Skip to main content
Log in

Spectrophotometric monitoring of lipoxygenase and cyclooxygenase pathway activity using ionophore stimulated whole blood

  • Mediators of Acute Inflammation
  • Published:
Agents and Actions Aims and scope Submit manuscript


We have employed clotting human blood stimulated with ionophore to develop a system for measuring cyclooxygenase, 5-lipoxygenase, and 12-lipoxygenase pathway products released into the serum fraction. In a single chromatographic run, 5-HETE, 12-HETE, 12-OH-heptadecatrienoic acid (HHT), LTB4, 20-OH-LTB4, and 20-COOH-LTB4 are quantitated by UV monitoring after separation by HPLC. The kinetics of product formation/release of all fatty acid products into the serum show an apparent lag of approximately 2 min, after which time the amounts of HHT, 5-HETE, and LTB4, respectively, plateau at 10 min while 12-HETE increases over a 60 min period. The system is responsive to standard cyclooxygenase and lipoxygenase inhibitors, and is of value of evaluating prospective blockers of AA metabolism in a whole blood setting.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others



arachidonic acid






dimethyl sulfoxide


hydroxyeicosatetraenoic acid


high performance liquid chromatography

IC50 :

concentration causing 50% inhibition



LTB4 :

leukotriene B4




nordihyroguaiaretic acid


nonsteroidal anti-inflammatory drug

PGB2 :

prostaglandin B2




trifluoroacetic acid


  1. F. Carey and R. A. Forder,Radioimmunoassay of LTB 4 and 6-trans-LTB 4:analytical and pharmacological characterization of immunoreactive LTB 4 in ionophore stimulated human blood. Prost. Leuk. Med.22, 57–70 (1986).

    Article  Google Scholar 

  2. P. Gresele, J. Arnout, M. C. Coene, H. Deckmyn and J. Vermylen,Leukotriene B 4 production by stimulated whole blood: comparative studies with isolated polymorphonuclear cells. Biochem. Biophys. Res. Commun.137, 334–342 (1986).

    Article  PubMed  Google Scholar 

  3. F. J. Sweeney, J. D. Eskra and T. J. Carty,Development of a system for evaluating 5-lipoxygenase inhibitors using human whole blood. Prostaglandins, Leukotrienes and Medicine (1987) in Press.

  4. J. D. Eskra, M. J. Pereira and M. J. Ernest,Solid phase extraction and high-performance liquid chromatography analysis of lipoxygenase products. Anal. Biochem.154, 332–337 (1986).

    Article  PubMed  Google Scholar 

  5. M. Hamberg and B. Samuelsson,Prostaglandin endoperoxides. Novel transformations of arachidonic acid in human platelets. Proc. Natl. Acad. Sci. (USA)71, 3400–3404 (1974).

    Google Scholar 

  6. P. Borgeat and B. Samuelsson,Arachidonic acid metabolism in polymorphonuclear leukocytes: Effects of ionophore A-23187. Proc. Natl. Acad. Sci. (USA)76, 2148–2152 (1979).

    Google Scholar 

  7. M. Koller, W. Schonfeld, J. Knoller, K.-D. Bremm, W. Konig, B. Spur, A. Crea and W. Peters,The metabolism of leukotrienes in blood plasma studied by high performance liquid chromatography. Biochim. Biophys. Acta833, 128–134 (1985).

    PubMed  Google Scholar 

  8. T. J. Carty, J. S. Stevens, J. G. Lombardino, M. J. Parry and M. J. Randall,Piroxicam, a structurally novel anti-inflammatory compound. Mode of prostaglandin synthesis inhibition. Prostaglandins19, 671–682 (1980).

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations


Rights and permissions

Reprints and permissions

About this article

Cite this article

Sweeney, F.J., Eskra, J.D., Ernest, M.J. et al. Spectrophotometric monitoring of lipoxygenase and cyclooxygenase pathway activity using ionophore stimulated whole blood. Agents and Actions 21, 393–396 (1987).

Download citation

  • Issue Date:

  • DOI: