雨生红球藻高效培养及其虾青素提取与纯化-efficient culture of haematococcus pluvialis and extraction and purification of astaxanthin.docxVIP

应器相连。(3) 探究了雨生红球藻中虾青素的最佳提取溶剂，结果表明，二甲基亚砜/ 丙酮（4/1, v/v）的混合溶剂最适宜虾青素的提取，其提取量为 34.73 mg/L，比 乙酸乙酯/乙醇（2/1, v/v）的混合溶剂提取下提高了 14.12%。并在单因素的基 础上，采用 Box-Behnken 对虾青素的提取条件进行优化，得到最优提取条件 为：提取温度，51.72℃；提取时间为 42.32 min；料液比为 1:6.33；虾青素的提 取量达 35.49 mg/L。(4) 通过薄层层析法和高效液相法检测从雨生红球藻中提取得到的总类胡萝 卜素，结果表明，总类胡萝卜素中主要含有虾青素单酯、虾青素双酯，游离虾 青素的含量较少，还有少量其它成分。并研究了正己烷与丙酮不同配比对虾青 素 TLC 展开效果的影响，结果表明，两者之间的最优配比（体积比）为 7:3， 在此展开剂的作用下，游离虾青素的 Rf 值为 0.21，而且，各种组分都能较好地 分开。(5) 对虾青素酯的皂化工艺进行了研究，以游离虾青素的产量为评价指标， 对影响虾青素酯皂化的碱浓度、温度和时间进行了探讨，并在单因素实验的基 础上，利用 Box-Behnken 对皂化的条件进行优化，结果表明，虾青素酯最优的 皂化条件为：碱浓度为 40.27 g/L，温度为 25.91 ℃，皂化时间为 26.56 min，游 离虾青素的产量最大，为 5.12 mg/L。然后，利用 TLC 和 HPLC 对皂化后的样 品进行了分析，结果表明，在皂化后的样品中，游离虾青素占大部分，还含有 很少量的虾青素酯和其它成分。(6)为进一步得到虾青素的纯化样品，对皂化后的样品进行硅胶(200 目)柱层 析，柱层析后的样品经 TLC 分析表明只有一个游离虾青素的色素斑点，HPLC 表明，游离虾青素的纯度在 90%左右。关键词： 雨生红球藻；虾青素；光谱；提取；分离纯化VHigh-efficient Culture of Haematococcus pluvialisand extraction and purification of astaxanthinAbstractAstaxanthin’s powerful antioxidant properties endow it many important physiologic functions, such as, improving animal immunity, being a foe of free radicals and reactive oxygen, inhibiting tumor and so on. Therefore, the natural astaxanthin has a widespread application and prospect. Furthermore, Haematococcus pluvialis is regarded as the best biological source of astaxanthin. In view of this, the research about increasing the cell concentration and the accumulation of astaxanthin, efficiently extracting astaxanthin from Haematococcus pluvialis and the purifyication of astaxanthin is of important significance. The contents and conclusions of this dissertation mainly include the following aspects:To explore the effects of spectrum on cell growth and astaxanthin production of Haematococcus pluvialis, absorption spectrum of cells on different growth phases (cell-growing and astaxanthin-producing stages), plus with the spectrum of different illuminants were measured. And the effects of different light intensity of diverse illuminants were studied on cell growth and accumulati