Standardized Procedures and Protocols for Starch

Preface
Contents
Contributors
Chapter 1: Importance of Starch Characterization
	1 Introduction
	2 Materials
		2.1 Importance of Starch in the Food Industry
			2.1.1 Starch as Thickening
			2.1.2 Starch as a Binding and Gelling Agent
			2.1.3 Starch as Fat Replacement
		2.2 Composition and Structure of Starch
	3 Methods
		3.1 Methods and Techniques for Starch Characterization
		3.2 Characterization Methods
			3.2.1 Differential Scanning Calorimetry
			3.2.2 X-Ray Diffraction
			3.2.3 Scanning Electron Microscopy
			3.2.4 Fourier-Transform Infrared Spectroscopy
		3.3 Starch Characterization in Industrial Applications
			3.3.1 Papermaking and Textiles
			3.3.2 Bioplastics and packaging Materials
		3.4 Starch Characterization in Nutritional Studies
			3.4.1 Digestibility of Starch and Glycemic Index
	4 Notes
	References
Chapter 2: Morphological Characterization of Starches
	1 Introduction
	2 Materials
		2.1 Optical Microscopy
		2.2 Scanning Electron Microscopy
		2.3 Transmission Electron Microscope
		2.4 Polarizing Microscope
		2.5 Confocal Laser Scanning Microscopy
		2.6 Atomic Force Microscopy
	3 Methods
		3.1 Optical Microscopy
			3.1.1 Starch Powder
			3.1.2 Liquid Starch Sample
		3.2 Scanning Electron Microscopy
			3.2.1 Powder Samples
			3.2.2 Block Samples
		3.3 Transmission Electron Microscope
		3.4 Polarizing Microscope
		3.5 Confocal Laser Scanning Microscopy
		3.6 Atomic Force Microscopy
	4 Notes
	5 Conclusion
	References
Chapter 3: Characterization Methods of Amylose and Amylopectin
	1 Introduction
		1.1 AM and AP: Characterization and Functional Role
			1.1.1 Overview of AM
			1.1.2 Characterization of AM
			1.1.3 Overview of AP
			1.1.4 Characterization of AP
	2 Materials and Methods
		2.1 Sample Preparation
			2.1.1 Starch Isolation from Wheat
			2.1.2 Starch Isolation from Barley Kernels
			2.1.3 Starch Extraction from Potato Flour
			2.1.4 Fractionation of AM and AP
			2.1.5 Separation and Purification of AM and AP [30]
		2.2 Determination of AM and AP Structure
			2.2.1 Spectrophotometric Method
			2.2.2 Potentiometric Titration Method
		2.3 Determination of AM Content
		2.4 Molecular Weight Determination of AM and AP
		2.5 Chain Length Distribution Analysis of AM and AP
		2.6 Rheological Analysis of AM and AP [37]
		2.7 Thermal Characterization of AM-AP Using Differential Scanning Calorimeter (DSC)
		2.8 Thermogravimetry Analysis (TGA)
		2.9 Morphological Characterization Using Scanning Electron Microscopy (SEM)
		2.10 Functional Characterization Using FTIR
		2.11 Pasting Properties of AM and AP
	3 Methods
		3.1 Protocol for Starch Isolation from Wheat
		3.2 Protocol for Starch Isolation from Barley Kernels
		3.3 Protocol of Starch Extraction from Potato
		3.4 Protocol Fractionation of AM and AP from Barley Starch
		3.5 Protocol for Separation and Purification of AM and AP from Rice Starch
		3.6 Protocol for the Determination of AM and AP Structure Using Spectrophotometer Method
		3.7 Protocol for the Determination of AM and AP Structure Using Potentiometric Titration Method
		3.8 Protocol for Determination of AM Content Using GPC
		3.9 Protocol for Molecular Weight Determination of AM and AP
		3.10 Protocol for Chain Length Distribution Analysis
		3.11 Protocol for Rheological Analysis of AM and AP
		3.12 Protocol for DSC Analysis of AM and AP
		3.13 Protocol TGA Analysis of AM and AP
		3.14 Protocol for Morphological Characterization Using Scanning Electron Microscopy (SEM)
		3.15 Protocol for Functional Characterization Using FTIR
		3.16 Protocol for Pasting Properties Evaluation of AM and AP
	4 Conclusion
	5 Notes
	References
Chapter 4: Molecular Structure Characterization of Starch
	1 Introduction
		1.1 Overview of Size Exclusion Chromatography (SEC) and Asymmetrical Flow Field-Flow Fractionation (AF4) for Molecular Charact...
