Analysis of polar lipids

Polar lipids are extracted from freeze dried cell material using a choroform:methanol:0.3% aqueous NaCl mixture, polar lipids are recovered into the chloroform phase (modified after Bligh and Dyer, 1959). When polar lipids and respiratory quinones should be analysed from the same biomass, a two stage method is used to first extract the respiratory quinones followed by the polar lipids (Tindall, 1990a,b).
Polar lipids are separated by two dimensional silica gel thin layer chromatography. The first direction is developed in chloroform:methanol:water, and the second in chloroform:methanol:acetic acid:water. Total lipid material is detected using molybdatophosphoric acid and specific functional groups detected using spray reagents specific for defined functional groups (Tindall et al., 2007). The labelled picture contributes the report.

Required material: For this method 200 mg freeze dried cells are needed. If the analysis of respiratory quinones is requested in parallel we need 200 mg freeze dried cells in total for both methods.

Dr. Susanne Verbarg

References

  1. Bligh, E.G. and Dyer, W.J. (1959). A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol., 37, 911-917
  2. Tindall, B.J. (1990a). A comparative study of the lipid composition of Halobacterium saccharovorum from various sources. Syst. Appl. Microbiol. 13, 128-130
  3. Tindall, B.J. (1990b). Lipid composition of Halobacterium lacusprofundi. FEMS Microbiol. Letts. 66, 199-202
  4. Tindall, B.J., Sikorski, J., Smibert ,R.M., and Kreig, N.R. (2007) Phenotypic characterization and the principles of comparative systematics. In Methods for General and Molecular Microbiology 3rd edn. Pp. 330-393.