Problems in organic UV-Visible spectroscopy

- Dr. A. Shunmugasundaram.
Retd. Professor of Chemistry,
VHNSN College, Virudhunagar

Beer- Lambert Law

A = absorbance or optical density

C = concentration in moles/litre or M or mol dm-3

l= path length in cm

Ɛ= molar absorptivity or molar extinction coefficient expressed in M-1 cm-1 or dm3 mol-1 cm-1

I0 = intensity of incident radiation

I = intensity of transmitted radiation

A 1 / A2 = C 1 /C2 at the same path length and wavelength

A1 / A2 = l1 / l2 at the same concentration and wavelength

Transmittance T = I / I0 % Transmittance %T = 100 X I / I0 Absorbance A = log 1 /T

Determination of concentration of components in a mixture

Absorbances of the mixture (A and B) are measured at two different wavelengths (λ1 and λ2) keeping the path
length the same.

Total absorbance of the mixture at λ1 At1 = ɛA1 CA l + ɛB1 CB l (1)

Total absorbance of the mixture at λ2 At2 = ɛA2 CA l + ɛB2 CB l (2)

 ɛA and ɛB are determined separately from the slope of the plot of absorbance vs concentration at the given
wavelength.

Use equation (1) to solve for CA

When the path length is the same,

At1 = ɛA1 CA + ɛB1 CB

At1 - ɛB1 CB = ɛA1 CA

CA = ( At1 - ɛB1 CB ) / ɛA1 (3)

Substitute CA in equation (2) to solve for CB

At2 = ɛA2 [ ( At1 - ɛB1 CB ) / ɛA1 ) ] + ɛB2 CB

Use the value of CB in equation (3) to solve for CA

Conjugated dienes and α ,β- unsaturated carbonyl compounds absorb in the region above 200 nm.

Woodward method to calculate λmax in diene system :

Base value for acyclic or heteroannular diene 215

Base value for homoannular diene 253

Extended conjugation +30

Alkyl or alkyl like substituent +5

Exocyclic double bond +5

OR alkoxy +6

Cl , Br +5

OAc +0

SR thioether +30

Woodward-Fieser rule to calculate λmax in α,β-unsaturated carbonyl compound

Base value for acyclic or 6 – membered cyclic α,β-unsaturated ketones 215 nm

Base value for 5 – membered cyclic α,β -unsaturated ketones 202 nm

Base value for α,β -unsaturated aldehydes 210 nm

Base value for α,β -unsaturated acids or esters 195 nm

Increment values

Extended conjugation +30

Second double bond homoannular +39

Exocyclic double bond +5

Substituent α β γ δ

Alkyl or alkyl like +10 +12 +18 +18

OH +35 +30 +30 +50

OR +35 +30 +17 +31

OAc +6 +6 +6 +6

Cl +15 +12 +12 +12

Br +25 +30 +25 +25

SR - +85 - -

NR2 - +95 - -

PROBLEMS

1) What is the wavelength in nanometer and centimeter corresponding to 5000 A 0 ?

2) If the transmittance of a sample in a quartz cell of the spectrometer is 0.6 what is the absorbance of the
sample ?

3) A light of 350 nm is passed through a solution. If the intensity of the incident light is 500 lumens and that
of the transmitted light is 100 lumens, what is the percentage of transmittance ?

4) In a spectrophotometer 300 lumens of light enter the sample cell and 200 lumens of light come out of the
cell. What is the absorbance of the sample ?
5) What is the absorbance of the 3 x 10-5 M solution taken in a cell of path length 3 cm. The molar
absorption coefficient of the solution is 8000 M-1 cm-1 and the wavelength of the radiation is 280 nm.

6) Calculate the absorbance of the 7.5 x 10-5 M solution taken in a quartz cell of path length 1 cm. The
molar absorption coefficient of the solution is 3000 M-1 cm-1 at 320 nm.

7) A solution of concentration 5 mM shows an absorbance 0.2 at 550 nm in a 1 cm cell. What is the value of
molar extinction coefficient ?

8) The absorbance is found to be 0.125 for a solution of concentration 0.0002 M at 350 nm. The path length
is 1cm. What is the molar absorption coefficient of the solution ?

9) A solution of an organic substance gives an absorbance 0.54 when measured at 500 nm in a 1 cm path
length cell. The molar absorption coefficient is 6000 M-1 cm-1. What is the concentration of the solution ?

10) The optical density of a solution is 1.0. What is the percentage transmittance ?

11) A solution of an organic substance gives an absorbance 0.6 at 300 nm in a quartz cell of 1 cm path length.
The molar absorption coefficient of the substance is 6 x 103 M-1cm-1. What is the concentration of the
solution ?

12) A solution of concentration 5 x 10-4 M in a 2 cm length quartz cell has an absorbance 1.5. What is the
molar absorption coefficient of the solution ?

