Chem 220

Tutorial #7
Chapter 18: Fundamentals of Spectrophotometry
Chapter 20: Spectrophotometers
 Quiz 6 Answer
Conditional formation constants for metal ion – EDTA titrations are
found from the formula K’f = αY4-Kf, where Kf values are tabulated
for many metal ions at 25oC, and αY4- depends on pH. At pH 9.0,
αY4- is 0.041. What is the minimum value of Kf needed for
quantitative reaction of metal ion with EDTA (99% complete reaction)
at pH 9.0, for a 0.01000 M solution of metal ion titrated with a 0.01000
M solution of EDTA?

At the equivalence point, the total concentration is 0.005000 M

For 99% complete reaction, K’f = 0.00495/(0.00005)2 = 1.98x106

For this K’f value, Kf = K’f/0.041 = 4.8x107

This is the minimum value of Kf needed for quantitative reaction under

these conditions.
 1) Fill in the blanks
a)  If you double the frequency of electromagnetic radiation,
you _________ the energy.
Double
E = hv
h = Planck’s constant
E = Energy
v = frequency
E (energy) and v (frequency) are directly related

Chapter 18, Problem 1, Page 456

1b) If you double the wavelength, you ______
the energy.

Half

E = (hc)/(λ)

h= Planck’s constant

c= speed of light

E (Energy) and λ(wavelength) are inversely related

1c) If you double the wavenumber, you
______ the energy.

Double
!
E = hc v

h= Planck’s constant

c= speed of light
! (wavenumber) are directly related to
E (energy) and v
one another
2) Calculate the frequency (Hz), wavenumber (cm-1), and energy
(J/photon and J/[mol of photons]) of visible light with a
wavelength of 562 nm.

•  v = frequency
= c/λ= (2.9979 x 108 m/s)/(562 x 10-9 m) = 5.33 x 1014 Hz
! = wavenumber
•  v

= 1 / λ=1/(562 x 109 m) = 1.78 x 106 m-1 = 1.78 x 104 cm-1

•  E = energy

= hv = (6.6262x 10-34 Js) (5.33 x 1014 s-1) = 3.53 x 10-19 J/photon

= (3.53 x 10-19 J/photon)(6.022 x 1023 )(1000kJ/1J)=213 kJ/mol

Chapter 18, Problem 3, Page 456

3) The absorbance of a 2.31 x 10-5 M solution of a compound is
0.822 at a wavelength of 266 nm in a 1.00 cm cell. Calculate the
molar absorptivity at 266 nm.

ε= molar absorptivity

= A/bc = (Absorbance)/(pathlength)(concentration of the sample)

= (0.822) / [(1.00cm)(2.31 x 10-5 M)]

= 3.56 x 104 M-1 cm-1

Chapter 18, Problem 12, Page 457

4) A compound with molecular mass 292.16 g/mol was dissolved in a 5-mL volumetric
flask. A 1.00 mL aliquot was withdrawn, placed in a 10-mL volumetric flask, and
diluted to the mark. The absorbance at 340 nm was 0.427 in a 1.000 cm cuvet. The
molar absorptivity at 340 nm is ε340=6130 M-1cm-1

a)  Calculate the concentration of compound in the cuvet.

C = A/εb = (0.427) / [(6130M-1cm-1)(1.000cm)]

= 6.97 x 10-5 M

b) What was the concentration of compound in the 5mL flask?

Sample had been diluted x 10 = 6.97 x 10-4 M

c) How many milligrams of compound were used to make the 5-mL

solution?
x(g)
= 6.97 x 10-4 M àx = 0.00102 g = 1.02 mg
(292.12g / mol)(5.00x10 −3 L)

Chapter 18, Problem 18, Page 457

 5) You have been sent to India to investigate the occurrence of goiter disease
attributed to iodine deficiency. As part of your investigation, you must make field
measurements of traces of iodide (I-) in groundwater. The procedure is to oxidize
I- to I2 and convert the I2 into an intensely colored complex with the dye brilliant
green in the organic solvent toluene.

a)  A 3.15 x 10-6 M solution of colored complex exhibited an absorbance of

0.267 at 635 nm in a 1.000 cm cuvet. A blank solution made from distilled
water in place of groundwater had an absorbance of 0.019. Find the
molar absorptivity of the colored complex.

