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The Colloidal Stability of Craft Beers Obtained with Gluten-Free Adjuncts: an Assessment of Aspects Related to Technology, Composition and Analysis

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The Colloidal Stability of Craft Beers Obtained with Gluten-Free Adjuncts: an Assessment of Aspects Related to Technology, Composition and Analysis The colloidal stability of craft beers obtained with gluten-free adjuncts: an assessment of aspects related to technology, composition and analysis A Ph.D. dissertation presented by Paolo Passaghe to the University of Udine for the degree of Ph.D. in the subject of Food Science (Cycle XXVI) Department of Food Science UNIVERSITY OF UDINE Italy March 2014 Coordinator: Mara Lucia Stecchini, Professor Department of Food Science University of Udine, Italy Supervisor: Dr. Stefano Buiatti Department of Food Science University of Udine, Italy Reviewers: Andrea Antonelli, Professor Department of Life Science University of Modena-Reggio Emilia, Italy Dr. Emanuele Boselli Department of Agricultural, Food and Environmental Sciences Polytechnic University of Ancona, Italy ABSTRACT It is generally known that haze in beer is directly related to the formation of complexes resulting from the interaction of haze active polyphenols and a protein fraction. This project aims to study issues related to the physical and chemical stability of craft beers. Specifically, the plan is to analyze how some variables in the malting process (i.e., pH of the first steeping water) and formulation (use of gluten-free adjuncts) can contribute to the colloidal stability (shelf-life) of the final beer. Polypeptides responsible for haze formation originate mainly from barley and are rich in the amino acid proline. Incidentally, these proteins are also responsible for the immune reaction experienced by coeliacs; therefore haze prevention in beer and rendering the beer “gluten-free” could be compatible practices. The primary aim of the study was to carry out micro malting tests in the laboratory on four gluten-free cereals/pseudocereals (millet, amaranth, buckwheat and quinoa) in order to identify the optimal conditions for obtaining malts suitable for the production of craft beers. The use of an alkaline solution in the first steeping may facilitate the extraction of proteinaceous and phenolic fractions from seeds; for this reason the pH of the first steeping water was modified. Quality of malts was checked analysing diastatic power, beta glucans content and Kolbach index (soluble nitrogen content as a percentage of total nitrogen). Nine beers with different formulations were produced in the laboratory (2 L, micro-brewing): beer made with 100% barley malt (reference sample), beers i with 40% substitute gluten-free malt using the best malts obtained from the micromalting tests and 60% of barley malt. The use of gluten-free substitutes was intended to reduce the base level of the protein fractions rich in proline. The evaluation of beer stability was carried out analyzing some indices validated by EBC (European Brewery Convention) (sensitive proteins and cold turbidity respectively), and two unconventional methods (gluten analysis and antioxidant activity, AA) to verify the possible correlation among these analyses and the official methods. Measurements of beer AA, which is mainly due to the polyphenol content, could give useful information on the colloidal stability of the final beer product. Subsequently, the experimental plan was transferred on a larger scale; three beers were produced at the pilot plant of University of Udine (capacity of 200 L) using only the malt with the highest diastatic power (buckwheat): the first with 100% malted barley (reference sample), the other two with increasing amounts of malted buckwheat (20 and 40%). The colloidal stability of beer samples was tested with the same four analyses used to verify physico-chemical stability of the laboratory produced samples. The results obtained from the micromalting tests were in line with the expectations: the four gluten-free cereal/pseudocereals were under-modified if compared to traditional malts, but potentially suitable as adjuncts (especially buckwheat). The unconventional methods used to study the physico-chemical stability of craft beers provided encouraging results: either gluten analysis or the crocin test (AA) data showed trends correlated with those obtained with the validated method (alcohol chill haze). Specifically, the gluten analysis discriminated the beers produced at the pilot plant better than sensitive proteins (EBC method). ii Based on the collected results, it can be concluded that the use of gluten- free adjuncts, combined with relevant process conditions (i.e. alkalinized first steeping water in the malting process), could lead to more stable final products with a gluten content less than 100 mg/L and so potentially suitable by coeliacs. The colloidal stability monitoring of the beers produced at the