Beer Analysis

Raw Material.- Barley and Malt Analysis.- 1 General Introduction.- 2 Barley Analysis.- 2.1 Visual Inspection.- 2.2 Mechanical Analysis.- 2.3 Germination Tests.- 2.4 Chemical Analysis.- 2.5 Varietal Identification.- 2.6 Micro-Malting.- 3 Malt Analysis.- 3.1 Wort Production and Analysis.- 3.2 Modification Analysis.- 3.3 Chemical Analysis.- 3.4 Enzyme Analysis.- 4 Further Reading.- References.- Reversed Phase HPLC as an Aid for the Identification of Barley Cultivars.- 1 Introduction.- 1.1 Reversed-Phase High Performance Liquid Chromatography.- 1.1.1 The RP-HPLC System.- 1.1.2 Sample Preparation.- 1.1.3 Hordeins.- 1.1.4 Albumins and Globulins.- 1.1.5 Glutelins.- 1.2 Hordein Identification.- 1.3 Repeatability of Hordein Elution.- 1.3.1 RP-HPLC Separations of Hordeins.- 1.3.2 RP-HPLC Separation of Albumins and Globulins.- 1.3.3 RP-HPLC Separation of Glutelins.- References.- The Application of Immunofluorescence for Detecting Specific Proteins in Barley Seeds.- 1 Introduction.- 2 Immunofluorescence.- 2.1 Fluorescent Dyes.- 2.2 Staining Methods.- 2.2.1 The Direct Method with Fluorochrome-Labelled Primary Antibodies.- 2.2.2 The Indirect Method with Fluorochrome-Labelled Secondary Antibodies.- 2.2.3 Use of Biotin-Labelled Primary Antibodies.- 2.2.4 Use of Biotin-Labelled Secondary Antibodies.- 3 Materials, Protocols and Comments.- 3.1 Production and Purification of Antibodies.- 3.2 Conjugation of Antibodies with Fluorochromes.- 3.2.1 Protocol for FITC Conjugation.- 3.2.2 Comments to the FITC Conjugation Procedure.- 3.2.3 Conjugation with Other Fluorochromes.- 3.3 Biotinylation.- 3.3.1 Protocol for Conjugation with Biotin.- 3.3.2 Comments to the Biotin Conjugation.- 3.4 Fixation.- 3.4.1 Protocol for PLP Fixation of Seeds to be Sectioned.- 3.4.2 Surface Fixation of Sanded Seeds for Retention of Germination Capacity.- 3.4.3 Comments to the Fixation.- 3.5 Preparation of Tissue.- 3.5.1 Thin Sections.- 3.5.2 Sanded Seeds.- 3.6 Staining Procedures.- 3.6.1 General Considerations.- 3.6.2 Direct Staining of Thin Sections.- 3.6.3 Indirect Staining of Thin Sections.- 3.6.4 Use of Biotin-Labelled Primary Antibodies.- 3.6.5 Use of Biotin-Labelled Secondary Antibodies.- 3.6.6 Two-Colour Staining of Thin Section.- 3.6.7 Direct Staining of Sanded Seeds.- 4 Concluding Remarks.- References.- Fluorimetric Methods for the Analysis of Malt Modification and Pre-Harvest Sprouting.- 1 Introduction.- 2 Modification and Homogeneity of Barley Malt.- 2.1 Field of Application.- 2.2 Sample Size.- 2.3 Reagents and Materials.- 2.4 Apparatus.- 2.5 Procedure.- 2.5.1 Fixation and Sanding.- 2.5.2 Staining.- 2.5.3 Examination.- 2.6 Expression of Results.- 2.7 Use of Malt Modification Analysis.- 3 Pre-Sprouting in Cereal Seeds.- 3.1 Field of Application.- 3.2 Sample Size.- 3.3 Reagents and Materials.- 3.4 Apparatus.- 3.5 Procedure.- 3.5.1 Fixation and Sanding.- 3.5.2 Staining.- 3.5.3 Examination.- 3.6 Expression of Results.- 3.7 Use of Pre-Sprouting Test.- 4 Further Applications of the Carlsberg Seed Fixation System.- References.- Hop Analysis.- 1 Introduction.- 2 Chemical Composition of Hops.- 3 Analysis.- 3.1 Resin Analysis of Hops.- 3.2 The Conductometric Lead Value of Hops.- 3.3 Verzele Conductometric Analysis of ?-Acids.- 3.4 Beer Bitterness Analysis.- 3.5 High Performance Liquid Chromatography (HPLC).- 4 Essential Oil of Hops.- 5 Polyphenols.- 6 Prospect.- References.- Determination of ?-Glucan in Barley, Malt, Wort and Beer.- 1 Introduction.- 2 The Nature of ?-Glucan.- 3 Quantification of Barley ?-Glucan.- 3.1 Methods Based on Extraction.