Fresenius' Journal of Analytical Chemistry

, Volume 347, Issue 8–9, pp 376–381 | Cite as

Analysis of leukotriene B4 as free acid and its δ-lactone by high-performance liquid chromatography: a study on the stability and formation of leukotriene B4 δ-lactone

  • Dimitrios Tsikas
  • Joachim Fauler
  • Jürgen C. Frölich
Original Papers Biochemical/Clinical Analysis
Received:
Revised:

Summary

Reversed-phase high-performance liquid chromatography (RP-HPLC) with UV detection (270 nm) was applied to study the stability of LTB4 δ-lactone in aqueous solutions and its chemical formation from LTB4. LTB4 δ-lactone was found to be unstable in aqueous buffered solutions (pH range 4 to 8) and was converted in a pH and time depending manner mainly to LTB4 and most probably to an isomer of LTB4 δ-lactone. This compound was spontaneously converted to LTB4 δ-lactone when incubated in the buffers and exists in equilibrium with LTB4 δ-lactone at a molar ratio of about 1:1. Incubation of LTB4 in buffers (pH 4, 6 and 8) and human plasma did not result in formation of detectable amounts of LTB4 δ-lactone. Conversion of LTB4 into its δ-lactone form in a yield of 62±6% (mean ± SD, n=3) was achieved by treatment of LTB4 with a mixture of acetic acid, acetic acid anhydride and granular calcium chloride (1/5/3, v/v/w %) in ethylacetate. Evidence for LTB4δ-lactone formation from LTB4 under these conditions was demonstrated by gas chromatography-mass spectrometry.

Keywords

Calcium Acetic Acid Ethylacetate Anhydride Human Plasma 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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References

  1. 1.
    Samuelsson B (1983) Science 220:568Google Scholar
  2. 2.
    Smith MJH, Ford-Hutchinson AW, Bray MA (1980) J Pharm Pharmacol 32:517Google Scholar
  3. 3.
    Schröder JM (1990) In: Ruzicka T (ed) Eicosanoids and the skin. CRC, Boca Raton 7:79Google Scholar
  4. 4.
    Boeynaems JM, Brash AR, Oates JA, Hubbard WC (1980) Anal Biochem 104:259Google Scholar
  5. 5.
    Serafin WE, Oates JA, Hubbard WC (1984) Prostaglandins 27:899Google Scholar
  6. 6.
    Strife RJ, Murphy RC (1984) J Chromatogr 305:3Google Scholar
  7. 7.
    Falardeau P, Oates JA, Brash AR (1981) Anal Biochem 115:359Google Scholar
  8. 8.
    Powell WS, Gravelle F (1988) J Biol Chem 263:2170Google Scholar
  9. 9.
    Powell WS, Gravelle F (1989) J Biol Chem 264:5364Google Scholar
  10. 10.
    Fauler J, Marx KH, Kaever V, Frölich JC (1989) Prostagl Leukotr Essent Fatty Acids 37:193Google Scholar
  11. 11.
    Tsikas D, Fauler J, Frölich JC (1992) Eicosanoids 5 [Suppl]:7Google Scholar
  12. 12.
    Hughes H, Mitchell JR, Gaskell SJ (1989) Anal Biochem 179:304Google Scholar
  13. 13.
    Carey F, Forder RA (1986) Prostagl Leukotr Med 22:57Google Scholar
  14. 14.
    Carey F, Forder RA, Gibson KH, Haworth D (1989) Prostagl Leukotr Essent Fatty Acids 36:57Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Dimitrios Tsikas
    • 1
  • Joachim Fauler
    • 1
  • Jürgen C. Frölich
    • 1
  1. 1.Department of Clinical PharmacologyHannover Medical SchoolHannoverGermany

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