Otero Paz, Saha Sushanta Kumar, Gushin Joanne Mc, Moane Siobhan, Barron John, Murray Patrick
Shannon Applied Biotechnology Centre, Limerick Institute of Technology, Moylish Park, Limerick, Ireland.
Reagecon Diagnostics Limited Shannon Free Zone, Shannon, Co. Clare, Ireland.
Anal Bioanal Chem. 2017 Jul;409(19):4659-4667. doi: 10.1007/s00216-017-0412-9. Epub 2017 Jun 7.
Microalgae have the potential to synthesize and accumulate lipids which contain high value fatty acids intended for nutrition and biodiesel applications. Nevertheless, lipid extraction methods for microalgae cells are not well established and there is not a standard analytical methodology to extract fatty acids from lipid-producing microalgae. In this paper, current lipid extraction procedures employing organic solvents (chloroform/methanol, 2:1 and 1:2, v/v), sodium hypochlorite solution (NaClO), acid-catalysed hot-water extraction and the saponification process [2.5 M KOH/methanol (1:4, v/v)] have been evaluated with two species of microalgae with different types of cell walls. One is a marine diatom, Phaeodactylum tricornutum, and the other a freshwater green microalga, Haematococcus pluvialis. Lipids from all types of extracts were estimated gravimetrically and their fatty acids were quantified by a HPLC equipped with Q-TOF mass spectrometer. Results indicated significant differences both in lipids yield and fatty acids composition. The chloroform and methanol mixture was the most effective extraction solvent for the unsaturated fatty acids such as DPA (C22:05), DHA, (C22:06), EPA (C20:05) and ARA (C20:04). While acid treatments improved the saturated fatty acids (SFAs) yield, especially the short chain SFA, lauric acid (C12:0), whose amount was 64% higher in P. tricornutum and 156% higher in H. pluvialis compared to organic solvent extractions. Graphical abstract ᅟ.
微藻有潜力合成并积累脂质,这些脂质含有用于营养和生物柴油应用的高价值脂肪酸。然而,微藻细胞的脂质提取方法尚未完善,且没有从产脂微藻中提取脂肪酸的标准分析方法。本文评估了目前使用有机溶剂(氯仿/甲醇,2:1和1:2,v/v)、次氯酸钠溶液(NaClO)、酸催化热水提取和皂化过程[2.5 M KOH/甲醇(1:4,v/v)]的脂质提取程序,使用了两种具有不同细胞壁类型的微藻。一种是海洋硅藻三角褐指藻(Phaeodactylum tricornutum),另一种是淡水绿微藻雨生红球藻(Haematococcus pluvialis)。通过重量法估算所有提取物类型中的脂质,并通过配备Q-TOF质谱仪的HPLC对其脂肪酸进行定量。结果表明脂质产量和脂肪酸组成均存在显著差异。氯仿和甲醇混合物是提取不饱和脂肪酸如二十二碳五烯酸(DPA,C22:05)、二十二碳六烯酸(DHA,C22:06)、二十碳五烯酸(EPA,C20:05)和花生四烯酸(ARA,C20:04)最有效的提取溶剂。而酸处理提高了饱和脂肪酸(SFA)的产量,尤其是短链SFA月桂酸(C12:0),与有机溶剂提取相比,其在三角褐指藻中的含量高64%,在雨生红球藻中的含量高156%。图形摘要ᅟ。