1. Background Co-Op research in college - Characterization of water soluble polymers PhD 1997, UF - Atomic Spectroscopy “Resonance Ionization Spectroscopy for the Ultratrace Detection of Mercury” 1997-2001Teaching, Undergraduate Research 2001-2004, Pfizer - Method Development for Drugs in Clinical Trials - esp. HPLC Co-Op research in college - Characterization of water soluble polymers PhD 1997, UF - Atomic Spectroscopy “Resonance Ionization Spectroscopy for the Ultratrace Detection of Mercury” 1997-2001Teaching, Undergraduate Research 2001-2004, Pfizer - Method Development for Drugs in Clinical Trials - esp. HPLC

2. Instrumental Analysis - things to keep in mind, or to question. Why do we do it that way? Why can’t we use this for that? Accuracy, LOD, acquisition time, S/N Technological Advances Examples: Home Pregnancy Tests, RP HPLC Why do we do it that way? Why can’t we use this for that? Accuracy, LOD, acquisition time, S/N Technological Advances Examples: Home Pregnancy Tests, RP HPLC

3. Is Light a Wave or a Particle? It ’ s both. Wave nature of light: c = in a vacuum Light travels in a wave c = 2.998 x 10 8 m/s Particle nature of light: E = h The energy of light is quantized - light travels in packets of energy called photons h = 6.626 x 10 -34 J·s 1 eV = 1.602 x 10 -19 J Wave nature of light: c = in a vacuum Light travels in a wave c = 2.998 x 10 8 m/s Particle nature of light: E = h The energy of light is quantized - light travels in packets of energy called photons h = 6.626 x 10 -34 J·s 1 eV = 1.602 x 10 -19 J Units

4. What makes a method spectrometric? Interaction between radiation and matter. Includes beams of particles. Interaction between radiation and matter. Includes beams of particles.

5. Review of Wave Properties of Light = 2.998 x 10 8 m/s

6. What happens to traveling light waves? Behavior of Light Waves Transmission: Transmission: Light traveling through a medium c =  v v = velocity of light (= c in a vacuum )  = refractive index of media  >1, light travels slower through media v = when light is traveling through a medium, it slows down, the wavelength changes Frequency is a fundamental property of radiation - doesn’t change based on medium. Transmission: Transmission: Light traveling through a medium c =  v v = velocity of light (= c in a vacuum )  = refractive index of media  >1, light travels slower through media v = when light is traveling through a medium, it slows down, the wavelength changes Frequency is a fundamental property of radiation - doesn’t change based on medium.

7. Behavior of Light Waves Diffraction: Diffraction: Light is bent when it encounters an obstacle as it passes a sharp barrier or through a narrow opening (slit). Diffraction: Diffraction: Light is bent when it encounters an obstacle as it passes a sharp barrier or through a narrow opening (slit). Coherent radiation: waves have same frequency and remains in phase. Example: laser. Incoherent radiation: Example: light bulb (W filament) Diffraction is a consequence of Interference.

8. Diffraction Pattern

9. Behavior of Light Waves Interference: waves of same frequency (coherent) experience interference. Constructive Add together - bright spots in diffraction pattern Destructive Cancel each other out - dark spots in diffraction pattern Polychromatic interference examples

10. Polychromatic Interference

11. Behavior of Light Waves Reflection and Refraction Refraction: Reflection: Snell’s law, 1600’s Reflection and Refraction Refraction: Reflection: Snell’s law, 1600’s

12. Ibn Sahl ~1000 AD Ibn Sahl ~1000 AD

13. Behavior of Light Waves Polarization With a polarizing filter, you can remove reflected light (“noise”) from your picture (“signal”) Polarization With a polarizing filter, you can remove reflected light (“noise”) from your picture (“signal”)

14. Particle Nature: The Photoelectric Effect Heinrich Hertz, 1887 Albert Einstein, 1905 Millikan, 1916 Heinrich Hertz, 1887 Albert Einstein, 1905 Millikan, 1916 a.k.a. Opto-Galvanic effect

16. How light interacts with matter Absorption An atom or molecule absorbs a photon of energy and is excited from the ground state. Emission and Luminescence An excited atom or molecule is unstable, returns to the ground state and gets rid of the excess energy by emitting a photon Scattering Photons bounce off of particles, either elastically (photon retains same E) or inelastically (change in E) Absorption An atom or molecule absorbs a photon of energy and is excited from the ground state. Emission and Luminescence An excited atom or molecule is unstable, returns to the ground state and gets rid of the excess energy by emitting a photon Scattering Photons bounce off of particles, either elastically (photon retains same E) or inelastically (change in E)

17. Energy States of Chemical Species The particle theory of light explains spectra in terms of energy states. We use the spectra to tell us something about the chemical species present in our sample.

18. Energy State Diagrams On the left is a diagram of an atom (Na) On the right is a diagram for a simple molecule On the left is a diagram of an atom (Na) On the right is a diagram for a simple molecule Spectra Least complex More complex

19. Applications of EM regions RadiationFrequency (Hz) WavelengthTransitionNon-Chemical Uses Radio< 3x10 11 >1 mmnuclear spin flips Signal Transmission MW3x10 11 -10 13 1 mm-25 ummolecular rotations, electron spin flips Cooking IR10 13 -10 14 25 um-2.5 ummolecular vibrations Heating VIS4-7.5x10 14 750 nm-400 nm valence electron Illumination UV10 15 -10 17 400 nm-1 nmvalence electron "Black" Lights X-Ray10 17 -10 20 1 nm-1 pminner electronImaging Gamma10 20 -10 24 <10-12 mnuclearCancer treatment

20. How we do quantitative analysis with spectroscopy TypeRadiant Power Measured Concentration Relationship Method EmissionEmitted, P e P e = kcAtomic Emission LuminescenceLuminescent, P L P L = kc Atomic and molecular fluorescence, phosphorescence, chemiluminescence ScatteringScattered, P sc P sc = kcRaman scattering, turbidimetry, and nephelometry AbsorptionIncident, P o & transmitted P -log P /P o = kcAtomic and molecular absorption