Table of Contents Preface to the New Edition... 3 Information about the MEBAK Website... 4 Contributors... 5 Table of Contents... 7 List of Abbreviations... 16 1 Barley... 14 1.1 Sample Collection... 16 1.2 Visual Inspection... 18 1.2.1 Odor... 18 1.2.2 Moisture... 18 1.2.3 Color and Luster... 18 1.2.4 Husk Character... 18 1.2.5 Degree of Contamination (Purity)... 19 1.2.6 Pests... 19 1.2.7 Damaged Barley Kernels... 20 1.2.7.1 Cracked Kernels... 21 1.2.7.1.1 Visual Inspection for Cracked Kernels... 21 1.2.7.1.2 Iodine Reaction with Starch... 21 1.2.7.2 Incomplete Lateral Husk Closure... 22 1.2.7.3 Husk Damage... 23 1.2.7.4 Immature Kernels from Secondary Growth... 24 1.2.7.5 Pre-germination... 25 1.2.8 Mycological Status... 26 1.2.8.1 Detection of Contamination by Fusarium graminearum... 26 1.2.8.2 Detection of Contamination by Fusarium culmorum... 28 1.2.8.3 Ergot... 29 1.2.9 Kernel Shape and Size... 29 1.2.10 Variety... 30 1.2.10.1 HCl Test... 30 1.2.11 Uniformity... 31 1.3 Physical Tests... 32 1.3.2 Thousand Kernel Weight (EBC)... 34 1.3.3 Hectoliter Weight... 36 1.4 Physiological Tests... 39 1.4.1 Germinative Capacity... 39 1.4.1.1 Rapid Staining Method (EBC)... 39 1.4.1.2 Hydrogen Peroxide Method (EBC)... 40 7
1.4.2 Germinative Energy... 42 1.4.2.1 Germinative Energy AUBRY Method (EBC)... 42 1.4.2.2 Germinative Energy SCHÖNFELD Method (EBC)... 45 1.4.2.3 Germinative Energy the Modified SCHÖNFELD Method... 46 1.4.2.4 Germinative Energy BRF Method (EBC)... 48 1.4.2.5 Germination Percentage and Germination Index (EBC)... 49 1.4.3 Water Sensitivity... 51 1.4.4 Water Uptake Capacity (Water-absorbing Power)... 53 1.4.5 Pre-germinated Barley Kernels... 56 1.4.5.1 Copper Sulfate Method... 56 1.4.5.2 Boiling Method... 57 1.4.5.3 Fluorescein Dibutyrate Method (EBC)... 57 1.5 Chemical-technical Tests... 60 1.5.1 Moisture Content... 60 1.5.1.1 Drying Oven Method (EBC)... 60 1.5.1.2 Rapid Methods... 63 1.5.1.2.1 Infrared Drying... 63 1.5.1.2.2 Near Infrared Reflectance Spectroscopy (NIR)... 64 1.5.2.1.3 Near Infrared Transmittance Spectroscopy (NIT)... 64 1.5.1.2.4 Microwave Drying... 64 1.5.1.2.5 Capacitance... 65 1.5.1.2.6 Conductivity Measurement... 66 1.5.2 Nitrogen (Raw Protein)... 66 1.5.2.1 KJELDAHL Method (EBC)... 66 1.5.2.2 DUMAS Combustion Method (EBC)... 70 1.5.2.3. Near Infrared Reflectance Spectroscopy (NIR) (EBC)... 73 1.5.2.4. Near Infrared Transmittance Spectroscopy (NIT) (EBC)... 75 1.5.3 Micromalting... 76 1.5.4 Husk Content (EBC)... 78 1.5.5 High Molecular Weight β-glucan... 80 1.5.5.1 Fluorimetric Method (EBC)... 80 1.6 Identification of Varieties... 87 1.6.1 Variety Identification by Means of Electrophoresis (EBC)... 87 1.6.2 Polymerase Chain Reaction Method... 89 2 Adjuncts... 92 2.1 Sample Collection... 92 2.2 Moisture (EBC)... 93 2.3 Extract... 94 2.3.1 DE CLERCK Method (EBC)... 94 2.3.2 ASBC Method (EBC)... 96 8
2.3.3 Enzymatic Method for Corn (EBC)... 99 2.3.4 ASBC Method for Liquid Adjuncts (EBC)... 101 2.4 Protein Content... 102 2.5 Fat Content (Free Crude Fat) (EBC)... 103 2.6 Color (EBC)... 106 2.7 Gelatinization Temperature... 107 3 Malt... 111 3.1 Barley Malt... 111 3.1.1 Sample Collection... 111 3.1.2 Visual Inspection... 111 3.1.2.1 Odor... 111 3.1.2.2 Flavor and Aroma... 111 3.1.2.3 Color and Luster... 111 3.1.2.4 Degree of Contamination (Purity)... 112 3.1.2.5 Shape and Size of the Kernels... 112 3.1.3 Physical and Physiological Tests... 112 3.1.3.1 Sieving Test... 112 3.1.3.2 Thousand Kernel Weight (EBC)... 113 3.1.3.3 Hectoliter Weight (HW)... 113 3.1.3.4 Floating Test (Sinker Test)... 114 3.1.3.5 Endosperm Character... 114 3.1.3.5.1 Glassy Kernels... 114 3.1.3.5.2 Color... 115 3.1.3.6 Friability... 115 3.1.3.6.1 Friabilimeter (EBC)... 115 3.1.3.7 Acrospire Development... 118 3.1.3.8 Modification and Homogeneity (Calcofluor Carlsberg Method EBC)... 120 3.1.3.9 Germinative Capacity... 124 3.1.4 Physico-chemical Analysis... 125 3.1.4.1 Moisture Content (EBC)... 125 3.1.4.2 Congress Mash... 128 3.1.4.2.1 DLFU Mill (Setting and Grinding) (EBC)... 128 3.1.4.2.2 Extract (EBC)... 132 3.1.4.2.3 Odor of the Mash (EBC)... 136 3.1.4.2.4 Iodine Test/Saccharification Rate (EBC)... 136 3.1.4.2.5 Filtration (EBC)... 137 3.1.4.2.6 Appearance... 137 3.1.4.2.7 ph (EBC)... 137 3.1.4.2.8 Wort Color... 140 9
3.1.4.2.8.1 Wort Color: Visual Determination (EBC)... 140 3.1.4.2.8.2 Malt Color: Spectrophotometric Measurement (EBC)... 142 3.1.4.2.9 Boiled Wort Color (EBC)... 144 3.1.4.2.10 Extract Difference... 148 3.1.4.3 Extract according to the Complete Extraction Procedure... 148 3.1.4.4 Viscosity (EBC) (Congress Wort, 65 C Mash)... 152 3.1.4.4.1 HÖPPLER Falling Ball Viscometer... 153 3.1.4.4.2 Micro Viscometer from Anton Paar... 156 3.1.4.4.3 Rotational Viscometer... 160 3.1.4.4.4 Capillary Viscometer... 160 3.1.4.5 Nitrogenous Substances... 164 3.1.4.5.1 Total Nitrogen... 164 3.1.4.5.1.1 KJELDAHL Method (EBC)... 164 3.1.4.5.1.2 DUMAS Combustion Method (EBC)... 164 3.1.4.5.1.3 Near Infrared Reflectance Spectroscopy (NIR)... 165 3.1.4.5.1.4 Near Infrared Transmittance Spectroscopy (NIT)... 165 3.1.4.5.2 Soluble Nitrogen... 165 3.1.4.5.2.1 KJELDAHL Method (EBC)... 165 3.1.4.5.2.2 Spectrophotometric Method (EBC)... 167 3.1.4.5.2.3 DUMAS Combustion Method (EBC)... 169 3.1.4.5.3 KOLBACH Index (EBC)... 169 3.1.4.5.4 Nitrogen Fractionation... 170 3.1.4.5.4.1 Precipitation with Phosphomolybdic Acid... 170 3.1.4.5.5 Low Molecular Weight Nitrogenous Substances... 172 3.1.4.5.5.1 Free Amino Nitrogen (FAN) (EBC)... 173 3.1.4.5.5.2 Amino Acids in Wort... 177 3.1.4.5.6 Near Infrared Transmittance Spectroscopy (NIT)... 183 3.1.4.6 Diastatic Power (EBC)... 183 3.1.4.7 α-amylase Activity... 187 3.1.4.7.1 International Method (EBC)... 187 3.1.4.7.2 Viscosimetric Method (Reference Method)... 191 3.1.4.8 β-glucan Activity... 194 3.1.4.9. High Molecular Weight β-glucan... 198 3.1.4.9.1 Fluorimetric Method (EBC)... 198 3.1.4.9.1.1 Malt... 198 3.1.4.9.1.2 Congress Wort... 199 3.1.4.9.2 Colorimetric Method (EBC)... 201 3.1.4.10 Fermentable Carbohydrates in Congress Wort... 202 3.1.4.10.1 Limit of Attenuation... 202 10
