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Plant metabolomics methods and protocols 1st Edition by Nigel W Hardy, Robert D Hall ISBN 1617795941 9781617795947

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Plant metabolomics methods and protocols 1st Edition Nigel W. Hardy Digital Instant Download

Author(s): Nigel W. Hardy, Robert D. Hall (editors)
ISBN(s): 9781617795947, 1617795941
Edition: 1
File Details: PDF, 6.11 MB
Year: 2012
Language: english
SKU: EB-4173540 Category: Tags: ,

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ISBN 10: 1617795941 
ISBN 13: 9781617795947
Author: Nigel W Hardy, Robert D Hall

Estimation of the metabolite complement of plant material involves a wide range of techniques and technologies and that breadth continues to increase.  Metabolomics research typically involves multiple sites for material preparation and analysis and most investigations are “high throughput”, meaning that chemical analysis of sample sets are inevitably carried out over an extended period of time.  In, Plant Metabolomics: Methods and Protocols expert researchers in the field detail many of the stages which are now commonly used to study plant metabolomics workflow. Stages of this workflow, up to and including the statistical analysis, accurate and detailed collection of meta-data are also essential for good process management, to satisfy reporting requirements and to ensure wider interpretability and reuse  results.Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Plant metabolomics methods and protocols 1st Table of contents:

Chapter 1: Introduction to Size Exclusion Chromatography (SEC)

  • Principle: SEC separates molecules based on size (hydrodynamic volume) in solution. Smaller molecules penetrate the pores of the stationary phase and elute later; larger molecules are excluded and elute first.

  • Applications: Polymer characterization, protein analysis, synthetic macromolecules, and biopolymers.

  • Advantages: Non-destructive, minimal interaction with stationary phase, reproducible molecular weight determination.

  • Limitations: Limited resolution for molecules of similar size; dependent on solvent compatibility and stationary phase.

Chapter 2: Stationary Phases for SEC

  • Cross-linked Polymers: Gel permeation chromatography (GPC) uses cross-linked polystyrene or agarose beads.

  • Silica-based Materials: Modified silica gels for high-pressure applications; limited pH range.

  • Pore Size Selection: Critical for target molecular weight range; multi-column setups can cover wide MW ranges.

Chapter 3: Molecular Weight Determination

  • Detectors:

    • Refractive Index (RI) – common, universal but insensitive.

    • UV-Vis – selective, requires chromophore.

    • Light Scattering (LS) – absolute molecular weight, independent of calibration.

    • Viscometry – hydrodynamic volume information, polymer conformation.

  • Calibration:

    • Standard polymers of known molecular weight.

    • Universal calibration: incorporates intrinsic viscosity to compare polymers of different chemistry.

Chapter 4: SEC Techniques for Polymers

  • High-Speed SEC: Faster analysis using smaller particles or higher pressure.

  • Low Molecular Weight SEC: Adjusted pore sizes for oligomers and small molecules.

  • Two-Dimensional Chromatography: SEC combined with other separation techniques (e.g., reversed-phase) for complex mixtures.

Chapter 5: Applications in Biochemistry

  • Proteins & Enzymes: Molecular weight distribution, oligomerization, aggregation studies.

  • Polysaccharides & Biopolymers: Characterization of natural macromolecules.

  • Purification: SEC as a mild, non-denaturing separation method.

Chapter 6: Practical Considerations

  • Column Selection: Match pore size and column length to target MW range.

  • Solvent & Temperature: Compatibility with analytes and stationary phase; prevent aggregation.

  • Sample Preparation: Dissolution, filtration, and avoiding particulate matter.

  • Troubleshooting: Peak tailing, baseline noise, and column fouling.

Chapter 7: Advanced SEC Techniques

  • High-Pressure SEC: Faster separations with higher resolution.

  • SEC-MALS: Multi-angle light scattering for absolute MW and radius of gyration.

  • SEC-Viscometry Coupling: Determines polymer branching and conformation.

Chapter 8: Emerging Trends

  • Integration with mass spectrometry.

  • Miniaturized, microfluidic SEC systems.

  • Combination with other chromatographic techniques for comprehensive molecular characterization.

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Tags: Nigel W Hardy, Robert D Hall, metabolomics