(567u) Development of Novel Method to Determine Triglyceride Content in Microalgae: Dunaliella Tertiolecta

Microalgal based biofuels have the potential to significantly supplement, if not completely replace, petroleum-derived transport fuels without adversely affecting the supply of food and other crop products. Microalgae can accumulate a large amount of intracellular lipid, especially non-polar triglycerides (TAGs), which are the best substrate for producing biodiesel. However, the method to determine triglyceride content in algal cells has not been well developed. In this research, fluorescence stain: Nile Red (NR) has been used to determine the lipid content in algal cells. Since NR can stain both triglyceride and phospholipids, we provide a novel model to specifically differentiate the triglyceride from phospholipids by measuring the NR fluorescence intensity at both 595nm and 635 nm. Two controls: triolein and phosphocholine have been used in this model. In this study, we combine 96-well flow cytometry and 96-well microplate fluorescence assay to quantify triglyceride content in marine microalgae: Dunaliella tertiolecta. Triglyceride content has been determined in various normal and stress growth conditions. We not only determine the triglyceride content but found the cell percentage that has triglyceride. Results of this research can be applied to monitor the lipid content and determine the growth condition to maximize production of microalgal triglyceride.