Biological Spectroscopy

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Biological Spectroscopy

• Introduction

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Objectives of course

• Understand the importance of spectroscopic
  techniques in identifying and quantifying
  molecules of biological interest.
• Outline the important criteria to be used in
  evaluating spectroscopic techniques.
• Focus, at this stage, on small (lt 1000 Da),
  molecules.
• Discuss the use of separation science approaches
  to characterise the complex mixtures found in
  biological systems.

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Objectives of course

• Discuss the instrumentation required and the
  experimental information provided by
• Infrared spectroscopy
• Nuclear magnetic resonance spectroscopy
• Mass spectrometry
• Interpret experimental data from each of the
  techniques both individually and in combination.
• Discuss relevant biological applications.

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Objectives of course

• A knowledge of the fundamentals of spectroscopic
  phenomena together with instrumentation is needed
  but a grasp of important concepts and their
  application will be presented without a detailed
  description using advanced mathematics and theory.

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Scope of course

• Vibrational spectroscopy
• Infrared
• Near infrared
• Raman
• Nuclear magnetic resonance spectroscopy
• 1H, 13C (1D only)
• Mass spectrometry
• Electron impact
• Chemical ionisation
• Electrospray ionisation
• Not including tandem mass spectrometry

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Scope of course

• Separation science
• Gas chromatography
• Liquid chromatography

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Spectroscopy

• Spectroscopy is the study of matter and its
  properties by investigating light, sound, or
  particles that are emitted, absorbed or scattered
  by the matter under investigation.

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Spectroscopy

• Spectroscopy may also be defined as the study of
  the interaction between light and matter.
  Historically, spectroscopy referred to a branch
  of science in which visible light was used for
  theoretical studies on the structure of matter
  and for qualitative and quantitative analyses.
  Recently, however, the definition has broadened
  as new techniques have been developed that
  utilize not only visible light, but many other
  forms of electromagnetic and non-electromagnetic
  radiation microwaves, radiowaves, x-rays,
  electrons, phonons (sound waves) and others.

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Spectroscopy

• Spectroscopy is often used in biological science
  for the identification of substances through the
  spectrum emitted from them or absorbed in them. A
  device for recording a spectrum is a
  spectrometer. Spectroscopy can be classified
  according to the physical quantity which is
  measured or calculated or the measurement process.

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Electromagnetic spectrum
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The electromagnetic spectrum
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Evaluation criteria for measurement approaches

• Sensitivity
• Speed
• Selectivity
• Sample preparation

• Information Content
• Ease of use
• Cost
• Availability

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Contributions of spectroscopic approaches

• Infrared spectroscopy (IR)
• Information regarding molecules structure
• Symmetry information
• Identification of functional groups
• Easy to use
• Unique, used as fingerprint
• Gas, liquid or solid

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Contributions of spectroscopic approaches

• Nuclear magnetic resonance spectroscopy (NMR)
• Types, numbers and connectivity of atoms.
• Confined to those nuclei with spin.
• Sensitivity (µg 1H, mg 13C)
• Predominately used for liquids

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Contributions of spectroscopic approaches

• Mass spectrometry (MS)
• Ions in the gas phase
• Molecular weight, nominal and exact
• Different isotopes can be recognised
• Fragment ions provide structural information
• Often combined with separation science approaches
• ng-pg sensitivity

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Isomer tree
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Positional Isomers
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Functional isomers
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Diastereoisomers
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Enantiomeric isomers (Chiral)
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Biological Spectroscopy

• Recommended books

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Recommended Texts

• Spectroscopy for the Biological Sciences
• Gordon G Hammes
• Wiley, 2005
• ISBN-13 978-0-471-71344-9
• Organic Structural Spectroscopy
• Joseph B Lambert, Herbert F Shurvell, David A
  Lightner, R Graham Cooks
• Prentice Hall, 1998
• ISBN 0-13-258690-8
• Spectrometric Identification of Organic Compounds
• R M Silverstein, G Clayton Bassler, Terence C
  Morrill
• Wiley, 1997
• ISBN 0-471-63404-2
• Spectroscopic Methods in Organic Chemistry
• Dudley H Williams, Ian Fleming
• McGraw Hill, 1995
• ISBN 0-07-709147-7