Research On Enzymatic Extraction And Antioxidant Activities On Neferine From Lotus Seed Core Powder

Lotus seed drill core is a bit of powder from the lotus stem end to the tip drill coring fruit formation,which is taken as a by-product when the shell lotus deep processing. Numerous studies showed that the powder alkaloids had strong free radical scavenging, antioxidant effect on human myocardial ischemia, arrhythmia, high blood pressure and better prevention and treatment effect.In this study, the cellulase-assisted ethanol extraction of neferine from lotus seed drill core powder as raw material were extracted and measured by its ability to clear free radical scavenging and total reducing power, which to study neferine’s vitro antioxidant activity. Meanwhile, adding neferine to the oil to study the antioxidant effect of rapeseed oil and lard oil by measuring the peroxide value of oils and fats to reflect neferine lipid oxidation inhibition.The main results are as follows:1.Neferine from lotus seed drill core powder was detected and analyzed by High performance liquid chromatography.Investigated by continuous injections from the linear relationship, precision, repeatability, stability and processing recoveries and other aspects of the feasibility of using RP-HPLC method to determine its chromatographic conditions:the Agilent TC-C18 column （4.6mm*250mm,5um） column; mobile phase ratio of methanol: aqueous solution of 0.1% triethylamine=64:36; and the flow rate and column temperature were controlled at 1.0 mL/min and 25℃; detection wavelength was 286nm, injection volume was 10 μL.Under these conditions, the average content of neferine from lotus seeds drill core powder was 0.415%.2.The cellulase-assisted extraction of neferine from lotus seed drill powder was optimized using response surface methodology on the basis of the results of single-factor experiments.The effects of cellulase dosage, hydrolysis pH, temperature and time were investigated to obtain the yield of neferine.The results showed that:cellulase dosage 3.1% （m/m）, pH 4.8 for 100 min, temperature 50 ℃, the yield of neferine reached to （7.02±0.22） mg/g under these optimal extraction conditions, higher than the ethanol extraction method （4.03±0.17） mg/g. The cellulase-assisted extraction significantly simplified process steps and milded extraction conditions, which could effectively improve the utilization of lotus seed drill powder.3.Using neferine calibrator to research the anti-oxidation test.The research showed that when VC or TBHQ as a positive control, there was not only some effect on-OH, O₂^-·and DPPH radical scavenging ability,but also had good overall capacity reduction.Moreover,its concentration had a dose-response relationship, and the maximum clearance rate could reach to 62.79%,76.23%,93.57%,respectively.4.Using the oven method and POV value index to study neferine oxidized fats in the main inhibition. The results showed:neferine of rapeseed oil and lard had a strong antioxidant effect, and both with neferine increased concentration showed a gradual enhancement effect.By using orthogonal experiment,the combination formula which contains rapeseed oil and lard oil,neferine, respectively VC and citric acid complex could significantly shorten the oxidation induction of fat.The combination were:0.075% neferine+ 0.030% citric acid+0.020% VC, lard oil:0.020% neferine+0.030% VC+0.010% citric acid.