Combined Reversed Phase HPLC, Mass Spectrometry, and NMR Spectroscopy for a Fast Separation and Efficient Identification of Phosphatidylcholines

Overview

Journal J Biomed Biotechnol

Specialty Biology

Date 2010 Sep 28

PMID 20871812

Citations 5

Authors

Jan Willmann

Herbert Thiele

Dieter Leibfritz

Affiliations

Soon will be listed here.

Abstract

In respect of the manifold involvement of lipids in biochemical processes, the analysis of intact and underivatized lipids of body fluids as well as cell and tissue extracts is still a challenging task, if detailed molecular information is required. Therefore, the advantage of combined use of high-pressure liquid chromatography (HPLC), mass spectrometry (MS), and nuclear magnetic resonance (NMR) spectroscopy will be shown analyzing three different types of extracts of the ubiquitous membrane component phosphatidylcholine. At first, different reversed phase modifications were tested on phosphatidylcholines (PC) with the same effective carbon number (ECN) for their applicability in lipid analysis. The results were taken to improve the separation of three natural PC extract types and a new reversed phase (RP)-HPLC method was developed. The individual species were characterized by one- and two-dimensional NMR and positive or negative ion mode quadrupole time of flight (q-TOF)-MS as well as MS/MS techniques. Furthermore, ion suppression effects during electrospray ionisation (ESI), difficulties, limits, and advantages of the individual analytical techniques are addressed.

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References

Silversand C, Haux C . Improved high-performance liquid chromatographic method for the separation and quantification of lipid classes: application to fish lipids. J Chromatogr B Biomed Sci Appl. 1998; 703(1-2):7-14. DOI: 10.1016/s0378-4347(97)00385-x. View

Olsson N, Salem Jr N . Molecular species analysis of phospholipids. J Chromatogr B Biomed Sci Appl. 1997; 692(2):245-56. DOI: 10.1016/s0378-4347(96)00507-5. View

Wang C, Kong H, Guan Y, Yang J, Gu J, Yang S . Plasma phospholipid metabolic profiling and biomarkers of type 2 diabetes mellitus based on high-performance liquid chromatography/electrospray mass spectrometry and multivariate statistical analysis. Anal Chem. 2005; 77(13):4108-16. DOI: 10.1021/ac0481001. View

Kan I, Melamed E, Offen D, Green P . Docosahexaenoic acid and arachidonic acid are fundamental supplements for the induction of neuronal differentiation. J Lipid Res. 2006; 48(3):513-7. DOI: 10.1194/jlr.C600022-JLR200. View

Sullentrop F, Moka D, Neubauer S, Haupt G, Engelmann U, Hahn J . 31P NMR spectroscopy of blood plasma: determination and quantification of phospholipid classes in patients with renal cell carcinoma. NMR Biomed. 2002; 15(1):60-8. DOI: 10.1002/nbm.758. View

Kim H, Wang T, Ma Y . Liquid chromatography/mass spectrometry of phospholipids using electrospray ionization. Anal Chem. 1994; 66(22):3977-82. DOI: 10.1021/ac00094a020. View

Ma H, Chou C, Wei S, Eaton D . Regulation of the epithelial sodium channel by phosphatidylinositides: experiments, implications, and speculations. Pflugers Arch. 2007; 455(1):169-80. DOI: 10.1007/s00424-007-0294-3. View

Bougnoux P, Chajes V, Lanson M, Hacene K, Body G, Couet C . Prognostic significance of tumor phosphatidylcholine stearic acid level in breast carcinoma. Breast Cancer Res Treat. 1992; 20(3):185-94. DOI: 10.1007/BF01834624. View

Adachi J, Yoshioka N, Sato M, Nakagawa K, Yamamoto Y, Ueno Y . Detection of phosphatidylcholine oxidation products in rat heart using quadrupole time-of-flight mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2005; 823(1):37-43. DOI: 10.1016/j.jchromb.2005.03.035. View

10.

Berridge M . Inositol trisphosphate and calcium signalling. Nature. 1993; 361(6410):315-25. DOI: 10.1038/361315a0. View

11.

Schiller J, Arnold K . Application of high resolution 31P NMR spectroscopy to the characterization of the phospholipid composition of tissues and body fluids - a methodological review. Med Sci Monit. 2002; 8(11):MT205-22. View

12.

Kafrawy O, Zerouga M, Stillwell W, Jenski L . Docosahexaenoic acid in phosphatidylcholine mediates cytotoxicity more effectively than other omega-3 and omega-6 fatty acids. Cancer Lett. 1999; 132(1-2):23-9. DOI: 10.1016/s0304-3835(98)00163-3. View

13.

TOUCHSTONE J . Thin-layer chromatographic procedures for lipid separation. J Chromatogr B Biomed Appl. 1995; 671(1-2):169-95. DOI: 10.1016/0378-4347(95)00232-8. View

14.

Silva J, Dasgupta S, Wang G, Krishnamurthy K, Ritter E, Bieberich E . Lipids isolated from bone induce the migration of human breast cancer cells. J Lipid Res. 2006; 47(4):724-33. DOI: 10.1194/jlr.M500473-JLR200. View

15.

Jeon E, Song H, Kim M, Moon H, Bae Y, Jung J . Sphingosylphosphorylcholine induces proliferation of human adipose tissue-derived mesenchymal stem cells via activation of JNK. J Lipid Res. 2005; 47(3):653-64. DOI: 10.1194/jlr.M500508-JLR200. View

16.

Stubbs C, Smith A . The modification of mammalian membrane polyunsaturated fatty acid composition in relation to membrane fluidity and function. Biochim Biophys Acta. 1984; 779(1):89-137. DOI: 10.1016/0304-4157(84)90005-4. View

17.

Meier S, Mjos S, Joensen H, Grahl-Nielsen O . Validation of a one-step extraction/methylation method for determination of fatty acids and cholesterol in marine tissues. J Chromatogr A. 2005; 1104(1-2):291-8. DOI: 10.1016/j.chroma.2005.11.045. View

18.

Brugger B, Erben G, Sandhoff R, Wieland F, Lehmann W . Quantitative analysis of biological membrane lipids at the low picomole level by nano-electrospray ionization tandem mass spectrometry. Proc Natl Acad Sci U S A. 1997; 94(6):2339-44. PMC: 20089. DOI: 10.1073/pnas.94.6.2339. View

19.

Merchant T, Kasimos J, Vroom T, de Bree E, Iwata J, de Graaf P . Malignant breast tumor phospholipid profiles using (31)P magnetic resonance. Cancer Lett. 2002; 176(2):159-67. DOI: 10.1016/s0304-3835(01)00780-7. View

20.

EXTON J . Phosphatidylcholine breakdown and signal transduction. Biochim Biophys Acta. 1994; 1212(1):26-42. DOI: 10.1016/0005-2760(94)90186-4. View