CHEM 3418.

001 Test 2 Review Questions

Chapter 13
1. Be able to answer the following questions at the end of Chapter 13: 13 (a-h), 16, 17, 18, 20,
23
13. (a) Hydrogen and deuterium lamps differ only in the gases that are used in the discharge.
Deuterium lamps generally produce higher intensity radiation.
(b) Filters provide low resolution wavelength selection often suitable for quantitative work, but
not for qualitative analysis or structural studies. Monochromators produce high resolution
(narrow bandwidths) for both qualitative and quantitative work.

(c) A phototube is a vacuum tube equipped with a photoemissive cathode and a collection anode.
The photo electrons emitted as a result of photon bombardment are attracted to the positively
charged anode to produce a small photocurrent proportional to the photon flux. A photovoltaic
cell consists of a photosensitive semiconductor sandwiched between two electrodes. An incident
beam of photons causes production of electron-hole pairs which when separated produce a
voltage related to the photon flux. Phototubes are generally more sensitive and have a greater
wavelength range. Photocells are in general simpler, cheaper and more rugged. Photocells do not
require external power supplies.

(d) A photodiode consists of a photo-sensitive pn-junction diode that is normally reverse-biased.

An incident beam of photons causes a photocurrent proportional to the photon flux. A
photomultiplier tube is a vacuum tube consisting of a photoemissive cathode, a series of
intermediate electrodes called dynodes, and a collection anode. Each photoelectron emitted by
the photocathode is accelerated in the electric field to the first positively charged dynode where it
can produce several secondary electrons. These are, in turn, attracted to the next positively
charge dynode to give rise to multiple electrons. The result is a cascade multiplication of 106 or
more electrons per emitted photoelectron. Photomultipliers are more sensitive than photodiodes,
but require a high voltage power supply compared to the low voltage supplies required by
photodiodes. Photomultipliers are larger and require extensive shielding. Photodiodes are better
suited for small, portable instruments because of their size and ruggedness.

(e) Both types of spectrophotometers split the beam into two portions. One travels through the
reference cell and one through the sample cell. With the double-beam-in-space arrangement,
both beams travel at the same time through the two cells. They then strike two separate
photodetectors where the signals are processed to produce the absorbance. With the double-
beam-in-time arrangement, the two beams travel at different times through the cells. They are
later recombined to strike one photodetector at different times. The double-beam-in-time
arrangement is a little more complicated mechanically and electronically, but uses one
photodetector. The double-beam-in-space arrangement is simpler, but requires two matched
photodetectors.

(f) Spectrophotometers have monochromators or spectrographs for wavelength selection.

Photometers generally have filters are use an LED source for wavelength selection. The
spectrophotometer can be used for wavelength scanning or for multiple wavelength selection.
The photometer is restricted to one or a few wavelengths.

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(g) A single-beam spectrophotometer employs one beam of radiation that irradiates one cell. To
obtain the absorbance, the reference cell is replaced with the sample cell containing the analyte.
With a double-beam instrument, the reference cell and sample cell are irradiated simultaneously
or nearly so. Double-beam instruments have the advantages that fluctuations in source intensity
are cancelled as is drift in electronic components. The double-beam instrument is readily adapted
for spectral scanning. Single-beam instruments have the advantages of simplicity and lower cost.
Computerized versions are useful for spectral scanning.

(h) Multichannel spectrophotometers detect the entire spectral range essentially simultaneously
and can produce an entire spectrum in one second or less. They do not use mechanical means to
obtain a spectrum. Conventional spectrophotometers use mechanical methods (rotation of a
grating) to scan the spectrum. An entire spectrum requires several minutes to procure.
Multichannel instruments have the advantage of speed and long-term reliability. Conventional
spectrophotometers can be of higher resolution and have lower stray light characteristics.

16.
In a deuterium lamp, the lamp energy from the power source produces an excited deuterium
molecule that dissociates into two atoms in the ground state and a photon of radiation. As the
excited deuterium relaxes, its quantized energy is distributed between the energy of the photon
and the energies of the two atoms. The latter can vary from nearly zero to the energy of the
excited molecule. Therefore, the energy of the radiation, which is the difference between the
quantized energy of the excited molecule and the kinetic energies of the atoms, can also vary
continuously over the same range. Consequently, the emission spectrum is a spectral continuum.

17.
Photons from the infrared region of the spectrum do not have enough energy to cause
photoemission from the cathode of a photomultiplier tube.

18. Tungsten/halogen lamps often include a small amount of iodine in the evacuated quartz
envelope that contains the tungsten filament. The iodine prolongs the life of the lamp and permits
it to operate at a higher temperature. The iodine combines with gaseous tungsten that sublimes
from the filament and causes the metal to be redeposited, thus adding to the life of the lamp.

