Autonomous University of the State of

Mexico
Faculty of Chemistry
Biological Pharmaceutical Chemistry

Subject: Instrumental Analysis.

Problem set
FOR THE FIRST MIDTERM

School Year 2020-B

Group 52
Instructor: Dr. Jorge Javier Ramírez
García.
18|Octubre|2020.

Alumna:
Saldivar Ahumada Alejandra
 PROBLEMATIC
for the First Partial

Nombre del alumno:Alejandra Saldivar Ahumada. Fecha:18|Octubre|[Link].

Spectrophotometry I
PROBLEM 1.
a. Indicate the absorbance value corresponding to a T value of 45.0%.

A = 2 - log%T
A = 2 - log(45.0%)
= 0.34675
b. If a solution with a concentration of 0.0100 M has a T = 45.0% at a given wavelength,
What will be the value of transmittance corresponding to a 0.0200 M solution of the same?
substance?
1
2= ( 2)
1= 1 ⁡⁡⁡⁡ 2= 2 1

1 2 ( 0.34675 )
= ⁡⁡⁡⁡⁡⁡⁡⁡⁡⁡ = 2= (0.0200M)
1 2 ( 0.0100M )

= A20.6935

1 2
T=10-0.6935
=
1 2 T=(0.20253) (100%)
1 2
= T= 20.253%
1 2

PROBLEM 2. 15 mg of a compound with a molar mass of 384.63 [Link]-1 is taken to form

5 mL of solution. Subsequently, an aliquot of 1.00 mL of that solution is taken to dilute it.
in a 10 mL volumetric flask up to the mark.
a. Find the concentration of the sample in the 5 mL flask.

1 1
15mg( )( ) = 3.8998x10-5
1000mg 384.63g

3.8998x10-5
= 7.7997x10-3
5x10-3
b. Determine the concentration of the substance in the 10 mL flask.

1 1
2=
2

(7.7997x10-3)(1 )
2=
(10 mL
C2=7.7997x10-4M
The 10 mL sample is placed in a cell with b = 0.5000 cm obtaining an absorbance of
0.634 at 495 nm. Determine the value of the molar absorptivity of the substance at this wavelength.
wave.

(0.634)
= (0.5000 )(7.7997 10 -4
)
=1625.7035cm-1M-1

PROBLEM 3. A quantity of compound with a molar mass of 292.16 g/mol is dissolved in a

5 mL volumetric flask. A 1.00 mL aliquot of the obtained solution is taken, diluting it.
up to 10 mL. The absorbance of this last solution measured in a cell of b = 1.00 cm at 340 nm
It is 0.427. The molar absorptivity of the compound at this wavelength is ε = 6130 M-1 cm-1.
a. Calculate the concentration of the solution placed in the cell.

0.427
= = 6.9657x10-5
(1.00cm)(6130cm−1 −1 )

b. What is the concentration of the solution initially prepared in the 5 mL flask?

1 1 = 2 2

2 2
1=
1

(10mL)(6.9657x10-5 )
1=
(1 )

1= 6.9657x10-4
c. How many milligrams of compound were used to prepare the 5 mL solution?

1 6.9657x10-4mol 292.16g1000mg
5 mL( )( )( )( ) = 1.0175 mg
1000mL 1 1 1
PROBLEM 4. During an experiment conducted to determine the amount of thiamine (vitamin
B1) in a pharmaceutical preparation, it was inadvertently read on the transmittance scale instead of
do it on the absorbance scale of the spectrophotometer. The first sample gives a reading of T1
= 82.2 % and the second T2 = 50.7 % at the wavelength of maximum absorbance. What is the relationship
of concentrations between the thiamine concentrations in both samples?
-Calculating A1y A2:

