Metabolism of cis- and trans-Resveratrol and Dihydroresveratrol in an Intestinal Epithelial Model

Trans-resveratrol, a well-known plant phenolic compound, has been intensively investigated due to its association with the so-called French paradox. However, despite its high pharmacological potential, trans-resveratrol has shown relatively low bioavailability. Trans-resveratrol is intensively metabolized in the intestine and liver, yielding metabolites that may be responsible for its high bioactivity. The aim of this study was to investigate and compare the metabolism of trans-resveratrol (tRes), cis-resveratrol (cRes) and dihydroresveratrol (dhRes) in an in vitro epithelial model using Caco-2 cell lines. Obtained metabolites of tRes, cRes and dhRes were analyzed by LC/MS Q-TOF, and significant differences in the metabolism of each compound were observed. The majority of tRes was transported unchanged through the Caco-2 cells, while cRes was mostly metabolized. The main metabolite of both cis- and trans-resveratrol observed as a result of colon microbial metabolism, dhRes, was metabolized almost completely, with only traces of the unchanged molecule being found. A sulphate conjugate was identified as the main metabolite of tRes in our model, while a glucuronide conjugate was the major metabolite of cRes and dhRes. Since metabolism of simple phenolics and polyphenols plays a crucial role in their bioavailability, detailed knowledge of their transformation is of high scientific value.

Fate of oestrogens during anaerobic blackwater treatment with micro-aerobic post-treatment

The fate of oestrone (E1), 17β-oestradiol (E2) and 17α-ethynyloestradiol (EE2) was investigated in a concentrated blackwater treatment system consisting of an UASB septic tank, with micro-aerobic post-treatment. In UASB septic tank effluent a (natural) total concentration of 4.02 μg/L E1 and 18.69 μg/L E2, comprising the sum of conjugated (>70% for E1 and >80% for E2) and unconjugated forms, was measured. During post-treatment the unconjugated oestrogens were removed to below 1 μg/L. A percentage of 77% of the measured unconjugated E1 and 82% of E2 was associated with particles >1.2 μm in the final effluent implying high sorption affinity of both compounds. When spiking the UASB septic tank effluent with E1, E2, EE2 and the sulphate conjugate of E2, removal in the micro-aerobic post-treatment was >99% for both E2 and EE2 and 83% for E1. The lower removal value for E1 was a result of (slow) deconjugation during the treatment, and in the final effluent still 40% of E1 and 99% of E2 was present in conjugated form. The latter was the result of incomplete deconjugation of the spiked E2(3S) in the post-treatment system.

Acetaminophen in the guinea pig: metabolite identification in blood, urine, and bile

The biotransformation of single acute oral doses of acetaminophen (100 mg/kg body weight) in adult male guinea pigs was studied by collecting serial blood, urine, and bile samples post-treatment and identifying and quantitating the concentrations of parent drug and excretory products by high performance liquid chromatography. The plasma half-life (βt/2) (mean ± SD) of acetaminophen was 1.87 ± 0.30 h, while that of the only metabolite detected in plasma, the glucuronide, was 2.41 ± 0.64 h. In 24-h urine samples, the predominant product was the glucuronide (90%) with a small amount of the sulphate conjugate (7.0%) and approximately 3.0% acetaminophen. In bile, the glucuronide was the major metabolite detected initially but, with time, this product decreased concomitantly with an increase in the cysteinyl conjugate. No sulphate was detected in bile but two unidentified metabolites were detected, having distinct column retention times and comprising approximately 6–10% of the total excretory products. The results demonstrated that glucuronidation is a high capacity biotransformation pathway for acetaminophen in this species, only small amounts of other conjugated products being detectable under usual circumstances.

Corticosterone storage within the adrenal cortex: evidence for a sulphate conjugate

ABSTRACT Incubation of homogenized rat adrenal glands with sulphatase for 18 h resulted in a significant increase in the amount of unconjugated corticosterone that could be extracted from the gland and which could not be attributed to de-novo synthesis of corticosterone during the incubation. Therefore corticosterone may be stored within the adrenal gland as a sulphate conjugate rather than as unconjugated hormone. Exposure of animals to footshock stress resulted in an increase in the amount of unconjugated corticosterone in the gland and a disappearance of the conjugate form. The rapid disappearance of the sulphate conjugate may reflect the activation of a steroid sulphatase by ACTH. However, the release of corticosterone from a storage form such as corticosterone sulphate could not explain entirely the initial increase in plasma corticosterone concentration following the stress. It was calculated that a pair of adrenal glands would have to store up to 20 nmole corticosterone in order to account for the initial increase in plasma corticosterone. This level was much greater than that extracted from a pair of glands (4·8 nmol) even after sulphatase pretreatment. J. Endocr. (1985) 104, 381–386

Assay of adenosine 3′-phosphate 5′-sulphatophosphate in hepatic tissues

A fluorimetric assay, based on the ability of boiled hepatic extracts to support the sulphO-conjugation of harmol, was used to demonstrate and quantify PAdoPS (adenosine 3′-phosphate 5′-sulphatophosphate) present in liver. A stoicheiometric relationship was established between the sulphate conjugate formed and the ‘active sulphate’ utilized. Guinea-pig, rat, mouse and rabbit livers contain 3.3, 2.9, 0.8 and 0.5 mumol of PAdoPS/100 G wet wt. respectively.

The availability of inorganic sulphate in blood for sulphate conjugation of drugs in rat liver in vivo. (35S)Sulphate incorporation into harmol sulphate

1. When Na235SO4 is injected intravenously in rats, it is immediately available for sulphate conjugation of the phenolic drug harmol (7-hydroxyl-1-methyl-9H-pyrido[3,4-b]indole) in the liver. This was established by following the time course of the biliary excretion of the sulphate conjugate of harmol, and the incorporation of [35S]sulphate into harmol sulphate. 2. During the 10min immediately after injection of Na235SO4 re-distribution of [35S]sulphate took place, which resulted in a rapid initial decrease in the plasma concentration of [35S]sulphate; a concomitant decrease in the amount of [35S]sulphate incorporated into harmol sulphate was observed, indicating that the co-substrate of sulphation, adenosine 3′-phosphate 5′-sulphatophosphate, equilibrates rapidly with [35S]sulphate in plasma. 3. The results suggest that the pool size of adenosine 3′-phosphate 5′-sulphatophosphate is very small; therefore the specific radioactivity of [35S]sulphate in plasma determines the specific radioactivity incorporated into sulphate esters at any time.