20. (a) The dark current is the small current that exists in a radiation transducer in the absence of
radiation. It has its origin in the thermal emission of electrons at the photocathode, in ohmic
leakage, and in radioactivity.
(b) A transducer is a device that converts a physical or chemical quantity into an electrical
signal.
(c) Scattered radiation is unwanted radiation that reaches the exit slit of a monochromator as a
result of reflections and scattering. Its wavelength is usually different from that of the radiation
reaching the slit directly from the dispersive device.
(d) Source flicker noise is caused by variations in experimental variables that control the source
intensity, such as power supply voltages and temperature. It can also be caused by mechanical
variations such as vibrations.
(e) Cell positioning uncertainty is caused by our inability to position the cell in the same exact
place each time. A random variation is introduced because the incident beam is imaged onto

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slightly different portions of the cell walls each time causing differences in the reflection,
transmission and scattering characteristics of the cell.
(f) A beam splitter is a device that causes an incident beam to divide into two beams at its output.
It can be made from mirrors, rotating choppers, or optical materials that cause a beam to be split
into two beams.

23.
Quantitative analysis can usually tolerate rather wide slits because measurements are often made
on an absorption maximum where there is little change in absorptivity over the bandwidth. Wide
slit widths are desirable because the radiant powers will be larger and the signal-to-noise ratio
will be higher. On the other hand, qualitative analysis requires narrow slit widths so that fine
structure in the spectrum will be resolved.

2. What are some of the factors that can lead to deviations in Beer’s Law and explain
why?
-chemical interactions: another species is interfering, example: oxidation reaction, an analyte
dissociates, associates, or reacts with a solvent to produce a product with a different
absorption spectrum than the analyte
-stray radiation: radiation from the instrument that is outside the nominal wavelength ban
chosen for the determination (from gratings, lenses, mirrors, filters, windows, etc) the
radiation may not pass through the sample and most likely differs from the chosen radiation.
This is a big problem at wavelength extremes where the sample doesn’t absorb well or the
detector doesn’t detect well
-non-monochromatic radiation(polychromatic: the relationship between absorption and
concentration may not be linear with polychromatic light
-high concentration: the particles are too close, self absorption may occur, dipole moments
may occur changing the distribution of electrons so the molecule absorbs light differently

3. Be able to discuss the effect of slit width on absorbance measurements.

The best spectrum depends on what you want. A more narrow slit width is necessary for
more spectral detail, qualitative. If quantitative data is desired a broader slit width should be
used.

4. What affect does stray (scattered) radiation have on spectra and explain why?
It can lead to deviation from the linear relationship between path length and absorbance. It
can also lead to negative absorbance errors. Or the instrument may be sensitive at that
wavelength but the sample is not. This will overwhelm the instrument and cause a false peak.
The detector may be sensitive to the stray light and cause a peak that shouldn’t be there.
Stray light limits the max absorbance. When absorbance is high the radiant power can be
comparable or lower than the stray light level. Absorption is smaller than the theoretical.

5. What are some of the differences in the atomic and molecular absorption spectra
and explain the observed differences.
Molecules have vibrational and rotational states along with the electronic states. Due to this
the spectra have lots of little lines that appear as a band. Atoms do not have the vibrational
and rotational states, the spectra exhibit strong individual peaks, narrow lines.

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6. Draw block diagrams identifying each of the major components of the following
types of instruments used for optical spectroscopy:

a) Emission
b) Absorption
c) Fluorescence
d) Single-beam (Figure 13.3 page 352)
e) Double-beam in space (Figure 13.3 page 352)
f) Double-beam in time (Figure 13.3 page 352)

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DB time
Single beam

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CHEM 3418.001 Test 2 Review Questions

DB space
Double beam

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7. Be able to match various types of sources, detectors, and optical materials with the
appropriate wavelength ranges they are most commonly used for.
Sources:
-Deuterium/Hydrogen Lamps: UV region, ouput range of 160-800 nm, in UV region (190-
400 nm)a continuum spectrum exists, above 400 nm the spectra consist of lines and bands
-Tungsten Filament Lamps: Visible and near-infrared, energy is similar to blackbody
radiation so is dependent on temperature, useful for wavelength region 350-2500 nm
-Light Emitting Diode: 375-1000 nm, white leds 400-800nm
-Xenon Arc Lamps: 2000-1000 nm, peak intensity around 500 nm

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8. What are the differences, advantages, and disadvantages between Single-Beam and
Double-Beam-Instruments?
The advantage of double beam instruments is that they compensate for all bt the most short-
term fluctuations in the radiant output of the source as well as the drift in the transducer and
amplifier. They also compensate for wide variations in source intensity with wavelength.
Double beam design is good for continuous recording of transmittance or absorbance spectra.
Double-beam instruments involve the splitting of a beam. The double beam in time is
preferred to double beam in space because the difficulty in matching the two detectors
needed for the double beam in space. The double-beam is less sensitive because of splitting
of light. The double beam in time the light spends part of the time in the sample and part of
the time in the blank. In the double beam in space the sample and blank receive only 50 % of
the light. The single beam has the potential to be the more sensitive instrument, but usually it
is cheaper material. The disadvantage of the single beam is that it takes more time. You
measure the sample and the blank at different times. You can’t compensate for fluctuation.
The single beam is more sensitive because your not splitting the light.

