Abstract
Hylak® forte is a postbiotic that inhibits the growth of pathogenic bacteria by reducing intestinal pH. It is assumed the potential presence of short-chain fatty acids (SCFAs) in Hylak® forte may contribute to this effect. In this current study, we analysed the composition of Hylak® forte, using a validated gas chromatography assay test method, to ascertain whether SCFAs are present in this postbiotic treatment. Hylak® forte was screened for C1 to C10 SCFAs by a gas chromatographic assay. In this assay, SCFAs were analysed as for their volatile ethyl ester derivatives in a 3.0 mL Hylak® forte sample. An additional screening procedure was conducted for the presence of vitamins and simple sugars. The gas chromatographic method for determining SCFAs was validated according to the requirements of ICH guideline Q2 (R1). Formic and acetic acids were identified in Hylak® forte at 27.92 ppm (90% confidence interval (CI), 26.90–28.94) and 306.17 ppm (90% CI, 277.11–335.22), respectively. Additional compounds were quantified in the solution, including vitamin B1 (0.029 mg/100 g), monosaccharides and disaccharides (2.767 g/100 g), as well as glutamic acid and glutamine (0.047 g/100 g). This study has identified formic acid in a range of 39.33 (90% CI, 36.50–42.17)–48.33 (90% CI, 45.91–50.76) ppm and acetic acid dropping down to 312.33 (90% CI, 295.32–329.35) from initial 415.67 ppm (90% CI, 385.93–445.41) in commercial Hylak® forte samples under forced degradation conditions. In addition, a range of other compounds in Hylak® forte was identified, including riboflavin and glutamine. Further studies are necessary to establish whether these compounds translate into tangible therapeutic effects.
References
Kamada N, Chen GY, Inohara N, Núñez G (2013) Control of pathogens and pathobionts by the gut microbiota. Nat Immunol 14:685–690. https://doi.org/10.1038/ni.2608
Neis EPJG, Dejong CHC, Rensen SS (2015) The role of microbial amino acid metabolism in host metabolism. Nutrients 7:2930–2946. https://doi.org/10.3390/nu7042930
Thursby E, Juge N (2017) Introduction to the human gut microbiota. Biochem J 474:1823–1836. https://doi.org/10.1042/BCJ20160510
Conlon MA, Bird AR (2014) The impact of diet and lifestyle on gut microbiota and human health. Nutrients 7:17–44. https://doi.org/10.3390/nu7010017
Ulluwishewa D, Anderson RC, McNabb WC, Moughan PJ, Wells JM, Roy NC (2011) Regulation of tight junction permeability by intestinal bacteria and dietary components. J Nutr 141:769–776. https://doi.org/10.3945/jn.110.135657
Trompette A, Gollwitzer ES, Yadava K, Sichelstiel AK, Sprenger N, Ngom-Bru C, Blanchard C, Junt T, Nicod LP, Harris NL, Marsland BJ (2014) Gut microbiota metabolism of dietary fiber influences allergic airway disease and hematopoiesis. Nat Med 20:159–166. https://doi.org/10.1038/nm.3444
Vieira AT, Fukumori C, Ferreira CM (2016) New insights into therapeutic strategies for gut microbiota modulation in inflammatory diseases. Clin Transl Immunol 5:e87. https://doi.org/10.1038/cti.2016.38
Adams CA (2010) The probiotic paradox: live and dead cells are biological response modifiers. Nutr Res Rev 23:37–46. https://doi.org/10.1017/S0954422410000090
Bermudez-Brito M, Plaza-Díaz J, Muñoz-Quezada S, Gómez-Llorente C, Gil A (2012) Probiotic mechanisms of action. Ann Nutr Metab 61:160–174. https://doi.org/10.1159/000342079
Madsen K, Cornish A, Soper P, McKaigney C, Jijon H, Yachimec C, Doyle J, Jewell L, De Simone C (2001) Probiotic bacteria enhance murine and human intestinal epithelial barrier function. Gastroenterology 121:580–591. https://doi.org/10.1053/gast.2001.27224
Tsilingiri K, Rescigno M (2013) Postbiotics: what else? Benefic Microbes 4:101–107. https://doi.org/10.3920/BM2012.0046