- 3.1.1 Viscosity Methods.- 3.1.2 Difference Methods.- 3.1.3 Selective Precipitation Methods.- 3.2 Non-Extraction Methods.- 3.2.1 Near Infrared Reflectance.- 3.2.2 Calcofluor Staining Used in Flour Suspensions.- 3.3 Enzymatic Methods.- 3.3.1 Methods Using Bacterial ?-Glucanase.- 3.3.2 Methods Based on Fungal Cellulases.- 4 Quantification of ?-Glucan in Malt, Wort and Beer.- 4.1 Total ?-Glucan Content of Malt.- 4.1.1 Non-Enzymatic Methods.- 4.1.2 Enzymatic Methods.- 4.2 ?-Glucan Content of Wort and Beer.- 4.2.1 Non-Enzymatic Methods.- 4.2.2 Enzymatic Methods.- 5 An Automatic ?-Glucan Analyzer System.- 5.1 Principle.- 5.2 Apparatus and Conditions.- 5.3 Quantification of ?-Glucan in Barley, Malt, Wort and Beer.- 5.3.1 Sample Preparation for Wort and Beer.- 5.3.2 Sample Preparation for Barley and Malt.- 6 Evaluation of Methods.- 6.1 Specificity.- References.- Analysis of Proanthocyanidins and Phenolic Acids in Barley, Malt, Hops and Beer.- 1 Introduction.- 2 Choice of Analytical Methods.- 3 Chromatographic Methods.- 3.1 High Performance Liquid Chromatography (HPLC).- 3.1.1 Preparation of Barley and Malt Samples.- 3.1.2 Preparation of Hop Samples.- 3.1.3 Preparation of Beer Samples.- 3.1.4 Choice of HPLC Columns.- 3.1.5 Mobile Phase.- 3.1.6 Standards.- 3.1.7 Quantitation.- 3.2 Thin Layer Chromatography (TLC).- 3.3 Column Chromatography.- 4 Colourimetric Methods.- 4.1 Determination of Proanthocyanidins in Barley, Malt, Wort and Beer.- 4.2 Measurement of Total Polyphenols in Beer and Wort.- 4.3 Analysis of Proanthocyanidins and Catechins Using Acidified Vanillin.- 4.4 Determination of Proanthocyanidins and Catechins in Beer with p-Dimethylaminocinnamaldehyde as a Reagent.- 5 Vanillin Test of Barley Grains.- References.- Yeast.- Breeding and Characterization of Brewer's Yeast Including Single Chromosome Transfer.- 1 Introduction.- 2 Sporulation, Isolation of Meiotic Progeny and Construction of Hybrids.- 2.1 Mass Spore Isolation.- 2.2 Isolation of Colonies and Spore Staining.- 2.3 Determination of Mating Type.- 2.4 Construction of Hybrids by Mating of Meiotic Segregants.- 2.5 Random Mating.- 3 Genetics of Diacetyl Formation.- 3.1 Isolation of Spontaneous Mutants Resistant to Sulfometuron Methyl.- 4 Transformation.- 5 Single Chromosome Transfer.- 5.1 Transfer of Chromosome V.- 5.2 Characterization of Chromosome V by Restriction Endonuclease Analysis.- References.- Genetic Analysis and Transformation of Distiller's Yeast.- 1 Introduction.- 2 Genetic Analysis.- 2.1 Sporulation and Mating Ability.- 2.1.1 Sporulation Test.- 2.1.2 Spore Isolation.- 2.1.3 Mating Type Tests.- 2.1.3.1 Induction of Respiratory-Deficient Mutations.- 2.1.3.2 Determination of Mating Type.- 2.1.3.3 Rare Mating on Solid Medium.- 2.1.3.4 Rare Mating in Liquid Medium.- 2.1.4 Characterization of Auxotrophic Spore Clones.- 2.1.5 Sporulation and Mating Ability of Strain A.- 2.2 Mutagenesis.- 2.2.1 Procedure.- 2.2.2 Mutants Obtained from Spore Clones of Strain A.- 2.3 Crossing and Tetrad Analysis.- 3 Molecular Characterization.- 3.1 Procedure.- 3.2 Analysis of Chromosomes III of Strain A.- 4 Genetic Transformation of an Industrial Yeast Strain.- 4.1 Selection of Transformants by Acquired Prototrophy.- 4.1.1 Transformation Procedure.- 4.1.2 Screening for Amylase Activity.- 4.1.3 Stability Test of Transformants.- 4.1.4 Integration of a Self-Replicating Plasmid.- 4.1.5 Breeding Through Transformation of Spore-Derived Strains.- 4.2 Integration Vector with Drug Resistance as Selective Marker.- 4.2.1 Transformation Procedure Using a Non-Replicating Plasmid.- 4.2.2 Screening for ?