___________________________________________________________________________
Polar to non polar = blue shift
Sigma to sigma* = blue shift
Non polar to polar = red shift
Pi to pi* = red shift

Chapter 14
1. How can nonabsorbing analytes be determined photometrically and what are some of
the requirements for successful application?
Nonabsorbing species can be determined photometrically if an absorbing complex is
formed (charge transfer absorption).
In order for it to be successful the color forming complex must be forced to near
completion. The procedural details: selection of the wavelength (wavelength corresponds
to absorption peak, at wavelength max is highest sensitivity, and a flat region to follow
beer’s law), control of variable that affect absorbance (pH, nature of solvent, temperature,
electrolyte concentration, and presence of interfering species), clean tubes( use matched,
clean, and non scratched, same tubes for blank and sample), calibration curve, and chose
solvent carefully (don’t use a wavelength lower than solvent limit or the solvent will
absorb)

Complex agent has to be in excess to be successful; force all molecules to bind =

completion

Ex. oxygen binds to hemoglobin as a complex

2. Why are electrons in double or triple bonds more easily excited by electromagnetic
radiation compared to single bonds?
The single bonds require sigma to sigma* transitions, this is a very large energy gap.

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A double\triple bond only requires a pi to pi* transition, this is a much lower energy gap.

Single bonds absorb high energy in vacuum/UV region

-problem: this breaks bonds, atmospheric components interfere (work in vacuum)

Double/triple bonds

3. What is the difference between the absorption spectra of inner transition elements (the
lanthanide and actinide series) to transition metals? What molecular process is
responsible for the absorption of UV/Vis radiation for these elements/compounds?
Which ones are more sensitive to environmental perturbations and why?

` The inner transition elements (lanthanides) produce very narrow well-defined and
characteristic peaks. If ligands are attached they have very little effect. The inner transition
elements deal with the inner electrons like the 4f and 5f which are shielded or screened from
external influences.

Transition metals absorb at one if not all of their oxidation states. The absorption peaks are
broad and strongly influenced by chimerical environmental factors. The d and f orbitals are
not as shielded. Ligands have drastic shift on the spectrum. The stronger the ligand the more
the spectrum will shift (to a shorter wavelength). There is a greater effect in transition metals.
D orbital is broad

Inorganic absorb light because of the splitting of the d and f orbitals. The splitting of the orbitals
from a lower energy to a higher energy orbital. The transition metals are more subject to
environmental perturbations because the electrons of the d orbital are farther from the
nucleus. The inner transition metals deal with electrons in the f orbital which is shielded and
closer to the nucleus.

-Electrons close to surface=affected by environment

-Electrons close to nucleus=shielded from environment

Energy splits in orbitals as ligands are attached for inorganic molecules = absorption

Organic compounds are going from sigma to sigma*, etc.

Inorganic =

4. Compare the purposes of UV/Vis and IR absorption spectroscopy.

UV/Vis-Quantitative, how much of something
IR-qualitative, functional groups

5. What is the main advantage of charge-transfer absorption over other types of

absorption phenomena?

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Charge-transfer absorption is particularly important because molar absorptivities are

unusually large (greater than 10,000) which leads to high sensitivity. Many inorganic and
organic complexes exhibit this type of absorption and are called charge-transfer complexes

6. What factors must be considered in the development of conditions that would yield
reproducible (preferably linear) results between absorbance and analyte concentration?
*Selection of the wavelength: a wavelength corresponding to an absorption peak, a flat
region, high sensitivity at wavelength max.
*Variables that affect absorbance:
pH-atom or species respond differently at different pH
nature of solvent: is it polar/not polar, cause red/blue shift
temperature: can change equilibrium
Electrolyte concentration: more electrolyte may cause interaction with analyte
Presence of interfering species
*Clean/matched cells/tubes
*Calibration curve (don’t use literature molar absorptivity)
*Solvent choice: don’t use a wavelength lower than solvent limit or it will absorb
If changes the distribution of electrons it will change how it interacts with light.
7. Why have single bond spectra of organic compounds not been widely exploited for
analytical purposes when studying absorption?
The excitation energy associated with electrons forming single bonds is sufficiently high so
absorption must occur in the vacuum-uv region. These transitions involve the excitation of
nonbonding n electrons to sigma* orbitals. The molar absorptivities of this transition is low
to intermediate. This is typically not done due to the experimental difficulty vacuum-uv
region. 1. Energy too high leading to bond rupture. 2. Have to work in a vacuum to get rid of
atomospheric componenets.
8. What variables influence absorbance?
*Variables that affect absorbance:
pH-atom or species respond differently at different pH
nature of solvent: is it polar/not polar, cause red/blue shift
temperature: can change equilibrium
Electrolyte concentration: more electrolyte may cause interaction with analyte
Presence of interfering species

