Development of an online-SPE-LC-MS method for the investigation of the intestinal absorption of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PHIP) and its bacterial metabolite PHIP-M1 in a Caco-2 Transwell system

Willenberg, Ina; Elsner, Leonie von; Steinberg, Pablo; Schebb, Nils Helge

doi:10.1016/j.foodchem.2014.06.038

Cited by 13 publications

(8 citation statements)

References 28 publications

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“…The toxicological relevance of the transformation of PhIP to PhIP‐M1 depends on uptake potential and relative toxic potencies. PhIP and PhIP‐M1 were shown to be transported at similar rates in several in vitro systems (Nicken et al ., 2010; 2015; Willenberg et al ., ). The mutagenic potency of PhIP‐M1 in the Ames test was only 5% that of PhIP, and the adverse response was speculated to be attributable to residual PhIP (Vanhaecke et al ., ).…”

Section: Resultsmentioning

confidence: 97%

The strict anaerobic gut microbe Eubacterium hallii transforms the carcinogenic dietary heterocyclic amine 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP)

Fekry

Engels

Zhang

et al. 2016

Environ Microbiol Rep

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2-Amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP) is the most abundant food-derived heterocyclic aromatic amine in well-cooked meats and may contribute to the recognized carcinogenicity of processed meats. In this study, a panel of human gut microbes was tested for their ability to convert PhIP to a conjugate PhIP-M1. Eubacterium hallii was newly identified to catalyse the conversion of PhIP to PhIP-M1 with high efficiency. The reaction was shown to involve the metabolism of glycerol to 3-hydroxypropionaldehyde as a key pathway. The proficiency of E. hallii in transforming PhIP in the presence of a complex intestinal microbiota was confirmed using batch fermentations inoculated with effluents from a continuous intestinal fermentation model mimicking human proximal and distal colon microbiota. In batch fermentations inoculated with proximal colon microbiota, PhIP-M1 transformation corresponded to an up to 300-fold increase of E. hallii. In contrast, PhIP transformation of distal colon microbiota was low but increased by 120-fold after supplementation with E. hallii. These findings indicate for the first time the relevance of the abundant commensal strict anaerobe E. hallii in the transformation of a dietary carcinogen that could contribute to its detoxification in the human colon.

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Section: Resultsmentioning

confidence: 97%

The strict anaerobic gut microbe Eubacterium hallii transforms the carcinogenic dietary heterocyclic amine 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP)

Fekry

Engels

Zhang

et al. 2016

Environ Microbiol Rep

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“…Up to now, it is unclear which transporter is involved in the PhIP efflux in the rat colon. Various research groups have shown that the breast cancer resistance protein, the multidrug resistance protein 2 and P-glycoprotein are involved in PhIP transport in various species (Walle and Walle 1999;Dietrich et al 2001;van Herwaarden et al 2003;Willenberg et al 2015), but their involvement could not be confirmed in previous Ussing chamber experiments with gut tissue samples of male Fischer 344 rats (Nicken et al 2013). …”

Section: Discussionmentioning

confidence: 99%

“…PhIP quantification was carried out by LC-MS with an automated sample preparation by on-line solid-phase extraction (SPE) in the backflush mode as described by Willenberg et al (2015). Briefly, two LC-10-ADvp LC systems (Shimadzu, Langenfeld, Germany) including 2 SCL10Avp controllers, 4 pumps (LC-10ADvp or LC-10AD), a column oven (CTO-10ASvp) set to 40 °C, two degassers (DGU-14A) and a high-pressure 2 position Cheminert sixport two-position valve (VICI, Schenkon, Switzerland) were used.…”

