Contents Series Preface ix Preface xi Acronyms, Abbreviations and Symbols xiii About the Author xvii 1 Introduction 1 1.1 Electromagnetic Radiation 2 1.2 Infrared Absorptions 5 1.3 Normal Modes of Vibration 6 1.4 Complicating Factors 11 1.4.1 Overtone and Combination Bands 11 1.4.2 Fermi Resonance 12 1.4.3 Coupling 12 1.4.4 Vibration–Rotation Bands 12 References 13 2 Experimental Methods 15 2.1 Introduction 15 2.2 Dispersive Infrared Spectrometers 16 2.3 Fourier-Transform Infrared Spectrometers 18 2.3.1 Michelson Interferometers 18 2.3.2 Sources and Detectors 19 2.3.3 Fourier-Transformation 20 2.3.4 Moving Mirrors 21 2.3.5 Signal-Averaging 22 vi InfraredSpectroscopy:Fundamentals and Applications 2.3.6 Advantages 23 2.3.7 Computers 23 2.3.8 Spectra 24 2.4 Transmission Methods 25 2.4.1 Liquids and Solutions 25 2.4.2 Solids 28 2.4.3 Gases 31 2.4.4 Pathlength Calibration 32 2.5 Reflectance Methods 33 2.5.1 Attenuated Total Reflectance Spectroscopy 33 2.5.2 Specular Reflectance Spectroscopy 35 2.5.3 Diffuse Reflectance Spectroscopy 36 2.5.4 Photoacoustic Spectroscopy 37 2.6 Microsampling Methods 38 2.7 Chromatography–Infrared Spectroscopy 41 2.8 Thermal Analysis–Infrared Spectroscopy 42 2.9 Other Techniques 43 References 44 3 Spectral Analysis 45 3.1 Introduction 45 3.2 Group Frequencies 46 3.2.1 Mid-Infrared Region 46 3.2.2 Near-Infrared Region 47 3.2.3 Far-Infrared Region 48 3.3 Identification 48 3.4 Hydrogen Bonding 49 3.5 Spectrum Manipulation 51 3.5.1 Baseline Correction 51 3.5.2 Smoothing 51 3.5.3 Difference Spectra 52 3.5.4 Derivatives 53 3.5.5 Deconvolution 54 3.5.6 Curve-Fitting 56 3.6 Concentration 57 3.7 Simple Quantitative Analysis 59 3.7.1 Analysis of Liquid Samples 59 3.7.2 Analysis of Solid Samples 62 3.8 Multi-Component Analysis 63 3.9 Calibration Methods 67 References 70 Contents vii 4 Organic Molecules 71 4.1 Introduction 71 4.2 Aliphatic Hydrocarbons 71 4.3 Aromatic Compounds 74 4.4 Oxygen-Containing Compounds 76 4.4.1 Alcohols and Phenols 76 4.4.2 Ethers 76 4.4.3 Aldehydes and Ketones 76 4.4.4 Esters 78 4.4.5 Carboxylic Acids and Anhydrides 79 4.5 Nitrogen-Containing Compounds 80 4.5.1 Amines 80 4.5.2 Amides 80 4.6 Halogen-Containing Compounds 82 4.7 Heterocyclic Compounds 83 4.8 Boron Compounds 83 4.9 Silicon Compounds 83 4.10 Phosphorus Compounds 84 4.11 Sulfur Compounds 85 4.12 Near-Infrared Spectra 86 4.13 Identification 88 References 93 5 Inorganic Molecules 95 5.1 Introduction 95 5.2 General Considerations 96 5.3 Normal Modes of Vibration 98 5.4 Coordination Compounds 102 5.5 Isomerism 104 5.6 Metal Carbonyls 105 5.7 Organometallic Compounds 107 5.8 Minerals 107 References 110 6 Polymers 113 6.1 Introduction 113 6.2 Identification 114 6.3 Polymerization 123 6.4 Structure 124 6.5 Surfaces 130 viii InfraredSpectroscopy:Fundamentals and Applications 6.6 Degradation 132 References 135 7 Biological Applications 137 7.1 Introduction 137 7.2 Lipids 138 7.3 Proteins and Peptides 141 7.4 Nucleic Acids 151 7.5 Disease Diagnosis 152 7.6 Microbial Cells 155 7.7 Plants 158 7.8 Clinical Chemistry 161 References 163 8 Industrial and Environmental Applications 167 8.1 Introduction 167 8.2 Pharmaceutical Applications 168 8.3 Food Science 174 8.4 Agricultural Applications 178 8.5 Pulp and Paper Industries 179 8.6 Paint Industry 180 8.7 Environmental Applications 183 References 185 Responses to Self-Assessment Questions 187 Bibliography 205 Glossary of Terms 211 SI Units and Physical Constants 215 Periodic Table 219 Index 221 Chapter 1 Introduction Learning Objectives • To understand the origin of electromagnetic radiation. • To determine the frequency, wavelength, wavenumber and energy change associated with an infrared transition. • To appreciate the factors governing the intensity of bands in an infrared spectrum. • To predict the number of fundamental modes of vibration of a molecule. • To understand the influences of force constants and reduced masses on the frequency of band vibrations. • To appreciate the different possible modes of vibration. • To recognize the factors that complicate the interpretation of infrared spectra. Infrared spectroscopy is certainly one of the most important analytical tech- niques available to today’s scientists. One of the great advantages of infrared spectroscopy is that virtually any sample in virtually any state may be studied. Liquids, solutions, pastes, powders, films, fibres, gases and surfaces can all be examined with a judicious choice of sampling technique. As a consequence of the improved instrumentation, a variety of new sensitive