9. Describe the difference in spectra observed in the gas, polar solvent, and nonpolar
solvent. Why are the spectra different?
-gas-phase: are used for volatile compounds. Can be obtained by allowing a drop or two
of the pure liquid to evaporate and equilibrate with the atmosphere in a stoppered cuvette.
Many lines are visible due to electronic, vibrational, and rotational transitions. There are
few collisions with a gas solvent, which means fewer changes in ground state so the
peaks are sharp
-polar solvents: like water, alcohols, esters, and ketones tend to obliterate spectral fine
structure arising from vibrational effects. The strong intermolecular forces cause
electronic peaks to blend giving a smooth absorption band. The polar solvent interacts

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with the nonbonding electrons. This causes greater distribution of energy states leading to
blurry spectrum.
-nonpolar solvent: like hydrocarbons, are similar to gas-phase, The electronic transitions
can be observed but the vibrational and rotational structure have been lost. The collisions
are sort of inbetween that of the gas and polar, so the spectrum is similar to the gas
solvent.

10. What factors lead to the absorption of UV/Vis radiation by organic molecules?
Organic molecules absorption deals with valence electrons that can be excited to higher
energy levels. Single bonds: are don’t in the VAC-UV region because the energy associated
with the electrons is so high. (n to sigma *) this is typically not performed because of the
difficulty with VAC-UV. Double bonds: (n/pi to pi*) The energy required (200-700) nm is
within UV-VIS region. Chromophores (unsaturated functional group as pi group) are
required.

11. What factors lead to the absorption of UV/Vis radiation by inorganic molecules?
Absorption in inorganic molecules involves transitions between filled and unfilled d/f-
orbitals (splitting of d and f orbital). The energy difference depends on the position on the
periodic table. Inner transition metals (lanthanide 4f, and actanid 5f) have electrons that are
shieled from external influences by electron orbitals with large principle quantum numbers.
So the bands are narrow and unaffected by species bound to outer electrons.

12. The absorption of UV or visible light by an atomic or molecular species can be

considered a two-step process. Describe that process in significant detail and explain
the differences in this process between atomic and molecular species.
Step 1. Electron Excitation, a collision has to occur and the molecule/atom must be able to
absorb that energy.
Step 2. Relaxation, the energy excitation must be lost. This could be done through heat,
formation of new species, fluorescence, or phosphorescence, etc…
Molecular species have vibrational and rotational states that excite and relax different
causing bands rather than peaks.
Atomic species don’t have vib./rot. States, only lines are observed.

13. What affect does solvent polarity have on UV/Vis spectra and why? Explain in terms of
types of transitions.
Polar solvent: cause blue shift in n to pi*/sigma*. Because the polar solvent interacts with the
nonbonding electrons lowering the energy of the nonbonding electrons. This increase the gap
between n and pi*/sigma*. This results in a blue shift(shift to shorter wavelengths) in order to
reach the pi*/sigma*
A polar solvent cause a red shift in pi to pi*. Because the attractive forces between the
solvent and the absorber lower the energy levels of the unexcited and excited states. It lowers
the energy of the excited state by more which decreases the energy gap. This results in a red
shift (longer wavelengths)

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14. What affect does conjugation have on UV/Vis spectra? Slide 28

Conjugations cause an increase in the wavelength max. Conjugation is more stable so shifts
to longer wavelengths. It also causes the molar absorptivity to double compared to only
single bonds because both double bonds absorb.

15. What is an auxochrome and what affect does it have on UV/Vis spectra and why?
An auxochrome is a functional group that does not absorb UV/VIS but shifts peak to longer
wavelengths(Red shift) and increases intensity. Auxochromic substituents have a pair of n
electrons capable of interacting with pi electrons which stabilizes the pi* state thereby
lowering its energy (red shift). Must have atleast 1 pair of nonbonding electrons that interact
with the pi electrons leading to stability and lowering of energy (red shift)

Ex. Benzene absorbs, a methyl group attached does not absorb (auxochrome); can effect the
absorbance of benzene

Auxochrome has 1 pair non bonding e- capable of interacting with pi bonds, lower energies
from pi to pi* = red shift

Going from polar to polar:

Non-polar to polar = red shift

Chapter 15
1. Be able to answer the following questions at the end of Chapter 15: 1-6.

15-1. Explain the difference between a fluorescence emission spectrum and a fluorescence
excitation spectrum. Which more closely resembles an absorption spectrum?
In a fluorescence emission spectrum, the excitation wavelength is held constant and the emission
intensity is measured as a function of the emission wavelength. In an excitation spectrum, the
emission is measured at one wavelength while the excitation wavelengths are scanned. The
excitation spectrum closely resembles an absorption spectrum since the emission intensity is
usually proportional to the absorbance of the molecule.