13) A solution shows a transmittance of 20% when taken in a cell of 2 cm thickness. The molar absorptivity of
the solution is 2 x 104 M-1 cm-1. What is the concentration of the solution ?

14) A sample of phenylalanine shows an absorbance 0.62 at a wavelength 260 nm. The path length of the
light is 2 cm and the molar absorption coefficient is 8800 M -1 cm-1. What is the concentration of the
sample ?

15) A food dye has a molar absorptivity of 20100 M-1 cm-1 at a wavelength of 480 nm. The solution of the dye
is taken in a cuvette of width 1 cm. The absorbance of the solution is 0.22. What is the concentration of
the solution ?

16) A solution contains 2 g/dm3 of a compound(Molecular mass 250). The solution in a cuvette transmits
75% of the incident light of wavelength 400 nm. What is the molar absorptivity of the substance in the
solution ?

17) A solution of concentration 1 x 10-4 M of a substance has an absorbance 0.18 at 400 nm in a 1 cm cuvette.
What is the absorbance when the concentration of the solution is doubled and measured at 400 nm in a
1 cm cuvette ?

18) A solution having concentration 1 x10-5 M has transmittance 70% at 300 nm in a 1 cm path length cell.
What is the transmittance of the solution in a 3 cm path length cell ?
19) 0.002 M solution of a light absorbing substance in a 1 cm cuvette transmits 60% of the incident light of
400 nm wavelength. What is the transmittance of a 0.006 M solution ?

20) A solution containing 2 x 10-5 M of a substance has 60% transmission at 300 nm in a 2 cm quartz cell.
Calculate the absorbance and transmittance of a 5 x10-5 solution in the same quartz cell of 2 cm path
length.

21) A solution of concentration 1 x 10-5 M has 50% transmittance at 450 nm in a 2 cm quartz cell. Calculate the
% of transmittance of a 3 x 10-5 M solution in the same quartz cell.

22) A mixture of two solutions A and B has a total absorbance of 1.22 at 380 nm. If the absorbance of A is 0.62
at 380 nm, what is the absorbance of B at the same wavelength ?

23) A compound of molecular mass 100 reduces the intensity of 300 nm radiation to 20% when passed
through a solution containing 20 mg of the compound in 200 cm3 of alcohol. The path length is 1.0 cm.
Calculate the molar absorption coefficient.

24) A solution of a mixture X and Y is taken in a cell of path length 3 cm. The molar absorption coefficient
and concentration of X are respectively 8000 M-1cm-1 and 3 x 10-5M and those of Y are 2500 M-1cm-1
and 7.5 x 10-5 M respectively at wave length 430 nm. What is the total absorbance of the mixture at that
wavelength ?

25) A compound (molecular mass 100) shows an absorption band at 260 nm. The molar absorption coefficient
is 1200 M-1cm-1. Calculate the %transmittance of the radiation when passed through a solution of 10 mg
in 100 ml of alcohol in a cell of path length 1 cm. What is the concentration of the solution if the
transmittance is required to be 30% ?

26) A mixture of two solutions A and B is analysed using UV –Vis spectroscopy at two different wavelengths.
There is no interaction between the species A and B. The path length of the cell is 1 cm. The following
spectral data are obtained:
Total absorbance at 550 nm = 0.72
Molar extinction coefficient of A at 550 nm = 2.2 M-1 cm-1
Molar extinction coefficient of B at 550 nm = 1.5 M-1 cm-1

Total absorbance at 500 nm = 0.77

Molar extinction coefficient of A at 500 nm = 1.5 M-1 cm-1
Molar extinction coefficient of B at 500 nm = 2.6 M-1 cm-1
Determine the concentration of A and B in the mixture.

27) A mixture of Fe3+ and Cu 2+ is treated with Ru(CN)64- to form a purple-blue complex with ferric ion ( λmax =
550 nm) and a pale green complex with cupric ion ( λmax =396 nm). The mixture is analyzed in a quartz cell
of path length 2 cm. The UV-Vis spectral data at two different wavelengths are given below:
Total absorbance at 550 nm = 0.366
Molar absorptivity of Fe3+ at 550 nm = 9970 M-1 cm-1
Molar absorptivity of Cu2+ at 550 nm = 34 M-1 cm-1
 Total absorbance at 396 nm = 0.218
3+
Molar absorptivity of Fe at 396 nm = 84 M-1 cm-1
Molar absorptivity of Cu2+ at 396 nm = 856 M-1 cm-1
3+ 2+
Determine the concentrations of Fe and Cu ions in the solution.

