Development and Validation of an in vitro Experimental GastroIntestinal Dialysis Model with Colon Phase to Study the Availability and Colonic Metabolisation of Polyphenolic Compounds^[*]

 

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Abstract

The biological effects of polyphenols depend on their mechanism of action in the body. This is affected by bioconversion by colon microbiota and absorption of colonic metabolites. We developed and validated an in vitro continuous flow dialysis model with colon phase (GastroIntestinal dialysis model with colon phase) to study the gastrointestinal metabolism and absorption of phenolic food constituents. Chlorogenic acid was used as model compound. The physiological conditions during gastrointestinal digestion were mimicked. A continuous flow dialysis system simulated the one-way absorption by passive diffusion from lumen to mucosa. The colon phase was developed using pooled faecal suspensions. Several methodological aspects including implementation of an anaerobic environment, adapted Wilkins Chalgren broth medium, 1.10⁸ CFU/mL bacteria suspension as inoculum, pH adaptation to 5.8 and implementation of the dialysis system were conducted. Validation of the GastroIntestinal dialysis model with colon phase system showed a good recovery and precision (CV < 16 %). Availability of chlorogenic acid in the small intestinal phase (37 ± 3 %) of the GastroIntestinal dialysis model with colon phase is comparable with in vivo studies on ileostomy patients. In the colon phase, the human faecal microbiota deconjugated chlorogenic acid to caffeic acid, 3,4-dihydroxyphenyl propionic acid, 4-hydroxybenzoic acid, 3- or 4-hydroxyphenyl acetic acid, 2-methoxy-4-methylphenol and 3-phenylpropionic acid. The GastroIntestinal dialysis model with colon phase is a new, reliable gastrointestinal simulation system. It permits a fast and easy way to predict the availability of complex secondary metabolites, and to detect metabolites in an early stage after digestion. Isolation and identification of these metabolites may be used as references for in vivo bioavailability experiments and for investigating their bioactivity in in vitro experiments.

^* Dedicated to Professor Dr. Dr. h. c. mult. Adolf Nahrstedt on the occasion of his 75th birthday.