15-2. Define the following terms: (a) fluorescence. (b) phosphorescence. (c) resonance
fluorescence. (d) singlet state. (e) triplet state. (f) vibrational relaxation, (g) internal
conversion, (h) external conversion, (i) intersystem crossing, (j) predissociation,
(k) dissociation, (1) quantum yield, (m) chemiluminescence.
(a) Fluorescence is the process in which a molecule, excited by the absorption of radiation,
emits a photon while undergoing a transition from an excited singlet electronic state to a lower
state of the same spin multiplicity (e.g., a singlet → singlet transition).

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(b) Phosphorescence is the process in which a molecule, excited by the absorption of

radiation, emits a photon while undergoing a transition from an excited triplet state to a lower
state of a different spin multiplicity (e.g., a triplet → singlet transition).
(c) Resonance fluorescence is observed when an excited species emits radiation of the same
frequency at used to cause the excitation.
(d) A singlet state is one in which the spins of the electrons of an atom or molecule are all
paired so there is no net spin angular momentum
(e) A triplet state is one in which the spins of the electrons of an atom or molecule are
unpaired so that their spin angular moments add to give a net non-zero moment.
(f) Vibrational relaxation is the process by which a molecule loses its excess vibrational
energy without emitting radiation.
(g) Internal conversion is the intermolecular process in which a molecule crosses to a lower
electronic state with emitting radiation.
(h) External conversion is a radiationless process in which a molecule loses electronic energy
while transferring that energy to the solvent or another solute.
(i) Intersystem crossing is the process in which a molecule in one spin state changes to
another spin state with nearly the same total energy (e.g., singlet → triplet).
(j) Predissociation occurs when a molecule changes from a higher electronic state to an
upper vibrational level of a lower electronic state in which the vibrational energy is great
enough to rupture the bond.
(k) Dissociation occurs when radiation promotes a molecule directly to a state with sufficient
vibrational energy for a bond to break.
(l) Quantum yield is the fraction of excited molecules undergoing the process of interest. For
example, the quantum yield of fluorescence is the fraction of molecules which have absorbed
radiation that fluoresce.
(m) Chemiluminescence is a process by which radiation is produced as a result of a chemical
reaction.

15-3. Why is spectrofluorometry potentially more sensitive than spectrophotometry?

For spectrofluorometry, the analytical signal F is proportional to the source intensity P0 and
the transducer sensitivity. In spectrophotometry, the absorbance A is proportional to the ratio
of P0 to P. Increasing P0 or the transducer sensitivity to P0 produces a corresponding increase
in P or the sensitivity to P. Thus the ratio does not change. As a result, the sensitivity of
fluorescence can be increased by increasing P0 or transducer sensitivity, but the that of
absorbance does not change.

15-4. Which compound in each of the pairs below would you expect to have a greater
fluorescence quantum yield" Explain.

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 CHEM 3418.001 Test 2 Review Questions

(a) Fluorescein because of its greater structural rigidity due to the bridging –O– groups. \
(b) o,o’-Dihdroxyazobenzene because the –N=N– group provides rigidity that is absent in the –
NH–NH– group.

15-5. Why do some absorbing compounds fluoresce but others do not?

Compounds that fluoresce have structures that slow the rate of nonradiative relaxation to the
point where there is time for fluorescence to occur. Compounds that do not fluoresce have
structures that permit rapid relaxation by nonradiative processes.

15-6. Discuss the major reasons why molecular phosphorescence spectrometry has not
been as widely used as molecular fluorescence spectrometry.
The triplet state has a long lifetime and is very susceptible to collisional deactivation. Thus, most
phosphorescence measurements are made at low temperature in a rigid matrix or in solutions
containing micelles or cyclodextrin molecules. Also, electronic methods must be used to
discriminate phosphorescence from fluorescence. Not as many molecules give good
phosphorescence signals as fluorescence signals. As a result, the experimental requirements to
measure phosphorescence are more difficult than those to measure fluorescence and the
applications are not as large.

2. Explain the similarities and differences between molecular fluorescence,

phosphorescence and chemluminescence.
All of them are the emission of radiation
-Fluorescence: a pathway by which an excited atom/molecule relaxes to its ground state,
emission of radiant energy, excitation brought on by absorption of a photon. (Singlet to
singlet)
-Phosphorescence: similar to fluorescence except has a sig. longer lifetime. Involves change
in electron spin (singlet to triplet)
-Chemiluminescence: the emission of energy as electromagnetic radiation during a chemical
reaction, excited species is formed during the course of a chemical reaction

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3. Compare resonance fluorescence to Stokes shift.

