The Preparation And Application Of Flavonoids From Changshan Huyou

Changshan Huyou has great medicinal value,especially the peel is rich in flavonoid compounds(Flavonoid Compound,FC),volatile oil and other active ingredients.How to develop the medicinal value of Changshan Huyou and promote the development of the agricultural industry is an important direction for experts,scholars and corporate entities.In this paper,the extraction,separation and purification process of Huyou flavonoids is studied,the content of effective ingredients of flavonoids in Changshan Huyou flavonoids is determined,and the anti-inflammatory effects of the effective ingredients are evaluated and the development of functional compound products is carried out.1.Study on the extraction and purification of flavonoids from Changshan Huyou.Taking Huyou Peel as raw material,the extraction optimization scheme of flavonoids from Changshan Huyou Peel was as follows : extraction twice,water : Huyou Peel 10:1,temperature 100 °C,time 1h.The yield of flavonoids from Changshan Huyou Peel was21.17 %.The purification experiment of flavonoids from Changshan Huyou by macroporous resin was carried out,and the optimum process parameters were obtained as follows : the sample solution was the crude extract of flavonoids from Changshan Huyou,the solubility was 3.68 mg / ml,the sample volume was 5.00 ml / g,75 % ethanol was used for elution,the p H value was 4.50,and the elution rate was 89.72 %;The average contents of naringin and neohesperidin in the crude extracts of flavonoids from Changshan Huyou were 6.24 % and4.50 % by HPLC-MS.The average contents of naringin and neohesperidin in refined dry powder of Changshan Huyou flavonoids were 50.02 % and 36.10 %.The total content of naringin and neohesperidin in refined dry powder of Changshan Huyou flavonoids was 8.03 times higher than that before purification.2.Evaluation of anti-inflammatory effect of refined dry powder with flavonoid compounds from Changshan Huyou.Lipopolysaccharide(LPS)induced transgenic neutrophils fluorescence labeling and transgenic macrophages fluorescence labeling inflammatory zebrfish models were established respectively.To investigate the anti-inflammatory effect and the effect of refined dry powder of flavonoids on inflammatory zebrafish,and to conduct histopathological analysis experiments The maximum tolerance solubility of zebrafish to the refined dry powder of flavonoids from Changshan Huyou was8000 μg / m L.A normal control group,a model control group,and a positive control group were set,and the concentration of indomethacin was 60 μg / m L.The concentrations of refined dry powder of flavonoids from Citrus Changshan Huyou in the three experimental groups were 889,2667 and 8000 μg / m L,respectively.The number of neutrophils in the inflammatory site of zebrafish : 3 in the normal control group,22 in the model control group,13 in the positive control group,17,14,15 in the three experimental groups,respectively,and the effects on neutrophil inflammation were 22.70 %,36.40 % and 31.80 %,respectively.In the positive control group,the fluorescence signal intensity of zebrafish inflammatory site(371000 pixels)was 51 %,and the fluorescence signal intensity of zebrafish inflammatory site was 520175,310845 and 435642 pixels,respectively.The improvement of macrophages was 31.4 %,59.0 % and 42.5 %,respectively.Fluorescence PCR experiment was carried out by extracting and determining the concentration of zebrafish RNA.The relative expression of TGFβ-1 and IL-1β in the experimental group was significantly lower than that in the model group(5.79),indicating that it can significantly inhibit gene expression.The relative expression of TGFβ-1 gene in the positive control group was 2.48,which was significantly different from that in the model control group,indicating that the expression of TGFβ-1 gene in the positive control group was significantly decreased.The relative expression levels of inflammatory related genes TGFβ-1 in the three experimental groups were 3.27,1.90 and 3.19,respectively.Compared with the model control group,the difference was extremely significant,indicating that the refined dry powder of Changshan Huyou flavonoids can significantly inhibit the expression of TGFβ-1 gene.The relative expression level of IL-1βgene in the model control group was 11.91,compared with that in the normal control group(1.10),and the relative expression level of IL-1β gene in the positive control group was 1.83,indicating that the IL-1β gene expression in the positive control group was significantly decreased.The relative expression levels of IL-1β were 4.28,5.80 and 6.21 in the three experimental concentrations of refined dry powder of Changshan Huyou flavonoids,respectively,which showed significant anti-inflammatory effects.3.Research and development of functional compound products.Preparation of tablet product by compounding refined dry powder of flavonoids from Changshan Huyou and American ginseng extract.