Study On High Effective Extraction And Purification Process Of Vinblastine From Catharanthus Roseus

Catharanthus roseus (L.) G Don is one of the Catharanthus in Apocynaceae. It widely grows in China. There are lots of alkaloids in it, of which vinblastine has a broad-spectrum anticancer activity, it can treat much kinds of cancer. In this paper we established RP-HPLC determination method and sample preparation method, and studied on the technology of vinblastine high effective extraction and purification, and finished the middle experiment from fresh catharanthus roseus. And finally we established the new vinblastine produce process. Here are the main conclusions:1. Established the HPLC detection method of four vinca alkaloids: vinblastine, vincristine, catharanthine, vindoline.These 4 ingredients were separated on a Diamonsil (C18 column 250x4.6mm i.d., 5 )with methanol: acetonitrile =4:1 as solvent A, and water: diethylamine =986:14 , H3PO4 adjust pH to 7.2, as solvent B, and the mobile phase was composed of 380ml A and 620ml B. The detection was performed at a flow rate of 2.0ml/min, and detected at 220nm within 30min.2. Established extraction and purification process of vinblastine. The process is:Extracted ExtractedFresh catharanthus roseus Extracted liquid Crude extractionConcentrated and dryedAcidifyed Column chromatographyCrude vinblastine Purified vinblastineExtracted Acidifyed VLB-sulfateRecrystalizedThe detailed parameters are:(1) Fresh material grinding extraction process: using 75% alcohol as extract solvent, sample and extraction solvent rate is 1:3, in 2min. Aggrandizement extraction process is: using 75% alcohol as extract solvent, sample and extraction solvent rate is 1:5, air flew rate is 2m3/h, and extraction time is 25min. In this process, vinblastine extraction rate is 97.3%. And it can get to 10% purity after extraction andconcentration process.(2) vinblastine purification process is: crude vinblastine: water: boiling alcohol= 100:200:4000, from this process we got crude vinblastine(40%). And then using silicon gel(300-500mu) to purify process, the sample quality is 3g, elution solvent is CHCl3:MeOH=100:2 i n 3 BV/h. Yield 1 .13g v inblastine(95%). I n 6 0C, using 90% alcohol as crystal solvent, then cooled slowly to 0癈, yield vinblastine crystalloid(98.2%).In this process, vinblastine utmost yield is 0.006% with purity of 98.2%. We established vinblastine quality criteria and also studied on the feasibility of produce 9kg vinblastine per year base on this process.