28) A mixture of two substances A and B is analysed at two different wavelengths (320 nm and 405 nm) using
a cell of path length 1 cm. The total absorbance and molar extinction coefficients of A and B at two
different wavelengths are given below:
Total absorbance at 320 nm = 0.1304
Molar extinction coefficient of A at 320 nm = 517 M-1 cm-1
Molar extinction coefficient of B at 320 nm = 279 M-1 cm-1
Total absorbance at 405 nm = 0.0695
Molar extinction coefficient of A at 405 nm = 85 M-1cm-1
Molar extinction coefficient of B at 405 nm = 738 M-1cm-1
Determine the concentrations of A and B in the solution.

29) From the following UV- visible spectral data of a mixture A and B at 400 nm, calculate the total
absorbance of the mixture.
ɛ (M-1cm-1) Concentration Path Length
A 67000 1x10-5M 1 cm
B 6000 2x10-4M 1 cm

Answer to the problems

1) Wavelength 500 nm ; 5 x 10-5 cm

2) A = 0.2217
3) T % = 20
4) A = 0.1761
5) A = 0.72
6) A = 0.225
7) Molar absorption coefficient = 40 M-1 cm-1
8) Molar absorption coefficient = 625 M-1 cm-1
9) Concentration = 9 x 10-5 M or mol L-1
10) T % = 10
11) Concentration = 1 x 10-4 M or mol L-1
12) A = 1500 M-1 cm-1
13) Concentration = 1.75 x 10-5 M
14) Concentration = 3.52 x 10-5 M
15) Concentration = 1.09 x 10-5 M
16) Molar absorptivity = 15.6 M-1 cm-1
17) A = 0.36
18) T = 0.3428 or T% = 34.28
19) T = 0.216 or T% = 21.6
 20) A = 0.5548 and T = 0.2791 or T% = 27.91
21) T% = 12.5
22) A = 0.6
23) Molar absorption coefficient = 699 M-1 cm-1
24) At = 1.2825
25) T% = 6.3 and Concentration = 4.358 x 10-4 M
26) Conc. of A = 0.2069 M and Conc. of B = 0.1766 M
27) Conc . of Fe3+ = 3.585 x 10-5 M and Conc. of Cu2+ = 2.511 x 10-4 M
28) Conc. of A = 2.148 X 10-4 M and Conc. of B = 6.944 x 10-5 M
29) 1.87

Calculate the λmax for the following conjugated dienes

:
Answer: λmax values for conjugated dienes

1) 230 nm (215 + 3X 5 = 230 )

2) 230 nm ( 215 + 2x 5 + 5 = 230 )
3) 263 nm ( 253 + 2x 5 = 263 )
4) 235 nm ( 215 + 4x 5 = 235 )
5) 235 nm ( 215 + 3x 5 + 5 = 235 )
6) 235 nm ( 215 + 3x 5 + 5 = 235 )
7) 323 nm (253 + 5x 5 +30 + 3x 5 = 323 )
8) 240 nm ( 215 + 4x 5 + 5 = 240 )
9) 278 nm (253 + 4x 5 + 5 = 278 )
10) 250 nm (215 + 5x 5 + 10 = 250 )
11) 241 nm ( 215 + 3x 5 + 5 + 6 = 241 )
12) 353 nm ( 253 + 5x 5 + 30 + 30 + 3x 5 = 353 )
13) 240 nm ( 215 + 3x 5 + 5 + 5 = 240 )
14) 240 nm ( 215 + 3x 5 + 2x 5 = 240 ) Cross conjugation
15) 290 nm ( 215 + 6x 5 + 30 + 3x 5 = 290 )
16) 283 nm ( 253 + 4x 5 + 2x 5 = 283 ) Cross conjugation

Calculate the λmax for the conjugated carbonyl compounds

Answer: λmax values for conjugated carbonyl compounds

1) 249 nm (215 + α10+ β2X 12 = 249 )

2) 227 nm ( 215 + β12 = 227 )
3) 244 nm ( 215 + β2x 12 + 5 = 244 )
4) 274 nm ( 215 + β2x 12 + OH 35 = 274 )
5) 236 nm ( 202 + α10 + β2 x 12 = 236 )
6) 249 nm ( 215 + α10 + β2x 12 = 249 )
7) 335 nm (215 + α10 + δ2x 18 + 39 + 30 +5 = 335 )
8) 280 nm ( 215 + β12 + δ18 + 30 + 5 = 280 )
9) 227 nm (215 + β12 = 227 )
10) 246 nm (202 + α10 + β2x 12 + 2 x 5 = 246 )
11) 231 nm ( 202 + β2x 12 + 5 = 231 )
12) 256 nm ( 202 + β2x 12 + 5 + 25 = 256 )
13) 267 nm ( 202 + β12 + δ18 + 30 + 5 = 267 )
14) 326 nm ( 202 + β12 + γ18 + δ18 + ζ18 + 30 + 2x 5 = 326 )
15) 254 nm ( 215 + α10 + β2x 12 + 5 = 254 ) Cross conjugation