	2 Materials
		2.1 Buffers and Solutions
		2.2 Enzymes and Standards
		2.3 Equipment
	3 Methods
		3.1 Sample Preparation Methods
			3.1.1 Milling of Grains
			3.1.2 Starch Isolation from Barley Flour
			3.1.3 Starch Extraction from Normal, High Amylose, and Waxy Corn Flour
			3.1.4 Starch Extraction from Millets
			3.1.5 Dimethyl Sulfoxide Treatment and Starch Solubilization
			3.1.6 Debranching
			3.1.7 Modified Starch
		3.2 Molecular Characterization of Starch Using SEC-PAD
		3.3 Molecular Characterization of Modified Starch Using SEC-MALS/MALLS-DRI
		3.4 Molecular Characterization of Starch Using HPSEC-MALS-RI System
		3.5 Molecular Characterization of Starch Using the AF4-MALS-RI System
		3.6 Conclusion
	4 Notes
	References
Chapter 5: Starch Paste Properties
	1 Introduction
	2 Materials
	3 Methods
		3.1 Rapid Viscosity Analyzer
		3.2 Protocol
		3.3 For RVA Analysis
		3.4 Analysis of Starch Pasting Properties
		3.5 Viewing Results During a  Test
		3.6 Interpretation of RVA  Data
		3.7 Factors Affecting the Starch Sticking Curve
	4 Notes
	5 Conclusion
	References
Chapter 6: Rheological Characterization
	1 Introduction
		1.1 Rheology Basics and Characterization
			1.1.1 Static Methods
			1.1.2 Dynamic Methods: Oscillatory Testing Methods
			1.1.3 Dynamic Methods: Wave Propagation Method
			1.1.4 Dynamic Methods: Steady Flow Method
			1.1.5 Determining Key Measurements
			1.1.6 Rheology Fitting Models
			1.1.7 Pasting Properties and Gel Strength
		1.2 Rheometry Experimental Methods
			1.2.1 Flow Viscometers used in Production Floor in Industrial Settings
			1.2.2 Rotational Instrument Methods
				Concentric Cylinder
				Cone and Plate
				Parallel Plate
				Selecting Geometry
				Rheometry Calibration and Considerations
			1.2.3 Capillary and Slit Viscometers
			1.2.4 Rheoscope
			1.2.5 Systemic Rheology
	2 Materials
		2.1 Starch Overview
			2.1.1 Starch Sources and Rheological Behaviors
			2.1.2 Investigating Phase Transition in Starch
			2.1.3 Materials to Minimize Evaporation Losses
			2.1.4 Advanced Equipment
	3 Methods
		3.1 Standard Procedures and Protocols
			3.1.1 Rheological Properties of Non-Newtonian Materials by Rotational Viscometer
			3.1.2 Cone/Plate Viscosity at a 500/s Shear Rate
			3.1.3 High-Shear Viscosity Using a Cone/Plate and Capillary Viscometer
			3.1.4 General Principles of Rotational and Oscillatory Rheometry
			3.1.5 Dynamic Mechanical Properties Melt Rheology
			3.1.6 Powder Rheology
			3.1.7 Dough Rheology
	4 Notes
	5 Conclusion
	References
Chapter 7: X-Ray Diffraction Analysis for Starch
	1 Introduction
		1.1 Application of XRD in the Characterization of Different Starches
			1.1.1 Cereal Starches
			1.1.2 Tuber Starches
			1.1.3 Legume Starches
		1.2 XRD Principle
			1.2.1 X-Ray Generation and Imaging
	2 Materials
		2.1 Sample
		2.2 Instrumentation
		2.3 Sample Preparation
	3 Methods
		3.1 For Powder XRD Analysis (Fig. 5)
			3.1.1 Relative Crystallinity of Starch Polymorphs
			3.1.2 Selection of the Peaks for the Fitting Procedure
			3.1.3 Selection of the Most Suitable Fitting Function
