Ambo University

Department of Chemistry
Chem. 572 Advanced Instrumental Analyses
Instrumentation and interpretation of spectra

Exercises in spectroscopic techniques

1. A 15.0-mg sample of a compound with a molecular mass of 384.63 was dissolved in a 5mL volumetric flask. A 1.00-mL aliquot was withdrawn, placed in a 10-mL volumetric
flask, and diluted to the mark. Then 10-mL sample was placed in a 0.500-cm cuvet and
gave an absorbance of 0.634 at 495 nm. Find the molar absorptivity (495, with units of M1

.cm-1) at this wavelength

Ans. M1 = n/v(L) = 15/(384.63*5) = 0.0078 mol/L and M1V1 = M2V2, M2 = 1*0.0078/10 =

0.00078 mol,

= A/cb = 0.634/(0.00078*0.5) = 1625.64M-1cm-1

2. During an assay of the thiamine (vitamin B 1) content of a pharmaceutical preparation, the

percent transmittance scale was accidentally read, instead of the absorbance scale of the
spectrophotometer. One sample gave a reading of 82.2% T, and a second sample gave a
reading of 50.7% T at a wavelength of maximum absorbance. What is the ratio of
concentrations of thiamine in the two samples?
Ans. A1 = c1b = - log T

= 2 - log %T

and A2 = c2b = - log T

= 2 - log %T

C1: C2 = (2 - log 82.2 )/(2 - log 50.7 ) = 0.288

3. A 1.28104M solution of potassium permanganate has a transmittance of 0.5 when

measured in a 1 cm cell at 525 nm.
1. Calculate the molar absorptivity coefficient for the permanganate at thiswavelength.
2. If the concentration is doubled what would be the absorbance and thepercentage
transmittance of the new solution?

4. A polluted water sample contains approximately 0.1 ppm of chromium, (M = 52 g mol1).

The determination for Cr(VI) based upon absorption by its diphenylcarbazide complex
(max= 540nm, max= 41 700 Lmol1 cm1) was selected for measuring the presence of the
metal. What optimum pathlength of cell would be required for a recorded absorbance of
the order of 0.4?
5. Paints and varnishes for use on exteriors of buildings must be protected from the effects
of solar radiation which accelerate their degradation (photolysis and photochemical
reactions). Given that: M = 500 g mol1; _max = 15 000 Lmol1 cm1 for _max =
350nm, what must be the concentration (expressed in gL 1) of a UV additive M such that
90% of the radiation is absorbed by a coating of thickness 0.3 mm?
6. Pyridine, a common laboratory compound that is known to cause sterility in men,
exhibits a UV transition at max 270 nm. When pyridine is reacted with dilute HCl, this
absorption band goes away. Explain this phenomenon using pictures and words. Does this
data explain what type of transition is occurring in the UV-absorption spectrum of
pyridine at 270 nm?
7. By employing the empirical rules of WoodwardFieser, predict the position of the
maximum absorption of the compounds whose structures are seen below

8. If chloroform (trichloromethane) exhibits an infrared peak at 3018cm 1 due to the CH

stretching vibration, calculate the wavenumber of the absorption band corresponding to
the CD stretching vibration in deuterochloroform (experimental value 2253cm1).
9. To determine the concentrations (mol/L) of Co(NO3)2 (A) and Cr(NO3)3 (B) in an
unknown sample, the following representative absorbance data were obtained.
Measurements were made in 1.0 cm glass cells.
1. Calculate the four molar absorptivities: A(510), A(575), B(510) and B(575).
2. Calculate the molarities of the two salts A and B in the unknown.
10. If T =300K, calculate the ratio of the populations NE1/NE2 for a proton in an NMR
spectrometer where the applied magnetic field B = 1_4T. Make the same calculation for
the case where the field B=7T, given that =2.6752108 rad T1 s1.
11. 1. Calculate the chemical shift, , in ppm of a proton ( 1H), whose NMR signal is
displaced by 220 Hz with respect to TMS (the field of the spectrometer is 1.879 T).
2. The resonance signal for a proton is displaced by 90 Hz with respect to TMS when
measured on a 60 MHz spectrometer. What would happen to this displacement if an
apparatus of 200 MHz was employed?
3. What would be the corresponding chemical shift of the same protonwith both of these
spectrometers?
12. Spectrophotometric analysis of phosphate can be performed by the following
procedure: Standard A. KH2PO4(FM 136.09): 81.37 mg dissolved in 500.0 mL of water
solutionsB. Na2MoO4 . 2H2O (sodium molybdate): 1.25 g in 50 mL of 5 M H2SO4
C. H3NNH2+ 3 SO2- 4 (hydrazine sulfate): 0.15 g in 100 mL of H2O
Procedure Place the sample (either an unknown or the standard phosphate solution, A) in
a 5-mL volumetric flask, and add 0.500 mL of B plus 0.200 mL of C. Dilute to almost 5
mL with water, and heat at 100C for 10 minutes to form a blue product
(H3PO4(MoO3)12, 12- molybdophosphoric acid). Cool the flask to room temperature,
dilute to the mark with water, mix well, and measure the absorbance at 830 nm in a 1.00cm cell.

