Home › Botany › Arabidopsis thaliana Polar Glycerolipid Profiling by Thin Layer Chromatography (TLC) Coupled with Gas-Liquid Chromatography (GLC)
Botany JoVE (Open Access) Citable · DOI

Arabidopsis thaliana Polar Glycerolipid Profiling by Thin Layer Chromatography (TLC) Coupled with Gas-Liquid Chromatography (GLC)

DOI: 10.3791/2518-v
What you'll learn
  • ✓ Extract and isolate polar glycerolipids from plant tissue using TLC separation
  • ✓ Convert fatty acyl groups to methyl esters via transmethylation
  • ✓ Quantify fatty acid composition using gas-liquid chromatography
  • ✓ Interpret lipid profiling results from combined TLC-GLC workflow
Protocol

Composition of polar lipid extracts and the fatty acid composition of individual glycerolipids are determined in a simple and robust lipid profiling experiment. For this purpose, glycerolipids are isolated by thin layer chromatography and subjected to transmethylation of their acyl groups. Fatty acyl methylesters are quantified by gas-liquid chromatography.

Difficulty
intermediate
Total time
~3–4 days (including extraction, chromatography runs, and analysis)
Model organism
Arabidopsis thaliana

Steps

1
Extract polar lipids from plant tissue

Prepare plant samples and perform solvent extraction of total polar lipids. Detailed procedures for tissue preparation and extraction solvent selection are outlined.

▶ 01:50
2
Separate glycerolipids by thin layer chromatography

Apply extracted lipids to TLC plates and run with appropriate solvent systems to resolve individual glycerolipid classes (e.g., PC, PE, PG, MGDG, DGDG). Identify lipid spots for downstream analysis.

▶ 02:49
3
Convert lipid acyl groups to fatty acid methyl esters

Scrape isolated TLC spots and perform transmethylation reaction to convert fatty acyl chains into volatile methyl ester derivatives suitable for GLC analysis.

▶ 06:21
4
Quantify fatty acids by gas-liquid chromatography

Inject fatty acid methyl ester samples into GLC and detect individual fatty acid components by flame ionization or mass spectrometry. Generate chromatograms for quantification.

▶ 08:29
5
Analyze and interpret lipid profiling results

Compare representative chromatograms and results showing polar lipid composition and fatty acid distributions across different glycerolipid classes in Arabidopsis samples.

▶ 10:21
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