• References

• 1 Crozier A, Jaganath IB, Clifford MN. Dietary phenolics: chemistry, bioavailability and effects on health. Nat Prod Rep 2009; 26: 1001-1043
  CrossrefPubMedGoogle Scholar
• 2 Garcia-Lafuente A, Guillamon E, Villares A, Rostagno MA, Martinez JA. Flavonoids as anti-inflammatory agents: implications in cancer and cardiovascular disease. Inflamm Res 2009; 58: 537-552
  CrossrefPubMedGoogle Scholar
• 3 Bosscher D, Breynaert A, Pieters L, Hermans N. Food-based strategies to modulate the composition of the intestinal microbiota and their associated health effects. J Physiol Pharmacol 2009; 60 (Suppl. 06) S5-S11
  PubMedGoogle Scholar
• 4 DʼArchivio M, Filesi C, Di Benedetto R, Gargiulo R, Giovannini C, Masella R. Polyphenols, dietary sources and bioavailability. Ann Ist Super Sanita 2007; 43: 348-361
  PubMedGoogle Scholar
• 5 Manach C, Williamson G, Morand C, Scalbert A, Remesy C. Bioavailability and bioefficacy of polyphenols in humans. I. Review of 97 bioavailability studies. Am J Clin Nutr 2005; 81 (Suppl. 01) 230S-242S
  CrossrefPubMedGoogle Scholar
• 6 Selma MV, Espin JC, Tomas-Barberan FA. Interaction between phenolics and gut microbiota: role in human health. J Agric Food Chem 2009; 57: 6485-6501
  CrossrefPubMedGoogle Scholar
• 7 Kemperman RA, Bolca S, Roger LC, Vaughan EE. Novel approaches for analysing gut microbes and dietary polyphenols: challenges and opportunities. Microbiology 2010; 156: 3224-3231
  CrossrefPubMedGoogle Scholar
• 8 Williamson G, Clifford MN. Colonic metabolites of berry polyphenols: the missing link to biological activity?. Br J Nutr 2010; 104 (Suppl. 03) S48-S66
  CrossrefPubMedGoogle Scholar
• 9 Shen LH, Luten J, Robberecht H, Bindels J, Deelstra H. Modification of an in-vitro method for estimating the bioavailability of zinc and calcium from foods. Z Lebensm Unters Forsch 1994; 199: 442-445
  CrossrefPubMedGoogle Scholar
• 10 Bosscher D, Van Caillie-Bertand M, Robberecht H, Van Dyck K, Van Cauwenbergh R, Deelstra H. In vitro availability of calcium, iron, and zinc from first-age infant formulae and human milk. J Pediatr Gastroenterol Nutr 2000; 32: 54-58
  PubMedGoogle Scholar
• 11 Bosscher D, Lu Z, Van Cauwenbergh R, Van Caillie-Bertrand M, Robberecht H, Deelstra H. A method for in vitro determination of calcium, iron and zinc availability from first-age infant formula and human milk. Int J Food Sci Nutr 2001; 52: 173-182
  CrossrefPubMedGoogle Scholar
• 12 De Boever P, Deplancke B, Verstraete W. Fermentation by gut microbiota cultured in a simulator of the human intestinal microbial ecosystem is improved by supplementing a soygerm powder. J Nutr 2000; 130: 2599-2606
  PubMedGoogle Scholar
• 13 Guyton AC, Hall JE. Propulsion and mixing of food in the gastrointestinal tract. Philadelphia: W.B. Saunders Company; 2006
  Google Scholar
• 14 Zhao Z, Moghadasian MH. Bioavailability of hydroxycinnamates: a brief review of in vivo and in vitro studies. Phytochem Rev 2010; 9: 133-145
  CrossrefPubMedGoogle Scholar
• 15 Manach C, Scalbert A, Morand C, Remesy C, Jimenez L. Polyphenols: food sources and bioavailability. Am J Clin Nutr 2004; 79: 727-747
  PubMedGoogle Scholar
• 16 Stalmach A, Mullen W, Barron D, Uchida K, Yokota T, Cavin C, Steiling H, Williamson G, Crozier A. Metabolite profiling of hydroxycinnamate derivatives in plasma and urine after the ingestion of coffee by humans: identification of biomarkers of coffee consumption. Drug Metab Dispos 2009; 37: 1749-1758
  CrossrefPubMedGoogle Scholar
• 17 Stalmach A, Steiling H, Williamson G, Crozier A. Bioavailability of chlorogenic acids following acute ingestion of coffee by humans with an ileostomy. Arch Biochem Biophys 2010; 501: 98-105
  CrossrefPubMedGoogle Scholar
• 18 Monteiro M, Farah A, Perrone D, Trugo LC, Donangelo C. Chlorogenic acid compounds from coffee are differentially absorbed and metabolized in humans. J Nutr 2007; 137: 2196-2201
  PubMedGoogle Scholar
• 19 Begley M, Hill C, Gahan CG. Bile salt hydrolase activity in probiotics. Appl Environ Microbiol 2006; 72: 1729-1738
  CrossrefPubMedGoogle Scholar
• 20 Khaleghi M, Kermanshahi RK, Yaghoobi MM, Zarkesh-Esfahani SH, Baghizadeh A. Assessment of bile salt effects on s-layer production, slp gene expression and some physicochemical properties of Lactobacillus acidophilus ATCC 4356. J Microbiol Biotechnol 2010; 20: 749-756
  PubMedGoogle Scholar