			3.1.4 Estimating Starch Crystallinity
			3.1.5 For Investigating the Gelatinization of Starch
			3.1.6 For Retrogradation of Starch
		3.2 Interpretation of XRD  Data
			3.2.1 Millet Starch Representation
	4 Notes
	5 Conclusion and Future Direction
	References
Chapter 8: Starch Digestibility Protocols
	1 Introduction
		1.1 Overview of Starch Hydrolysis and Digestibility
		1.2 Starch Structure and Its Influence on Digestibility
	2 Starch Digestibility and Protocols
		2.1 Overview and Significance of Estimation of In Vivo Digestibility of Starch
		2.2 Overview and Significance of Estimation of In Vitro Digestibility of Starch
	3 Materials
		3.1 For In Vivo Studies
			3.1.1 The Human Ileostomy Model
			3.1.2 Combined CO2/H2 Breath Test
			3.1.3 In Vivo Glycemic Index Method
			3.1.4 In Situ Mobile Nylon Bag Method
			3.1.5 Starch Digestibility Using In Vivo Digesta
			3.1.6 In Vivo Glycemic Response Using Mice
		3.2 For In Vitro Studies
			3.2.1 Using Pepsin Enzyme
			3.2.2 Using Amylase Enzyme
			3.2.3 Using Combination of Enzymes Pepsin: Amylase
	4 In Vivo Starch Digestibility Methods and Protocols
		4.1 Human Model
			4.1.1 The Human Ileostomy Model
			4.1.2 Combined CO2/H2 Breath Test
			4.1.3 In Vivo Glycemic Index Method
		4.2 Animal Model
			4.2.1 In Situ Mobile Nylon Bag Method
			4.2.2 Starch Digestibility Using In Vivo Digesta
			4.2.3 In Vivo Glycemic Response Using Mice
	5 In Vitro Starch Digestibility Methods and Protocols
		5.1 Using Pepsin Enzyme
			5.1.1 Processed Foods
			5.1.2 Whole Grains (Rice and Barley)
			5.1.3 For Pasta Products
		5.2 Using Amylase
		5.3 Using Combination of Enzymes Pepsin-Amylase
	6 Conclusion
	7 Notes
	References
Chapter 9: Physically Modified Starch
	1 Introduction
	2 Materials
	3 Methods
		3.1 Heat-Moisture Treatment (HMT)
			3.1.1 Principle
			3.1.2 Procedure
			3.1.3 Effect on Starch Properties
		3.2 Annealing (ANN)
			3.2.1 Principle
			3.2.2 Procedure
			3.2.3 Effect on Starch Properties
		3.3 Osmotic Pressure Treatment (OPT)
			3.3.1 Principle
			3.3.2 Procedure
			3.3.3 Effect on Starch Properties
		3.4 Microwave Heating (MWH)
			3.4.1 Principle
			3.4.2 Procedure
			3.4.3 Effect on Starch Properties
		3.5 Extrusion
			3.5.1 Principle
			3.5.2 Procedure
			3.5.3 Effect on Starch Properties
		3.6 Ultrasonication
			3.6.1 Principle
			3.6.2 Procedure
			3.6.3 Effect on Starch Properties
		3.7 High Hydrostatic Pressure (HHP)
			3.7.1 Principle
			3.7.2 Procedure
			3.7.3 Effect on Starch Properties
		3.8 Pulse Electric Field (PEF)
			3.8.1 Principle
			3.8.2 Procedure
			3.8.3 Effect on Starch Properties
	4 Notes
	5 Conclusion
	References
Chapter 10: Chemically Modified Starch
	1 Introduction
		1.1 Etherification of Starch
		1.2 Esterification of Starch
		1.3 Cross-Linking of Starch
		1.4 Oxidation of Starch
		1.5 Hydrolysis of Starch
		1.6 Dual Chemical Modification of Starch
	2 Materials
		2.1 Butyl Etherification
		2.2 Hydroxypropylation of Corn Starch and Cassava Starch
		2.3 Double Etherification with CHPTMA and CHPS-Na [56]
		2.4 Benzyl Chloride in Ethanol (Solvent-Based Etherification)