Hylak® forte User Information. https://www.google.de/search?q=hylak+user+forte+user+infomration&rlz=1C1EJFA_enDE781DE781&oq=hylak+user+forte+user+infomration&aqs=chrome..69i57j33.7326j1j4&sourceid=chrome&ie=UTF-8. Accessed 20 Aug 2002, last revision March 2016
Ríos-Covián D, Ruas-Madiedo P, Margolles A, Gueimonde M, de los Reyes-Gavilán CG, Salazar N (2016) Intestinal short chain fatty acids and their link with diet and human health. Front Microbiol 7:185. https://doi.org/10.3389/fmicb.2016.00185
Macfarlane GT, Macfarlane S (2012) Bacteria, colonic fermentation, and gastrointestinal health. J AOAC Int 95:50–60. https://doi.org/10.5740/jaoacint.SGE_Macfarlane
(2005) ICH harmonised tripartite guideline. Validation of analytical procedures: text and methodology Q2(R1). https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf. Accessed 2016
Metcalfe LD, Schmitz AA (1961) The rapid preparation of fatty acid esters for gas chromatographic analysis. Anal Chem 33:363–364. https://doi.org/10.1021/ac60171a016
Shenderov BA (2013) Metabiotics: novel idea or natural development of probiotic conception. Microb Ecol Health Dis 24:20399. https://doi.org/10.3402/mehd.v24i0.20399
Belousova EF, Nikitine YV, Mishurovskaya NC, Zlatkina AR (2005) Possibilities of microbial metabolite preparations for intestinal microbiota restoration. Cons Medicum 7:9–13
Hiller E, Schach H (1955) Clinical and experimental experiences on the possibility of the influence of yoghurt metabolites on pathological conditions of intestinal flora. Arzneimittelforschung 5:592–599
Konig H, Manz W, Singer U (1991) Influence on intestinal bacteria. Basic principles of treatment using bacterial metabolic products. Der Kassenarzt 18:52–61
Krieger D, Vanek E, Manz W, Elsasser R (1991) Influence of bacterial metabolic products on the fecal flora of healthy volunteers and on added pathogenic bacteria. Nb Med 4:169–172
den Besten G, van Eunen K, Groen AK, Venema K, Reijngoud DJ, Bakker BM (2013) The role of short-chain fatty acids in the interplay between diet, gut microbiota, and host energy metabolism. J Lipid Res 54:2325–2340. https://doi.org/10.1194/jlr.R036012
Fukuda S, Toh H, Hase K, Oshima K, Nakanishi Y, Yoshimura K, Tobe T, Clarke JM, Topping DL, Suzuki T, Taylor TD, Itoh K, Kikuchi J, Morita H, Hattori M, Ohno H (2011) Bifidobacteria can protect from enteropathogenic infection through production of acetate. Nature 469:543–547. https://doi.org/10.1038/nature09646
Cummings JH (1981) Short chain fatty acids in the human colon. Gut 22:763–779. https://doi.org/10.1136/gut.22.9.763
Topping DL, Clifton PM (2001) Short-chain fatty acids and human colonic function: roles of resistant starch and nonstarch polysaccharides. Physiol Rev 81:1031–1064. https://doi.org/10.1152/physrev.2001.81.3.1031
Morrison DJ, Preston T (2016) Formation of short chain fatty acids by the gut microbiota and their impact on human metabolism. Gut Microbes 7:189–200. https://doi.org/10.1080/19490976.2015.1134082
Miller TL, Wolin MJ (1996) Pathways of acetate, propionate, and butyrate formation by the human fecal microbial flora. Appl Environ Microbiol 62:1589–1592
Flint HJ, Duncan SH, Scott KP, Louis P (2015) Links between diet, gut microbiota composition and gut metabolism. Proc Nutr Soc 74:13–22. https://doi.org/10.1017/S0029665114001463
Macfarlane S, Macfarlane GT (2003) Regulation of short-chain fatty acid production. Proc Nutr Soc 62:67–72. https://doi.org/10.1079/PNS2002207
Louis P, Hold GL, Flint HJ (2014) The gut microbiota, bacterial metabolites and colorectal cancer. Nat Rev Microbiol 12:661–672. https://doi.org/10.1038/nrmicro3344
Pomare EW, Branch WJ, Cummings JH (1985) Carbohydrate fermentation in the human colon and its relation to acetate concentrations in venous blood. J Clin Invest 75:1448–1454. https://doi.org/10.1172/JCI111847
Pessione E (2012) Lactic acid bacteria contribution to gut microbiota complexity: lights and shadows. Front Cell Infect Microbiol 2:86. https://doi.org/10.3389/fcimb.2012.00086
