Intestinal absorption of rutin in free and conjugated forms.

Quercetin is one of the most common flavonoids in nature, occurring mainly in glycosidic forms such as rutin. Rutin has been reported to exert numerous biochemical and pharmacological activities, though information about its absorption and metabolism is scarce. The aim of this study was to investigate intestinal handling of luminally administered rutin in an isolated preparation of luminally and vascularly perfused rat small intestine. A synthetic perfusate free from blood components was used as vascular medium, with a perfluorocarbon as oxygen carrier. Luminal media consisted of a bicarbonate-buffered sodium chloride solution spiked with rutin (40.5 +/- 1.8 micromol/L). Viability was maintained during the entire perfusion; no differences between rutin and control perfusions for perfusion pressure, lactate-pyruvate ratio, oxygen uptake, and acid-base homeostasis were observed. About 10% of the administered rutin appeared at the vascular side, chiefly as free rutin (5.6%), but some rutin sulfate (2.5%) and glucuronide (2.0%) were also detected. The conjugates were preferentially absorbed to the vascular side, while only traces of the glucuronide (0.2%) were found in the luminal perfusate. Minute amounts of the rutin administered were located in the intestinal tissue (1.1%) in the form of unchanged rutin and its glucuronide and sulfate conjugates. The model used serves as a valuable tool for understanding intestinal handling of the bioactive flavonol glycoside rutin, and the obtained results confirm uptake of rutin in the rat small intestine.

Isoflavones from tofu are absorbed and metabolized in the isolated rat small intestine.

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.

A novel efficient method to identify beta-glucuronidase activity in rat small intestine.

BACKGROUND: Recent epidemiologic studies promote the notion that high intake of food rich in phytochemicals protects against degenerative diseases such as coronary heart diseases and cancer. Phytochemicals are detoxified in mammalian tissues by conjugation with glucuronic acid yielding less active glucuronide conjugates. However, in several tissues glucuronide conjugates are reactivated by the cleaving enzyme beta-glucuronidase. The aim of the present study was to develop a routinely manageable, rapid technique to localize the beta-glucuronidase activity in the small intestinal tissue. METHODS: Histologic slices of rat duodenum, jejunum, and ileum were incubated with a specific chromogenic beta-glucuronidase substrate, 5-bromo-4-chloro-3-indolyl-beta-D-glucuronide (X-GlcU). After enzymatic cleavage, X-GlcU yields 5-bromo-4-chloro-3-indol, a dark blue crystalline precipitate easily monitored by light microscopic technique. RESULTS: The number and intensity of the crystals were highest in the jejunum and lowest in the ileum. In all three sections of the small intestine, the highest activity was observed at the villar tip and in the tela submucosa and only moderate activity in other layers of the intestinal tissue. CONCLUSIONS: By using the X-GlcU-technique, it could be demonstrated convincingly that beta-glucuronidase exists in all three segments of the rat small intestine. The proposed method is an efficient, simple, and convenient method to visualize beta-glucuronidase activity.

Influence of the acetification process on phenolic compounds.

Little is known about the change of phenolic compounds and total phenolic content by the acetification process. The aim of this study was to assess the contents of selected phenolic compounds of cider and red and white wines in comparison to phenolic profiles in corresponding vinegars by using a new HPLC method for the simultaneous separation and quantification of polar phenolic acids and less polar flavonoids. Identifications were made by retention times and by means of mass spectra. Additionally, total phenolic contents of wines and vinegars were determined photometrically. The decrease in total phenol content by the acetification process was highest for cider vinegars (40%) and lower for red and white wine vinegars (13 and 8%, respectively). A decrease in the contents of individual phenolic compounds of vinegars from white white and ciders was not observed. In contrast, the contents of individual phenolic compounds in red wine vinegar decreased approximately 50%.

Assessment of resveratrol bioavailability in the perfused small intestine of the rat.

