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Simultaneous analysis of polar and non-polar components of cell membrane phospholipids by GC-MS

  • Václav M. Bierhanzl EMAIL logo , Miroslava Bursová , Martin Ston , Radomír Čabala and Gabriela Seydlová
Published/Copyright: June 25, 2016
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Chemical Papers
From the journal Chemical Papers Volume 70 Issue 10

Abstract

A rapid and selective GC-MS method was optimised for the simultaneous analysis of fatty acids, phosphoethanolamine, phosphoglycerol and phosphoserine, which comprise the polar headgroups of phospholipids. Enzymatic hydrolysis was used for sample preparation to selectively release fatty acids and prevent cleavage of the phosphoester bonds of the polar headgroups. A two-step consecutive derivatisation of the fatty acids and polar headgroups was applied to enable the simultaneous analysis of the selected analytes. The method was tested on samples of cell membrane phospholipids of Bacillus subtilis, a microbe with a broad spectrum of fatty acids. This approach can accelerate and simplify biotechnological research and quality control. The experimental conditions were optimised using a chemometric approach denoted as experimental design.

Keywords: bacterial membrane; branched-chain fatty acids; GC-MS; phospholipids; lipidomics; silylation

Acknowledgements

This work receive financial support from grant nos. SVV260317 and 13-18051P from the Czech Science Foundation.

Supplementary data

Supplementary data associated with this article can be found in the online version of this paper (DOI: 10.1515/chempap-2016-0073).

References

Armstrong, R. A., Eperjesi, F., & Gilmartin, B. (2002). The application of analysis of variance (ANOVA) to different experimental designs in optometry. Ophtalmic and physiological optics, 22, 248–256. 10.1046/j.1475-1313.2002.00020.x.Search in Google Scholar

Bierhanzl, V. M., Čabala, R., Ston, M., Kubinec, R., Henger-ics Szabó, A., & Podolec, P. (2015). Gas chromatography with mass spectrometry analysis of phosphoserine, phospho-ethanolamine, phosphoglycerol, and phosphate. Journal of Separation Science, 38, 67–72. 10.1002/jssc.201400657.Search in Google Scholar

Boumann, H. A., Hopmans, E. C., van de Leemput, I., Op den Camp, H. J. M., van de Vossenberg, J., Strous, M., Jetten, M. S. M., Sinninghe Damsté, J. S., & Schouten, S. (2006). Lad-derane phospholipids inanammox bacteria comprise phos-phocholine and phosphoethanolamine headgroups. FEBS Microbiological Letters, 258, 297–304. 10.1111/j.1574-6968.2006.00233.x.Search in Google Scholar

Butovich, I. A., Uchiyama, E., Di Pascuale, M. A., & McCul-ley, J. P. (2007). Liquid chromatography–mass spectrometric analysis of lipids present in human meibomian gland secretions. Lipids, 42, 765–776. 10.1007/s11745-007-3080-2.Search in Google Scholar

Caboni, M. F., & Lercker, G. (1984). Analysis of phospholipids in cow’s milk by high-temperature injection gas chromatography and high-performance liquid chromatography. Journal of Chromatography A, 315, 223–231. 10.1016/s0021-9673(01)90739-3.Search in Google Scholar

Costa, N. R., & Pereira, Z. L. (2010). Multiple response optimization: a global criterion-based metod. Journal of Chemometrics, 24, 333–342. 10.1002/cem.1312.Search in Google Scholar

Fuchs, B., Schiller, J., Su¨ß, R., Schu¨renberg, M., & Suckau, D. (2007). A direct and simple method of coupling matrix-assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS) to thin-layer chromatography (TLC) for the analysis of phospholipids from egg yolk. Analytical and Bioanalytical Chemistry, 389, 827–834. 10.1007/s00216-007-1488-4.Search in Google Scholar

García-Cañas, V., Simó, C., Herrero, M., Ibá˜nez, E., & Ci-fuentes, A. (2012). Present and future challenges in food analysis, foodomics. Analytical Chemistry, 84, 10150–10159. 10.1021/ac301680q.Search in Google Scholar

Hibbert, D. B. (2012). Experimental design in chromatography: A tutorial review. Journal of Chromatography B, 910, 2–13. 10.1016/j.jchromb.2012.01.020.Search in Google Scholar

Kawai, F., Shoda, M., Harashima, R., Sadaie, Y., Hara, H., & Matsumoto, K. (2004). Cardiolipin domains in Bacillus subtilis Marburg membranes. Journal of Bacteriology, 186, 1475–1483. 10.1128/jb.186.5.1475-1483.2004.Search in Google Scholar

Li, Y., Wu, S., Wang, L., Li, Y., Shi, F., & Wang, X. (2010). Differentiation of bacteria using fatty acids profiles from gas chromatography-tandem mass spectrometry. Journal of Science of Food and Agriculture, 90, 1380–1383. 10.1002/jsfa.3931.Search in Google Scholar

Myers, R. H., Montgomery, D. C., & Anderson-Cook, C. M. (2009). Response surface methodology, process and product optimization using designed experiment. New York, NY, USA: John Wiley & Sons.Search in Google Scholar

