Basic Protocols in Enology and Winemaking

Preface
Contents
Contributors
Chapter 1: Total and Volatile Acidity: Traditional and Advanced Methods
	1 Introduction
	2 Methods
		2.1 Total Acidity by International Organization of Vine and Wine (OIV-MA-AS313-01)
			2.1.1 Chemicals
			2.1.2 Apparatus
			2.1.3 Procedure
				Sample Preparation
				Potentiometric Titration
				Titration with Indicator (Bromothymol Blue)
			2.1.4 Calculation
		2.2 Total Acidity by Adolfo Lutz Institute (IAL 235/IV)
			2.2.1 Chemicals
			2.2.2 Apparatus
			2.2.3 Procedure
			2.2.4 Calculation
		2.3 Volatile Acidity by International Organization of Vine and Wine (OIV-MA-AS313-02)
			2.3.1 Chemicals
			2.3.2 Apparatus
			2.3.3 Procedure
			2.3.4 Calculation
		2.4 Volatile Acidity by Adolfo Lutz Institute (IAL 236/IV)
			2.4.1 Chemicals
			2.4.2 Apparatus
			2.4.3 Procedure
			2.4.4 Calculation
		2.5 Malic and Tartaric Acids Using High-Performance Liquid Chromatography (HPLC)
			2.5.1 Chemicals
			2.5.2 Apparatus
			2.5.3 Analytical Conditions
			2.5.4 Procedure
	3 Conclusion
	4 Notes
	References
Chapter 2: Alcohol Content: Traditional and Advanced Methods
	1 Introduction
	2 Wine Alcohol Determinations
		2.1 Alcohol Determination by Ebulliometry
			2.1.1 Materials
			2.1.2 Method
				Determination of the Water Boiling Point
				Determination of the Wine Boiling Point
		2.2 Alcohol Content Determination by Distillation Followed by Density Determination by Hydrometry
			2.2.1 Materials
			2.2.2 Methods
				Wine Sample Distillation
				Determination of the Alcohol Content of the Distillate by Hydrometry
		2.3 Gas Chromatography (GC) Analysis of Ethanol
			2.3.1 Materials
			2.3.2 Method
		2.4 Analysis of Ethanol by Liquid Chromatography (LC)
			2.4.1 Materials (Adapted from)
			2.4.2 Method (Adapted from)
		2.5 Enzymatic Analysis
			2.5.1 Preparation of the Following Solutions (Adapted from Ough and Amerine)
		2.6 Infrared Spectroscopy Methods
			2.6.1 Materials
				Near-Infrared Analyzers
					Methods
				Mid-Infrared Analyzers
					Methods
	3 Notes
	References
Chapter 3: Total and Reducing Sugars: Traditional and Advanced Methods
	1 Introduction
	2 Refractometer and Hydrometer in Grapes
	3 Chemical Method to Determine Reducing Substances
	4 Glucose and Fructose by the Enzymatic Method
	5 High-Performance Liquid Chromatography (HPLC)
		5.1 Detectors
			5.1.1 Refractive Index Detector (RI)
			5.1.2 Evaporative Light Scattering Detector (ELSD)
			5.1.3 Electrochemical Detector (PAD)
		5.2 Sample Preparation for Sugar Analysis in  Wine
		5.3 Filtration: An Indispensable Unit Operation for Samples and Mobile Phases
	6 Case Study: HPLC Method to Determine Glucose and Fructose in Grape  Must
		6.1 Method Validation
		6.2 Analysis
	7 Other Methods
	8 Conclusion
	References
Chapter 4: Total Phenolic Content: Traditional Methods
	1 Introduction
	2 Total Phenolic Using Folin-Ciocalteu Reagent
		2.1 Chemicals
		2.2 Apparatus
		2.3 Method
		2.4 Calculation
	3 Total Phenolic Content by Optical Density at 280 nm (OD 280 Value)
		3.1 Chemicals
		3.2 Apparatus
		3.3 Method
	4 Tannin Determination by Spectrophotometry
		4.1 Chemicals
		4.2 Apparatus
		4.3 Standard Curve Preparation
		4.4 Procedure
		4.5 Calculation
	5 Notes
	References
Chapter 5: Color Indexes: Traditional and Advanced Methods
	1 Introduction
	2 Materials
	3 Methods
		3.1 Color Intensity
			3.1.1 Analytical Procedure
		3.2 CIELAB
			3.2.1 Analytical Procedure
	References
Chapter 6: Anthocyanin Identification and Quantitation by High Performance Liquid Chromatography Coupled with Mass Spectrometr...
	1 Introduction
	2 Total and Non-bleaching Anthocyanins Using Spectrophotometry
		2.1 Chemicals
		2.2 Apparatus
		2.3 Method
		2.4 Calculation
	3 Anthocyanin Identification and Quantitation Using HPLC-DAD-ESI-MS/MS
		3.1 Chemicals
		3.2 Solutions and Solvents
		3.3 Apparatus
		3.4 Sample Preparation
		3.5 Analysis of Anthocyanins by High Performed Liquid Chromatography Coupled with Mass Spectrometry (HPLC-MSn)
	4 Notes
	References
Chapter 7: Flavonol Identification and Quantitation by High Performance Liquid Chromatography Coupled with Mass Spectrometry (...
	1 Introduction
	2 Materials
		2.1 Chemicals
		2.2 Solutions and Solvents
		2.3 Equipment
	3 Methods
		3.1 Phenolic Compound Extraction from Grapes
		3.2 Flavonol Isolation
		3.3 Sample Preparation
		3.4 Analysis of Flavonols by High Performed Liquid Chromatography Coupled with Mass Spectrometry (HPLC-MSn)
	4 Notes
	References
8: Flavan-3-ol (Flavanol) Identification and Quantitation by High Performance Liquid Chromatography Coupled with Mass Spectrom...
	1 Introduction
	2 Flavan-3-ol in Grapes and Wines by Liquid Chromatography Coupled with Mass Spectrometry
	3 Electrospray Source Ionization (ESI)
	4 Types of LC-MS Instrument That Can Be Used for Quantitation (Mass Analyzer)
	5 Flavan-3-Ols Concentrations in Grapes and Wines by Liquid Chromatography Associated with Mass Spectrometry
	6 Conclusion
	References
Chapter 9: Hydroxybenzoic and Hydroxycinnamic Acid Derivatives (HCAD) Identification and Quantitation by High Performance Liqu...
	1 Introduction
	2 Materials
		2.1 Chemicals
		2.2 Solutions and Solvents
		2.3 Equipment
	3 Methods
		3.1 Phenolic Compound Extraction from Grapes
		3.2 Phenolic Acid Isolation
		3.3 Sample Preparation
		3.4 Analysis of Hydroxybenzoic and Hydroxycinnamic Acid Derivatives by High Performed Liquid Chromatography Coupled with Mass ...
	4 Notes
	References
Chapter 10: Stilbene Identification and Quantitation by High-Performance Liquid Chromatography Coupled with Mass Spectrometry ...
	1 Introduction
	2 Materials
	3 Method
		3.1 Preparation of Grape Samples: Extraction Method
		3.2 Determination of Stilbenes in Grapes and Wine: HPLC-DAD Method
		3.3 Stilbene Identification: HPLC-MS
	4 Notes
	References
Chapter 11: Analysis of the Free and Bound Fraction of Volatile Compounds in Musts and Wines by GC/MS: Results Interpretation ...
	1 Introduction
		1.1 Wine Mayor Volatile Compounds
		1.2 Minor Volatile Compounds
		1.3 Odor Activity Values (OAVs)
	2 Materials
	3 Methods
		3.1 Analysis of Major Volatile Compounds
		3.2 Analysis of Minor Volatile Compounds
			3.2.1 Extraction
			3.2.2 GC-MS Analysis
		3.3 The Odor Activity Value (OAV) Method
	4 Notes
	References
Chapter 12: Identification of Wine Compounds by Nuclear Magnetic Resonance
	1 Introduction
	2 Materials
		2.1 Sample Preparation
		2.2 NMR Experiments
		2.3 Software
	3 Methods
		3.1 Sample Preparation
		3.2 NMR Spectra Acquisition
			3.2.1 1D 1H NOESY Parameters (See Note 4)
			3.2.2 2D 1H-13C HSQC Parameters
		3.3 1D Spectrum Processing and Analysis
			3.3.1 1D Spectrum Processing
			3.3.2 Identification of Molecules in the Wine Sample by 1D 1H Peak Assignment by Chenomx
			3.3.3 1D 1H NOESY Spectra Assignment Using HMDB
		3.4 2D Spectrum Processing and Analysis
			3.4.1 2D Spectrum Processing
			3.4.2 Identification of Molecules in the Wine Sample by 2D 1H-13C Peak Assignment by COLMARm
	4 Notes
	References
Chapter 13: Ethanol Suppression on Wine Analysis Using Nuclear Magnetic Resonance (NMR)
	1 Introduction
	2 Materials
		2.1 Sample Preparation
		2.2 Acquisition and Processing of Spectrum
	3 Methods
		3.1 Sample Preparation
		3.2 1D Spectrum Acquisition and Shaped Pulse Design
		3.3 2D 1H-1H Spectrum Acquisition with Ethanol Suppression
		3.4 1H-13C HSQC Pulse Sequence Modification for Ethanol Suppression
	4 Notes
	References
Chapter 14: Methods to Determine Biogenic Amines in Wine by RP-HPLC
	1 Introduction
		1.1 Method of Biogenic Amines Determination in Wine by High-Performance Liquid Chromatography with Photodiode Array Detection ...
			1.1.1 Reagents, Materials, and Equipment
			1.1.2 Sample Preparation
			1.1.3 Operating Conditions (as an Example)
			1.1.4 Biogenic Amines Identification
			1.1.5 Calculation
		1.2 Dispersive Solid Phase Extraction (dSPE) Clean-up/Concentration Method for Biogenic Amine Determination in Wines Using Ben...
			1.2.1 Reagents, Materials, and Equipment
			1.2.2 Sample Preparation
			1.2.3 Derivatisation Step (According to Özdestan and Üren with Some Modifications)
			1.2.4 HPLC Analysis and Quantitation
				Eluents
				Elution Gradient
				Quantitation
		1.3 Analysis of Biogenic Amines in Musts and Wines Using HPLC: Orto-Phthalaldehyde (OPA) Derivatisation (Adapted from Resoluti...
			1.3.1 Reagents, Materials, and Equipment
			1.3.2 Sample Preparation
			1.3.3 Derivatisation
			1.3.4 Mobile Phase
			1.3.5 Internal Calibration
			1.3.6 Calculation of Concentrations
	References
Index