Resveratrol, which is present in grapes, wine and peanuts, is believed to possess chemoprotective properties such as anticarcinogenic effects and to provide protection against cardiovascular diseases. Little is known, however, about its intestinal absorption. We investigated the absorption and metabolism of resveratrol by using an isolated preparation of luminally and vascularly perfused rat small intestine. A synthetic perfusate free from blood components was used as vascular medium with a perfluorocarbon as oxygen carrier. Luminal media consisted of a bicarbonate buffered sodium chloride solution spiked with resveratrol in physiological, nutritionally relevant concentrations (28, 34 and 57 micromol/l, respectively). Viability was maintained during the entire perfusion and no significant differences between resveratrol and control perfusions for oxygen consumption, arterial pressure, lactate-pyruvate ratio and acid-base homeostasis were observed. Vascular uptake of luminally administered resveratrol was 20.5%. The majority of the absorbed resveratrol was conjugated to yield resveratrol glucuronide (16.8%), which was also the main luminal metabolite (11.2%). Lesser amounts of resveratrol sulfate, 3.0% and 0.3%, were found on the luminal and vascular side, respectively, while only minute amounts of resveratrol and resveratrol conjugates (1.9%) were found in the intestinal tissue. The structures of the resveratrol conjugates were verified by liquid chromatography coupled with mass spectometry (LC-MS). The results demonstrate an ample uptake and metabolic conversion of resveratrol. The proposed perfusion model serves as a tool to evaluate intestinal absorption and metabolic handling of phytochemicals, a pertinent input to the ongoing discussion about their health benefits.

Absorption and metabolism of genistin in the isolated rat small intestine.

Uptake and intestinal metabolism of physiologically active genistin were studied in an ex vivo intestinal perfusion model; luminally applied concentrations were 5.9, 12.0, and 23.8 micromol/l. The intestinal absorption of genistin was 14.9% (+/-2.3, n=9), irrespective of the amounts applied. The majority of the absorbed genistin appeared as genistein glucuronide (11.6%), also recovered as the main metabolite on the luminal side (19.5%). Minor amounts of genistin (1.3%) and genistein (1.9%) were found on the vascular side, whereas 15.4% of applied genistin was luminally cleaved to yield genistein. Sulfate derivatives of genistein or genistin were not observed.

Absorption and metabolism of genistein in isolated rat small intestine.

Studies suggest a variety of biological effects for the isoflavonoid genistein, but there is little information regarding small intestinal absorption and metabolism. The aim of this study was to investigate the absorption and metabolism of luminally administered genistein in an isolated preparation of luminally and vascularly perfused rat small intestine. A synthetic perfusate free from blood components was used as vascular medium, with a perfluorocarbon as oxygen carrier. Luminal media consisted of a bicarbonate buffered sodium chloride solution spiked with genistein (12 micromol/L). Viability was maintained during the entire perfusion as indicated by no significant differences between genistein and control perfusions for perfusion pressure, lactate-pyruvate ratio, oxygen uptake and acid-base homeostasis. Luminal disappearance rate of genistein did not change throughout the entire perfusion time. After a significant increase until about 30 min, vascular appearance rate of total genistein reached an equilibrium. The intestinal absorption of luminally administered genistein was 40.6%, corresponding to an average uptake of 2.9 nmol. min(-1). g dry intestine(-1). The majority (31.3%) of the absorbed genistein appeared as genistein glucuronide, also recovered as the main metabolite on the luminal side (13.3%). Only small amounts of genistein (2.6%) and genistein glucuronide (2.9%) were found in the intestinal tissue. The results demonstrate a favorable uptake of genistein, a pertinent addition to the ongoing discussion about health benefits of isoflavones.

Determination of selected phytochemicals by reversed-phase high-performance liquid chromatography combined with ultraviolet and mass spectrometric detection.

A robust, routinely manageable and sensitive RP-HPLC method combined with UV (270 nm) and ESI-MS detection was established for the determination of abundant pertinent phenolic compounds (phytochemicals) from various biological matrices. Phytochemicals were extracted by aqueous methanol (80%), extracts were analysed without further purification. Baseline separation was achieved within 30 min for 19 phytochemicals and excellent sensitivity (6-42 pmol at S/N = 3) was obtained. The identity of the phytochemicals was confirmed with standard compounds and with LC-MS. The repeatabilities for the majority of the phytochemicals ranged between 3% and 6%. The practicability of the method was shown in complex biological matrices by analysing onion and soybean extracts. This generally applicable technique may serve as a valuable tool for a rapid screening and a specific measurement of phytochemicals in food extracts and biological fluids and serve as analytical instrument for future biochemical and physiological studies.

Chemoprevention--a novel approach in dietetics.

Chemoprotective potential of naturally occurring phytochemicals in food is a major area of scientific interest. Results acquired from epidemiologic studies suggest a reduced risk of degenerative diseases with high phytochemical consumption. Bioavailability of phytochemicals is a critical issue, though their significant absorption has been demonstrated. Phytochemicals possess an array of biochemical and pharmacological qualities like antioxidative, anticarcinogenic, antimicrobial, cholesterol-lowering and antithrombotic activities.