nmr analysis of beer - particularly sour beers

Download NMR Analysis of Beer - Particularly Sour Beers

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NMR presentation on Sour Beer Analysis and other Beer Brewing Processes

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  • 1. When Beer Goes Sour: An NMR InvestigationJOHN EDWARDS AND ADAM DICAPRIOPROCESS NMR ASSOCIATESMESTRELAB MNOVA USERS MEETINGSMASH ATLANTA, GA, SEPTEMBER 7, 2014

2. What is Sour Beer?Wikipedia says:Sour beer is a beer style characterized by an intentionally acidic, tart, sour taste.Wild Brews: Beer beyond the Influence of Brewers YeastCategory 17 of the Beer Judge Certification ProgramEncompasses: Berliner Weisse, Flanders Red Ale, Flanders Brown Ale, Lambic, Fruit Lambic, Gueuze 3. What goes in to a sour beer?GrainMalted Barley, Unmalted Wheat, Specialty MaltsDextrins, Dextrins, DextrinsSikaru beer (3000 B.C.) 62.5% Barley Malt + 37.5% Raw Wheat2Modern Lambic Brasserie Cantillon recipe 65% Barley Malt + 35% Raw Wheat2HopsAged & OxidizedAging Vessels A sour beers home for up to a full centuryOak Barrels (French & American)Oak FoudreStainless Steel Tankhttp://www.newbelgium.com/Community/Blog/12-03- 23/Who-wants-more-sour-beer.aspxhttp://www.belgianbeermagazine.com/oud-beersel-brewery/ 4. Who goes in to a sour beer?Dozens of organismsBacteriaEnterobacteriaceaeCitrobacter spp., Enterobacter spp., Klebsiella spp., Hafnia spp.LactobacillaceaePediococcus spp., Lactobacillus spp.,Acetobacter spp.Klebsiella spp.YeastsKloeckera apiculataSaccharomyces spp.Brettanomyces spp.Pichia spp.Candida spp.Hansenula spp.Cryptococcus spp.Typical Coolship design. 5. Why is Sour Beer Sour?Straight Lambic, Flanders Ales, Gueuze, Berliner WeisseLactic, Acetic, Succinic Acid185% - 10% - 5%Fruit Lambics1Cherries, Grapes Malic AcidRaspberries Citric Acid 6. Chemistry of Sour BeersApplication of Quantitative NMR to Biologically Acidified MashesQuantitative NMR and Descriptive Chemistry of American Wild Ales and genuine Belgian Lambic 7. Berliner Weisse & Biological AcidificationReinheitsgebot of 1516Beer can contain only malt, hops & waterUnmalted wheat and yeast added in the Provisional Law of 19964Artificial alteration of pH is illegal5Development of Biological Acidification/Sour MashingUtilization of native microbes for pH adjustment6Perform Starch ConversionCool to ~120FInnoculateHold Temperature (120F) 8. Temperature Dependence of the Sour Mash TechniqueGoal:Does the magic temperature of 120F have a chemical significance?Record and quantify sour metabolites & contaminant products as a function of sour mash temperatureDetermine wt% of metabolites using Maleic Acid internal standardmanual integration, and Mestrelab GSD SMA plug-inLactic AcidAcetic Acid (contaminant)Succinic AcidEthanol-Amino Butyric Acid (contaminant) 9. Typical beer chemistry observed by 1H NMR 10. Absolute Metabolite Proportions0.002.004.006.008.0010.0012.0014.0016.009095100105110115120125130135140 Metabolite Weight % vs. TemperatureLactic AcidAcetic AcidSuccinic AcidPoly. (Lactic Acid)Poly. (Acetic Acid)Poly. (Succinic Acid)00.20.40.60.811.21.490110130Percent by MassTemperature (F)Weight % Ethanol 11. Relative ProportionsR = 0.96760.050.0100.0150.0200.0250.0300.095105115125135WT % LACTIC / WT % SUCCINICTEMPERATURELactic:SuccinicR = 0.99410.002.004.006.008.0010.0012.0014.0016.0090100110120130140WT % LACTIC / WT % ACETICTEMPERATURE (F) Lactic:Acetic 12. Conclusions120F is sub-optimal for acid productionAround 120F Lactic acid reaches a relative maximumLactic good, Acetic badAim is pH adjustment, not flavor adjustment 13. Lambics of Belgium & Lambic-Styles of the USAWhats the difference?Lambic From the Pajottenland / Senne River Valley Region of BelgiumAmerican likenesses styled as American Wild Ale (AWA) or American Coolship Ale (ACA)Different Microbial CommunityFollow same general successionACA involves a more diverse community of Lactic Acid Bacteria and Minority Yeasts 14. The Chemistry of Sour BeersGoalsComparative analysis of organic acids using quantitative NMRManually integrated and SMA analysis against a known mass of Maleic AcidLactic Acid, Acetic Acid, Succinic Acid, Citric