Resonance fluorescence is typical of atomic species but rarely occurs in molecular species. In
resonance fluorescence the same photons that are absorbed are emitted (same energy)
Molecules rarely undergo resonance fluorescence because the lose energy with the molecules
in vib/rot states or transfer to other areas of the molecule
Stokes shift if typical of molecular species. The energy emitted is less than that absorbed
because some energy is lost within the molecule.

4. What effect does temperature, concentration, and solvent viscosity have on fluorescence
and why?
-Temperature: Lower temperatures decrease the movement of the molecules. This decreases
collisions of the molecules. Lower temperature increases the probability of fluorescence.
Higher temperatures allow movement of the molecules or collision of molecules. Higher
temperatures decrease the quantum efficiency of fluorescence, because of collisions
transferring energy (collisional quenching)
-Concentration: at low concentrations fluorescence is linear but is starts to lose linearity at
high concentrations. Linearity is lost due to self quenching () and self absorption which are
greater at higher concentrations.

-Solvent viscosity: high viscosity decreases the movement of molecules which decrease the
collision of the molecules. Therefore higher solvent viscosity promotes fluorescence. Low
viscosity solvents increase the number of collisions which decreases the probability of
fluorescence

5. List and explain the various ways an excited molecule can lose its energy (relaxation or
deactivation processes).
-emission of a photon via fluorescence or phosphorescence
(Fluorescence always occurs from ground excited state because vibrational relaxation occurs
so fast)
Radiationless processes;
*Vibrational relaxation: have a short lifetime, emitted at longer wavelengths,
fluorescence always involves a transition from the lowest vibrational level of an excited
electronic state. Relaxation from higher vibrational state to a lower vibrational state
(cause of stokes shift) Always occurs first.
*Internal Conversion: intermolecular process, a molecule passes to a lower energy
electronic state without the emission of radiation. More efficient when 2 electronic
energy levels are sufficiently close for there to be an overlap in vibrational energy. May
results in predissociation(breaking of bonds) (cause of stokes shift)
*External Conversion: collisional quenching, interaction and energy transfer between the
excited molecule and the solvent or other solutes. Increase solvent viscosity and
temperature decrease collisional quenching by decreasing the movement of the molecules
(this increase fluorescence) This is the way most energy is lost and why very few
molecules fluoresce.
*Intersystem crossing: the spin of the electron is reversed. Transition is enhanced if the
vibrational levels of the two states overlap. Most common in atoms containing heavy
atoms-halogens-heavy atom effect.

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Internal conversion, external conversion, and intersystem crossing compete with fluorescence
and phosphorescence. The longer is stays in the excited state the higher the probability of losing
energy in nonradiative fashion.

6. Why is fluorescence seldom observed from sigma*  sigma transitions?

The transition is too energetic. If occurs it often causes bond ruptures and the absorption of
atmospheric components.

7. Describe some of the structural features of a molecule that affect fluorescence.

Most unsubstituted aromatic compounds fluoresce in solution. Efficiency of fluorescence
increases with the number of rings and their condensation. Fused ring structures also exhibit
fluorescence. Substitutions on the benzene ring cause shifts in wavelength of fluorescence
and also the efficiency of fluorescence. The greater the number of resonance structures (??)
the greater the probability of fluorescence. Rigidity increases the probability of fluorescence.
Rigidity and the number of resonance structures increase the probability of fluorescence.
Hetero atoms(non carbon and hydrogen) change/decrease the fluorescence

A hetero atom added to a cyclic structure decreases fluorescence.

Another cyclic structure added to an existing cyclic structure will increase fluorescence.
The more vibrations/moving = less fluorescence (need rigidity)

8. Discuss the various factors that affect fluorescence and phosphorescence.

Transitions:
Structure: unsubstituted aromatic compounds increase fluorescence with the increasing
number of rings. Double bonds increase fluorescence. The number of resonance structures
increase fluorescence. Increased rigidity increases fluorescence.
pH: The fluorescence of an aromatic compound with acidic or basic ring substituents is
usually pH dependent. Both the wavelength and the emission intensity arc likely to be
different for the protonated and unprotonated forms of the compound. The additional
resonance forms lead to a more stable first excited state; fluorescence in the ultraviolet region
is the consequence.
Temperature: Low temperature results in a higher probability of fluorescene. At higher
temperatures there are a greater number of collisions.
Solvent: Increased solvent viscosity increases probability of fluorescence by limiting the
number of collisions.
Heavy atoms: Decrease fluorescence due to increase in the rate of triplet formations. This
same reason causes an increase in phosphorescence.
Dissolved Oxygen: will interfere with fluorescence may cause intersystem crossing and
conversion to triplet state. Get rid of oxygen to increase fluorescence.
Concentration: High concentration will cause a loss of linearity in fluorescence due to self
quenching and self absorption.

