Rapid detection and quantification of triacylglycerol by HPLC-ELSD in chlamydomonas reinhardtii and chlorella strains

Naoko Kobayashi, Eric A. Noel, Austin Barnes, Julian Rosenberg, Concetta Dirusso, Paul Black, George A. Oyler

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1035-1049
Number of pages15
JournalLipids
Volume48
Issue number10
DOIs
StatePublished - Oct 2013

Keywords

  • Chlamydomonas reinhardtii
  • Chlorella
  • FAME
  • GC-MS
  • HPLC-ELSD
  • Lipid
  • Lipid class
  • TAG quantification
  • TLC

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Cell Biology

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