Public Laboratory of Bioenergy and Biofuels, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao 266101, China.
J Chromatogr A. 2011 Nov 11;1218(45):8289-93. doi: 10.1016/j.chroma.2011.09.043. Epub 2011 Sep 21.
Simple and rapid quantitative determination of fatty-acid-based biofuels is greatly important for the study of genetic engineering progress for biofuels production by microalgae. Ideal biofuels produced from biological systems should be chemically similar to petroleum, like fatty-acid-based molecules including free fatty acids, fatty acid methyl esters, fatty acid ethyl esters, fatty alcohols and fatty alkanes. This study founded a gas chromatography-mass spectrometry (GC-MS) method for simultaneous quantification of seven free fatty acids, nine fatty acid methyl esters, five fatty acid ethyl esters, five fatty alcohols and three fatty alkanes produced by wild-type Synechocystis PCC 6803 and its genetically engineered strain. Data obtained from GC-MS analyses were quantified using internal standard peak area comparisons. The linearity, limit of detection (LOD) and precision (RSD) of the method were evaluated. The results demonstrated that fatty-acid-based biofuels can be directly determined by GC-MS without derivation. Therefore, rapid and reliable quantitative analysis of fatty-acid-based biofuels produced by wild-type and genetically engineered cyanobacteria can be achieved using the GC-MS method founded in this work.
简单快速地定量测定基于脂肪酸的生物燃料对于研究微藻生产生物燃料的基因工程进展非常重要。理想的生物燃料应在化学性质上与石油相似,如基于脂肪酸的分子,包括游离脂肪酸、脂肪酸甲酯、脂肪酸乙酯、脂肪醇和脂肪烷烃。本研究建立了一种气相色谱-质谱联用(GC-MS)方法,用于同时定量测定野生型集胞藻 PCC 6803 及其基因工程菌株产生的七种游离脂肪酸、九种脂肪酸甲酯、五种脂肪酸乙酯、五种脂肪醇和三种脂肪烷烃。使用内标峰面积比较对 GC-MS 分析获得的数据进行定量。评估了该方法的线性、检测限(LOD)和精密度(RSD)。结果表明,脂肪酸基生物燃料无需衍生即可直接通过 GC-MS 进行测定。因此,本研究建立的 GC-MS 方法可用于快速可靠地定量分析野生型和基因工程蓝藻产生的基于脂肪酸的生物燃料。