转基因棉子中棉酚的提取条件及含量测定

Extracting Conditions and Content Measurement of Gossypol in Transgenic Cotton Seeds

采用乙醇为提取液,对影响棉酚提取率的5种因素进行了考察。结果表明,提取液体所占的体积分数、提取温度和提取时间3种因素对提取率的影响较为显著。进一步进行3因素3水平中心组合试验,运用SAS软件建立了棉酚提取的二元回归方程,并通过岭脊分析得到棉酚最优提取条件为提取温度26.7℃,乙醇体积分数76.3%,提取时间4.5h,棉酚得率预测值为0.109%。在优化条件下,对转基因与非转基因棉子中的棉酚进行提取,苯胺法测得转基因棉子中棉酚的提取率为0.103%,非转基因棉子中棉酚的提取率为0.104%。

The extracting of gossypol by ethanol and determination by aniline was investigated. Through single factor experiments, three levels of the factors including extraction concentration, extraction temperature and extraction time had the marked influence on the yield of gossypol. So these three factors were selected to the central composite design experiment. The SAS software was used to established the quadric regression model for the yield of gossypol. Through ridge analysis the optimum combination was obtained： When the extraction parameters are composed of extraction temperature 26.7 ℃, ethanol concentration 76.3%, extraction time 4.5 h, the predicted maximum yield of gossypol is 0.109%. Under these optimization extraction parameters, the gossypol in transgenic and non-transgenic cottonseeds were extracted, the yield were 0.103% and 0.104% respectively.