techniques have now been developed in order to examine formerly intractable samples. Infrared spectrometers have been commercially available since the 1940s. At that time, the instruments relied on prisms to act as dispersive elements, InfraredSpectroscopy:FundamentalsandApplications B.Stuart 2004JohnWiley&Sons,Ltd ISBNs:0-470-85427-8(HB);0-470-85428-6(PB) 2 InfraredSpectroscopy:Fundamentals and Applications but by the mid 1950s, diffraction gratings had been introduced into disper- sive machines. The most significant advancesin infraredspectroscopy, however, have come about as a result of the introduction of Fourier-transform spectrom- eters. This type of instrument employs an interferometer and exploits the well- established mathematical process of Fourier-transformation. Fourier-transform infrared (FTIR) spectroscopy has dramatically improved the quality of infrared spectra and minimized the time required to obtain data. In addition, with con- stant improvements to computers, infrared spectroscopy has made further great strides. Infrared spectroscopy is a technique based on the vibrations of the atoms of a molecule. An infrared spectrum is commonly obtained by passing infrared radiationthroughasampleanddeterminingwhatfractionoftheincidentradiation isabsorbedataparticularenergy.Theenergyatwhichanypeakinanabsorption spectrumappearscorrespondstothefrequencyofavibrationofapartofasample molecule. In this introductory chapter, the basic ideas and definitions associated with infrared spectroscopy will be described. The vibrations of molecules will be looked at here, as these are crucial to the interpretation of infrared spectra. Once this chapter has been completed, some idea about the information to be gained from infrared spectroscopy should have been gained. The following chapter will aid in an understanding of how an infrared spectrometer produces a spectrum. After working through that chapter, it should be possible to record a spectrum and in order to do this a decision on an appropriate sampling tech- nique needs to be made. The sampling procedure depends very much on the type of sample to be examined, for instance, whether it is a solid, liquid or gas. Chapter 2alsooutlinesthevarioussamplingtechniquesthatarecommonlyavail- able. Once the spectrum has been recorded, the information it can provide needs to be extracted. Chapter 3, on spectrum interpretation, will assist in the under- standing of the information to be gained from an infrared spectrum. As infrared spectroscopy is now used in such a wide variety of scientific fields, some of the many applications of the technique are examined in Chapters 4 to 8. These chapters should provide guidance as to how to approach a particular analytical problem in a specific field. The applications have been divided into separate chapters on organic and inorganic molecules, polymers, biological applications and industrial applications. This book is, of course, not meant to provide a com- prehensive review of the use of infrared spectroscopy in each of these fields. However, an overview of the approaches taken in these areas is provided, along withappropriatereferencestotheliteratureavailableineachofthesedisciplines. 1.1 Electromagnetic Radiation Thevisiblepartoftheelectromagneticspectrumis,bydefinition,radiationvisible to the human eye. Other detection systems reveal radiation beyond the visi- ble regions of the spectrum and these are classified as radiowave, microwave, Introduction 3 infrared, ultraviolet, X-ray and γ-ray. These regions are illustrated in Figure 1.1, together with the processes involved in the interaction of the radiation of these regions with matter. The electromagnetic spectrum and the varied interactions between these radiations and many forms of matter can be considered in terms of either classical or quantum theories. ThenatureofthevariousradiationsshowninFigure 1.1havebeeninterpreted by Maxwell’s classical theory of electro- and magneto-dynamics – hence, the term electromagneticradiation. According to this theory, radiation is considered as two mutually perpendicular electric and magnetic fields, oscillating in single planes at right angles to each other. These fields are in phase and are being propagatedasasinewave,asshowninFigure 1.2.Themagnitudesoftheelectric and magnetic vectors are represented by E and B, respectively. A significant discovery made about electromagnetic radiation was that the velocityofpropagationinavacuumwasconstantforallregionsofthespectrum. Thisisknownasthevelocityoflight,c,andhasthevalue2.997925×108 