Limit of detection: 80% B/B₀: 5.5 pg/ml · Sensitivity: 50% B/B₀: 32 pg/ml · Prostaglandin D₂ (PGD₂) is the major eicosanoid released from stimulated mast cells. PGD₂ is an unstable compound which is rapidly metabolized and eliminated from the circulation. This makes quantitation of PGD₂ an unreliable indicator of its in vivo production. 11β-PGF_2α is the primary plasma metabolite of PGD₂ in vivo, the levels of which can increase from 6 pg/ml in a normal healthy volunteer to 490 ng/ml in patients with systemic mastocytosis.¹ The normal human urinary excretion of 11β-PGF_2α is about 11 ng/mmol creatinine (~400 ng/24 hr), which is increased nearly 3-fold upon allergen-induced bronchoconstriction in asthmatics.^2,3 Unlike most prostaglandin metabolites, 11β-PGF_2α retains potent biological activity. 11-β-PGF_2α is equipotent to PGF_2α in inducing human bronchial smooth muscle contractions and inhibition of adipose differentiation.^4,5 11β-PGF_2α was also shown to inhibit ADP or thrombin-induced human platelet aggregation at concentrations of 0.14 to 2.8 µM. ⁶ The Cayman 11β-PGF_2α Assay is a competitive assay that provides accurate measurements of 11β-PGF_2α within the range of 7.8-1,000 pg/ml. Inter and intra-assay CV’s of less than 15% can be achieved at most concentrations. This assay allows sensitive detection of 11β-PGF_2α in the most common sample matrix, which is urine. Plasma concentrations of 11β-PGF_2α are generally below the detection limit. For plasma and other complex sample matrices, we recommend purification of the sample prior to analysis; a purification protocol is included in the kit booklet. Measurements of 11β-PGF_2α in urine using our EIA gives values in the same range, but slightly higher than those obtained by GC-MS. This appears to be due to measurement of 11β-PGF_2α plus the 2,3-dinor metabolite of 11β-PGF_2α.²

1 Liston, T.E., and Roberts, L.J. Transformation of prostaglandin D₂ to 9α,11β-(15S)-trihydroxy-prosta-(5Z,13E)-dien-1-oic acid (9α,11β-prostaglandin F2): A unique biologically active prostaglandin produced enzymatically in vivo in humans. Proc Natl Acad Sci USA 82 6030-6034 (1985).

2 O'Sullivan, S., Mueller, M.J., Dahlén, S., et al. Analyses of prostaglandin D₂ metabolites in urine: Comparison between enzyme immunoassay and negative ion chemical ionization gas chromatography-mass spectrometry. Prostaglandins Other Lipid Mediat 57 149-165 (1999).

3 O'Sullivan, S., Dahlén, B., Dahlén, S., et al. Increased urinary excretion of the prostaglandin D₂ metabolite 9α,11β-prostaglandin F2 after aspirin challenge supports mast cell activation in aspirin-induced airway obstruction. J Allergy Clin Immunol 98 421-432 (1996).

4 Coleman, R.A., and Sheldrick, R.L.G. Prostanoid-induced contraction of human bronchial smooth muscle is mediated by TP-receptors. Br J Pharmacol 96 688-692 (1989).

5 Lepak, N.M., and Serrero, G. Inhibition of adipose differentiation by 9α, 11β-prostaglandins F_2α. Prostaglandins 46 511-517 (1993).

6 Pugliese, G., Spokas, E.G., Marcinkiewicz, E., et al. Hepatic transformation of prostaglandin D₂ to a new prostanoid, 9α,11β-prostaglandin F2, that inhibits platelet aggregation and constricts blood vessels. J Biol Chem 260 14621-14625 (1985).

Liston, T.E., and Roberts, L.J. Transformation of prostaglandin D₂ to 9α,11β-(15S)-trihydroxy-prosta-(5Z,13E)-dien-1-oic acid (9α,11β-prostaglandin F2): A unique biologically active prostaglandin produced enzymatically in vivo in humans. Proc Natl Acad Sci USA 82 6030-6034 (1985).

Pugliese, G., Spokas, E.G., Marcinkiewicz, E., et al. Hepatic transformation of prostaglandin D₂ to a new prostanoid, 9α,11β-prostaglandin F2, that inhibits platelet aggregation and constricts blood vessels. J Biol Chem 260 14621-14625 (1985).

Coleman, R.A., and Sheldrick, R.L.G. Prostanoid-induced contraction of human bronchial smooth muscle is mediated by TP-receptors. Br J Pharmacol 96 688-692 (1989).

Maxey, K.M., Maddipati, K.R., and Birkmeier, J. Interference in enzyme immunoassays. J Clin Immunoassay 15 116-120 (1992).

Pradelles, P., Grassi, J., and Maclouf, J.A. Enzyme immunoassays of eicosanoids using acetylcholinesterase as label: An alternative to radioimmunoassay. Anal Chem 57 1170-1173 (1985).

Lepak, N.M., and Serrero, G. Inhibition of adipose differentiation by 9α, 11β-prostaglandins F_2α. Prostaglandins 46 511-517 (1993).

O'Sullivan, S., Dahlén, B., Dahlén, S., et al. Increased urinary excretion of the prostaglandin D₂ metabolite 9α,11β-prostaglandin F2 after aspirin challenge supports mast cell activation in aspirin-induced airway obstruction. J Allergy Clin Immunol 98 421-432 (1996).

Maclouf, J., Grassi, J., and Pradelles, P. Development of enzyme-immunoassay techniques for the measurement of eicosanoids. 355-364 (1987).

O'Sullivan, S., Mueller, M.J., Dahlén, S., et al. Analyses of prostaglandin D₂ metabolites in urine: Comparison between enzyme immunoassay and negative ion chemical ionization gas chromatography-mass spectrometry. Prostaglandins Other Lipid Mediat 57 149-165 (1999).

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