		2.5 Etherification with Dimethyl Sulfoxide (DMSO)
		2.6 1-Allyloxy-2-Hydroxy-Propyl-Starches
		2.7 Etherification with Methyl Methacrylate
		2.8 Hydroxypropyl Banana Starch
		2.9 Hydroxypropylation of Maize Starch
		2.10 Esterification with Citric Acid
		2.11 Citrate Esterification
		2.12 Esterification with Octenyl Succinic Anhydride
		2.13 Esterification with Saturated Fatty Acid Chlorides
		2.14 Octenyl Succinic Anhydride Modification
		2.15 Maleic Anhydride Esterified Starch by the Dry Method
		2.16 Esterification with Ferulic Acid Chloride
		2.17 Esterification with Glutaric Acid
		2.18 Starch Cross-Linked Phosphate
		2.19 Cross-Linked Oat Starch with POCl3 [61]
		2.20 Cross-Linked Starch with Sodium Trimetaphosphate (STMP)
		2.21 Cross-Linked Starch with Epichlorohydrin
		2.22 Cross-Linking with STMP/STPP of Corn Starch
		2.23 Cross-Linked Sago Starch with POCl3
		2.24 Cross-Linked Tapioca Starch with STMP/STPP
		2.25 Cross-Linking with Phosphoryl Chloride (POCl3) of Taro Starch
		2.26 Oxidation of Hybrid Maize Starch [64]
		2.27 Oxidation of Sweet Potato Starch [65]
		2.28 Oxidation of Mucuna Seed Beans Starch
		2.29 Oxidation of Cassava Starch
		2.30 Oxidation of Breadfruit Starch
		2.31 Oxidation of Fermented Cassava Starch
		2.32 Oxidation by Ozone
		2.33 Acid Hydrolysis of Corn Starch
		2.34 Starch Nanocrystals with Acid Hydrolysis
		2.35 Acid Hydrolysis of Sweet Potato Starch [65]
		2.36 Alkali Hydrolysis of Rice Starch
		2.37 Alkali Hydrolysis of Pea Seed Starch
		2.38 Dual Modification Using Cross-Linking/Acetylation/Hydroxypropylation Methods of Rice Starch [69]
		2.39 Dual Modification Using Cross-Linking/Hydroxypropylation (HPT) Methods of Sweet Potato Starch [70]
		2.40 Dual Modification of Wheat Starch Oxidation/Cross-Linking
		2.41 Dual Modification of Tapioca Starch Using Succinylation/Cross-Linking Methods
		2.42 Dual Modification of Waxy Cassava Starch Using Acid Hydrolysis/Succinylation Method [73]
		2.43 Acetylated Distarch Adipate
		2.44 Acetic Acid and Ultrasound
		2.45 Microwave and Other Chemical Modifications
		2.46 Succinylation and Annealing
	3 Methods
		3.1 Butyl Etherification
		3.2 Hydroxypropylation of Corn Starch and Cassava Starch
		3.3 Double Etherification with CHPTMA and CHPS-Na
		3.4 Benzyl Chloride in Ethanol (Solvent-Based Etherification)
		3.5 Etherification with Dimethyl Sulfoxide (DMSO)
		3.6 1-Allyloxy-2-Hydroxy-Propyl-Starches
		3.7 Etherification with Methyl Methacrylate
		3.8 Hydroxypropyl Banana Starch
		3.9 Hydroxypropylation of Maize Starch
		3.10 Esterification with Citric Acid
		3.11 Citrate Esterification
			3.11.1 Debranching of Waxy Maize Starch
			3.11.2 Preparation of Ester-Modified Starch
		3.12 Esterification with Octenyl Succinic Anhydride
		3.13 Esterification with Saturated Fatty Acid Chlorides
		3.14 Octenyl Succinic Anhydride Modification
		3.15 Maleic Anhydride Esterified Starch by the Dry Method
		3.16 Esterification with Ferulic Acid Chloride
			3.16.1 Preparation of Ferulic Acid Chloride
			3.16.2 Preparation of Potato Starch Ester