m s−1. If one complete wave travelling a fixed distance each cycle is visualized, it may be observed that the velocity of this wave is the product of the wavelength, λ (thedistancebetweenadjacentpeaks),andthefrequency,ν(thenumberofcycles Change of spin Change of Change of Change of Change of Change of orientation configuration electron electron nuclear distribution distribution configuration Radiowave Microwave Infrared Visible and X-ray γ-ray ultraviolet 10 103 105 107 109 Energy (J mol−1) Figure 1.1 Regionsoftheelectromagneticspectrum.FromStuart,B.,BiologicalApplica- tionsofInfraredSpectroscopy,ACOLSeries,Wiley,Chichester,UK,1997.University of Greenwich, and reproduced by permission of the University of Greenwich. λ E E B B Direction of propagation B B E E Figure 1.2 Representation of an electromagnetic wave. Reproduced from Brittain, E. F. H., George, W. O. and Wells,C. H. J., Introduction to Molecular Spectroscopy, Academic Press, London,Copyright (1975), with permission from Elsevier. 4 InfraredSpectroscopy:Fundamentals and Applications per second). Therefore: c =λν (1.1) The presentation of spectral regions may be in terms of wavelength as metres or sub-multiples of a metre. The following units are commonly encountered in spectroscopy: 1 A˚ =10−10 m 1 nm=10−9 m 1 µm=10−6 m Another unit which is widely used in infrared spectroscopy is the wavenumber, ν, in cm−1. This is the number of waves in a length of one centimetre and is given by the following relationship: ν=1/λ=ν/c (1.2) This unit has the advantage of being linear with energy. During the 19th Century, a number of experimental observations were made which were not consistent with the classical view that matter could interact with energyinacontinuousform.WorkbyEinstein,PlanckandBohrindicatedthatin many ways electromagnetic radiation could be regarded as a stream of particles (or quanta) for which the energy, E, is given by the Bohr equation, as follows: E =hν (1.3) where h is the Planck constant (h=6.626×10−34 J s) and ν is equivalent to the classical frequency. Processesof change, including those of vibration and rotation associated with infrared spectroscopy, can be represented in terms of quantized discrete energy levels E , E , E , etc., as shown in Figure 1.3. Each atom or molecule in a sys- 0 1 2 tem must exist in one or other of these levels. In a large assembly of molecules, therewillbeadistributionofallatomsormoleculesamongthesevariousenergy levels.Thelatterareafunctionofaninteger(thequantumnumber)andaparam- eter associated with the particular atomic or molecular process associated with thatstate.Wheneveramoleculeinteractswithradiation,aquantumofenergy(or E 3 E 2 E 1 E0 Figure 1.3 Illustrationofquantizeddiscreteenergylevels. Introduction 5 photon) iseitheremittedorabsorbed.Ineachcase,the energyof thequantum of radiation must exactly fit the energy gap E −E or E −E , etc. The energy 1 0 2 1 of the quantum is related to the frequency by the following: (cid:1)E =hν (1.4) Hence, the frequency of emission or absorption of radiation for a transition between the energy states E and E is given by: 0 1 ν=(E −E )/h (1.5) 1 0 Associatedwiththeuptakeofenergyofquantizedabsorptionissomedeactivation mechanismwherebytheatomormoleculereturnstoitsoriginalstate.Associated with the loss of energy by emission of a quantum of energy or photon is some priorexcitationmechanism.Bothoftheseassociatedmechanismsarerepresented by the dotted lines in Figure 1.3. SAQ1.1 Caffeinemoleculesabsorbinfraredradiationat1656cm−1.Calculatethefollow- ing: (i) wavelengthofthisradiation; (ii) frequencyofthisradiation; (iii) energychangeassociatedwiththisabsorption. 1.2 Infrared Absorptions Foramoleculetoshowinfraredabsorptionsitmustpossessaspecificfeature,i.e. anelectricdipolemomentofthemoleculemustchangeduringthevibration.This is the selection rule for infrared spectroscopy. Figure 1.4 illustrates an example of an ‘infrared-active’ molecule, a heteronuclear diatomic molecule. The dipole momentofsuchamoleculechangesasthebondexpandsandcontracts.Bycom- parison,anexampleofan‘infrared-inactive’moleculeisahomonucleardiatomic molecule because its dipole moment remains zero no matter how long the bond. Anunderstandingofmolecularsymmetryandgrouptheoryisimportantwhen initiallyassigninginfraredbands.Adetaileddescriptionofsuchtheoryisbeyond the scope of this book, but symmetry and group theory are discussed in detail in other texts [1, 2]. Fortunately, it is not necessary to work from first principles each time a new infrared spectrum is obtained.