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探索红酵母所产虾青素的分离与纯化工艺,为其特性研究和开发利用提供重要参考依据。首先采用经萃取、浓缩后的虾青素粗提液,通过NaOH-乙醇溶液进行皂化,再经过聚酰胺柱层析纯化虾青素,最后用TLC法(Thin Layer Chromatography)和HPLC法(High Performance Liquid Chromatography)检测其纯化结果。结果表明:虾青素最适皂化条件为将样品置于20g/L的NaOH-乙醇皂化液中,15℃,皂化10min,再以聚酰胺为固定相,依次按正己烷、正己烷:二氯甲烷=4:1、正己烷:二氯甲烷=2:1、二氯甲烷为流动相对虾青素样品进行梯度洗脱,收集游离虾青素馏分置于35℃,0.08MPa真空条件下浓缩,检测其纯度为92.9%。
To explore the isolation and purification of astaxanthin produced by Rhodotorula, which provides an important reference for the research and development of its properties. First, the crude extract of astaxanthin extracted and concentrated was saponified by NaOH-ethanol solution and purified by polyamide column chromatography. Finally, the astaxanthin was purified by TLC and High Performance Liquid Chromatography. The results showed that the optimum saponification conditions for astaxanthin were as follows: the sample was placed in NaOH-ethanol saponification solution of 20g / L, saponified at 15 ℃ for 10min, then polyamide was used as the stationary phase, followed by n-hexane, n-hexane: Methane = 4: 1, n-hexane: dichloromethane = 2: 1, dichloromethane as mobile phase of astaxanthin samples gradient elution, the collection of free astaxanthin fraction was placed at 35 ℃, 0.08MPa concentrated under vacuum, Its purity was 92.9%.