-Galactosidase Activity.- References.- Automated Testing of Brewer's Yeast Strains for Fermentation Characteristics (Multiferm, System Carlsberg).- 1 Introduction.- 2 Testing Techniques.- 2.1 Small-Scale Fermentations.- 2.2 MULTIFERM Fermentations.- 2.2.1 Description of the MULTIFERM System.- 2.2.2 Working Procedures.- 2.2.2.1 Propagation of Yeast.- 2.2.2.2 Wort Collection and Handling.- 2.2.2.3 Start of Fermentations.- 2.2.2.4 Sampling During Fermentations.- 2.2.2.5 End of Fermentations.- 2.2.2.6 Cleaning in Place of the MULTIFERM.- 2.2.2.7 Re-Use of Yeast.- 2.2.3 Analyses Performed.- 2.2.3.1 Yeast Cell Size and Yeast Cell Number.- 2.2.3.2 Pitching Rate.- 2.2.3.3 Extract Determination.- 2.2.3.4 Yeast Yield.- 2.2.3.5 Vicinal Diketones and Ester/Alcohol (Aroma Profile) Analyses.- 2.2.4 Data.- 2.2.4.1 Data Handling and Storage.- 2.2.4.2 Data Evaluation.- 3 Conclusion.- References.- The Use of Nucleotide Sequence Polymorphisms and DNA Karyotyping in the Identification of Brewer's Yeast Strains and in Microbiological Control.- 1 Introduction.- 1.1 Restriction Fragment Length Polymorphisms.- 1.2 DNA Karyotypes.- 2 Storage and Propagation of Yeast Strains.- 3 Restriction Site Analysis of Yeast Genomic DNA.- 3.1 Isolation of Yeast Genomic DNA for Restriction Site Analysis.- 3.2 Analysis of Restriction Fragment Length Polymorphisms (RFLP).- 3.2.1 RFLP Level A.- 3.2.2 RFLP Level B.- 4 Southern Transfer and Molecular Hybridization of Radioactively Labelled Probes.- 4.1 Transfer of DNA Molecules from Agarose Gels onto Nitrocellulose Membranes by Southern Transfer.- 4.2 Molecular Hybridization.- 4.2.1 Hybridization Buffers.- 4.2.2 Nick Translation.- 4.2.3 Post Hybridization Wash.- 5 Isolation of Yeast Genomic DNA for Chromosome Separation.- 5.1 Orthogonal Field Alternation Gel Electrophoresis.- 5.2 Preparation of Chromosomal DNA for OFAGE Karyotypes.- References.- Screening and Testing New Distillers' Yeasts for Their Potential in Molasses Ethanol Fermentations.- 1 Introduction.- 2 Screening and Testing Techniques.- 2.1 Small-Scale Batch Fermentations.- 2.1.1 Description of Apparatus.- 2.1.2 Propagation of Yeast.- 2.1.3 Fermentation and Analyses.- 2.2 Small-Scale Continous Fermentations.- 2.2.1 Propagation of Yeast.- 2.2.2 Fermentation and Analysis.- 2.3 Full-Scale-Fermentation.- 2.3.1 Propagation of Yeast.- 2.3.2 Fermentation Equipment.- 2.3.3 Analyses Performed.- 2.3.3.1 Oxidation Method of Ethanol Determination.- 2.3.3.2 Sugar Concentration Determined by Fermentation.- 2.3.4 Regulation of Fermentation.- 3 Evaluation of Results.- 3.1 Small-Scale Fermentations.- 3.2 Production-Scale Fermentations.- References.- Genetic Analysis and Manipulation of Mucor Species by DNA-Mediated Transformation.- 1 Introduction.- 1.1 Taxonomy of Mucor.- 1.2 Industrial Use of Mucor.- 1.3 Genetic Analysis and Manipulation.- 2 General Methods.- 2.1 Media.- 2.2 Growth Conditions.- 2.3 Harvesting, Storage and Germination of Sporangiospores.- 2.4 Replica Plating of Colonies.- 3 DNA-Mediated Transformation of Mucor.- 3.1 Plasmids pMCL006 and pMCL1302.- 3.2 Method for Transformation of Mucor with Plasmid DNA.- 3.3 Properties of Transformants.- 4 Isolation, Regeneration and Fusion of Protoplasts from Mucor.- 4.1 Formation of Protoplasts from Mucor.- 4.2 Preparation of a Lytic Enzyme from Streptomyces sp. no. 6 (Streptozyme).- 4.3 Regeneration of Protoplasts.- 4.4 Fusion of Protoplasts.- 5 Isolation of Marker Genes for Use in DNA-Mediated Transformation.- 5.1 Selectable Marker Genes Coding for Resistance to Antibiotics.- 5.2 Selectable Marker Genes Coding for Metabolic Enzymes.