Stability And Scale-up Of Separation On Anti-diabetic Compounds From Chinese Bayberry(Morella Rubra Sieb.et Zucc.)

Chinese bayberry(Morella rubra Sieb.et Zucc.)is rich in bioactive compounds such as anthocyanins and flavonols.Preliminary studies have shown that its extracts have hypoglycemic activity.But whether direct consumption of bayberry fruit has an effect on blood glucose regulation remains to be studied.The fruit ripening season of bayberry is quite limited,usually from June to July.Freeze drying can effectively maintain the bioactive compounds of fruit.However,the stability of the anti-diabetic compounds in freeze-dried powder remains unclear.In addition,scale-up preparation of the anti-diabetic compounds in the bayberry deserves further investigation,which is of great significance for the development of bayberry-related products.In the present study,hypoglycemic effects of freeze-dried fruit powder on KK-A~y diabetic mice and high-fat diet C57BL/6 mice were evaluated.Stability of its anti-diabetic compounds was analyzed and scale-up preparation of anthocyanin was optimized.The main results are as follows:1.Freeze-dried powder of‘Biqi'fruit showed hypoglycemic activity in KK-A~ydiabetic mice.Feeding of bayberry freeze-dried powder(9 weeks)relieved the diabetic symptoms and reduced random blood glucose.Also,feeding of bayberry freeze-dried powder(19 weeks)in high-fat diet C57BL/6 mice reduced both fasting blood glucose and random blood glucose.Meanwhile,bayberry freeze-dried powder also significantly alleviated liver lipid accumulation in high-fat diet C57BL/6 mice.2.The stability of bioactive compounds in freeze-dried powder of‘Biqi'and‘Dongkui'fruit stored at room temperature and light-proof pakage was investigated.High performance liquid chromatography(HPLC)analysis showed that the content of C3G would gradually decrease with the prolonged storage time.In‘Biqi',the initial content of C3G was 5.77 mg/g DW,and it was 4.86 mg/g DW after 12 months'storage.In‘Dongkui',the initial content of C3G was 1.70 mg/g DW,and it was 1.46 mg/g DW after 12 months'storage.In general,freeze-drying maintained high content of C3G and flavonols,and there was no significant loss of antioxidant activities,as shown by the ABTS,DPPH and FARD activity data.3.Macroporous resin and HSCCC technology were combined to establish a scale-up preparation process of crude extracts and anthocyanins from bayberry fruit.After a step of D101 macroporous resin purification,the purity of anthocyanins increased from0.56%to 25.02%in the crude extract.Further HSCCC purification with two-phase solvent system composed of methyl tert-butyl ether-n-butanol-acetonitrile-water-trifluoroacetic acid(2:2:1:5:0.01,v/v)resulted in final purity of more than 98%for the anthocyanins.During scale-up,with amplifying of the macroporous resin column and sample loading,large amount of crude extract was prepared.There was no significant difference in the composition and contents of anthocyanins and flavonols,antioxidant activity,and?-glucosidase activity of the crude extracts between different batches,indicating that the scale-up preparation process is stable and feasible.Overall,bayberry freeze-dried powder has hypoglycemic effects on diabetic mice.Research on the stability and scale up preparation of the anti-diabetic compounds can provide foundation for development of hypoglycemic products based on bayberry fruit resource.