(0.267 − 0.019)
ε= A/bc = = 7.87 x 104 M-1 cm-1
(3.15x10 −6 M )(1.000cm)

Chapter 18, Problem 20, Page 457

 5b) The absorbance of an unknown solution
prepared from groundwater was 0.175. Find
the concentration of the unknown.

(0.175 − 0.019)
c= A/εb =
(7.87x10 −4 M −1cm −1 )(1.000cm)

c = 1.98 x 10-6 M
6) Nitrite ion, NO2- , is a preservative for bacon and other foods,
but it is potentially carcinogenic. A spectrophotometric
determination of NO2- includes a number of steps. The following
solutions were analyzed:

A.  50.0 mL of food extract known to contain no

nitrite; final absorbance = 0.153

B.  50.0 mL of food extract suspected of containing

nitrite; final absorbance = 0.622

C.  Same as B, but with 10.0 µL of 7.50 x 10-3 NaNO2

added to the 50.0 mL sample; final absorbance =
0.967

Chapter 18, Problem 22, Page 458

 Number 6 continued…
a)  Calculate the molar absorptivity, ε, of the colored product. A 5.00 cm cell
was used

Note: Sample C contains colored product from nitrite added, absorbance = 0.977

First, determine absorbance due to added colored product:

0.977 – 0.622 (From sample B, suspected of containing nitrite) = 0.345

Now, calculate concentration of colored product with a total volume of 54.0 mL:

((7.50 x 10-3 M)(10.0 x 10-6 L)) / (0.054 L) = 1.98 x 10-6 M

Now, solve for molar absorptivity:

ε=A/bc = (0.345) / ((5.00)(1.98 x 10-6) = 4.97 x 104 M-1cm-1

 Number 6 continued…
b) How many micrograms of NO2- (FM 46.01 g/mol) were
present in 50.0 mL of food extract?
Note: Sample C gives an absorbance of 0.345 and Sample A (no
nitrite) gives an absorbance of 0.153
Determine the new absorbance of Sample B in relation to A:
0.622 – 0.153 = 0.469
Determine the moles of Sample B in relation to Sample C:
(x moles)/(7.50 x 10-8 moles) = (0.469) / (0.345)
x = 1.020 x 10-7 moles NO2- = 4.69 µg
7) 0.10 mM KMnO4 has an absorbance maximum of 0.26 near
525 nm in a 1.00 cm cell. Find the molar absorptivity and the
concentration of a solution whose absorbance is 0.52 at 525 nm
in the same cell.

•  First, find molar absorptivity from Beer’s Law:

Molar Absorptivity =ε
= A/bc = (Absorbance)/(pathlength)(concentration of the sample)

ε= (0.26) / [(1.0cm)(0.10 x 10-3 M)]

ε= 2600 M-1 cm-1

8th edition, Chapter 17, Section 17-4 Example, Page 401

 Number 7 Continued…
•  Now find concentration using the molar absorptivity:

c = A/εb

c = (0.52)/(2600 M-1 cm-1)(1.0cm)

c = 0.0002 M or 0.20 mM
 8) The transmittance of a 0.010M solution of a
compound in a 0.100 cm pathlength cell is T=8.23%.
Find the absorbance and the molar absorptivity.
•  First convert T into a decimalà 8.23% = 0.0823

•  Now calculate absorbance (A)

-A = log (T)
-A = log (0.0823)
-A = -1.0846
A = 1.08

8th edition, Chapter 17, Section 17-2 Example, Page 397

 Now calculate molar
absorptivity

ε= A/(b)(c)
ε = (1.08)/[(0.10cm)(0.01M)]
ε = 1084.6
ε = 1.08 x 103 M-1 cm-1
 9) What is the wavelength, wavenumber and
name of radiation with an energy of 100kJ/
mole?
100 kJ/mole = 100000 J/mole
100000J 1mole −19
x 23
= 1.661x10 J / molecule
mole 6.022x10 molecules
ε = 1.661 x 10-19 J/molecule
hc hc
ε= ⇒λ=
λ ε
(6.626x10 −34 J ⋅ s)(3x108 m / s)
λ= −19 = 0.00000119 m = 1.20 μm
(1.66x10 J / molecule)

λ= 1.19 x 10-6 m
8th edition, Chapter 17, Section 17-2 Example, Page 396
Now calculate wavenumber…

1 1 1
ν! = = =
λ 0.00000119m 0.000119cm

! 3
ν = 8.33x10 cm −1

Recall, the wavelength was 1.19 x 10-6 m…….