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 CHEM 3418.001 Test 2 Review Questions

9. Discuss the advantages and disadvantages of molecular luminescence over absorption

techniques.
Molecular luminescence has higher sensitivity(can go to much lower concentrations),
large linear concentration range, more specificity, there is also less background noise.
The disadvantage is that with the higher specificity only a few compounds can be measured
via molecular luminescence and is subject to serious interference effects from the matrix at
lower concentrations. Molecular luminescence is less applicable since fewer species
fluoresce.

10. Discuss how the Pauli Exclusion Principle is related to fluorescence and
phosphorescence.
The Pauli exclusion principle says that only 2 electrons can occupy an orbital at one time and
the 2 electrons must be opposite in spin. Fluorescence deals with the excited single state,
while phosphoresce deals with the excited triplet state. In the excited triplet state the
electrons are in opposite spin. The one electron cannot come back down until it changes spin
(because of the Pauli Exclusion Principle)

11. Draw a Jablonski diagram (Figure 15-1 page 357). Label and discuss each of the
diagram. Be as thorough as possible. DO NOT NEED TO DRAW

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12. Define the following terms:

a. self quenching: collision between excited molecules. Ex; benzene collides with
benzene causing a transfer of energy
b. self absorption: cause of nonlinearity, wavelength of emission overlaps an
absorption peak, excited photon may be absorbed by another before it exits the
cell
c. excitation spectrum: obtained by measuring luminescence intensity at a fixed
wavelength while the excitation wavelength is varied. Is often identical to an
absorption spectra taken under the same conditions. Shows which wavelengths
absorbed actually cause excitation. Is a subset of the absorption spectra. No every
wavelength absorbed causes excitation
To collect keep the emission monochrometer steady at a known fluorescence
wavelength. Vary the excitation monochrometer at a shorter wavelength (higher
energy)
d. emission spectrum: fluorescence and phosphorescence spectra involve excitation
at a fixed wavelength while recording the emission intensity as a function of
wavelength. Shows wavelengths that are emitted by the sample
To collect keep the excitation monochrometer steady at a wavelength that causes
excitation. Vary the emission monochrometer at a higher wavelength(less energy).
e. fluorescence lifetime : Time that passes from excitation to emission, varies
within different species, can be used to distinguish species.
f. heavy atom effect: intersystem crossing(spin of electron is reversed) is most
common in molecules that contain heavy atoms such as iodine or bromine. The
presence of heavy atoms makes a change in spin more favorable

13. Explain the relationship between vibrational relaxation and fluorescence.

Vibrational relaxation is a nonradiative form of relaxation, while fluorescence is radiative
relaxation. There is no emission of a photon in vibrational relaxation, it is relaxation to a
lower vibrational state. This vibrational energy loss always occurs first and very fast
compared to the emission of a photon. Due to this fluorescence always occurs from the
excited ground state.

14. Discuss the effect of concentration on fluorescence intensity.

Fluorescence is also related to absorbance, at low concentrations is linear, but as
concentrations rises it begins to lose linearity. Higher concentration causes self-quenching
and self-absorption which case a loss of linearity

15. Discuss two fluorometric methods for the determination of inorganic species.
*Direct method: formation of a fluorescing chelate (complex) and the measurement of its
emission. The complex fluoresces.

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 CHEM 3418.001 Test 2 Review Questions

*Indirect method: Diminution of fluorescence resulting from the quenching action of the
substance being determined. Have a species that fluoresces with something that quenches
fluorescence. You look at the decrease in fluorescence.

Ex. Bromine quenches fluorescence

16. Why is it not necessary to have a wavelength selector when performing a

chemluminescence experiment.
The only source of radiation is the chemical reaction between the analyte and the reagent.

17. Why are fluorescence methods more sensitive and selective than absorption techniques?
In absorption your looking at everything coming in, if only a few absorb you have very high
background noise. Multiple species in the solution may absorb. In fluorescence you are not
seeing the source light. You are looking into the darkness for a few photons. There is no
source interference. Can go much lower and stay linear leading to single photon detection.
Can look at multiple species in a solution and only one may fluoresce. It is very selective to
that one species.

18. Why is chemluminescence such a sensitive technique?

There is a very low noise level because there is no light source. Excitation is from the
chemical reaction. Every single photon hitting the detector is from the reaction.

19. How are fluorescence and phosphorescence different in terms of intensity, lifetime, and
energy?
Fluorescence is higher energy, and a shorter lifetime. Phosphorescence is lower energy, and
longer lifetime. Intensity varies or depends based on conditions that have been setup.

Chapter 16
1. Compare the purposes of UV/Vis and IR absorption spectroscopy.
UV/Vis- quantitative, how much IR- qualitative, functional groups.
IR- organic material absorbs IR because of the covalent bonds. Ionic materials do not
have covalent bonds so they do not absorb IR. Salts are typically used.

