| Radix Puerariae is the dried root of Leguminous plants of Pueraria lobata (Willd.) Ohwi, known as the "thousand year ginseng" reputation, is the national Ministry of Health approved medicinal and edible plant. Have improve immunity, antipyretic, protect liver, anti-oxidation, fall blood sugar, improve the effect such as disease of heart head blood-vessel. Numerous studies show that the main active ingredient in the Radix Puerariae was the flavonoids, of which puerarin was the highest. The content of puerarin and flavonoids of Radix Puerariae is usually used as evaluation standard of quality.This article taking Radix Puerariae as the raw material, mainly includes the following aspects. Respectively using ethanol refluxing extraction, papain enzyme assisted extraction method to study the extraction process of active ingredients of Radix Puerariae; In addition, the purification technology of the effective components of Radix Puerariae was researched, explored a mature route of separation techology. Finally, compared the antioxidant capacity of extract of Radix Pueraria.1. The extraction of effective components from Radix Puerariae with ethanol refluxing method:With the ratio of puerarin and flavonoids in Radix Puerariae as index, the content of puerarin and flavonoids were determined by UV spectrophotometer and high performance liquid chromatography respectively. The effect of the different extraction conditions on the ratio of puerarin and flavonoids were evaluated comprehensively. The extraction process was:with 20 times the amount of 50% ethanol solution as extraction solvent, the extraction time was 120 min, and extraction for 2 times, the ratio of puerarin was 1.998%, the ratio of flavonoids was 8.914%.2. The extraction of effective components from Radix Puerariae with papain enzyme hydrolysis method:Single factor was studied on the effect of the ratio of puerarin and flavonoids, such as:enzymolysis time, concentration of enzyme, enzymolysis pH value and enzymolysis temperature; Fuether by using the orthogonal test, with the ratio of Puerarin and Flavonoids as comprehensive evaluation index to obtain the optimum extraction conditions. The results show that:the enzymolysis time was 4 h, enzyme concentration was 2%, enzymolysis temperature was 60℃, enzymolysis pH=6. The ratio of puerarin was 2.201%, the ratio of flavonoids was 10.613%. The ratio of Papain enzyme assisted extraction of active ingredients of radix puerariae is better than ethanol refluxing extraction.3. By macroporous resins static experiment, compared the adsorption and desorption of HPD-750、FL-2、AB-8、X-5 and NKA-Ⅱ five kinds of macroporous adsorption resins, screened AB-8 macroporous resin as the absorption resin for purifying Radix Puerariae. Through the single factor experiment studied the effect of the pH, the saturated adsorption quantity, the sample liquid concentration, the ratio of diameter to height, the eluent agent concentration and dosage, obtain the optimum purification conditions. Radix Puerariae alcohol extract directly on the sample after enrichment, the concentration of sample was 7.592 mg/mL, and the ratio of diameter to height was 1:6, after absorption,3 BV water was used to elute impurity, then,3 BV 40% ethanol elution was used to elute. The content of Puerarin was 29.59%, the content of flavonoids was 64.39%.4. The antioxidant activity of puerarin and flavonoids were determined by DPPH free radical assay and 2- deoxyribose oxidation degradation. The experimental results show that:the pure puerarin can not eliminate DPPH· effectively, but also can not very good protect the degradation of 2-deoxyribose induced by Fenton reaction. However, pueraria extract could eliminate DPPH· effectively, but also could protect the degradation of 2-deoxyribose induced by Fenton reaction. The results shows that the Radix Puerariae contains the chemical composition of stronger antioxidant ability, these ingredients may be other more hydroxyl of flavonoids. |
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