Spectroscopy

MARK F. VITHA is a Windsor Professor of Chemistry at Drake University. He received his Ph.D. from the University of Minnesota. He is the editor of the Chemical Analysis Series (Wiley), the author of Chromatography: Principles and Instrumentation (Wiley 2017), and a co-editor of the books High Throughput Analysis for Food Safety (Wiley, 2014) and Interfaces and Interphases in Analytical Chemistry (ACS, 2011). He has received three teaching awards, including the Levitt Teacher of the Year Award, and has been named a Ronald D. Troyer Research Fellow at Drake University.

• ABOUT THE COVER ixPREFACE xi1. Fundamentals of Spectroscopy 11.1 Properties of Electromagnetic Radiation 11.1.1 Speed, c 21.1.2 Amplitude, A 21.1.3 Frequency, υ 31.1.4 Wavelength, λ 31.1.5 Energy, E 31.1.6 Wavenumber, 61.2 The Electromagnetic Spectrum 71.2.1 Radio‐Frequency Radiation (10−27 to 10−21 J/photon) 81.2.2 Microwave Radiation (10−23 to 10−22 J/photon) 101.2.3 Infrared Radiation (10−22 to 10−19 J/photon) 111.2.4 Ultraviolet and Visible Radiation (10−19 to 10−18 J/photon) 121.2.5 X‐Ray Radiation (10−15 to 10−13 J/photon) 131.2.6 Alpha, Beta, and Gamma Radiation (10−13 to 10−11 J/photon and Higher) 131.3 The Perrin–Jablonski Diagram 151.3.1 Timescales of Events 181.3.2 Summary of Radiative and Nonradiative Processes 191.4 Temperature Effects on Ground and Excited State Populations 191.5 More Wave Characteristics 211.5.1 Adding Waves Together 211.5.2 Diffraction 211.5.3 Reflection 251.5.4 Refraction 281.5.5 Scattering 291.5.6 Polarized Radiation 311.6 Spectroscopy Applications 341.7 Summary 34Problems 34References 36Further Reading 382. UV‐Visible Spectrophotometry 392.1 Theory 402.1.1 The Absorption Process 402.1.2 The Beer–Lambert Law 432.1.3 Solvent Effects on Molar Absorptivity and Spectra 492.2 UV‐Visible Instrumentation 522.2.1 Sources of Visible and Ultraviolet Light 542.2.2 Wavelength Selection: Filters 582.2.3 Wavelength Selection: Monochromators 612.2.4 Monochromator Designs: Putting It All Together 752.2.5 Detectors 792.3 Spectrophotometer Designs 852.3.1 Single‐Beam Spectrophotometers 852.3.2 Scanning Double‐Beam Instruments 892.3.3 Photodiode Array Instruments 932.4 The Practice of Spectrophotometry 982.4.1 Types of Samples That Can Be Analyzed 992.4.2 Preparation of Calibration Curves 1002.4.3 Deviations from Beer’s Law 1032.4.4 Precision: Relative Concentration Error 1112.4.5 The Desirable Absorbance Range 1142.5 Applications and Techniques 1162.5.1 Simultaneous Determinations of Multicomponent Systems 1162.5.2 Difference Spectroscopy 1172.5.3 Derivative Spectroscopy 1182.5.4 Titration Curves 1192.5.5 Turbidimetry and Nephelometry 1212.6 A Specific Application of UV‐Visible Spectroscopy: Enzyme Kinetics 1222.6.1 Myeloperoxidase, Immune Responses, Heart Attacks,and Enzyme Kinetics 1222.6.2 Possible Mechanism for Myeloperoxidase Oxidation of LDL via Tyrosyl Radical Intermediates 1232.7 Summary 127Problems 127References 132Further Reading 1343. Molecular Luminescence: Fluorescence, Phosphorescence, and Chemiluminescence 1353.1 Theory 1353.1.1 Absorbance Compared to Fluorescence 1363.1.2 Factors That Affect Fluorescence Intensity 1413.1.3 Quenching 1463.1.4 Quantum Yield and Fluorescence Intensity 1473.1.5 Linearity and Nonlinearity of Fluorescence: Quenching and Self-Absorption 1493.2 Instrumentation 1533.2.1 Instrument Design 1543.2.2 Sources 1543.2.3 Filters and Monochromators 1573.2.4 Component Arrangement 1583.2.5 Fluorometers 1583.2.6 Spectrofluorometers 1593.2.7 Cells and Slit Widths 1643.2.8 Detectors 1663.3 Practice of Luminescence Spectroscopy 1673.3.1 Considerations and Options 1673.3.2 Fluorescence Polarization 1683.3.3 Time‐Resolved Fluorescence Spectroscopy 1723.4 Fluorescence Microscopy 1733.4.1 Fluorescence Microscopy Resolution 1753.4.2 Confocal Fluorescence Microscopy 1753.5 Phosphorescence and Chemiluminescence 1773.5.1 Phosphorescence 1773.5.2 Chemiluminescence 1773.6 Applications of Fluorescence: Biological Systems and DNA Sequencing 1793.7 Summary 186Problems 186References 190Further Reading 1924. Infrared Spectroscopy 1934.1 Theory 1934.1.1 Bond Vibrations 1964.1.2 Other Types of Vibrations 1984.1.3 Modeling Vibrations: Harmonic and Nonharmonic Oscillators 2004.1.4 The 3N−6 Rule 2074.2 FTIR Instruments 2094.2.1 The Michelson Interferometer and Fourier Transform 2104.2.2 Components of FTIR Instruments: Sources 2244.2.3 Components of FTIR Instruments: DTGS and MCT Detectors 2264.2.4 Sample Handling 2274.2.5 Reflectance Techniques 2314.3 Applications of IR Spectroscopy, Including Near‐IR and Far‐IR 2344.3.1 Structure Determination with Mid‐IR Spectroscopy 2354.3.2 Gas Analysis 2354.3.3 Near‐Infrared Spectroscopy (NIR) 2364.3.4 Far‐Infrared Spectroscopy (FIR) 2454.4 Summary 248Problems 248References 251Further Reading 2545. Raman Spectroscopy 2555.1 Energy-Level Description 2555.2 Visualization of Raman Data 2585.3 Molecular Polarizability 2595.4 Brief Review of Molecular Vibrations 2615.5 Classical Theory of Raman Scattering 2625.6 Polarization of Raman Scattering 2655.6.1 Depolarization Ratio 2665.7 Instrumentation and Analysis Methods 2665.7.1 Filter Instruments 2675.7.2 Dispersive Spectrometers 2705.7.3 Fourier Transform Raman Spectrometers 2715.7.4 Confocal Raman Instruments 2715.7.5 Light Sources 2735.8 Quantitative Analysis Methods 2745.8.1 Calibration Curves 2745.8.2 Curve Fitting 2745.8.3 Ordinary Least Squares 2755.8.4 Classical Least Squares 2775.8.5 Implicit Analytical Methods 2775.9 Applications 2775.9.1 Art and Archeology 2775.9.2 Pharmaceuticals 2785.9.3 Forensics 2795.9.4 Medicine and Biology 2795.10 Signal Enhancement Techniques 2825.10.1 Resonance Raman Spectroscopy 2835.10.2 Surface-Enhanced Raman Spectroscopy 2835.10.3 Nonlinear Raman Spectroscopy 2845.11 Summary 286Problems 286References 288Further Reading 289SOLUTIONS 291INDEX 315