• 21 Zarate G, Gonzalez S, Chaia AP, Oliver G. Effect of bile on the B-galactosidase activity of dairy propionibacteria. Lait 1999; 80: 267-276
  PubMedGoogle Scholar
• 22 Zavaglia AG, Kociubinski G, Perez P, Disalvo E, De Antoni G. Effect of bile on the lipid composition and surface properties of bifidobacteria. J Appl Microbiol 2002; 93: 794-799
  CrossrefPubMedGoogle Scholar
• 23 Lafay S, Gil-Izquierdo A, Manach C, Morand C, Besson C, Scalbert A. Chlorogenic acid is absorbed in its intact form in the stomach of rats. J Nutr 2006; 136: 1192-1197
  PubMedGoogle Scholar
• 24 Lafay S, Morand C, Manach C, Besson C, Scalbert A. Absorption and metabolism of caffeic acid and chlorogenic acid in the small intestine of rats. Br J Nutr 2006; 96: 39-46
  CrossrefPubMedGoogle Scholar
• 25 Gonthier MP, Verny MA, Besson C, Remesy C, Scalbert A. Chlorogenic acid bioavailability largely depends on its metabolism by the gut microflora in rats. J Nutr 2003; 133: 1853-1859
  CrossrefPubMedGoogle Scholar
• 26 Hollman PC, de Vries JH, van Leeuwen SD, Mengelers MJ, Katan MB. Absorption of dietary quercetin glycosides and quercetin in healthy ileostomy volunteers. Am J Clin Nutr 1995; 62: 1276-1282
  PubMedGoogle Scholar
• 27 Olthof MR, Hollman PC, Katan MB. Chlorogenic acid and caffeic acid are absorbed in humans. J Nutr 2001; 131: 66-71
  CrossrefPubMedGoogle Scholar
• 28 Erk T, Williamson G, Renouf M, Marmet C, Steiling H, Dionisi F, Barron D, Melcher R, Richling E. Dose-dependent absorption of chlorogenic acids in the small intestine assessed by coffee consumption in ileostomists. Mol Nutr Food Res 2012; 56: 1488-1500
  CrossrefPubMedGoogle Scholar
• 29 Farah A, Monteiro M, Donangelo CM, Lafay S. Chlorogenic acids from green coffee extract are highly bioavailable in humans. J Nutr 2008; 138: 2309-2315
  CrossrefPubMedGoogle Scholar
• 30 Gonthier MP, Remesy C, Scalbert A, Cheynier V, Souquet JM, Poutanen K, Aura AM. Microbial metabolism of caffeic acid and its esters chlorogenic and caftaric acids by human faecal microbiota in vitro . Biomed Pharmacother 2006; 60: 536-540
  CrossrefPubMedGoogle Scholar
• 31 Rechner AR, Smith MA, Kuhnle G, Gibson GR, Debnam ES, Srai SK, Moore KP, Rice-Evans CA. Colonic metabolism of dietary polyphenols: influence of structure on microbial fermentation products. Free Radic Biol Med 2004; 36: 212-225
  CrossrefPubMedGoogle Scholar
• 32 De Boever P, Wouters R, Vermeirssen V, Boon N, Verstraete W. Development of a six-stage culture system for simulating the gastrointestinal microbiota of weaned infants. Microb Ecol Health D 2001; 13: 111-123
  CrossrefPubMedGoogle Scholar
• 33 Possemiers S, Verthe K, Uyttendaele S, Verstraete W. PCR-DGGE-based quantification of stability of the microbial community in a simulator of the human intestinal microbial ecosystem. FEMS Microbiol Ecol 2004; 49: 495-507
  CrossrefPubMedGoogle Scholar
• 34 Gao K, Xu A, Krul C, Venema K, Liu Y, Niu Y, Lu J, Bensoussan L, Seeram NP, Heber D, Henning SM. Of the major phenolic acids formed during human microbial fermentation of tea, citrus, and soy flavonoid supplements, only 3,4-dihydroxyphenylacetic acid has antiproliferative activity. J Nutr 2006; 136: 52-57
  PubMedGoogle Scholar
• 35 Minekus M, Smeets-Peeters M, Bernalier A, Marol-Bonnin S, Havenaar R, Marteau P, Alric M, Fonty G, Huis inʼt Veld JH. A computer-controlled system to simulate conditions of the large intestine with peristaltic mixing, water absorption and absorption of fermentation products. Appl Microbiol Biotech 1999; 53: 108-114
  CrossrefPubMedGoogle Scholar
• 36 Bosscher D, Van Caillie-Bertand M, Deelstra H. Optimalisation of in vitro methods to the intraluminal digestive conditions of children up to 3 years of age in order to determine bioavailability of minerals and trace elements from diets for children. Gastroenterol 1998; 36: 17-27
  PubMedGoogle Scholar
• 37 Coles LT, Moughan PJ, Darragh AJ. In vitro digestion and fermentation methods, including gas production techniques, as applied to nutritive evaluation of foods in the hindgut of humans and other simple-stomached animals. Anim Feed Sci Technol 2005; 123–124: 421-444
  PubMedGoogle Scholar
• 38 Aura AM, OʼLeary KA, Williamson G, Ojala M, Bailey M, Puupponen-Pimia R, Nuutila AM, Oksman-Caldentey KM, Poutanen K. Quercetin derivatives are deconjugated and converted to hydroxyphenylacetic acids but not methylated by human fecal flora in vitro . J Agric Food Chem 2002; 50: 1725-1730
  CrossrefPubMedGoogle Scholar