pilot plant has already been planned, as well as the descriptive sensory analysis of the same samples. iii iv RIASSUNTO È noto che l'intorbidamento nella birra sia la diretta conseguenza della formazione di complessi risultanti dall'interazione tra frazioni polifenoliche e proteiche cosiddette attive. Questo progetto ha lo scopo di studiare le problematiche legate alla stabilità chimico fisica delle birre artigianali. Nello specifico, l'obiettivo è stato quello di analizzare come alcune variabili del processo di maltazione (ad esempio il pH dell'acqua utilizzata nella prima bagnatura) o come la formulazione (utilizzo di succedanei privi di glutine), possano contribuire alla stabilità colloidale (shelf-life) del prodotto finale. I polipeptidi responsabili dell'intorbidamento derivano principalmente dall'orzo e sono ricchi nella loro sequenza amminoacidica dell'amminoacido prolina. Queste frazioni proteiche sono inoltre responsabili delle reazioni di intolleranza nelle persone affette da celiachia; da ciò deriva l'idea che abbassare il contenuto di glutine nella birra possa stabilizzare la stessa verso i fenomeni di intorbidamento. Quindi, il primo obiettivo di questo lavoro è stato individuare le condizioni ottimali di maltazione di quattro cereali/pseudocereali privi di glutine (miglio, amaranto, grano saraceno e quinoa) al fine di ottenere malti idonei per la produzione della birra artigianale, attraverso delle prove di micro maltazione effettuate in laboratorio. L'utilizzo di una soluzione alcalina nella prima bagnatura può facilitare l'estrazione dai semi di frazioni proteiche e polifenoliche; per questo motivo è stato modificato il pH dell'acqua impiegata nella prima bagnatura. La qualità dei malti ottenuti è stata controllata attraverso le analisi del potere diastatico, dei beta glucani e dell'indice di Kolbach (rapporto tra azoto solubile e totale). v Sono state, quindi, prodotte nove birre in laboratorio (micro-birrificazioni da 2 L): un riferimento ottenuto con solo malto d'orzo (100% p/p), e birre prodotte con il 40% p/p di succedaneo senza glutine (ottenuto dalle prove di micro maltazione), e il restante 60% p/p di malto d'orzo. L'impiego di succedanei senza glutine aveva l'obiettivo di ridurre il livello di base delle frazioni proteiche con un alto contenuto dell'amminoacido prolina. La valutazione della stabilità colloidale delle birre è stata effettuata attraverso due indici dell'EBC (European Brewery Convention) (rispettivamente proteine sensibili e torbidità a freddo), e due metodi non convenzionali (analisi del glutine e dell'attività antiossidante, AA), ed è stato valutato il grado di correlazione tra le diverse metodiche. La misura dell'AA dei campioni di birra, principalmente determinata dai composti polifenolici contenuti nei medesimi campioni, potrebbe fornire informazioni utili sulla stabilità colloidale della birra. Successivamente, il piano sperimentale è stato trasferito su scala maggiore; sono state prodotte tre birre presso l'impianto pilota universitario (micro-birrificazioni da 200 L): la prima con il 100% di malto d'orzo, e le altre due con quantità crescenti di grano saraceno maltato (20 e 40%). La valutazione della stabilità colloidale delle birre è stata realizzata con le medesime analisi utilizzate per i campioni di birra prodotti in laboratorio. I risultati ottenuti con le prove di micro-maltazione sono in linea con le attese: i quattro cereali/pseudocereali senza glutine sono risultati sotto-modificati rispetto ad un tradizionale malto, ma potenzialmente impiegabili come succedanei per la produzione di birra (in modo particolare il grano saraceno). Le due metodiche non convenzionali, utilizzate per valutare la stabilità chimico fisica delle birre artigianali, hanno fornito risultati incoraggianti: sia i dati ottenuti con l'analisi del glutine che quelli ottenuti con il test della crocina (AA) vi hanno evidenziato andamenti correlabili a quelli ottenuti con il metodo validato (test della torbidità a freddo). Nello specifico, l'analisi del glutine discriminava meglio le birre prodotte presso l'impianto pilota rispetto al test delle proteine sensibili (metodo EBC). Sulla base dei risultati raccolti, può essere dedotto che l'impiego di succedanei privi di glutine, affiancato ad adeguate condizioni di processo (ad esempio l'alcalinizzazione dell'acqua impiegata nella prima bagnatura del processo di maltazione), potrebbe portare alla produzione di birre più stabili e con un contenuto di glutine inferiore a 100 mg/L e, quindi, idonee per le persone affette da celiachia. Inoltre, è stato pianificato sia il monitoraggio della stabilità colloidale che l'analisi sensoriale delle birre prodotte presso l'impianto pilota. vii viii TABLE OF CONTENTS 1. OVERVIEW OF THE BREWING PROCESS ................................................................ 1 1.1 Beer .......................................................................................................................
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