When referencing the radiation diagram on page
395, 10-6 corresponds with Infrared
 10) Find the absorbance and transmittance of a 0.00240 M
solution of a substance with a molar absorptivity of 313
M-1cm-1 in a cell with a 2.00 cm pathlength.

Solve for absorbance:

A=εbc=(313 M-1cm-1)(2.00cm)(0.00240 M)=1.50

Solve for transmittance:

log T = -A

T = 10logT = 10-A = 10-1.50 = 0.0316

Chapter 18, Section 18-2 Example, Page 436

 11a) Pure hexane has negligible UV absorbance above a wavelength of 200 nm. A
solution prepared by dissolving 25.8 mg of benzene (FM 78.11) in hexane and dilution
to 250 mL had an absorption peak at 256 nm and an absorbance of 0.266 in a 1.0 cm
cell. Find the molar absorptivity of benzene at this wavelength.

[C6H6] = {(0.0258 g)(78.11g/mol)}/(0.250 L) = 1.32 x 10-3 M

Find Molar absorptivity from Beer’s Law:

ε= A/(bc) = (0.266)/(1.0)(1.32x10-3)

= 201.3 M-1cm-1

Chapter 18, Section 18-4 Example, Page 440

 11b) A sample of hexane contaminated with benzene had an
absorbance of 0.070 at 256 nm in a cuvet with a 5.0 cm
pathlength. Find the concentration of benzene in mg/L.

•  Know that ε=201.3 M-1cm-1 from part A

[C6H6] = (A)/(εb)=(0.070)/(201.3)(5.0)

= 6.95 x 10 -5 M

•  Convert to correct units:

[C6H6] = (6.95 x 10 -5 mol/L)(78.11x103mg/mol)

= 5.4 mg/L
12a) What resolution is required for a diffraction
grating to resolve wavelengths of 512.23 and
512.26 nm?

•  Resolution = ability to distinguish two closely spaced peaks

•  Resolution = (λ/Δλ)

= (512.245/(512.26-512.23))

= (512.245/0.03)

=1.7 x 104

Chapter 20, Problem 11, Page 525

 12b) With a resolution of 104, how close in nm
is the closest line to 512.23 nm that can barely be
resolved?
•  Rearrange equation from part a
•  Resolution = (λ/Δλ)

Δλ= (λ)/Resolution
= (512.23)/(104)
=0.05nm
 12c) Calculate the fourth-order resolution of a
grating that is 8.00 cm long and is ruled at 185
lines/mm.
•  Resolution=(λ/Δλ)=nN
•  n= diffraction order
•  N=number of grooves of the grating that are illuminated

•  Resolution = nN

=(4)(8.00cm x 185 lines/mm)

Note: Need to convert lines/mm into lines/cm)

=(4)(8.00cm x 1850 cm-1)

= 1.5 x 104
12d) Find the angular dispersion (ΔΦ) between light rays with
wavelengths of 512.23 and 512.26 nm for first-order diffraction
(n=1) from a grating with 250 lines/mm and Φ=3.0o

•  Dispersion: measures the ability to separate wavelengths differing

by Δλthrough the difference in angle, ΔΦ(radians)
Δφ n
•  Dispersion of grating: =
Δλ d cos φ
•  First, calculate for first order diffraction:

Given:
n= 1

Φ=3o

Δλ=(512.26-512.23)=0.03nm=3x10-5um
d= (250 lines/mm)(1mm/1000um) = 0.25 lines/um = 4 um/line
 12d continued
Δφ n
=
Δλ d cos φ
Δφ (1) radians o
= 0
= 0.250 = 14.3 / µ m
Δλ (4µ m)cos3 µm
o −5 −4
Δφ = (14.3 / µ m)(3x10 µ m) = 4.3x10 deg rees
 13a) The true absorbance of a sample is 1.500, but 0.50% stray
light reaches the detector. Find the apparent transmittance and
apparent absorbance of the sample.