(a) When 0.140 mL of solution A was analyzed, an absorbance of 0.829 was recorded. A
blank carried through the same procedure gave an absorbance of 0.017. Find the
molar absorptivity of blue product.
(b) A solution of the phosphate-containing iron-storage protein ferritin was analyzed by
this procedure. The unknown contained 1.35 mg of ferritin, which was digested in a
total volume of 1.00 mL to release phosphate from the protein. Then 0.300 mL of this
solution was analyzed by the procedure above and found to give an absorbance of
0.836. A blank carried through this procedure gave an absorbance of 0.038. Find the
weight percent of phosphorus in the ferritin.
13. You are a supervisor in a laboratory which focuses on the determination of trace and
ultra-trace levels of toxic substances in water samples. Usually, for the determination of
Pb in water, a Graphite Furnace Atomic Absorption Spectrophotometer (GFAAS) is
used but, since it was down for repairs, one of the technicians that you supervise
decided to use the operable Flame AAS instrument. After establishing that absorbance
varied linearly with the amount of Pb, the following absorbance value data (for 5
replicate measurements) were obtained for a blank and for a sample submitted for
analysis: submitted for analysis:
a) Based on the average absorbance value obtained for the sample, the technician
reported that the water sample contained 500 ppb Pb (15 ppb Pb is the maximum Pb
level allowed in drinking water!). As the lab supervisor, you are ultimately
responsible for the analysis results that are reported. Should this result be reported?
EXPLAIN. (NOTE: your response should be based on the above data. HINT: you
may wish to calculate the S/N for the sample data . . . )
b) If a 5000 ppb Pb standard gave an average absorbance value of 0.024, what would the
detection limit (ppb Pb) be for the determination of Pb using this instrument?
14. a)

The

Pd

4,4'-bis(dimethylamino)thiobenzophenone

complex,

in

aqueous

solution,absorbs very strongly in the UV/Vis spectral region, with a molar absorptivity

of 2.12 x 105. Assuming that the minimum measurable absorbance is 0.010 and that a
cell with a 2.50-cm pathlength is available, what is the lowest possible concentration
(ppb) of Pd that can be determinedspectrophotometrically
b) Assuming that the readout uncertainty of the spectrophotometer is 0.10 %-T, what is
the %-relative uncertainty in making an absorbance measurement of 0.010?
c) Assuming that the Pd 4,4'-bis(dimethylamino)thiobenzophenone system follows
Beer's Law in this concentration range, calculate the detection limit for Pd with this
system (i.e., the concentration of Pd, in ppb, that wouldgive a S/N = 3).
d) Assuming that the Pd 4,4'-bis(dimethylamino) thiobenzophenone absorption
bandwidth is 50 nm and the monochromatoreffis 5 nm, what effect would a 10x
increase in the slitwidth of the monochromator have on theapplicability of Beers
Law with this system? Illustrate with a Beers Law plot.
15. a) The most common detector used in FT-IR is the so-called TGS (Triglycine Sulfate)
detector which is a thermal detector and has poor sensitivity. Explain why a
photoemissive detector (like the photomultiplier tube commonly used in UV/Vis
spectrometry) cannot be used in FT-IR.
b) Arrange the following stretches in order of increasing vibration frequency: C-H, C-C,
CC, C=O, C=C. Briefly justify/explain your answer.
c) How many fundamental vibrational modes are predicted for water (H2O)? Diagram
one vibrational mode that you would expect to be IR active.
16. Elemental analysis at ppb levels can be performed using ICP atomic emission
spectrometry (ICP-AES), graphite furnace atomic absorption spectrometry (GFAAS),
and anodic stripping voltammetry (ASV). Which one of these methods would be best
for the daily determination of carcinogenic Cr(VI) in water discharged from a
manufacturing facility? Briefly justify your choice.