- 5.3 Isolation and Enrichment of Auxotrophic Mutants of Mucor.- 6 Analysis of Transformants.- 6.1 Isolation of RNA.- 6.2 Isolation of DNA.- 6.3 Recovery of Plasmid DNA.- 7 Genetic Manipulation of Mucor by DNA-Mediated Transformation. Concluding Remarks.- References.- Final Product.- Malt and Hop Flavanoids in Pilsner Beer.- 1 Importance.- 2 Identification of Flavanoids in Barley, Malt, Hops and Beer.- 2.1 Barley and Malt Flavanoids.- 2.2 Hop Flavanoids.- 2.3 Beer Flavanoids.- 3 Analysis of Beer Flavanoids.- 3.1 Analysis of Beer Anthocyanogens.- 3.1.1 Introduction.- 3.1.2 Method.- 3.2 Analysis of Total Polyphenolics.- 3.2.1 Introduction.- 3.2.2 Method.- 3.3 Analysis of Flavanoids.- 3.3.1 Introduction.- 3.3.2 Method.- 3.4 Comparison of the Methods.- 4 Significance of the Analytical Results.- 5 Analysis of Beer Flavanoids by High-Performance Liquid Chromatography.- 5.1 Gas Chromatography.- 5.2 High Performance Liquid Chromatography.- 5.2.1 Isolation of Proanthocyanidins from Beer.- 5.2.2 High Performance Liquid Chromatography.- References.- Analytical Measurement of Volatile Sulphur Compounds in Beer.- 1 Introduction.- 2 Volatile Sulphur Compounds in Beer.- 3 Analytical Methods for Determining Volatile Sulphur Compounds in Beer.- 3.1 General Remarks.- 3.2 Non-Chromatographic Methods.- 3.2.1 Sulphur Dioxide.- 3.2.1.1 The Monier-Williams Distillation/Titrimetric Procedure.- 3.2.1.2 The Para-Rosaniline Colorimetric Procedure.- 3.2.1.3 The DTNB Method.- 3.2.2 Hydrogen Sulphide.- 3.3 Chromatographic Methods.- 3.3.1 Analyte Detection.- 3.3.2 Chromatographic Separation.- 3.3.3 Sample Preparation/Introduction into Gas Chromatograph.- 3.3.3.1 Static Headspace Sampling.- 3.3.3.2 Dynamic Headspace Sampling (Purge and Trap Methods).- 3.3.3.3 Methods for Concentrating Sulphur Compounds of Lower Volatility.- 4 Conclusions.- References.- Short Introduction to Headspace Analysis.- 1 Introduction.- 2 The Static Headspace Method.- 3 The Dynamic Headspace Method.- 4 Sampling Methods.- 5 Factors Affecting Choice of Equilibrium Conditions.- 6 Quantitative Headspace Analysis.- 7 Final Remarks.- References.- The Use of Principal Components Analysis in Monitoring the Quality of Beer.- 1 Introduction.- 2 The Principles of PCA.- 2.1 The Objectives.- 2.2 Conceptualizing the Multivariate Problem.- 2.3 A Stepwise Explanation of PCA.- 3 The Use of PCA in Brewing Science.- 3.1 A Detailed Example of the Use of PCA in Brewing Science.- 3.2 Other Examples of the Use of PCA in Brewing Science.- 4 Conclusions.- References.- German Beer Purity Law and its Influences on the Properties and Analysis of Beer.- 1 The German Beer Purity Law.- 2 Complaint of the EEC Commission at the European Court of Law Against the Federal Republic of Germany.- 3 Arguments for Retaining the Beer Purity Law.- 4 Beers with Adjuncts.- 5 Analytical Analysis of Additives and Harmful Substances.- 5.1 Immunological and Immunochemical Methods.- 5.1.1 Qualitative and Half-Quantitative Indication Methods.- 5.1.1.1 Double Diffusion in Agar.- 5.1.1.2 Immunoelectrophoresis.- 5.1.1.3 Counter Current Electrophoresis.- 5.1.2 Quantitative Identification Methods.- 5.1.2.1 Basic Radial Immunodiffusion.- 5.1.2.2 Electroimmunodiffusion.- 5.1.2.3 Two-Dimensional Immunoelectrophoresis.- 5.1.3 Highly Sensitive Immunochemical Methods.- 5.1.3.1 Radio Immuno Assay.- 5.1.3.2 ELISA of EIA (Enzymelinked Immunosorbent Assay).- 5.2 Gas Chromatography and High Performance Liquid Chromatography (HPLC).- 5.3 Electrophoresis.- 5.4 Isotachophoresis.- 5.5 Atomic Absorption Spectrometry.- 6 Summary.- References.