•  True transmittance, T=10-A =P/Po

T=10-1.500 = 0.0316

Po=1, P=0.0316, S=0.005

P+S
•  Apparent transmittance =
Po + S

0.0316 + 0.0050
= = 0.0364
1+ 0.0050
•  Apparent absorbance = - log(T)

= - log (0.0364) = 1.439

Chapter 20, Problem 12, Page 525

 13b) How much stray light can be tolerated if the
absorbance error is not to exceed 0.001 at a true
absorbance of 2?
•  Apparent absorbance = (2)-(0.001) = 1.999

•  Apparent transmittance= 10-1.999 =0.01002305

P+S
=
Po + S

(0.010) + S
= = 0.01002305
1+ S

S = 2.328 x 10-5 = 0.002328%

13c) A research quality spectrophotometer has a stray light level
of <0.00005% at 340 nm. What will be the maximum
absorbance error for a sample with a true absorbance of 2? Or 3?

•  True absorbance = 2
P+S (0.010) + (0.0000005)
•  Apparent transmittance = =
Po + S 1+ (0.0000005)

= 0.01000049

•  Apparent absorbance = -log (T)

= -log (0.01000049) = 1.999978

•  Absorbance error = 2-1.999978 = 0.000022

 13c continued
•  True absorbance = 3
0.001+ 0.0000005
•  Apparent transmittance = = 0.001000495
1+ 0.0000005
•  Apparent absorbance = -log(T)

=-log(0.001000495)

=2.999785

•  Absorbance error = 3 – 2.999785

= 0.000215
 14) If the true absorbance of a sample is 2.00
and there is 1.0% stray lights, find the apparent
absorbance.
Apparent transmittance = (P + S)/(Po + S)

Po = Irradiance passing through the reference

P = Irradiance passing through the sample

S = stray light

•  First, relate absorbance and transmittance:

T = 10-A = 10-2.00 = 0.010 = 1.0%

T = P/Po = 0.010

Chapter 20, Section 20-2 Example, Page 500

 Number 14 continued…
Apparent transmittance = (P + S)/(Po + S)
•  Given:
•  S = 1.0% = 0.010
•  P/Po = 0.010 = 0.010/1

•  Now solve for apparent transmittance:

Apparent transmittance = (0.010 + 0.010)/(1 + 0.010)

Apparent transmittance = 0.0198

•  Now solve for apparent absorbance:

A = -log (T) = -log (0.0198) = 1.70
15) What level of stray light gives an absorbance error
of 0.01 at an absorbance of 2? That is, what value of S
gives an apparent absorbance of 1.99?
•  Given:
•  A = 2
•  Apparent absorbance = 1.99
•  Absorbance error = 0.01

•  First, solve for apparent transmittance:

-  log (T) = A

-  log (T) = 1.99

T = 0.010233

8th edition, Chapter 19, Section 19-2 Example, Page 454

 Number 15 continued…
•  Now solve for S given that apparent transmittance =
0.010233 :

Apparent transmittance = (P + S)/(Po + S)

0.010233 = (0.01 + S)/(1 + S)

0.010233 + 0.010233S = 0.01 + S

0.000233 = 0.989767S

S = 0.000235 = 0.024%
16) Find the minimum angle θi for total reflection in the optical
fiber in Figure 20-21 if the index of refraction of the cladding is
1.400 and the index of refraction of the core is (a) 1.600 or (b)
1.800.

