Determination, Extraction, Antitumor Activity Of Oleanolic Acid And Ursolic Acid From Chinese Quince

Chinese quince contains an abundance of nutrition components, which is a traditional Chinese plant used as herb and food. The paper investigated the extraction, separation and structures of oleanolic acid (OA) and ursolic acid (UA) from Chinese quince with the advanced methods. The antitumor activity and immunodulatory effects on tumor-bearing mice were studied at the same time. These researches laid an useful foundation for studying Chinese quince and functional product. The main results are as followed:1. Established a new analysis method for OA and UA presented inChinese quince by HPLCHubei province is mainly one of producing areas for Chinese quince. UA existed in Chinese quince was identified by HPLC. A high performance liquid Chromatographic method for the analysis of OA and UA has been established. The optimum conditions were: column, ODS Hypersil-C18 (5μm, 250mm×4.6mm i.d.) ;mobile phase, methanol: water: acetic acid=85: 15: 0.01, (v/v/v); temperature, 25℃; wavelength of UV detector, 210nm; flowrate, 1.0mL/min; sensitivity setting, 0.01AUFS; injection volume, 10μL. The chromatogram is better and the Rs of OA and UA is 1.4. The contents of OA in Chinese quince is 0.59%, UA is 0.12%.2. Extracting technics of OA and UA of Chinese quinceThe extraction conditions of OA and UA from Chinese quince were discussed, by the orthogonal tests the optimum technics were confirmed as follows, extract OA from Chinese quince is, materials granularity 40, 60℃, extracting 2 times, material: solvent= 1:8(w/v), 95% ethanol, 3h per time; extract UA from Chinese quince is, materials granularity 60, 60℃, extracting 2 times, material: solvent= 1:6(w/v), 95% ethanol, 3h per time.3. Purification and Identification of OA of Chinese quinceThe OA were prepared and purified with recrystallizing method, the purity of crystal is 98.6% after recrystallizing operation several times. The experimental results showed that this method is simple and convenient. HPLC/MS/MS, HPLC, IR, UV-VIS and MP were introduced to identify the crystal; the results showed that it's physical and chemic characters accorded with literature.4. Antitumor activity of extract from Chinese quince and effect on immune function of H₂₂-bearing mice in vivoFemale Kun-Ming mice were divided into 6 groups for experiment. The result indicated that the inhibiting rate of H₂₂ tumor were 26.47% by highest dose (300mg/kg·d). The pathology observation of the tumor tissue showed that the treated groups with extract have inhibiting effects on H₂₂ tumor, contrasting with ttunor-bearing controlled group. The delayed-type of hypersensitivity, serum hemolysin content, macrophage phagocytosis, nature killer cells activity of tumor-bearing mice could be upgraded significantly by 75～300 mg/kg·d of extract, which indicated that they could increase immune function in tumor-bearing mice. Meanwhile, the extract, could enhance the content of SOD and decrease the content of MDA in tumor-bearing mice serum, so they also have antioxidant effect on tumor-bearing mice.