溶剂法提取微拟球藻脂质的工艺优化 及其脂质组学分析Optimization of extraction of Nannochloropsis sp. lipid with solvent and its lipidomics analysis
2022
张美兰1，赵康宇1，张立伟1,2，何东平1,2，郑竟成1,2 ZHANG Meilan1, ZHAO Kangyu1, ZHANG Liwei1,2, HE Dongping1,2， ZHENG Jingcheng1,2
研究了溶剂法提取微拟球藻脂质的工艺，对比了氯仿-甲醇（体积比1∶ 1）、氯仿、异丙醇、石油醚、正己烷-乙醇（体积比1∶ 1）、正己烷、乙醇7种溶剂对微拟球藻脂质得率和二十碳五烯酸（EPA）含量的影响，确定乙醇作为提取溶剂。采用单因素试验研究了乙醇体积分数、料液比、提取温度和提取时间对微拟球藻脂质提取的影响，并通过正交试验对工艺条件进行优化，最后采用UPLC-Q-TOF-MS对所得微拟球藻脂质进行脂质组学分析。结果表明，微拟球藻脂质最佳提取工艺条件为乙醇体积分数80%、料液比1∶ 17.5、提取温度65 ℃、提取时间2 h。在最佳工艺条件下，微拟球藻脂质得率为（1587±0.21）%，EPA含量为（14.49±0.14）g/100 g。微拟球藻脂质中检出17种脂质，其中12种脂质含有EPA，含有EPA的脂质占总脂质的45.24%。The extraction of Nannochloropsis sp. lipid by solvent was studied.The effects of seven systems including chloroform-methanol(volume ratio 1∶ 1), chloroform, isopropanol, petroleum ether, n-hexane-ethanol (volume ratio 1∶ 1), n-hexane and ethanol on the yield and EPA content of Nannochloropsis sp. lipid were compared. Ethanol was determined as the extraction solvent. The effects of ethanol volume fraction, material-to-liquid ratio, extraction temperature and extraction time on the extraction of Nannochloropsis sp. lipid was studied by single factor experiment and the process conditions were optimized by orthogonal experiment. Moreover, the lipidomics of the Nannochloropsis sp. lipid were analyzed by UPLC-Q-TOF-MS. The results showed that the optimal extraction conditions were obtained as follows:ethanol volume fraction 80%, material-to-liquid ratio 1∶ 17.5, extraction temperature 65 ℃, and extraction time 2 h. Under these conditions, the yield of Nannochloropsis sp.lipid was (15.87±0.21)%, and EPA content was (14.49±014)g/100 g. Seventeen kinds of lipids were determined in Nannochloropsis sp.lipid, in which only twelve kinds of lipid contained EPA. The lipids containing EPA accounted for 45.24% of the total lipids.
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