1= ( =)0.0851
2 − log82.2

2= ( = )0.2949
2 - log50.7
Obtaining the relationship:
 1 2
=
1 2

2 2
=
1 1

20.2949
= = 3.4652
1 0.0852

PROBLEM 5. Ammonia can be determined spectrophotometrically through its reaction

with phenol in the presence of hypochlorite, resulting in a blue substance that has its absorption
maximum at 625 nm. A sample of 4.37 mg of protein is chemically digested to convert into
ammonia all the nitrogen present, and at the end of the treatment the volume of the sample is 100.00
A 10.00 mL aliquot of this solution is treated with 5.00 mL of phenol and 2 mL of hypochlorite.
of sodium, and the sample is diluted to 50 mL, measuring its absorbance at 625 nm in a 1.00 cell.
cm thick after 30 minutes. A reference standard solution is also prepared with
1.00 x 10 -2 g de cloruro de amonio disueltos en un litro de agua; una alícuota de 10 mL de esta
pattern dissolution is treated the same way as problem dissolution. The blank is prepared
using distilled water instead of the problem.
PM (NH4Cl) = 53.50 [Link]-1 PA (N) = 14.006 [Link]-1
SAMPLE Absorbance
White 0.140
Reference 0.380
Problem 0.582

PM (NH4Cl) = 53.50 [Link]-1 PA (N) = 14.006 [Link]-1

[Link] the molar absorptivity of the blue product.

Obtaining the Co of the colored product:

1NH4Cl 1 1 -5
= 10 mg NH4Cl( )( )( ) (10 mL )= 3.7383x10 M
53.50 mg NH4Cl1000mL 50mL

Calculating the molar absorptivity:

0.308 - 0.140
= 4′ 494.3820 cm−1 −1
(1cm)(3.7383x10-5 )

[Link] the mass percentage of nitrogen in the protein.

Calculating Nitrogen Co in 50mL:
0.582 - 0.140
= = 9.8345x10-5
4′ 494.3820cm−1 −1

Calculating Nitrogen Co in 10mL:

 9.8345x10-5M(50mL)
= = 4.9172x10-4
(10mL
Calculating the percentage:
14.006 mg 100%
4.9172x10-4 ( 10 mL() )( ) = 15.7599%
1 4.37 mg
PROBLEM 6. The copper (I) ion forms a colored complex with neocuproine which presents
a maximum absorbance at 454 nm. This complex can be extracted with isoamyl alcohol, which
it is not soluble in water. Suppose you apply the following procedure:
A rock that contains copper is crushed and the metals are extracted with a strong acid. The solution
acid is neutralized with a base, and the resulting solution is brought to 250 mL. 2) an aliquot of 10
mL of this is treated with 10 mL of reducing agent to convert all the copper to cuprous ion, added
10 mL of buffer to maintain the pH at a suitable value for the formation of the complex. 3) It
Take 15 mL of this solution, add 10 mL of neocuprein and 20 mL of isoamyl alcohol.
After shaking vigorously, the two phases separate and the copper complex is entirely in
the organic phase. The absorbance of the organic phase is measured at 454 nm in a 1.00 cm cell. The
The prepared blank shows an absorbance of 0.056.
If the rock sample has one milligram of copper, what is the concentration of it present?
in the organic phase?
10 mL15 mL
1 ( )( ) 0.02 mg of the organic compound Copper(II) in 20 mL of the dilating phase
250mL 30mL
0.02 mg C
= = 1.5736x10-5 ⁡ ⁡ ⁡ ⁡ ⁡ á
(63.546 mg/mmol)(20 mL)
[Link] ε = 7.90 x 10^3 M-1 cm-1 for the complex, what will be the measured absorbance value?

= ε

= 1( )(1.5736x10 -5M )( 3
7.90x10 cm −1−1 ) = 0.12431

A = 0.12431+ 0.056 = 0.1803

If a different rock is analyzed and an uncorrected absorbance of 0.874 is obtained.