		3.17 Esterification with Glutaric Acid
		3.18 Starch Cross-Linked Phosphate
		3.19 Cross-Linked Oat Starch with POCl3 [61]
		3.20 Cross-Linked Starch with Sodium Trimetaphosphate (STMP)
		3.21 Cross-Linked Starch with Epichlorohydrin
		3.22 Cross-Linking with STMP/STPP of Corn Starch
		3.23 Cross-Linked Sago Starch with POCl3
		3.24 Cross-Linked Tapioca Starch with STMP/STPP
		3.25 Cross-Linking with Phosphoryl Chloride (POCl3) of Taro Starch
		3.26 Oxidation of Hybrid Maize Starch [64]
		3.27 Oxidation of Sweet Potato Starch [65]
		3.28 Oxidation of Mucuna Seed Beans Starch
		3.29 Oxidation of Cassava Starch
		3.30 Oxidation of Breadfruit Starch
		3.31 Oxidation of Fermented Cassava Starch
		3.32 Oxidation by Ozone
			3.32.1 Set Up for the Treatment
			3.32.2 Method for Treatment of Starch
		3.33 Acid Hydrolysis of Corn Starch
		3.34 Starch Nanocrystals with Acid Hydrolysis
		3.35 Acid Hydrolysis of Sweet Potato Starch [65]
		3.36 Alkali Hydrolysis of Rice Starch
		3.37 Alkali Hydrolysis of Pea Seeds Starch
		3.38 Dual Modification Using Cross-Linking/Acetylation/Hydroxypropylation Methods of Rice Starch [69]
		3.39 Dual Modification Using Cross-Linking/Hydroxypropylation (HPT) Methods of Sweet Potato Starch [70]
		3.40 Dual Modification of Wheat Starch Oxidation/Cross-Linking
		3.41 Dual Modification of Tapioca Starch Using Succinylation/Cross-Linking Methods
		3.42 Dual Modification of Waxy Cassava Starch Using Acid Hydrolysis/Succinylation Method [73]
		3.43 Acetylated Distarch Adipate
		3.44 Acetic Acid and Ultrasound
			3.44.1 Ultrasonic Treatment
			3.44.2 Acetylation
			3.44.3 Dual Modification
		3.45 Microwave and Other Chemical Modifications
		3.46 Succinylation and Annealing
	4 Notes
	5 Conclusion
	References
Chapter 11: Enzymatically Modified Starch
	1 Introduction
		1.1 Enzymes and Their Principles for Starch Modification
		1.2 Role of Enzymes in Starch Modification
		1.3 Types of Enzymes Used in Starch Modification
			1.3.1 Endoamylases
			1.3.2 Exoamylases
			1.3.3 Debranching Enzymes
			1.3.4 Transferases
	2 Protocols for Different Enzymatic Modifications of Starch
		2.1 Materials
			2.1.1 Alpha-Amylase
			2.1.2 Beta-Amylase
			2.1.3 Pullulanase
			2.1.4 Branching Enzyme
			2.1.5 Protocol for the Determination of Starch with Glucoamylase
		2.2 Method for Different Enzymatic Modifications of Starch
			2.2.1 Procedure for Alpha-Amylase Enzymatic Hydrolysis of Starch
			2.2.2 Procedure for Beta-Amylase Enzymatic Hydrolysis of Starch
			2.2.3 Procedure for Alpha-Glucosidase Enzymatic Hydrolysis of Starch
			2.2.4 Procedure for Isoamylase Enzymatic Hydrolysis of Starch
			2.2.5 Procedure for Pullulanase Enzymatic Hydrolysis of Starch
			2.2.6 Procedure for Branching Enzyme for Starch Hydrolysis
			2.2.7 Procedure for Cyclodextrin Glucosyltransferase Enzymatic Hydrolysis of Starch
			2.2.8 Procedure for Alpha-Glucanotransferases/Amylomaltase Enzymatic Hydrolysis of Starch
			2.2.9 Determination of Liberated Glucose
	3 Effect of Starch Composition Analysis on EMS
	4 Conclusion
	References
Index