BE132P Instrumentation in Biological Engineering 1

Name: CUEVAS, Bernadette V. Date: June 8, 2020

Student Number: 2015102318 Program/Year: BE/4

EXERCISE 1: SPECTROCHEMICAL ANALYSIS

Objectives:
Calculate concentrations of samples and unknowns, and identify structures using
spectrochemical methods

Introduction:
Measurements based on light and other forms of electromagnetic radiation are widely used
throughout analytical chemistry. Spectroscopy is an analytical technique that determine the
interactions of radiation and matter. Spectroscopic analytical methods are based on measuring
the amount of radiation produced or absorbed by molecular or atomic species of interest.
Spectroscopic methods are classified according to the region of the electromagnetic spectrum
used or produced in the measurement. The gamma ray, X-ray, ultraviolet (UV), visible, infrared
(IR), microwave, and radio-frequency (RF) regions have been used.
Spectroscopy has played a vital role in the development of modern atomic theory. In addition,
spectrochemical methods have provided perhaps the most widely used tools for the elucidation
of molecular structure as well as the quantitative and qualitative determination of both inorganic
and organic compounds.

In this exercise, students will practice solving problems involving measurements made with
absorption of UV, visible, and IR radiation, and calculation involving atomic absorptions.
BE132P Instrumentation in Biological Engineering 1

Answer the following chronologically. Show complete solution for each problem.

1. Describe how you would use a serial dilution to prepare 100 mL each of a series of standards with
concentrations of 1.00×10–5, 1.00×10–4, 1.00×10–3, and 1.00×10–2 M from a 0.100 M stock solution.

A 10mL volumetric pipet and 100mL of volumetric flask will be used for each steps for a serial dilution. By
using the formula, CorigVorig=CnewVnew, Hence,

Corig, M Cnew, M Vorig, mL Vnew, mL

-2
0.100 1.00x10 10 100
1.00x10-2 1.00x10-3 10 100
-3 -4
1.00x10 1.00x10 10 100
1.00x10-4 1.00x10-5 10 100
From a 0.100M stock solution, transfer 10mL of the solution to another flask and dilute it to 100mL to
make 1.00x10-2M concentration. This will serve as an initial concentration for the next dilution. To another
flask, pipet another 10mL of the 1.00x10-2M of the solution and dilute it to 100mL thus, making the
concentration to 1.00x10-3M. Repeat the same procedure until a concentration of 1.00x10-5M is obtained.

2. A compound of formula weight 180 has an absorptivity of 286 cm−1 g−1 L. What is its molar absorptivity?

GIVEN: SOLUTION:

F.wt=180 ε= (a)(F.wt)

a= 286 cm-1g-1L ε=(286 cm-1g-1L)(180 g mol-1)

REQUIRED: molar absorptivity ε=51,480 cm-1 mol-1 L

3. A solution in the lab has an absorbance of 0.663 in a 1.0 cm cell, too concentrated for an accurate
measurement. The sample is diluted from 1 to 5 ml and then measured in a 2.0 cm cell. If the
measurement is linear, what is the expected absorbance?

SOLUTION:

A=abc A=0.265
A=(0.663)(2.0cm)(0.2)

4. If the molar absorptivity of the analyte in Problem #3 is ε = 8.32 l/mole cm, what is the concentration
of the solution?

GIVEN: A= εbc

ε =8.32 l/mole cm 0.663= (8.32 l/mol cm)(1.0cm)(c)

REQUIRED: concentration 0.663

c=(8.32)(1.0)
SOLUTION: c= 0.0796M
5. The absorbance of a 0.855 M standard of a complex between magnesium and ASN (mg-ASN) was
collected from 320 to 490 nm in 10 nm steps. Plot the data using Excel and determine the best wavelength
to collect them.
BE132P Instrumentation in Biological Engineering 1

Absorbance of Mg-ASN
0.35
0.2750.2880.2810.267
0.3 0.244 0.255
0.221 0.2310.217
0.25
Absorbance

0.199 0.189
0.1580.174 0.163
0.2 0.1460.131
0.125 0.114
0.15
0.1
0.05 0
0

Wavelength (nm)

The best wavelength is at 390nm since the graph showed a peak at this wavelength due to obtaining the
highest absorbance.

a. What is the molar absorptivity, ε, of the mg-ASN complex if the solution was 0.855 M in a 1.0
cm cell?
GIVEN: A= εbc
c= 0.855M ε = A/bc
b=1.0cm ε= 0.288/(1.0cm)(8.55M)
REQUIRED: Molar absorptivity, ε ε = 0.337 cm l-1 mol-1
SOLUTION:
b. What is the concentration of an mg-ASN solution that has an absorbance of 0.198 at 288 nm
in a 0.5 cm cell?

