Xiaowan Periwinkle Medicinal Ingredients Vincamine Separation And Purification Process And Its Tissue Culture Studies

Vinca minor L. contains a variety of similar monoterpene indole alkaloids, vincamine is one of the major monoterpenoid indole alkaloids. As a cerebral vasodilator, vincamine make coronary blood flow, and the role of selective small blood vessels and capillaries in the brain, cardiovascular and cerebrovascular insufficiency caused by the cycle of disease and dysfunction have all kinds of treatment and improvement.Further research optimizated the process of extraction, purification,refining and HPLC determination system for vincamine extraction form V. minor L. To optimization and establish the production process for vincamine extraction formⅤ. minor L. While, expand the tissue culture ofⅤ. minor L. to seek the vincamine production way by plant tissue culture.The results are as follows:1. Utilized diode array detector, on the basis of retention time comparison, the spectrum to be confirmed and the mobile phase systems for determination vincamine was optimized.And established the best mobile phase for determination of vincamine extraction form V. minor L.,in the present. The best mobile phase was methanol-1% diethylamine(pH7.5) (65:35 v/v).2. According to alkaloid extraction methods reported in the literature, the vincamine extraction process was optimized,and established the optimal extraction method currently.In the same time,determinated the content of vincamine extraction formⅤ. minor L.,that from different areas and different harvesting season. The cultivation had a significant effect on the content of vincamine inⅤ. minor L.. The content of vincamine inⅤ. minor L. had a significant difference in the same harvest of different ages and the same between different harvest age.3. Utilized AB-8 macroporous resin as the separation of the carrier, ethanol as eluting system, for preliminary separation and purification vincamine. The optimum elution process was established:distilled water elution 5 BV(columu volume),60% ethanol elution 5 BV, 90% ethanol elution 5 BV, combined 90% ethanol eluate concentrated to a certain volume of use.4. While combined with high-speed counter-current chromatography, further purification and refining vincamine that had been preliminary purified by AB-8 macroporous resin. The two-phase solvent system for HSCCC composed of chloroform methanol and 0.3mol/L HCL (4:3:2 in volume ratio),photogenic as the stationary phase, lower phase as mobile phase, with the flow rate of 2 mL/min and the detection wavelength of 280 nm.The vincamine purity after purification was 80.2%.5. Sterile culture system was established, the optimal hormone combination for callus initiation was MS+2,4-D 1.0 mg/L+6-BA 0.5 mg/L+NAA 0.5 mg/L. Callus subculture medium was B5+2.0 mg/L NAA+2.0 mg/L 2,4-D。...