Very low vitamin C activity of orally administered L-dehydroascorbic acid.

The biological activity of L-dehydroascorbic acid (DHA), which is easily formed from L-ascorbic acid (ASC) during storage and cooking processes, has been considered to be equivalent to that of ASC on the basis of studies made several decades ago, when a specific method to determine ASC was not available. The nutritional activity of orally ingested DHA has now been evaluated by comparing ASC concentrations in 12 tissues of rats administered four different doses of ASC. Determinations were made by using the specific and sensitive method, which had been developed by us. Here it is shown that the efficiency of DHA was almost 10% of that of ASC on a molar basis, based on animal experiments using the inherently scorbutic ODS rat, which is a convenient human model animal to investigate the metabolism of vitamin C. On the basis of these findings, it is proposed that it is necessary to reevaluate the nutritional requirement of vitamin C based on both ASC and DHA contents of foods.     

Antihypertensive effects of Lactobacillus-fermented milk orally administered to spontaneously hypertensive rats

Products fermented with lactic acid bacteria may show antihypertensive effects via substances such as angiotensin I-converting enzyme inhibitor (ACEI) and γ-aminobutyric acid (GABA). It was previously found that milk fermented with Lactobacillus paracasei subsp. paracasei NTU 101 (101FM) or Lactobacillus plantarum NTU 102 (102FM) has ACEI and GABA activities. This study aimed to investigate the antihypertensive effects of 101FM and 102FM orally administered to spontaneously hypertensive rats (SHRs). Eight hours after a single oral administration or after 8 weeks of weekly (chronic) administration, 101FM and 102FM significantly decreased systolic and diastolic blood pressures in the SHRs. Microscopic examination of aortic tissue demonstrated that 101FM and 102FM reduced the disorganization of the media layer. These findings suggest that orally administered 101FM and 102FM have antihypertensive effects, possibly via ACEI and GABA activity, in SHRs. Therefore, 101FM and 102FM may be useful ingredients in physiologically functional foods to prevent hypertension.     

Occurrence of orally administered mulberry 1-deoxynojirimycin in rat plasma

1-deoxynojirimycin (DNJ), a potent glucosidase inhibitor, is a characteristic constituent of the mulberry leaf. Dietary mulberry DNJ may be beneficial for the suppression of abnormally high blood glucose levels, thereby preventing diabetes mellitus. Although there is considerable interest in the effects of mulberry DNJ, the intestinal absorption and pharmacokinetic profile of orally administered mulberry DNJ have never been characterized. In this study, we developed a method for determining the level of plasma DNJ by hydrophilic interaction chromatography coupled to a mass spectrometric detector (HILIC-MS) to investigate the absorption and metabolism of orally administered mulberry DNJ in rats. DNJ was separated from plasma extract on a TSK gel Amide-80 column, a representative column for HILIC. At postcolumn, DNJ was concurrently detected and identified by MS. The plasma DNJ concentration in fasted rats was below the detection limit [<1 microg (6 nmol)/mL]; however, the concentration reached a maximum [15 microg (92 nmol)/mL] 30 min after the administration of mulberry DNJ (110 mg/kg of body weight), and the DNJ concentration decreased rapidly thereafter. When the rats received different amounts of mulberry DNJ (1.1, 11, and 110 mg/kg of body weight), dose-dependent incorporation of DNJ into the plasma was confirmed. We did not detect any DNJ metabolites in the plasma. These findings indicate that orally administered mulberry DNJ is absorbed as an intact form from the alimentary tract and then is quickly excreted from the body. The developed HILIC-MS method could be applied in determining levels of DNJ in urine and tissues, and therefore, the method would be a powerful tool for studying the metabolic fate of mulberry DNJ as well as its bioavailability.     

Combination of lipids and emulsifiers enhances the absorption of orally administered quercetin in rats

The effects of lipids, emulsifiers, and ethanol on the absorption of orally administered quercetin in rats were investigated for its efficient intestinal absorption. Rats were administered 150 micromol/kg quercetin in water supplemented with lipids and/or emulsifiers, or ethanol, and blood was collected from the tail for 6 h after administration. Co-administration of lipids such as lecithin and soybean oil or emulsifiers including sucrose fatty acid ester, polyglycerol fatty acid ester, and sodium taurocholate had no statistically significant effects on quercetin absorption, although these constituents rather increased the accumulation of conjugated forms of quercetin and those of isorhamnetin in rat plasma. However, the combination of lipids and emulsifiers enhanced the absorption of quercetin significantly. Thirty and fifty percent (v/v) of the ethanol in the vehicle raised the efficiency of quercetin absorption in a concentration-dependent manner. Quercetin absorption-enhancing effects of these constituents seemed to be affected by quercetin's solubility in respective vehicles used for the administration. Ethanol is not helpful for the effective absorption of quercetin, as a high concentration is required. In conclusion, a combination of lipids and emulsifiers is necessary for enhancing quercetin absorption.     

