Extraction And Anti-allergic Activity Of Citri Reticulatae Pericarpium Total Flavonoids

Citri Reticulatae Pericarpium is dried fruit peel of Citrus reticulata Blanco and its cultivars, which is one of traditional Chinese herbal medicine. It has vomiting and stomach, dampness and phlegm and other effects. Citri Reticulatae Pericarpium contains flavonoids, volatile oils, alkaloids, polysaccharides and trace elements and other chemical constituents. The flavonoids of Citri Reticulatae Pericarpium have activities of antioxidant, anti-inflammatory, anti-allergic and antitumor. In this study,extraction process of total flavonoids in Citri Reticulatae Pericarpium was optimized,and anti-allergic activity of total flavonoids was studied. Details are shown as follows:1. Extraction process of Citri Reticulatae Pericarpium total flavonoids. Reflux extraction method was applied. The extraction fields were detected by HPLC. The solvents, concentration, dosage and extraction times on the extraction of total flavonoids were studied. When the ethanol concentration was 60%, and the votage was 10 times of amount of material were chosen as the optimal single factors, the flavonoids can get a high extraction fields. Orthogonal test method was used to investigat the best conditions. As the results shown, the ethanol concentration was60%, the votage was 8 times of amount of material and the extraction times were three times(each time was 1.5,1.0,0.5h) were chosen. The total flavonoids were enriched by polyamide resin. The type of eluent, concentration and dosage were studied. As the results shown, ethanol was used as eluent(concentration 60%) and the votage of it was set at 3BV.Optimum extraction conditions of the total flavonoids are: Citri Reticulatae Pericarpium, crushed and sieved, was accurately weighed 5.000 g. The ethanol was used as solvent(concentration 60%), the votage of it was 8 times of amount ofmaterial, the extraction times were three times(each time was 1.5,1.0,0.5h). And the ethanol(concentration was 60%) was used as elunent, the votage of it was 3BV of polyamide resin. Collected eluent was the total flavonoids extract, and the total flavonoids were obtained in a rotary evaporator.2. Anti-allergic activity of total flavonoids was done by animal experiments. In this part, the OVA-induced BALB/c mouse allergic model was used. Intraperitoneal and subcutaneous injections were combinated to sensitized BALB/c mouse to establish the animal allergic model. Intranasal was applied as excitation mode. The serum and bronchoalveolar lavage fluid were collected and lung tissue sections were prepared. Contents of Ig E of serum and IL-4, IL-13 of lavage fluid cytokine were detected by ELISA. Lung tissue sections were stained with H&E treated.It was found that different concentration of the total flavonoids(10 mg·kg-1,30mg·kg-1 and 50 mg·kg-1) were able to reduce contents of Ig E levels of mice serum and IL-4, IL-13 of lavage fluid cytokine significantly. And it was also found that inflammatory infiltration of lung tissue was significantly inhibited by H&E strained of lung sections. In short, the total flavonoids extracted by this method has anti-allergic effects.