Research of the Composition and Antioxidant Activities of Flavonoids from Elaeagnus Angustifolia Blossoms in Ning Xia
|School||Ningxia Medical University|
|Keywords||Elaeagnus angustifolia Blossoms flavonoid extraction separation free radical Acute hepatic injury|
We studied the content determination and extraction of the total flavonoids from Elaeagnus angustifolia Blossoms in Ningxia （NX-EAB）, and separated 5 kinds of monomer from it and had determined 2 kinds. We also studied the scavenging activities of the different extracts and monomer of NX-EAB on 1,1-diphenyl-2-DPPH （DPPH·）, and studied the protective effects of substances in （AE-NX-EAB） on acute hepatic injury induced by carbon tetrachloride （CCl4） in mice. These will provide experimental data for the application of the total flavonoids in NX-EAB. As follows:1.The determination of the total flavonoids of NX-EAB. Referring to the Technical Standard For Chinese Pharmacopoeia （2005 version）, the total flavonoids in NX-EAB were determined by ultraviolet and visible spectrophotometry with NaNO2-Al （NO3） 3-NaOH as chromogenic agent and rutin as standard substance, in average, the content rate of the total flavonoids in NX-EAB extracted by ligroin was 2.70%,the content rate of that not extracted by ligroin was 1.02%. High performance liquid chromatography （HPLC） was applied to determine rutin, the content rate is 0.029mg/g.2.The extraction method of total flavonoids of NX-EAB was studied. The supersonic technique is the best extraction method we had studied. Through the single factor experiment, we found that Mass （NX-EAB） /Volume （solvent）, density of ethanol, extraction period and extraction times of NX-EAB influenced the extraction of flavonoids significantly, and the best extraction solvent was ethanol. Through the L9（34） orthogonal design,we confirmed that the best extracting technology of total flavonoids of NX-EAB was A2B2C1D3, as following: The density of ethanol: 70%, Mass （NX-EAB） /Volume （solvent） ratio: l: 20, the ultrasonic extraction period: 1.0 h, the extracting times: 3 times.3.Separation、purification and identification of monomer from NX-EAB. We had separated 5 kinds of monomer from NX-EAB. One kind of flavonoid monomer BIII（5,7,4′-trihydroxyflavonol-3-O-（6′′-O-E-p-coumaroyl）-β-D-glucopyranoside）and one kind of glycoside CcII （2-phenylethyl-β-D-glucopyranoside） were separated and purified, They were the first isolated substances from NX-EAB。4.The study on scavenging free radical active component of NX-EAB. The scavenging activities of different compositions of NX-EAB were detected by ultraviolet and visible spectrophotometry. （1） ASE-NX-EAB and several extractions of the ASE-NX-EAB had scavenging activity on DPPH?. The scavenging activity was as following: water left of ASE-NX-EAB extracted by n-butanol > ASE-NX-EAB > ASE-NX-EAB extracted by n-butanol>ASE-NX-EAB extracted by acetidin>ASE-NX-EAB extracted by ligroin. （2） The different scavenging activities of SC-I-NX-EAB, SC-II-NX-EAB and components separated by other solvents on DPPH?, The scavenging activity of SC-I-NX-EAB and SC-II-NX-EAB had some characteristics, as following: SC-NX-EAB > water left of SC-NX-EAB extracted by ligroin>water left of SC-NX-EAB extracted by n-butanol>SC-NX-EAB extracted by acetidin≈SC-NX-EAB extracted by n-butanol。（3） The monomer BIII of the ASE-NX-EAB also has scavenging activity on DPPH·. And IC50 was 0.209 mg/ml.5 Study on the protective effects of AE-NX-EAB on acute hepatic injury induced by carbon tetrachloride （CCl4） in mice. The content of ALT, AST in mice serum and SOD, MDA and GSH in liver was determined by visible spectrophotometry, and observed the pathological changes of liver in mice by hematoxylin-eosin staining. The results showed that AE-NX-EAB could reduce the levels of ALT and AST in mice who had the CCl4-induced acute liver injury, It was significantly different from model group （P<0.05）. And MDA level in liver was also decreased significantly. The activity of SOD and GSH was enhanced,And also significantly different from model group （P<0.05）. In pathology, the liver cells from model group had bigger volume and more fragile. The liver marge was thicker and the surface was yellow brown and had many blooding points. Using microspectacle, we found that the hepatic lobules were destroyed and nucleus of liver cells disappeared. The liver cells degenerated, necrosed and lots of inflammatory cells infiltrated at the local sites aroud liver central veins. The therapy groups had more significant improvement than the model group.