Absorption, disposition, metabolism, and excretion of [3-(14)C]caffeic acid in rats

Male Sprague-Dawley rats ingested 140 × 10(6) dpm of [3-(14)C]trans-caffeic acid, and over the ensuing 72 h period, body tissues, plasma, urine, and feces were collected and the overall levels of radioactivity determined. Where sufficient radioactivity had accumulated, samples were analyzed by HPLC with online radioactivity and tandem mass spectrometric detection. Nine labeled compounds were identified, the substrate and its cis isomer, 3'-O- and 4'-O-sulfates and glucuronides of caffeic acid, 4'-O-sulfates and glucuronides of ferulic acid, and isoferulic acid-4'-O-sulfate. Four unidentified metabolites were also detected. After passing down the gastrointestinal tract, the majority of the radiolabeled metabolites were excreted in urine with minimal accumulation in plasma. Only relatively small amounts of an unidentified (14)C-labeled metabolite were expelled in feces. There was little or no accumulation of radioactivity in body tissues, including the brain. The overall recovery of radioactivity 72 h after ingestion of [3-(14)C]caffeic acid was ～80% of intake.     

Identification and quantification of the conjugated metabolites derived from orally administered hesperidin in rat plasma

Hesperidin is a biologically effective flavonoid. Several studies have reported that dietary hesperidin was converted to conjugated metabolites, such as hesperetin-glucuronides and sulfoglucuronides, during absorption and metabolism. However, the chemical structures of the conjugated metabolites, especially the sites of glucuronidation and sulfoglucuronidation in plasma, were unconfirmed. Therefore, the concentrations of the metabolites conjugated at various sites in plasma could not be individually quantified. In the present study, we identified the chemical structures and concentrations of the major conjugated metabolites in rat plasma after oral administration of hesperidin. Two hesperetin-glucuronides were prepared and identified as hesperetin-7-O-beta-D-glucuronide and hesperetin-3'-O-beta-D-glucuronide. Using these authentic compounds, the concentrations of hesperetin-7-O-beta-D-glucuronide and hesperetin-3'-O-beta-D-glucuronide in rat plasma were individually determined by liquid chromatography-mass spectrometry. In rat plasma, hesperetin-glucuronides were primarily comprised of hesperetin-7-O-beta-D-glucuronide and hesperetin-3'-O-beta-D-glucuronide. The concentration of hesperetin-7-O-beta-D-glucuronide was slightly higher than that of hesperetin-3'-O-beta-D-glucuronide. Furthermore, not only hesperetin conjugates but also homoeriodictyol conjugates were observed in rat plasma. The present study is the first report elucidating the chemical structures and changes in individual concentrations in rat plasma of glucuronides derived from orally administered hesperidin.     

Fractionation of radioactivity in the milk of goats administered 14C-aflatoxin B1

A detailed fractionation of radioactivity in the milk of goats administered 14C-aflatoxin B1 at low doses was performed. The milk collected in the first 24 h following dosing contained radioactivity equivalent to 0.45-1.1% of the dose given. The radioactivity in each sample was partitioned into 4 fractions: ether, protein, dichloromethane, and water-alcohol. Over 80% of the radioactivity was detected in the dichloromethane fraction, of which over 95% was attributable to aflatoxin M1. No aflatoxin B1 or other known aflatoxin metabolites were detected in any fraction. The results indicate that the major metabolite of aflatoxin B1 in goat milk is aflatoxin M1 and that other metabolites, including conjugates, are of minor significance.     

Distribution of total 14C residue in egg yolk, albumen, and tissues following oral [14C]sulfamethazine administration to hens

The distribution of total 14C residues was studied in egg yolk and albumen after administration of either single or multiple oral dosages of [14C]sulfamethazine (SMZ). One day after a single dose of [14C]SMZ (121 mg of sulfamethazine, 2.42 x 10(7) dpm), the 14C residue concentration peaked in egg albumen and egg yolk with the concentration in the former >4-fold greater than in the latter. Three days postdose, the 14C residue concentration in the yolk was approximately 7-fold higher than in the egg albumen. A multiple dose of [14C]SMZ containing sulfamethazine mass equivalent of an average therapeutic dose (282 mg, 2.9 x 10(7) dpm) for chickens was also administered orally for six consecutive days to hens. A significantly reduced level of egg production was observed during the medication, and most of the hens stopped laying eggs after the last dose. The 14C residue concentrations peaked on the last day (sixth) of medication in egg albumen and yolk. The 14C residue concentrations were also measured in liver, muscle, blood, and plasma of chickens sacrificed at 1, 24, 48, and 72 h after the last dose. Highest concentrations of 14C residue were accumulated in liver followed by, in decreasing order, blood, plasma, and muscle.