Section: Quantification Of Phipmentioning

confidence: 99%

Impact of dextran sulphate sodium-induced colitis on the intestinal transport of the colon carcinogen PhIP

et al. 2015

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Colorectal cancer is one of the most frequent cancers in Western countries. Chronic intestinal diseases such as Crohn's disease and ulcerative colitis, in which the intestinal barrier is massively disturbed, significantly raise the risk of developing a colorectal tumour. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a genotoxic heterocyclic aromatic amine that is formed after strongly heating fish and meat. In this study, the hypothesis that PhIP uptake in the gut is increased during chronic colitis was tested. Chronic colitis was induced by oral administration of dextran sulphate sodium (DSS) to Fischer 344 rats. The transport of PhIP in eight different rat intestinal segments was examined in Ussing chambers. The tissues were incubated with 10 µM PhIP for 90 min, and the concentration of PhIP was determined in the mucosal and serosal compartments of the Ussing chambers as well as in the clamped tissues by LC-MS. Although chronic colitis was clearly induced in the rats, no differences in the intestinal transport of PhIP were observed between control and DSS-treated animals. The hypothesis that in the course of chronic colitis more PhIP is taken up by the intestinal epithelium, thereby increasing the risk of developing colorectal cancer, could not be confirmed in the present report.

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“…24, 25 Cell monolayers that exceeded a resistance of 300 Ω cm −2 were incubated with either 1 μM or 10 μM of TPPU solution in DMSO on the apical side. Medium samples on the apical and basolateral side were collected and frozen immediately after 1, 3 and 6 hours.…”

Section: Methodsmentioning

confidence: 99%

“…The apparent permeability coefficient (P app ) was calculated for t = 1 h as described. 24–27 In order to assure that TPPU does not compromise the integrity of the monolayer additional experiments were performed in the presence of Lucifer yellow (LY; 100 μM in the apical compartment), a marker for paracellular diffusion and no LY could be detected in the basolateral chamber.…”

Section: Methodsmentioning

confidence: 99%

Oral treatment of rodents with soluble epoxide hydrolase inhibitor 1-(1-propanoylpiperidin-4-yl)-3-[4-(trifluoromethoxy)phenyl]urea (TPPU): Resulting drug levels and modulation of oxylipin pattern

Ostermann¹,

Herbers²,

Willenberg³

et al. 2015

Prostaglandins & Other Lipid Mediators

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Epoxides from polyunsaturated fatty acids (PUFAs) are potent lipid mediators. In vivo stabilization of these epoxides by blockade of the soluble epoxide hydrolase (sEH) leads to anti-inflammatory, analgesic and normotensive effects. Therefore, sEH inhibitors (sEHi) are a promising new class of drugs. Herein, we characterized pharmacokinetic (PK) and pharmacodynamic properties of a commercially available potent sEHi 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea (TPPU). Cell culture studies suggest its high absorption and metabolic stability. Following administration in drinking water to rats (0.2, 1, and 5 mg TPPU/L with 0.2% PEG400), TPPU’s blood concentration increased dose dependently within the treatment period to reach an almost steady state after 8 days. TPPU was found in all the tissues tested. The linoleic epoxide/diol ratios in the tissues were dose dependently increased, indicating significant sEH inhibition. Overall, administration of TPPU with the drinking water led to systemic distribution as well as high levels and thusmakes chronic sEH inhibition studies possible.

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Development of an online-SPE-LC-MS method for the investigation of the intestinal absorption of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PHIP) and its bacterial metabolite PHIP-M1 in a Caco-2 Transwell system

Cited by 13 publications

References 28 publications

The strict anaerobic gut microbe Eubacterium hallii transforms the carcinogenic dietary heterocyclic amine 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP)

The strict anaerobic gut microbe Eubacterium hallii transforms the carcinogenic dietary heterocyclic amine 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP)

Impact of dextran sulphate sodium-induced colitis on the intestinal transport of the colon carcinogen PhIP

Oral treatment of rodents with soluble epoxide hydrolase inhibitor 1-(1-propanoylpiperidin-4-yl)-3-[4-(trifluoromethoxy)phenyl]urea (TPPU): Resulting drug levels and modulation of oxylipin pattern

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