| Sweet cherry is rich in nutrients,it is a good source of polyphenols,flavonoids,melatonin and other natural active ingredients,it is very good in antioxidant and anti-aging activities.However,it is easily affected by environmental factors to form a large number of imperfect fruit,and abandoned in the field,resulting in a large amount of waste of resources.At present,the active substances in sweet cherry fruit have not been developed effectively.In this paper,the extraction and purification process of free phenol was optimized by using imperfect sweet cherry as raw material.The composition and structure of total phenol in sweet cherry were determined.The antioxidant activity of free phenol in sweet cherry and the antiaging activity of C.elegans were investigated.(1)The response surface method was used to optimize the extraction process of free phenol from the damaged fruit of sweet cherry by ethanol reflux method.The results showed that the optimal process was: solid-liquid ratio is 1:20,ethanol concentration is 40%,extraction temperature is 50 ?,extraction time is 15 min,number of time is 2,under the extraction conditions,the extraction rate of free phenol of sweet cherry could reach 1.50±0.03 mg/g.(2)By comparing the adsorption and desorption rates of 10 kinds of resins,the HPD-600 macroporous resin which is more suitable for purifying free phenol of sweet cherry was screened.By optimizing the HPD-600 macroporous resin adsorption and desorption conditions,it is concluded that the best technology of purification of sweet cherry free phenol: the sample liquid concentration is 15.09?g/m L,p H of 4,the temperature is 25 ?,the velocity is 2 BV/h,volume is 3 BV,the desorption liquid concentration is 80%,p H of 3,the desorption time is 2 hours,the desorption flow rate is 2 BV/h,and the elution volume is 3.5 BV.(3)The structure of total phenol in sweet cherry was analyzed by Ultraviolet Absorption Spectrum and Fourier Infrared Spectroscopy.The results showed that the main substances of total phenol in sweet cherry before and after purification are all polyphenol compounds,and also contain a small amount of glycosides,flavonoids are the main components of polyphenols.The crude extract contains a special absorption peak of-CH2,and the structure of benzene ring substituents is also different.(4)6 kinds of antioxidant indicators in vitro were applied for studying the radical scavenging activity of free phenol in sweet cherry.The results showed that the free phenol of sweet cherry had better scavenging ability on ABTS+·,DPPH·and NO2-·,the IC50 values are 0.27 mg/m L,1.9 mg/m L,2.45 mg/m L;the scavenging ability of ·OH,O2-·and total reducing force was not obvious.(5)The aging model of the model organism C.elegans was established to determine the effects of free phenol crude extract from sweet cherry on physiological indexes and pressure resistance of the C.elegans.The results showed that the concentration of 100 g/m L,200 g/m L and 300 g/m L of free phenol crude extract from sweet cherry could improve the movement ability of C.elegans,the movement of nematodes was strongest at a polyphenol concentration of 200 g/m L.The concentration of 200 g/m L of sweet cherry polyphenols could improve the movement ability,reproductive ability and life span of nematodes,and had no effect on the body development of C.elegans;it has obvious effects on the resistance of nematodes to walnut quinone damage,hydrogen peroxide damage,and can significantly improve the survival rate of C.elegans.However,it had no significant effect on the ability of nematode to resist ultraviolet radiation and high temperature stress. |
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