•  Figure 20-21

Chapter 20, Problem 23, Page 526

 16a) Find the minimum angle for total
reflection when the index of refraction of
the core is 1.600
ncoresinθi=ncladdingsinθr

For total reflection: sinθr ≥1

ncladding
sinθi≥ (Page 462)
ncore
Given:
ncladding=1.400
ncore=1.600

1.400
sinθi ≥ à θi ≥ 61.04o
1.600
16b) Index of refraction of the
core is 1.800
ncoresinθi=ncladdingsinθr
For total reflection: sinθr ≥1
ncladding
sinθi≥ (Page 462)
ncore
Given:
ncladding=1.400
ncore=1.800

sinθi ≥
1.400 à θi ≥ 51.06o
1.800
17a) If a diffraction grating has a resolution of 104, is
it possible to distinguish two spectral lines with
wavelengths of 10.00 and 10.01 µm?
•  Resolution = λ/Δλ

•  Known resolution = 104

•  For a λ =10.00 µm, find Δλ:

Δλ= 10.00 µm – 10.01 µm = 0.01 µm

•  Solve for resolution of spectral lines:

= 10.00 µm / 0.01 µm = 103

•  Resolution given greater, so the spectral lines will be resolved

Chapter 20, Exercise A, Page 524

 17b) With a resolution of 104, how close in
wavenumbers (cm-1) is the closest line to 1000
cm-1 that can barely be resolved?
•  Given resolution = 104

λ= 1 / ν!

λ= 1 / (1000 cm-1)(10-4 cm/µm) = 10 µm

•  Solve for Δλ

Δλ= λ/Resolution

Δλ= (10 µm) / (104) = 10-3 µm

•  10.001 µm could be resolved from 10.000 µm

 17c) Calculate the resolution of a 5.0 cm long
grating ruled at 250 lines/mm for first order (n=1)
diffraction and tenth-order (n=10) diffraction.
•  How many lines are present?

(5.0 cm) (2500 lines/cm) = 12,500 lines

•  Now solve for resolution for both orders of diffraction:

Resolution = n N= (diffraction order)(number of grooves of grating)

•  First Order
Resolution = (1)(12,5000) = 12,5000
•  Tenth Order
Resolution = (10)(12,500) = 125,000
17d) Find the angular dispersion (ΔΦ, in radians and degrees)
between light rays with wavenumbers of 1000 and 1001 cm-1 for
second-order diffraction (n=2) from a grating with 250 lines/
mm and Φ=30o.
Δφ n
•  Dispersion of grating: =
Δλ d cos φ
Δφ 2 radians radians
= = 577 = 0.577
Δλ ( 1mm )cos(30 o ) mm µm
250lines

•  Now convert radians to degrees = (radians/π) x 100

[(0.577 radians/µm)/(π)] x 100 = 33.1 degrees/µm

•  Now solve for Δφ :

Δλ =1001 – 1000 = 1 cm-1 = 0.01 µm
Δφ =(0.577 radians/µm)(0.01µm) = 6 x 10-3 radians = 0.3o
 18) A measurement with a signal-to-noise ratio of
100/1 can be thought of as a signal, S, with 1%
uncertainty, e. That is, the measurement is S ±e = 100 ±
1
a)  Use the rules for propagation of uncertainty to show that, if
you add two such signals, the result is total signal = 200 ±
√2, giving a signal-to-noise ratio of 200/√2 = 141/1

Rules for Propagation of Uncertainty can be found on pages 57-64.

For this problem, we refer to the rules for addition and subtraction:

e4= e12 + e22 + e32

(100 ± 1) + (100 ± 1) = 200 ± e

e=√(e12+e22) = √(12+12) = √(2)

Chapter 20, Problem 34, Page 527

 18b) Show that if you add four such
measurements, the signal-to-noise ratio
increases to 200/1.

(100 ± 1) + (100 ± 1) + (100 ± 1) +(100 ± 1) = 400 ± e

e= √(12+12+12+12) = 2

400 ± 2 = 400:2 = 200:1

18c) Show that averaging n measurements increases the
signal-to-noise ratio by a factor of √n compared with
the value of one measurement.
•  Initial measurement signal/noise = 100/1
•  Averaging n measurements gives:
average signal = n(100) = 100
n

average noise = n
=
1
n n

avgsignal 100
= = 100 n
avgnoise 1 / n
which is √n times greater than the original value of signal/noise
 Quiz Time
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