How many milligrams of copper are there in the rock? PA (Cu) = 63.546 [Link]-1

= 0.874 - 0.056 = 0.818

0.818
= = 1.0354x10-4
(7.90x103 −1 −1 )(1 )

1.0354x10-4 ( 20 mL(63.546
) ) = 0.13159mg C₁₁

30 mL 250 mL
0.13159mgC( )( ) = 6.5795 mg de C₁
15 mL10 mL
PROBLEM 7. The nitrite ion is used as a preservative for bacon and other foods.
generating a controversy regarding its potential carcinogenic effect. In a determination
spectrophotometric of nitrite, a series of reactions are carried out that conclude with the
formation of a colored product with maximum absorbance at
520 nm. The procedure followed to develop color can be summarized as follows:

1) 50.00 mL of the problem solution containing nitrite is added to 1.00 mL of solution of

sulfamido acid (reaction 1:1).

After 10 minutes, 2.00 mL of 1-aminonaphthalene solution (reaction 1:1) and 1.00

mL of buffer solution.

3) 15 minutes later, the absorbance is read at 520 nm in a cell with b = 5.00 cm. With this technique
Three solutions are analyzed:

DISSOLUTION VOLUME AND CHARACTERISTICS ABSORBANCE AT 520nm

A 50mL of food extract with 0.153
negligible amount of nitrites
B 50mL of food extract of 0.622
which is suspected to have nitrite
C The same as B, with the addition of 0.967
10µL of NaNO solution27 times 10-3M

a. Calculate the molar absorptivity of the colored product.

( + 2mL+ 1)
= 50mL + 1mL Calculating CSodium nitrite
:
+1
2 2
= 54.01 2
= =

− = 0.622 − 0.153 = 0.469 7.5x10-3

[ ( 1 10-5 L) ]
− = 0.967 − 0.153 = 0.814 = = 1.3886x10-6
2
0.05401
− = − + 2 -Calculating to :

0.814 = 0.469 - A 0.814 − 0.469

2 = = 49′ 3355cm−1 −1
(5cm)1.3886x10-6 )
0.814 = 0.469 - dc

b. How many micrograms of nitrite are present in the 500 mL of food extract B?
PM (NO2-46.004 [Link]-1

−
=

0.469 0.05401L 46.004g 1

= -6 ( )( )( ) 4.6002
( 5cm1.3886x10
)( M ) 1 1 1 10-6
PROBLEM 8. The spectrophotometric analysis of phosphates can be carried out using the following
procedure:
1) The sample is placed in a 5 mL volumetric flask and 0.500 mL of solution is added.
sodium molybdate and sulfuric acid and 0.200 mL of hydrazine sulfate solution, and
dilute almost to the fill line with distilled water.
2) The diluted solution is heated for 10 minutes at 100 ºC, forming a blue compound (acid 1,2
molybdophosphoric.
3) the flask is cooled, filled with distilled water, and the absorbance of the solution is measured.
resulting at 830 nm using a cell of 1.00 cm.
When analyzing 0.140 mL of phosphate standard solution - KH2PO4 (MW 136.09 g/mol) - prepar
By dissolving 81.37 mg of this in 500.00 mL of water, an absorbance of 0.829 is obtained.
A blank prepared in the same way has an absorbance of 0.017. Find the molar absorptivity of the
colored product.
Obtaining the Co of the colored product:
1 1 1 -5
= 81.37mg Fosfato( )( )( (
) 0.14mL )= 3.3483x10 M
136.09 mg of fat in 500 mL 5 mL

-Obtaining the molar absorptivity:

0.829 − 0.017
= = 24′251.1125cm−1 −1
(1cm)(3.3483x10-5 )
0.300 mL of ferritin solution (iron storage protein that contains phosphate)
obtained by digestion of 1.35 mg of protein in 1.00 mL of solvent is analyzed with this
procedure, obtaining an absorbance of 0.836. The blank gives an absorbance of 0.038.
Find the mass percentage of phosphate in ferritin.
PM (PO43-) = 94.972 [Link]-1
-Calculating phosphate Co:
0.836 − 0.038
= = 3.2905x10-5
24251.1125cm−1 −1

