摘要
本文采用反相高效液相色谱纯化制备了从玉米小斑病病菌C小种(HelminthospriummaydisRaceC)分离出来的毒素ToxinI,经NMR的结构分析证实此制备物纯度较高,达到结构分析要求。用氯仿提取含有HMC的毒素培养滤液,经过多次TLC展层层析(CHCl_3:MeOH=9:1),刮取具有侵染活性的部分(R_f=0.4)溶解在小体积甲醇中备用。色谱柱为Waters(制备型250mm×10mm)填料为YWG-C18反相柱,粒度10um,流动相选用甲醇:水55:45,流速为0.8ml/min,检测波长272nm,进样量每次100ul,当主要峰出现时,收集合并冻干保存(保留时间9.5min)。
A reversed-phase HPLC method has been successfully used to separate and prepare the HMC-Toxin fromHelminthosprium maydis Race C It is evidenced that the separated sample by HYLC is highly purified and suitable for theNMR structural analysis. The filtered toxin culture solution containing HMC-toxin was extracted by chloroform andmult-separated through TLC by using the development reagent of CHCL3:MeOH(9:1). Then bioactive fraction of toxin toC-cytoplasm corn was collected and dissolved in 500ul methanol for HPLC analysis and preparation. The optimum conditions were observed by using a 10um YMG-C18, analytical column (250mm×4.6mm) and methanol;water eluent in PrimaryexperimentS. A preparative columns 250nun× 10mm, with the same pekings and methanol;water(55:45) as mobile phase at aflow-rate of 0.8ml / min and UV detector at 272nm were used for preparation. 100ul of sample were injected into HPLCsystem with 100ul loop of Aheodyne injection valve every time, The eluate containing purified sample was collected whenthe main peak is appeared. The retention time of main componemt (Toxin I). from HMC-toxin is 9.5min.This method couldbe used for preparation for structure analysis of natural products.
出处
《微生物学通报》
CAS
CSCD
北大核心
1999年第1期47-49,共3页
Microbiology China
关键词
HMC毒素
制备
高效液相色谱法
High performance liquid chromatography , Preparation, HMC toxin