Institute of Biomolecular Chemistry, National Research Council, Via Campi Flegrei 34, Pozzuoli 80078, Naples, Italy.
Mar Drugs. 2013 Sep 30;11(10):3742-53. doi: 10.3390/md11103742.
Accurate characterization of biomass constituents is a crucial aspect of research in the biotechnological application of natural products. Here we report an efficient, fast and reproducible method for the identification and quantitation of fatty acids and complex lipids (triacylglycerols, glycolipids, phospholipids) in microalgae under investigation for the development of functional health products (probiotics, food ingredients, drugs, etc.) or third generation biofuels. The procedure consists of extraction of the biological matrix by modified Folch method and direct analysis of the resulting material by proton nuclear magnetic resonance (¹H NMR). The protocol uses a reference electronic signal as external standard (ERETIC method) and allows assessment of total lipid content, saturation degree and class distribution in both high throughput screening of algal collection and metabolic analysis during genetic or culturing studies. As proof of concept, the methodology was applied to the analysis of three microalgal species (Thalassiosira weissflogii, Cyclotella cryptica and Nannochloropsis salina) which drastically differ for the qualitative and quantitative composition of their fatty acid-based lipids.
准确描述生物质成分是天然产物生物技术应用研究的一个关键方面。在这里，我们报告了一种高效、快速和可重复的方法，用于鉴定和定量研究中的微藻中的脂肪酸和复杂脂质（三酰基甘油、糖脂、磷脂），这些微藻被开发用于功能性保健品（益生菌、食品成分、药物等）或第三代生物燃料。该方法包括通过改良的 Folch 法提取生物基质，然后通过质子核磁共振（¹H NMR）直接分析所得材料。该方案使用参考电子信号作为外部标准（ERETIC 方法），允许在高通量筛选藻类收集物和遗传或培养研究期间的代谢分析中评估总脂质含量、饱和度和类别的分布。作为概念验证，该方法应用于三种微藻（威氏海链藻、隐甲藻和盐生杜氏藻）的分析，这三种微藻的脂肪酸基脂质的定性和定量组成有很大差异。