Elshaghabee FMF, Bockelmann W, Meske D, de Vrese M, Walte HG, Schrezenmeir J, Heller KJ (2016) Ethanol production by selected intestinal microorganisms and lactic acid bacteria growing under different nutritional conditions. Front Microbiol 7:47. https://doi.org/10.3389/fmicb.2016.00047
Enjalbert B, Millard P, Dinclaux M, Portais JC, Létisse F (2017) Acetate fluxes in Escherichia coli are determined by the thermodynamic control of the Pta-AckA pathway. Sci Rep 7:42135. https://doi.org/10.1038/srep42135
Torino MI, Mozzi F, Font de Valdez G (2005) Exopolysaccharide biosynthesis by Lactobacillus helveticus ATCC 15807. Appl Microbiol Biotechnol 68:259–265. https://doi.org/10.1007/s00253-004-1865-2
Tedelind S, Westberg F, Kjerrulf M, Vidal A (2007) Anti-inflammatory properties of the short-chain fatty acids acetate and propionate: a study with relevance to inflammatory bowel disease. World J Gastroenterol 13:2826–2832. https://doi.org/10.3748/wjg.v13.i20.2826
Brown AJ, Goldsworthy SM, Barnes AA, Eilert MM, Tcheang L, Daniels D, Muir AI, Wigglesworth MJ, Kinghorn I, Fraser NJ, Pike NB, Strum JC, Steplewski KM, Murdock PR, Holder JC, Marshall FH, Szekeres PG, Wilson S, Ignar DM, Foord SM, Wise A, Dowell SJ (2003) The orphan G protein-coupled receptors GPR41 and GPR43 are activated by propionate and other short chain carboxylic acids. J Biol Chem 278:11312–11319. https://doi.org/10.1074/jbc.M211609200
Maslowski KM, Vieira AT, Ng A, Kranich J, Sierro F, di Yu, Schilter HC, Rolph MS, Mackay F, Artis D, Xavier RJ, Teixeira MM, Mackay CR (2009) Regulation of inflammatory responses by gut microbiota and chemoattractant receptor GPR43. Nature 461:1282–1286. https://doi.org/10.1038/nature08530
Hill MJ (1997) Intestinal flora and endogenous vitamin synthesis. Eur J Cancer Prev 6(Suppl 1):S43–S45. https://doi.org/10.1097/00008469-199703001-00009
Rowland I, Gibson G, Heinken A, Scott K, Swann J, Thiele I, Tuohy K (2018) Gut microbiota functions: metabolism of nutrients and other food components. Eur J Nutr 57:1–24. https://doi.org/10.1007/s00394-017-1445-8
Magnúsdóttir S, Ravcheev D, de Crécy-Lagard V, Thiele I (2015) Systematic genome assessment of B-vitamin biosynthesis suggests co-operation among gut microbes. Front Genet 6:148. https://doi.org/10.3389/fgene.2015.00148
Said HM, Ortiz A, Moyer MP, Yanagawa N (2000) Riboflavin uptake by human-derived colonic epithelial NCM460 cells. Am J Phys Cell Phys 278:C270–C276. https://doi.org/10.1152/ajpcell.2000.278.2.C270
Shin R, Suzuki M, Morishita Y (2002) Influence of intestinal anaerobes and organic acids on the growth of enterohaemorrhagic Escherichia coli O157:H7. J Med Microbiol 51:201–206. https://doi.org/10.1099/0022-1317-51-3-201
Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, Morelli L, Canani RB, Flint HJ, Salminen S, Calder PC, Sanders ME (2014) Expert consensus document. The international scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat Rev Gastroenterol Hepatol 11:506–514. https://doi.org/10.1038/nrgastro.2014.66
Acknowledgements
This study was sponsored by Ratiopharm GmbH, which is an affiliate of Teva Pharmaceutical Industries Ltd. Medical writing support was provided by Matthew Gunther, PhD, of Zoetic Science, an Ashfield company, part of UDG Healthcare plc, and was funded by Ratiopharm GmbH (Ulm, Germany).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of Interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Patil, S., Sawant, S., Hauff, K. et al. Validated Postbiotic Screening Confirms Presence of Physiologically-Active Metabolites, Such as Short-Chain Fatty Acids, Amino Acids and Vitamins in Hylak® Forte. Probiotics & Antimicro. Prot. 11, 1124–1131 (2019). https://doi.org/10.1007/s12602-018-9497-5
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12602-018-9497-5