Acid, Malic AcidAnalyze linear and branched dextrin ratios among multiple stylesUtilize multivariate analysis to discriminate multiple styles of sour beer 15. Acid Differences77%20%3%American Geuze-StyleLacticAceticSuccinic85%11%4%Belgian GeuzeLacticAceticSuccinic43%18%4%35%American Kriek-StyleLacticAceticSuccinicMalic35%27%3%35%American Framboise-StyleLacticAceticSuccinicCitricBeerLA (mg/L)AA (mg/L)SA (mg/L)CA (mg/L)MA (mg/L)American Geuze-Style5386.01410.0238.500American Framboise- Style3896.72972.1394.63890.70American Kriek-Style4682.81965.7423.403777.8Boone Mariage Parfait 20094506.5488.1217.000Oude Geuze Vieille3497.8454.1175.200Geuze Fond Tradition6807.8698.6218.800Drie Fonteinen A5137.6865.9234.600Drie Fonteinen B5389.9917.7228.900 16. BreweryBeerLacticAcidAcetic AcidSuccinic AcidCitric AcidLactic AcidAcetic AcidSuccinic AcidCitricAcidAllagashConfluence850.5402.4167.4248.2901.3437.6184.3353.2BudweiserBud Light93.938.135.882.7117.117.734.262.6Crooked StaveSurette4699.2564.9195.7265.54247.9549.2185.6262.4Drie FonteinenOude Gueuze5137.6865.9234.60.04935.2787.1204.60.0Drie FonteinenOude Gueuze5389.9917.7228.90.05180.0775.2194.40.0LoverBeerD'Uva Beer3071.4781.5876.0358.03302.8759.9842.4377.3BooneMariage Parfait 20094506.5488.1216.90.04591.8403.2193.10.0Comparison of results obtained by manual integration and by automatic Mnova SMA. Units=mg/LManual IntegrationMnova SMA 17. Absolute concentrations of major components found in AWA (Acids: left y-axis, EtAc: right y-axis). The age of the beers refers to the fermentation time (in years) for the respective barrels sampled, as all samples came from separate batches. The 1 & 3 year old barrels were brewed in the Winter (November) and the 2 year old barrel was brewed in the Spring (May). Blend refers to the finished Gueuze-style AWA.0200400600800100012001400160018000100020003000400050006000700080009000100001123BlendConcentration (mg/L)Age (years)Citric AcidSuccinic AcidAcetic AcidLactic AcidEthyl Acetate 18. Anomeric 1H region of the beer spectrum maltose, dextrins and reduced sugars 19. Dextrin Ratios01234567(1-4) MALTODEXTRIN / (1-6) MALTODEXTRINSTYLERatio of (1-4) Maltodextrin to (1- 6) Maltodextrin by StyleDegree of FermentationDependent on:StrainStyleIngredients 20. Multivariate Analysis Principal Component AnalysisSegregate based largely on sugar detailAmericanCoolshipBeersBelgianGeuzesSaisonsBelgianFoudreBeerIPA + Lagers100%BrettBeersAmericanFoudreBeerHybrid BeerSaisonsLambicACAIPA + LagerBel. Foudre100% Brett.Am. Foudre 21. PCA on Small Acid Region Only Binned Data 22. Aromatic Region of Beer Spectrum 23. Poor Discrimination of BeerStyles from aromatic chemistry 24. ConclusionsACAs & Belgian Lambics have differing acid profilesRatio of linear and branched maltodextrins can be used to differentiate stylesMultivariate analysis can differentiate between ACAs, Belgian Lambics and 6 other styles 25. MaltLovibondAmerican MaltsU.S. two-row1.4 - 1 8U.S. six-row1.5 - 1.9Pale Malt2Canadian two-row1.3 - 1.7Canadian six-row1.4 - 1.9Wheat Malt2.2Vienna Malt4Munich Malt10Carapils2Light Crystal10Pale Crystal40Medium Crystal60Dark Crystal120Victory Malt25Special Roast50Chocolate Malt350Roast Barley300Black Barley450-500Black Patent500Malt ChemistryMaillard Chemistry Glucose + Amino Acid -- Schiff Base 26. 13C NMR Malt/Hot Water Extracts 27. 1H NMR Malt/Hot Water Extracts 28. Multivariate Analysis andChemistry Identification 29. Hop Chemistry 30. Alpha acids (humulone, adhumulone, cohumulone)Beta acids (primarily humulene, myrcene, caryophyllene and farnesene)Hops in beer are the acids and essential oils ofHops Bittering, Flavor, Aroma, Foam StabilityAlpha acids isomerization toiso-alpha acids over the course ofboiling the beer wort (60+ minutes) 31. More Ongoing Fermentation ProjectsYards Brewing Complete analysis of the brewing process from mash through finished bottled products Thomas Jefferson AleNMR and Portable MS analysis of beer production process at Mill Street Brewery, PoughkeepsieExpand to include Mead and Cider 32. AcknowledgmentsAdam DiCaprio needs a job in the Raleigh-Durham AreaAllagash Brewing Company,Portland MEYards Brewing Company, Philadelphia PA