols has strong activity of scavenging free radica,antimutagenic,anti-aging and antibacterial etc.It has strong effect for pharmaceutical industry,commodity and foodstuff.We have studied on the qualitative and quantitative analysis method of Pomegranate peel polyphenols, extraction process, the technology of isolation and purification of polyphenols in pome-granate peel with macroporous absorption resin and the antioxidative effect. It provide many references for researches of functional foods.The present study is composed of the following four parts.1.Optimal Microwave-assisted extracting conditions for polyphenols of pomegranate peel by Response Surface MethodologyObjective In this design,we extraction polyphenols of pomegranate peel with microwave assisted extraction. Methods Through the single factor and Box-Behnken design experiments. Results the comminution degree of pomegranate peel powder, the extraction temperature, the ratio of pomegranate peel powder to the solution, extraction time and times were>100 Mesh,75℃,1:30,140s and 3 times, respectively. In this condition, the best process conditions yield of polyphenols was 22.06g/100g.Conclusion This method is feasible compare with the most Traditional methods.2.Study on the technology of isolation and purification of polyphenols in pome- granate peel with macroporous absorption resinObjective By researching to resin of D101,DI40,AB-8,D160,HPD7225,we want to get Their differences of performance for adsorption and find the best resin, Simultaneously, Find the best conditions for isolation and purification. Methods The research elect static and dynamic experiment for elect adsorption and desorption. Results In these five resins,D101 resin have a good adsorption and desorption effect.So, there elect D101 resin to purify polyphenols of pomegranate peel.additionly,By researching to resin,we obtain the best conditions.It is electing D101 resin, velocity of flow is 2BV/h, density is lOmg/mL, capacity of clerning agent is 5BV,density of ethanol eluent is 70% and it consumption is 5.5BV。 Conclusion This method is effective and feasible to separate and purify polyphenols of pomegranate peel.3.Study on the activities of polyphenols in pomegranate peelObjective Antioxidative activity of Pomegranate Peel extract in vitro was studied. Methods The effect of free radicals scavenging was investigated with different special chemical systems of hydroxyl radical, superoxide radicals, DPPH radical and inhibitor lipid-peroxidation with homogenates of mouse liver.Results Ability of Pomegranate Peel extract for scavenging hydroxyl radical and superoxide radicals and DPPH radicals of was as follows:IC5o=71.37mg/mL, SC50=557.55mg/mL, DC5o=31.98mg/mL, respectively. Gallic acid was IC5o=51.85mg/mL, SC5o=289.2mg/mL, DC5o=37.65mg/mL, respectively. Green tea polyphenols was IC50=66.67mg/mL, SC50=1043.91mg/mL, DC5o=22.48mg/mL, Inhibitive effects of Pomegranate Peel extract on lipid-peroxidation of homogenates of mouse liver EC50 was 5.21mg/mL, Gallic acid EC50 was 22.82mg/mL, Green tea polyphenols EC50 was 3.47mg/mL. And half-inhibitory concentration of DPPH·,·OH,O2 and anti-lipid peroxidation were 26.33mg/L,57.86 mg/L,416.31 mg/L and 3.69 mg/L for purification of polyphenols in pomegranate peel.Conclusion The extract of Pomegranate Peel can effectively scavenge free radical and inhibit lipid-peroxidation significantly.4.Establishment of the RP-HPLC method of ingredient analysis for polyphenols in pomegranate peelObjective A method was established to determine the contents of Chlorogenic acid, Epicatechin, Ellagic acid and Quercetin in pomegranate peel Polyphenol. Methods reversed-phase HPLC was used to separate and determine the contents of Chlorogenic acid, Epicatechin, Ellagic acid and Quercetin in the pomegranate peel Polyphenol.The chromatographic conditions were as follows:Hyspersil ODS2 (250mm×4.6mm,5um) at 30℃ with UV detector at 310nm, mobile phase of acetonitrile-0.4% phosphoric acid(17:83),and flow rate was 1.Oml/min. Simultaneously, there establish a method to measure the content of pomegranate glycosides.The chromatographic conditions were as follows:Hyspersil ODS2 (250mm×4.6mm,5um) at 25℃ with UV detector at 232nm, mobile phase of methanol-0.4% glacial acetic acid(7:93),and flow rate was 1.0ml/min. Results Chlorofenic acid, Epicatechin, Ellagic acid, Quercetin and pomegranate glycosides showed a good Linear relationship with Peak area in a Appropriate concentration range, and the average recoveries were 100.66% (RSD=2.06%),100.05%(RSD=0.58%),100.05%(RSD=0.51%),99.51%(RSD=1.22%) and 99.79%(RSD=0.52%).Conclusion This method was simple,accurate, reliable and has good reproducibility.