Calculating the percentage:

5mL 94.972 mg 0.3 mL100%
3.2905x10-5 ( )( )( )( ) = 1.15745%
0.3mL 1 1 1.35mg
 Spectrophotometry II
PROBLEM 1. A 25.00 mL sample containing iron is analyzed.
treating it with nitric acid and reacting it with an excess of potassium cyanide to form
a red complex. The solution is diluted to 100.00 mL and an aliquot of it is placed in a
variable optical path cell. A reference sample of 10.00 mL with [Fe3+] = 6.80 x 10-4
It is about the same substance being diluted to 50.00 mL and placing a aliquot of it in a.
cell of 1.00 cm. The problem solution gives the same absorbance as the reference solution.
When placed in a cell of 2.48 cm. What is the concentration of iron in the sample problem?
( + )
=

6.80x10-4
( )(10 mL
= = 1.36x10-4
50 mL
50 = 100

1 1 1= 2 2 2

(1 ) -4F)
1 (1.36x10 = (2.48 cm) 2 2

How it's the same:

(1 )( 1.36x10 -4 M100
) mL
2= = ( ) = 2.1935x10-4
2.48cm 25mL
PROBLEM 2. The absorbance of cobalt nitrate (Co(NO3)2) and chromium nitrate (Cr(NO3)3)
additives on the visible spectrum. It is decided to analyze a solution that contains both
compounds. For this, two wavelengths are chosen: 400 and 505 nm and a 1 cm cell is used.
cm for the essay. The results are shown in the table. Calculate the chromium concentrations.
and cobalt in the mixture problem.
A400= 1.167 Co2+ Cr3+
A5050.674 400 0.530 15.2
505 5.07 5.60
At 400nm:

400⁡ = 2++ 2+

400⁡ = 1 1 1+ 2 2 2

400⁡ = (1(0.530)C1+ (1 )(15.2)C2

1.167 = (1(cm)(0.530)C1+ (1 )(15.2)C2
Clearing C2:
1.167 − 0.530C1
2=
15.2
At 505nm:
 505⁡ = 2++ 2+

505⁡ = 1 1 1+ 2 2 2

505⁡ = (1(5.07)C1+ (1(cm)(5.60)C2

0.674 = (1(5.07)C1+ (1(5.60)C2
-Substituting C2:
1.167 - 0.530C1
0.674 = 5.07C1+ 0.560
15.2
Solving for C1in calculator:

10.050064 M = Co2+

2= 0.07503M = Cr3+
PROBLEM 3. A sample is desired to be analyzed that
TABLE 1: SAMPLE
it contains analytes A and B. In the laboratory is available
Wavelength Absorbance
425 0.095 of the standard solutions of both analytes of
520 0.301 exactly known concentrations. After a
preparation process for the analysis in which the
the sample is diluted to one tenth, 1 mL of this is measured at 425 nm and at 580 nm in a 1.00 cm cuvette
optical path, obtaining the data from Table I. Laboratory standards are subjected to
same procedure. The obtained results are shown in Table II. Determine the concentration
from A and B in the sample.

TABLE 2: PATTERN SOLUTION

Analytical MolarityWavelength Absorbance
A 0.0922 425nm 0.545
580nm 0.125
B 0.1023 425nm 0.227
580nm 0.823
Creating a new table and plottingB/AXlike 'x' yAM/AAas 'y':

AA AB AM AM/AA AB/AX
425 0.545 0.227 0.095 0.17431193 0.41651376
520 0.125 0.823 0.301 2.408 6.584
3
2.5
y = 0.3622x + 0.0235
2 R² = 1
1.5
1
0.5
0
0 2 4 6 8
We have the equation and values:

= 0.3622 + 0.0235

= (0.3622∗ 0.102310)( =)0.3705M

(
0.0235∗ 0.0922 10 )(= 0.0216M
)