Ostrowska, J., Skrzydlewska, E., & Figaszewski, Z. A. (2000). Isolation and analysis of phospholipids. Chemia Analyticzna, 45, 613–629.Search in Google Scholar

Podolec, P., Hengerics Szabó, A., Blaško, J., Kubinec, R., Górová, R., Višňovský, J., Gnipová, A., Horváth, A., Bier-hanzl, V., Hložek, T., & Čabala, R. (2014). Direct silylation of Trypanosoma brucei metabolites in aqueous samples and their GC–MS/MS analysis. Journal of Chromatography B, 967, 134–138. 10.1016/j.jchromb.2014.07.023.Search in Google Scholar

Ran-Ressler, R. R., Lawrence, P., & Brenna, J. T. (2012). Structural characterization of saturated branched chain fatty acid methyl esters by collisional dissociation of molecular ions generated by electron ionization. Journal of Lipid Research, 53, 195–203. 10.1194/jlr.d020651.Search in Google Scholar

Rastogi, A., Hutchinson, T. E., & Pereira, B. M. J. (2005). Phospholipase C from two bacterial strains acts differently on pure phospholipids and membrane bound glycosylphos-phatidylinositol (GPI) anchors. Indian Journal of Biochemistry and Biophysics, 42(2), 92–99.Search in Google Scholar

Ryan, T. P. (2007). Modern experimental design. New York, NY, USA: John Wiley & Sons.Search in Google Scholar

Seppänen-Laakso, T., Laakso, I., & Hiltunen, R. (2002). Analysis of fatty acids by gas chromatography, and its relevance to research on health and nutrition. Analytica Chimica Acta, 465, 39–62. 10.1016/s0003-2670(02)00397-5.Search in Google Scholar

Seydlová, G., & Svobodová, J. (2008). Development of membrane lipids in the surfactin producer Bacillus subtilis. Folia Microbiologica, 53, 303–307. 10.1007/s12223-008-0047-5.Search in Google Scholar

Seydlová, G., Halada, P., Fišer, R., Toman, O., Ulrych, A., & Svobodová, J., (2012). DnaK and GroEL chaperones are recruited to the Bacillus subtilis membrane after short-term ethanol stress Journal of Applied Microbiology, 112, 765–774. 10.1111/j.1365-2672.2012.05238.x.Search in Google Scholar

Seydlová, G., Fišer, R., Čabala, R., Kozlík, P., Svobodová, J., & Pátek, M. (2013), Surfactin production enhances the level of cardiolipin in the cytoplasmic membrane of Bacillus subtilis. BBA Biomembranes, 1828, 2370–2378. 10.1016/j.bbamem.2013.06.032.Search in Google Scholar

Youseff, M. F. M., Xu, X., & Guo, Z. (2012). Comparison of fatty acid methyl and ethyl esters as biodiesel base stock, a review on processing and production requirements. Journal of the American Oil Chemists Society, 91, 525–531. 10.1007/s11746-014-2443-0.Search in Google Scholar

Received: 2015-9-17
Revised: 2016-3-10
Accepted: 2016-3-11
Published Online: 2016-6-25
Published in Print: 2016-10-1

© 2016 Institute of Chemistry, Slovak Academy of Sciences

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https://doi.org/10.1515/chempap-2016-0073
Keywords for this article
bacterial membrane; branched-chain fatty acids; GC-MS; phospholipids; lipidomics; silylation

Articles in the same Issue

  1. Original Paper
  2. Simultaneous analysis of polar and non-polar components of cell membrane phospholipids by GC-MS
  3. Original Paper
  4. Cloud point extraction of disulfiram for its HPLC-MS/MS determination in synthetic urine
  5. Original Paper
  6. Revealing the seed proteome of the health benefitting grain amaranth (Amaranthus cruentus L.)
  7. Original Paper
  8. Possible role of hydrolytic enzymes (Sap, Kex2) in Candida albicans response to aromatic compounds bearing a sulfone moiety
  9. Original Paper
  10. Using nutritional and oxidative stress to increase content of healthbeneficial fatty acids in oleaginous and non-oleaginous yeasts
  11. Original Paper
  12. Fatty acids and amino acids of entomopathogenic fungus Conidiobolus coronatus grown on minimal and rich media
  13. Original Paper
  14. Promotional effect of cobalt addition on catalytic performance of Ce0.5Zr0.5O2 mixed oxide for diesel soot combustion
  15. Original Paper
  16. Microwave-assisted continuous reactive distillation process for preparation of ethyl acetate
  17. Original Paper
  18. Wall-retardation effects on particles settling through non-Newtonian fluids in parallel plates
  19. Original Paper
  20. Microwave-assisted decomposition of fgd gypsum in the presence of magnetite and anthracite
  21. Original Paper
  22. Effect of PHB on the properties of biodegradable PLA blends
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  25. Short Communication
  26. UV-induced reduction of Ag+ by diazene sulphonates: new method of metallisation of surfaces
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