2. What are the advantages of a Fourier transform infared spectrometer compared to

a dispersive instrument?
A Fourier transform infared spectrometer is faster because all wavelengths can be done at
one time. A dispersive instrument can scan only 1 wavelength at a time. A fourier has
only 1 detector and doesn’t need a monochrometer.

3. What type of detectors are used in IR spectroscopy? (sensitive thermometer)

*Thermal transducer: heat capacity of absorbing element must be low, thermal noise is a
major issue. Cooling the detector decreases noise. The beam is chopped.
*Thermocouples: 2 pieces of metal fused to each end of a dissimilar metal.
Dissimilar(difference in electron transfer ability makes it more sensitive) Results in a
current from one to the other

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*Bolometer: 2 thin strips of metal, resistance changes with temp.

*Pyroelectric transducer: crystal between 2 electrodes. 1 electrode is IR transparent. The
crystal is heated and the charge changes, charge detected as a current. More sensitive than
thermocoupler. Common for FTIR
*Photoconducting transducer: film of semiconductor material on nonconducting surface.
Decreases the electrical resistance. Measure change in resistance. Hg/Cd/Te offer
superior response time (need to be cooled with liquid nitrogen)

4. What types of molecular transitions are associated with IR absorption?

Vibrational and Rotational transitions

Chapter 20 Molecular Mass Spectrometry

1 Discuss the advantages and disadvantages of electron impact sources.
Advantages: Convenient/easy, high currents resulting in good sensitivity, reproducible
fragmentation pattern allows identification of analyte. Very reproducible, good for
qualitative, can compare to a library frag pattern.
Disadvantages: Extensive fragmentation may cause loss of molecular ion peak, limited to
easily “volatized” samples if it can’t go into the gas phase then you cant use electron impact,
limited to analytes with molecular weights less than 10^3 daltons (1000 amu). If the
molecular weight gets too high it is no longer volatile

2 Discuss the advantages and disadvantages of chemical ionization sources.

Advantage: gives a high response for molecular ion, can get molecular weight of species, it
can be more sensitive
Disadvantage: Can’t use for fragmentation pattern, not reproducible so can’t compare with
library record. The line production is dependent on the reagent and sample reagent pressure
which both vary too much. You lose information that could be obtained from the
fragmentation.

3 Describe the purpose of the quadrupole mass spectrometer and how it works.
The analyzer consists of four rods. The rods operate in pairs each carrying a voltage. Ions of
the proper M/Z value successfully traverse the entire filter. High pass rods filter out ions with
too low of an M/Z. The low pass rods filter out ions with too high of an M/Z. At a set of
conditions only ions of a specific M/Z successfully travel through, the rest are drawn into the
rods. Mass to charge ratio determines stable trajectory.

4 What is the purpose of chemical ionization and how is this done?

A soft ionization method. Relies on the charge being transferred from a reagent molecule to
the sample. Results is reduced fragmentation and increased production of molecular ion. A
reagent gas is introduced directly into the source. The reagent is most ionized.

5 How do the spectra for Electron Impact (EI) and Chemical Ionization (CI) sources
differ from one another?
EI spectra: less molecular ion present, lots of fragmentation, reproducible fragmentation
pattern.

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CI spectra: molecular ion peak, less fragmentation, also reduced amount of structural info,
not reproducible, secondary collisions, part of “finger print”, can give you better detection
limits

6 Why is 70 eV commonly used for EI sources?

70 eV is a good compromise. It causes fragmentation but it won’t destroy it. Lower energies
won’t ionized, and higher energies will completely destroy it.

7 List the various components of electron impact source and discuss each in detail.
*Filament: made of Re, source of 70 eV electrons. It creates the ion
*Target: anode used in association with the filament to produce electrons. Ion travels toward
*Repeller: + charge electrode used to push positive ions out of the ionization source (same
charge as ion)
*Lens stack: series of increasingly more negative electrodes used to accelerater ions to
constant kinetic energy. Accelerate and push toward detector. Need all to have same amount
of kinetic energy so can determine mass charge. Small will hit before the heavier.

8 What is the detector for a GC/MS?

An electron multiplier ….. NOT a mass analyzer or mass spec

Be able to draw a theoretical mass spectra for a simple compound such as water, carbon
dioxide methane, etc. and discuss how this spectra was generated in the mass spectrometer
and discuss each of the peaks in your theoretical spectra.
 Mass spectra are typically represented as a ‘bar’ type graph.
 Each line is used to represent a specific M/Z. (mass/charge)
 X axis best equates to mass
 Y axis – abundance
 Bas peak – the largest line, the most abundant
Types of Information from Graph
 Mass of fragments
 Presence of isotope lines
 Gaps between lines

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 18 is the mass of water

 What shows fragmentation
where it can lose 1 or 2 H
which represent lines 16 and
17

 The line at 44 is the MW of CO2

 The very short lines at 45 and 46
could be attributed to Isotopes…

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