1 Gas chromatography-mass spectrometry demonstrated the presence of arachidonic acid (AA), 6-keto-prostaglandin Fl, and thromboxane B2 (TxB2) in all extracts of homogenized muscle or mucosa from human stomach, terminal ileum or sigmoid colon. Prostaglandin D2 (PGD2), PGE2 or PGF2. were usually found more often in the mucosal extracts. The 12-hydroxy-derivative of AA (12-HETE) was detected in all extracts of the colon but in only some of the other tissues. 2 Most prostanoids tested contracted the longitudinal muscle, the order of potency being U-466 19(an epoxymethano analogue of PGH2)>PGE2>PGF2,,>PGD2; PG12 usually caused relaxation, whereas its breakdown products or TxB2 had weak and variable effects.3 U-46619 or, less potently, PGF2a contracted the circular muscle, whereas PGI2 and usually PGE2 caused relaxation. PGD2, 6-keto-PGF1,,, 6,15-diketo-PGFir. or TxB2 usually had little or no effect.4 PGI2 antagonized contractions to some excitatory prostanoids, without greatly affecting contractions to acetylcholine. 5 For both muscle layers there was a gradient in sensitivity to prostanoids along the gastrointestinal tract. The sensitivities were stomach> distal ileum>sigmoid colon. 6 The results are discussed in relation to gastrointestinal physiology and pathophysiology.