Triacylglycerol profiling of microalgae Chlamydomonas reinhardtii and Nannochloropsis oceanica

Triacylglycerols (TAGs) from microalgae can serve as feedstock for the production of biofuels. To gain a comprehensive understanding of TAG metabolism in algae through genetic and molecular approaches, and to improve algal biofuel production, efficient and quantitative phenotyping methods focusing on TAGs are required. Towards this end, a facile ultrahigh performance liquid chromatography–mass spectrometry protocol was developed for TAG profiling, achieving identification and quantification of intact TAG molecular species in two algae. TAG profiling was performed in Chlamydomonas reinhardtii and Nannochloropsis oceanica grown in nitrogen (N)-replete or N-depleted medium. For the quantification of algal TAGs and fatty acids, two sets of internal standards were developed by taking advantage of the presence of pheophytin and specific fatty acids in algal samples. Comparison of algal TAG levels was simplified by using these internal standards for TAG analysis, paving the way for high-throughput mutant screening.