PROBLEM 4. Transferrin (Molecular Weight 81000 g/mol) and desferrioxamine B (Molecular Weight 650 g/mol) are
colorless compounds capable of binding to Fe3+ to form colored complexes in a 1:2 ratio and
1:1 with maximum absorption wavelengths of 470 nm and 428 nm respectively. The
The molar absorbance of these two compounds forming complexes with iron is given at two.
different wavelengths:

MOLECULAR ABSORPTIVITY (M-1cm-1)

A Transferrin-2 Fe(III) Deferoxamine-Fe(III)
420 3540 2730
470 4170 2290

A transferrin solution has an absorbance of 0.463 at 470 nm in a 1.0 cm cuvette.

Calculate the concentration of transferrin in [Link]-1 and that of iron in μ[Link]-1.

-For transferrin:
=

0.463
= = 1.1103x10-4 ⁡ ⁡
(1(cm)(4170M−1 −1 )

1.1103x10-4mold complex 81′ 000g1000mg 1

= ( )( )( ) 8.9935 mg/mL
⁡ ⁡ 1 1000mL

For iron:
1.1103x10-4mold complex 55.847g 1 1
ℎ =( )2( )( )( )( ) = 12.4013 /
⁡ ⁡ 1 10-6 1000mL

b. Shortly after adding desferrioxamine (which dilutes the sample), the absorbance at 470
At 424 nm it is 0.424 and at 428 nm it is 0.401. Calculate the percentage of iron that is complexed.
with transferrin and desferrioxamine.
PA (Fe) = 55.847 [Link]-1
At 400nm:

428⁡ = +

428⁡ = 1 1 1+ 2 2 2

428⁡ = (1cm)(3540)C1+ (1(cm)(2730)C2

 0.401= (1cm)(3540)C1+ (1cm)(2730)C2
Clearing C2:
0.401-3540C1
2=
2730
At 470nm:

470⁡ = +

470⁡ = 1 1 1+ 2 2 2

505⁡ = 1 1 1+ 2 2 2

0.424 = (1cm)(4170)C1+ (1cm)(2290)C2

Substituting C2:
0.401− 3540°C1
0.424 = 4170°C1+ 2290
2730
Solving for C1in calculator:

7.2991 10-5

= 5.2237x10-5
Calculating the percentage of iron in T and D:

7.2991 10-5 1.4598x10-4

1= ( )
2( )= ⁡

2= 5.2237x10-5 ⁡
= 1.9821 10-4
(1.4598x10-4 )(100%)
% = = 73.6458% delivered
1.9821 10-4
(5.2237x10-5 )(100%)
% = 26.5141%⁡ ⁡
1.9821 10-4
PROBLEM 5. The spectra shown in the figure correspond to solutions of MnO.4-1.0 x
2-
10-4 M, Cr2O71.00 x 10-4 M and a mixture of both with unknown composition.
The table shows the absorbances obtained at different wavelengths, find the
concentration of each species in the mixture.
A Manganese oxide4-Pattern
Cr2O72- Pattern Mix
266 0.042 0.410 0.756
288 0.082 0.283 0.571
320 0.168 0.158 0.422
350 0.125 0.318 0.672
360 0.056 0.181 0.366
Creating a new table and graphingB/AX as 'x' yAM/AAhow 'y'
MnO4-Cr2O72-
A Mix AM/AA AB/AA
Pattern Pattern
266 0.042 0.41 0.756 18 9.76190476
288 0.082 0.283 0.5716.96341463 3.45121951
320 0.168 0.158 0.4222.51190476 0.94047619
350 0.125 0.318 0.672 5.376 2.544
360 0.056 0.181 0.3666.53571429 3.23214286

We have the equation and values:

= 1.7537x + 0.8873
( -4 ) -4
= 1.7537 *1.00x10 M= 1.7537x10 M
(
0.8873∗ 1.00x10 M-4= 8.873x10
) -5