title: Rapid detection and quantification of triacylglycerol by HPLC-ELSD in Chlamydomonas reinhardtii and Chlorella strains.
abstract: Triacylglycerol (TAG) analysis and quantification are commonly performed by first obtaining a purified TAG fraction from a total neutral lipid extract using thin-layer chromatography (TLC), and then analyzing the fatty acid composition of the purified TAG fraction by gas chromatography (GC). This process is time-consuming, labor intensive and is not suitable for analysis of small sample sizes or large numbers. A rapid and efficient method for monitoring oil accumulation in algae using high performance liquid chromatography for separation of all lipid classes combined with detection by evaporative light scattering (HPLC-ELSD) was developed and compared to the conventional TLC/GC method. TAG accumulation in two Chlamydomonas reinhardtii (21 gr and CC503) and three Chlorella strains (UTEX 1230, CS01 and UTEX 2229) grown under conditions of nitrogen depletion was measured. The TAG levels were found to be 3-6 % DW (Chlamydomonas strains) and 7-12 % DW (Chlorella strains) respectively by both HPLC-ELSD and TLC/GC methods. HPLC-ELSD resolved the major lipid classes such as carotenoids, TAG, diacylglycerol (DAG), free fatty acids, phospholipids, and galactolipids in a 15-min run. Quantitation of TAG content was based on comparison to calibration curves of trihexadecanoin (16:0 TAG) and trioctadecadienoin (18:2 TAG) and showed linearity from 0.2 to 10 μg. Algal TAG levels >0.5 μg/g DW were detectable by this method. Furthermore TAG content in Chlorella kessleri UTEX 2229 could be detected. TAG as well as DAG and TAG content were estimated at 1.6 % DW by HPLC-ELSD, while it was undetectable by TLC/GC method.
authors: Department of Biochemistry, University of Nebraska-Lincoln, 1901 Vine Street, Lincoln, NE, 68588, USA.
authors: Naoko
authors: N
authors: Kobayashi
authors: Eric A
authors: EA
authors: Noel
authors: Austin
authors: A
authors: Barnes
authors: Julian
authors: J
authors: Rosenberg
authors: Concetta
authors: C
authors: DiRusso
authors: Paul
authors: P
authors: Black
authors: George A
authors: GA
authors: Oyler
mesh_terms: D016825:Chlamydomonas reinhardtii / Q000737:chemistry*; D002708:Chlorella / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D008401:Gas Chromatography-Mass Spectrometry; D008027:Light; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D013997:Time Factors; D014280:Triglycerides / Q000032:analysis* / Q000737:chemistry
chemical_list: D014280:Triglycerides
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