3.1.4.10.1.1 Limit of Attenuation of Congress Wort (Fermentation Tube Method)... 203 3.1.4.10.1.2 Limit of Attenuation of Congress Wort (Reference Method EBC)... 205 3.1.4.10.1.3 Limit of Attenuation of Congress Wort (Rapid Method EBC)... 207 3.1.4.10.2 Determination of Fermentable Sugars by HPLC... 209 3.1.4.10.3 Fermentable Carbohydrates by HPLC (EBC)... 212 3.1.4.11 HARTONG-KRETSCHMER Mash Method, VZ 45 C, Isothermic 65 C mash... 215 3.1.4.12 Iodine Value of Laboratory Spent Grains... 218 3.1.4.13 Steam-volatile Phenols for the Determination of Substances Responsible for a Smoky Flavor... 222 3.1.4.14 Thiobarbituric Acid Index (TBI)... 225 3.1.4.15 Variety Identification by Means of Electrophoresis... 227 3.1.4.16 Mycological Status... 227 3.1.4.17 Dimethyl Sulfide and Its Precursors in Malt... 229 3.1.4.18 Nitrosamines in Malt... 232 3.1.4.19 Organic Acids in Congress Wort... 237 3.1.4.20 Minerals... 240 3.1.4.20.1 Ash Content... 240 3.1.4.20.2 Atomic Absorption Spectrophotometry... 241 3.1.4.20.2.1 Iron... 242 3.1.4.20.2.2 Sodium... 244 3.1.4.20.2.3 Potassium... 247 3.1.4.20.2.4 Calcium (EBC)... 249 3.1.4.20.2.5 Manganese... 252 3.1.4.20.2.6 Zinc (EBC)... 254 3.1.4.20.2.7 Copper... 257 3.1.4.20.2.8 Magnesium... 259 3.1.4.20.2.9 Aluminum... 262 3.1.4.20.2.10 Tin... 263 3.1.4.20.3 Detection of Metals by Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES/ ICP-OES)... 265 3.1.4.21 Gushing... 266 3.1.4.21.1 Rapid Test for Gushing for Malt and Adjuncts... 267 3.2 Specialty Malts... 270 3.2.1 Water (EBC)... 270 3.2.2 Extract (in Roasted and Caramel Malt) (EBC)... 270 3.2.3 Color (in Roasted and Caramel Malt) (EBC)... 272 11
3.2.4 Sour Malt... 273 3.2.4.1 ph... 273 3.2.4.2 Titration Acidity... 273 3.2.4.3 L- Lactic Acid (EBC)/D-Lactic Acid (Lactate)... 274 3.2.5 The Color of Liquid Malt Coloring Agent... 280 3.2.5.1 Visual Method... 280 3.2.5.2 Spectrophotometric Method... 281 3.3 Wheat Malt... 282 4 Hops and Hop Products... 283 4.1 Whole Hops and Pellets... 283 4.1.1 Sample Collection... 283 4.1.2 Manual Evaluation of Whole Hops... 284 4.1.2.1 Stem and Leaf Material in Harvested Hops... 284 4.1.2.2 Color and Luster... 285 4.1.2.3 Cone Condition... 285 4.1.2.4 Aroma... 285 4.1.2.5 α-acids... 286 4.1.2.6 Moisture Content... 286 4.1.2.7 Diseases, Pests and Seed Content... 286 4.1.2.8 Improper Handling... 286 4.1.2.9 Overall Evaluation... 287 4.1.3 Sample preparation... 290 4.1.4 Moisture Content (EBC Method)... 292 4.1.5 Bitter Substances... 294 4.1.5.1 Bitter Substances in Hops and Hop Products: Conductometric Value and Total Resins, Soft and Hard Resins (EBC Method)... 294 4.1.5.2 Determination of α-acids and β-acids by HPLC (EBC)... 304 4.2 Hop Extract... 310 4.2.1 Sample Collection... 310 4.2.2 Sample preparation... 310 4.2.3 Moisture (KARL FISCHER Method)... 310 4.2.4 Bitter Substances... 314 4.2.4.1 Bitter Substances in Hop Extract: Conductometric Value and Total Resin, Soft Resins and Hard Resins (EBC Method)... 314 4.2.4.2 Determination of α- and β-acids Using HPLC (EBC)... 317 4.3 Isomerized Hop Extract... 318 4 3.1 Sample Collection... 318 4.3.2 Sample Preparation... 318 12
4.3.3 Bitter Substances... 318 4.3.3.1 Determination of iso-α-acids, α-acids and β-acids in Hop Extracts and Isomerized Hop Extracts (EBC Method)... 318 5 Implementation and Validation of Analysis Methods... 324 5.1 General Information... 324 5.2 Terms, Definitions... 324 5.3 Implementation of New Methods: Approach... 327 5.4 Monitoring/Quality Assurance... 327 5.5 Basic Equations Used in Statistics... 328 5.5.1 Regression Analysis... 328 5.5.2 MANDEL Goodness of Fit Test... 330 5.5.3 Outlier test via F-test of residual variances... 330 5.6 Sources... 331 Index...... 342 List of Advertisers... 347 13