IV. ANALYTICAL TECHNIQUES
7. Ultraviolet/Visible Spectroscopy (UV/VIS)
- What kind of electrons must a compound have to display UV/Vis activity ?
- In UV/VIS, why does the absorption spectrum of a molecule appear as a broad band?
- What is the intensity of absorption in UV/VIS dependent upon?
- What is the usual source for UV radiation?
- What is the usual source for visible radiation?
- What criteria should be considered when choosing a solvent to dissolve your sample in prior to obtaining an ultraviolet absorption spectrum of the solution?
- What is a chromophore?
- What is an auxochrome?
- In addition to auxochromes, what other things can affect the UV absorption of a molecule?
- Can UV spectrophotometry be used for quantitative analysis?
- What characterizes a good chromophore?
- When referring to electromagnetic (EM) radiation, how do frequency and wavelength relate to each other?
- What is a wavenumber? How does it relate to frequency and wavelength?
- What is the Beer-Lambert Law?
- What is є? What units does є use?
- What is the order of the Beer-Lambert equation and how does this impact on its applicability in quantitative analyses.
- Why is UV-Vis not considered as discriminating as GC-MS or IR?
- Traditionally, what is the typical solvent used for UV-Vis in drug analysis? Can other solvents be used? If so, what are the benefits and drawbacks of their use?
- What scientific law governs UV-Vis as a quantitative technique?
- What are the benefits of quantitation with UV-Vis, compared to other drug analysis techniques?
- Can multiple analytes be quantitated in a single sample? What conditions have to be satisfied for this to occur?
- What materials can the cuvette be made from? What are the most common materials?
- What’s the difference between a single-beam and a double-beam UV-Vis spectrometer? How does this difference manifest itself on a practical level?
- How are solid samples prepped for UV-Vis analysis?
- How are solvent samples prepped for UV-Vis analysis?
- List some of the common preventive maintenance practices for a UV-Vis spectrometer.
- What is the most common cause of poor spectra with a UV-Vis spectrometer?
- List some circumstances in which UV-Vis spectrometry is a superior analysis technique.
- What are the limitations of UV-Vis spectrometry?
- Can the UV-Vis spectrum of a pure compound be related to its electronic structure and the chromophores present?
- Describe the transition of a pi electron from its ground state molecular orbital to its excited state.
- How does the radiation energy in the form of frequency or wavelength interact with the molecular orbitals states?
- Describe the components of UV-VIS spectrometer (detector, source, sample introduction) and their limitations.
- How does a change in solution pH affect the change in the transitions between molecular orbital energy states?
- What are the strengths of the UV-VIS experiment in drug analysis in differentiating different abused street drugs? What are the limitations?
- If a compound has a strong chromophore in the UV range but is a white crystalline material, what can be said about the visual spectrum. In theory, what is required to change on the original chromophore to produce a visual spectrum?
- Why is a diode-array UV detector commonly used in LC experiments?
- What role does the choice of solvent play in a UV/Vis experiment (i.e., if toluene, hexane, acetone where possible choices)?
- Is there high specificity in your UV/VIS experiment to differentiate two similar compounds?
- Describe the use and limitations of a library of UV spectra from the diode-array experiment for drugs of abuse.
- Define the following terms: single beam, double beam, diffraction grating, monochromator, photomultiplier, scan speed, slit width, resolution, frequency wavelength amplitude, intensity, absorbance, transmittance, electronic transitions, hyperchromic shift, bathochromic shift, hypochromic shift, auxochrome, chromophore, R-bands, K-bands, diode array, radiation source.
- Draw a schematic of a diode array UV spectrophotometer. Label each part and explain what each does.
- Explain ultraviolet spectroscopy to a jury. You may pick a particular drug to explain, if it helps.
- Explain ultraviolet spectroscopy to a group of your peers. This should be a more technical explanation.
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