A rapid method for the estimation of prostaglandin E2 in intestinal tissues using fluorescence derivatization.
A sensitive spectrofluorimetric method is described to determine small quantities of prostaglandin E2 in complex biological systems as intestinal tissues. The method is based on a solid phase extraction combined with a coupling with a fluorescent marker and measuring the derivatization product by fluorescence densitometry. After mixing the tissue with an ice-cold perchloric acid solution, adjusting the pH, centrifugation and filtration steps, the prostaglandins are retained on a solid phase extraction C18 disposable column. They are eluted with diethylether, derivatized with 4-bromomethyl-7-methoxy-coumarin using potassium carbonate as condensating agent and finally analysed using fluorescence densitometry on silica gel TLC plates. Applying this method, amounts down to 5 ng (per gram wet tissue) could be measured in intestinal tissues, the s.e.m. for replicated total analysis being less than 15%. The foregoing method is applied for the determination of PGE2 released in the intestinal wall under the influence of laxatives.
Nijs
G
G
Laboratory of Pharmaceutical Biology, Katholieke Universiteit Leuven, Belgium.
de Witte
P
P
Lemli
J
J
D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D000880:Anthraquinones / Q000494:pharmacology; D002855:Chromatography, Thin Layer; D003720:Densitometry; D015232:Dinoprostone / Q000032:analysis*; D007421:Intestine, Small / Q000737:chemistry*; D008297:Male; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling; D000095322:Anthrones
D000873:Anthracenes; D000880:Anthraquinones; D015232:Dinoprostone; C055876:rhein-9-anthrone; D000095322:Anthrones; C020491:rhein