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Evaluation of densitometric quantitation of ether phospholipids by high performance thin layer chromatography

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This paper reports on the use of uni-dimensional high performance thin layer chromatography on silica gel for the separation and detection of ether phospholipids involving in situ reflectance densitometry. Dipping reagents for charring (cupric sulfate-phosphoric acid and manganese chloride-sulfuric acid) or fluorescence enhancement (1-anilino-8-naphthalene sulfonate) were evaluated. Two lipophilic fluorescence reagents (2,5-bis-5′-tert-butylbenzoxazolyl-(2′))thiophene and 4-(N,N-dihexadecyl)amino-7-nitrobenz-2-oxa-1,3-diazole) which move in the mobile phase were tested. The latter reagent gave the more sensitive and stable measurements: nanogram amounts of ether phospholipids could be detected in a reproducible way. Since alkyl lysophospholipids are highly polar molecules, high performance thin layer chromatography is a challenge. Characteristics of sorbents and influence of mobile phases on detection and separation are discussed for the NBD-dihexadecylamine fluorescence detection method.

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Abbreviations

Thio-ALP:

DL-alfa-Phosphatidyl-choline-beita-O-methoxy-methyl-gamma-S-hexadecyl

Et-18-0Me:

DL-alfa-Phosphatidyl-choline-beita-O-methyl-gamma-O-octadecyl

Et-16-0Me:

DL-alfa-Phosphatidyl-choline-beita-O-methyl-gamma-O-hexadecyl

PAF:

DL-alfa-phosphatidyl-choline-beita-acetyl-gamma-O-hexadecyl

PALM:

DL-alfa-phosphatidyl-choline-beita-palmitoyl-gamma-O-hexadecyl

ALP:

Alkyl lysophospholipid

8-ANSA:

1-anilino-8-naphthalene sulfonate

BBOT:

(2,5-bis-(5ø-tert-butylben-zoxazolyl-(2′))thiophene

NBD-dihexadecylamine:

4-(N,N-dihexadecyl)amino-7-nitrobenz-2-oxa-1,3-diazole

TEA:

triethylamine

HPTLC:

High Performance Thin Layer Chromatography

References

  1. CRC Handbook of Chromatography. Lipids, Volume I.H. K. Mangold, G. Zweig, J. Sherma (Eds.). CRC Press, Boca Raton, Florida, USA, 1984.

    Google Scholar 

  2. Lipid Chromatographic Analysis. Volume I.G. V. Marinetti (Ed.). Marcel Dekker Inc., New York, Basel, 1976.

    Google Scholar 

  3. Journal of Chromatographic Library. Volume 37. Chromatography of lipids in biomedical research and clinical diagnosis.A. Kuksis (Ed.). Elsevier, Amsterdam (1987).

    Google Scholar 

  4. W. W. Christie, Lipid Analysis. 2nd Edition. Pergamon, Oxford, 1982.

    Google Scholar 

  5. W. W. Christie, Z. Lebensm. Unters. Forsch.,181, 171 (1985).

    CAS  Google Scholar 

  6. J. C. Touchstone, J. G. Alvarez, J. Chromatogr. Biomed. Appl.,429, 359 (1988).

    CAS  Google Scholar 

  7. F. B. Banks, J. Chromatogr.,254, 247 (1983).

    Article  CAS  Google Scholar 

  8. L. Kolarovic, H. Cousin, H. Traitler, J. High. Res. Chrom. & Chrom. Com.,8, 838 (1985).

    CAS  Google Scholar 

  9. A. Kuksis, J. J. Myher, J. Chromatogr. Biomed. Appl.,379, 57 (1986).

    CAS  Google Scholar 

  10. K. Nakamura, S. Handa, Anal. Biochem.,142, 406 (1984).

    Article  CAS  Google Scholar 

  11. L. Zhou, H. Shanfield, A. Zlatkis, J. Chromatogr.,239, 259 (1982).

    CAS  Google Scholar 

  12. D. Wollbeck, E. v. Kleist, I. Elmadfa, W. Funk, J. High Res. Chrom. & Chrom. Com.,7, 473 (1984).

    CAS  Google Scholar 

  13. G. Schmitz, G. Assmann, J. Chromatogr.,307, 65 (1984).

    CAS  Google Scholar 

  14. L. J. Macata, R. K. Yu, S. Ando, J. Lipid Res.,24, 1243 (1983).

    Google Scholar 

  15. L. Kolarovic, N. C. Fournier, Anal. Biochem.,156, 244 (1986).

    Article  CAS  Google Scholar 

  16. L. Kolarovic, H. Traitler, J. High Res. & Chrom. Com.,8, 341 (1985).

    CAS  Google Scholar 

  17. J. K. Yao, G. M. Rastetter, Anal. Biochem.,150, 111 (1985).

    Article  CAS  Google Scholar 

  18. A. A. Entezami, B. J. Venables, K. E. Daugherty, J. Chromatogr.,387, 323 (1987).

    Article  CAS  Google Scholar 

  19. I. Rustenbeck, A. Trümmer, S. Lenzen, J. Planar Crom.,2, 207 (1989).

    CAS  Google Scholar 

  20. R. A. Heynemann, D. M. Bernard, R. E. Vercauteren, J. Chromatogr.,68, 285 (1972).

    Google Scholar 

  21. H. F. Larsen, A. G. Trostmann, J. Chromatogr. Biomed. Appl.,226, 484 (1981).

    CAS  Google Scholar 

  22. H. G. Schaefer, P. Rohdewald, Clin. Chem.,35,821 (1989).

    CAS  Google Scholar 

  23. R. Kraus, J. Wuthe, R. Rüfer, J. Chromatogr. Biomed. Appl.,413, 257 (1987).

    Article  CAS  Google Scholar 

  24. L. Colarow, J. Planar Chromatogr.,2, 19 (1989).

    CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratory of Pharmaceutical Chemistry and Drug Quality Control, State University of Ghent, Harelbekestraat 72, B-9000, Ghent, Belgium

    I. Coene, M. E. Ramos, E. Van den Eeckhout & W. Van den Bossche

  2. Laboratory for Organic Chemistry, State University of Ghent, B-9000, Ghent, Belgium

    P. Sandra

Authors
  1. I. Coene
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  2. M. E. Ramos
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  3. E. Van den Eeckhout
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  4. P. Sandra
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  5. W. Van den Bossche
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Coene, I., Ramos, M.E., Van den Eeckhout, E. et al. Evaluation of densitometric quantitation of ether phospholipids by high performance thin layer chromatography. Chromatographia 30, 414–420 (1990). https://doi.org/10.1007/BF02328508

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  • DOI: https://doi.org/10.1007/BF02328508

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