Studies suggest a variety of biological effects of soybean isoflavones, but there is little information regarding small intestinal absorption and metabolism. The aim of this study was to investigate intestinal handling of luminally administered soybean-based tofu in an isolated preparation of the luminally and vascularly perfused rat small intestine (male Sprague-Dawley, approximately 45 d old). A synthetic emulsion free from blood components was used as vascular medium, with a perfluorocarbon as oxygen carrier. Luminal media consisted of tofu, predigested with pepsin and pancreatin and emulsified with bile acids, containing 39. 5 micromol/L genistein compounds and 19.1 micromol/L daidzein compounds. Viability of the organ preparation was maintained during the entire perfusion, confirmed by lack of significant differences between tofu and control perfusion experiments for arterial pressure, glucose consumption, oxygen uptake, lactate-pyruvate ratio and acid-base homeostasis. Daidzein (8.9%) and genistein (8.0%) compounds from tofu exhibited almost the same (P: > 0.05) absorption rate during small intestinal passage. The majority of the absorbed genistin appeared vascularly as genistein (4.4%), in addition to minor amounts of unchanged genistin (2.1%) and genistein glucuronide (1.5%). In the luminal effluent, a considerable increase of genistein (338%) as well as daidzein (190%) as cleavage products of the glucosides and malonyl-glucosides was observed. The distribution of daidzein compounds in the small intestine was not different from that of genistein compounds (P: > 0.05), except for the blood vessels, which had extremely low total amounts. Sulfate derivatives of genistein and daidzein compounds were not detectable. An effect of tofu ingredients was observed on absorption rate of genistin, on glucuronidation and on distribution of genistein glucuronide in the intestine.