17. Which of the following types of analysis involves interaction of the sample with the
least amount of energy? IR, NMR, UV-Vis, MS ,
18. Deduce the structure that corresponds to this spectral data. Mass spectrum: M: m/z 148
(100 %); M + 1: m/z 149 (11.25 %); M + 2: m/z 150 (0.54 %).

H-NMR: 7.5 - 8.0 ppm (multiplet, integral = 2.5), 3.0 ppm (triplet, integral = 1), 1.8 ppm

(sextet, integral = 1), 1.0 ppm (triplet, integral = 1.5).

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C-NMR (DEPT): 200.4 ppm

(singlet), 137.0 ppm (singlet), 132.8 ppm (doublet), 128.5 ppm (doublet), 128.0 ppm
(doublet), 40.4 ppm (triplet), 17.7 ppm (triplet), 13.8 ppm (quartet).
19. Provide a molecular structure that is consistent with the spectral data. Mass spectrum:
M: m/z 132,( 100%); M + 1: m/z 133, 96.89%); M + 2: m/z 134, (1.42%).

1H-NMR: 4.0 ppm (triplet, integral = 1.0); 3.9 ppm (singlet, integral = 1.5); 3.0 ppm
(singlet, integral = 1.0); 1.3 ppm (sextet, integral = 1.0); 0.9 ppm (triplet, integral = 1.5).
13C-NMR (DEPT): 171 ppm (singlet), 70 ppm (triplet), 56 ppm (triplet), 52 ppm
(quartet), 30ppm (triplet), 18 ppm (quartet).
20. Propose a structure for the compound C5H10O based on the following spectroscopic data

21. Propose a structure for the compound C 6H12O2 based on the following spectroscopic
data

22. For each of the following compounds, calculate the number of multiplets for each band
and their relative areas: (a) C1(CH2)3CI; (b) CH3CHBrCH3; (c) CH3CH2OCH3.

23. The proton NMR spectra shown in Figure below are for colorless, isomeric liquids
containing only carbonand hydrogen. Identify the two compounds.

FIGURE NMR spectra for two organic Isomers in COCI]. (Courtesy of Varian Inc
...Palo Alto. CA.)
24. The two mass spectra belong to the isomers of C 6H12O below. The peak at m/z 71 is
C4H7O+, which comes from loss of an ethyl group (CH 3CH2., 29 Da). What is the
expected intensity of m/z 72 relative to that of m/z 71?

Mass spectra of isomeric ketones with the composition C 6H12O from NIST/EPA/NIH
MassSpectral Database.
25. Predict the splitting patterns (number of lines for each signal) you would predict for
each type of carbon and proton in the molecule below

26. Which bond has a higher (= larger) infrared vibrational frequency: C=N or C=S?
Explain briefly.
27. Which one of the following bonds has the highest/largest infrared vibrational
frequency?
a. C-N
b. C-O
c. C-F
d. C-C
e. C-B
28. How many signals would you see in the

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C and 1H NMR spectra of the following

Compound?

29. Describe how you would quantitatively determine the following drugs using IR.

30. What do you understand by NMR-Spectroscopy? Discuss the NMR-phenomenon

including the following vital aspects:
(i) Spinning Nucleus, (ii) Effect of an External Magnetic Field,
(iii) Precessional Motion, (iv) Precessional Frequency, and (v) Energy Transitions.
31. Explain the following terminologies usually encountered in NMR-Spectroscopy :
(a) Spin quantum number, (b) Chemical shift, (c) Spin-spin interactions, and
(d) Pascals triangle.
32. What essential steps you would consider in order to interpret a NMR-spectrum?
Explain.
33. The spectra shown in Figure 1 (a)(d) were obtained for a compound of composition
C 67.0%, H 7.3%, N 7.8%; melting at 135C. Propose a plausible structure.