GIVEN: b= 0.5cm

A=0.198 ε = 0.337 cm l-1 mol-1

BE132P Instrumentation in Biological Engineering 1

REQUIRED: Concentration, c c= A/εb

SOLUTION: c= 0.198/(0.337)(0.5)

A= εbc c= 1.18M
6. Molar absorptivity data for the cobalt and nickel complexes with 2,3-quinoxalinedithiol are εCo = 36,400
and εNi= 5520 at 510 nm and εCo = 1240 and εNi = 17,500 at 656 nm. A 0.425-g sample was dissolved and
diluted to 50.0 mL. A 25.0-mL aliquot was treated to eliminate interferences; after addition of 2,3-
quinoxalinedithiol, the volume was adjusted to 50.0 mL. This solution had an absorbance of 0.446 at 510
nm and 0.326 at 656 nm in a 1.00-cm cell. Calculate the concentration in parts per million of cobalt and
nickel in the sample.

GIVEN: 0.425-g sample

εCo = 36,400 0.446 at 510 nm and 0.326 at 656 nm

εNi= 5520 at 510 nm 1.00-cm cell

εCo = 1240 REQUIRED: CNi and CCo in ppm

εNi = 17,500 at 656 nm SOLUTION:

BE132P Instrumentation in Biological Engineering 1

7. Estimate the frequencies of the absorption maxima in the IR spectrum of given compound below. From
these frequencies, assign molecular vibrations of corresponding to each of the bands.

C-O= 400-1600 C-C= 400-1600

C=C= 1600-1850 O-H= 2700-4000

C=O= 1600-1850 H-C=O= 2700-4000

C-H= 2700-4000

8. A 5.00-mL sample of blood was treated with trichloroacetic acid to precipitate proteins. After
centrifugation, the resulting solution was brought to pH 3 and extracted with two 5-mL portions of methyl
isobutyl ketone containing the lead-complexing agent APCD. The extract was aspirated directly into an
air/acetylene flame and yielded an absorbance of 0.502 at 283.3 nm. Five-milliliter aliquots of standard
solutions containing 0.400 and 0.600 ppm of lead were treated in the same way and yielded absorbance
of 0.396 and 0.599. Find the concentration of lead in the sample in ppm assuming that Beer’s law is
followed.

GIVEN: SOLUTION:

A283.3= 0.502 0.600−0.400

m= 0.599−0.396=0.985
C1= 0.400ppm y=mx+b
C2= 0.600ppm 0.396=(0.985)(0.400)+b
A1= 0.396 b= 2x10-3
A2=0.599 0.502=(0.985)(X)+2x10-3
REQUIRED: Concentration
X=0.504 ppm Pb
BE132P Instrumentation in Biological Engineering 1

9. Amines (weak base) form salts with picric acid (trinitrophenol), and all amine picrates exhibit an
absorption maximum at 359 nm with a molar absorptivity of 1.25 × 10 4. A 0.200-g sample of aniline, is
dissolved in 500 mL of water. A 25.0-mL aliquot is reacted with picric acid in a 250-mL volumetric flask and
diluted to volume. A 10.0-mL aliquot of this is diluted to 100 mL and the absorbance read at 359 nm in a
1-cm cell. If the absorbance is 0.425, what is the percent purity of the aniline?

GIVEN: 1-cm cell

molar absorptivity of 1.25 × 104 REQUIRED: % Purity of aniline

0.200-g sample of aniline SOLUTION:

the absorbance is 0.425

BE132P Instrumentation in Biological Engineering 1

10. Compounds A and B absorb in the ultraviolet region. Compound A exhibits an absorption maximum at
267 nm (a = 157) and a trailing shoulder at 312 nm (a = 12.6). Compound B has an absorption maximum
at 312 nm (a = 186) and does not absorb at 267 nm. A solution containing the two compounds exhibits
absorbances (using a 1-cm cell) of 0.726 and 0.544 at 267 and 312 nm, respectively. What are the
concentrations of A and B in mg/L?

GIVEN: 0.726 and 0.544 at 267 and 312 nm, respectively

A=267 nm (a = 157) REQUIRED: Concentrations of A and B.

A= 312 nm (a = 12.6) SOLUTION:

B=312 nm (a = 186)