Abstract
We present a method for the determination of triacylglycerol (TAG) profiles of oleaginous saltwater microalgae relevant for the production of biofuels, bioactive lipids, and high-value lipid-based chemical precursors. We describe a technique to remove chlorophyll using quick, simple solid phase extraction (SPE) and directly compare the intact TAG composition of four microalgae species (Phaeodactylum tricornutum, Nannochloropsis salina, Nannochloropsis oculata, and Tetraselmis suecica) using MALDI time-of-flight (TOF) mass spectrometry (MS), ESI linear ion trap-orbitrap (LTQ Orbitrap) MS, and ¹H NMR spectroscopy. Direct MS analysis is particularly effective to compare the polyunsaturated fatty acid (PUFA) composition for triacylglycerols because oxidation can often degrade samples upon derivatization. Using these methods, we observed that T. suecica contains significant PUFA levels with respect to other microalgae. This method is applicable for high-throughput MS screening of microalgae TAG profiles and may aid in the commercial development of biofuels.
Highlights
We present a method for the determination of triacylglycerol (TAG) profiles of oleaginous saltwater microalgae relevant for the production of biofuels, bioactive lipids, and high-value lipid-based chemical precursors
Microalgae have been identified as an attractive source for the production of biofuels in addition to bioactive lipids and high-value, lipid-based chemical precursors based on their ability to produce 20% or more of their mass as oil
We describe the use of 1H nuclear magnetic resonance (NMR) spectroscopy as a complementary method to monitor lipid extraction, the relative TAG purity, and the relative polyunsaturated fatty acid (PUFA) ratio based on integration and chemical shift of diagnostic TAG protons
Summary
Olive oil, used as a positive control for triacylglycerols, as well as 2,5-dihydroxybenzoic acid (DHB), was purchased from ACROS Organics. Solvents (ACS reagent-grade hexanes, methanol, acetic acid, diethyl ether, and chloroform), and acetate salts (LiOAc and NaOAc; >99% purity) were purchased from Sigma-Aldrich. SiliaPrep silica solid phase extraction (SPE) columns (Silicycle 500 mg, 3 ml capacity) were selected because they provided optimal separation of chlorophyll a from lipid samples. Deuterated chloroform (99.8%) was purchased from Cambridge Isotope Laboratories
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