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Exp 7 Spectrophotometry (Memo) - Questions and Calculations

This document contains the results of 6 spectrophotometry experiments where various concentrations of a solution were prepared and their absorbances measured. It also includes 6 multiple choice questions about spectrophotometry and 3 sample calculations involving molar concentrations, percent solutions, and preparing a buffer solution.

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Emeka Louis
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57 views

Exp 7 Spectrophotometry (Memo) - Questions and Calculations

This document contains the results of 6 spectrophotometry experiments where various concentrations of a solution were prepared and their absorbances measured. It also includes 6 multiple choice questions about spectrophotometry and 3 sample calculations involving molar concentrations, percent solutions, and preparing a buffer solution.

Uploaded by

Emeka Louis
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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Experiment 7: Spectrophotometry memo (questions and calculations)

Tube 1: Tube 4: (5)

C1V1 = C2V2 C1V1 = C2V2

( 1M) (0 ml) = C2 (3 ml) ( 1M) (1.5 ml) = C2 (3 ml)

C2 = 0 C2 = 0.5 M

Tube 2: Tube 5:

C1V1 = C2V2 C1V1 = C2V2

( 1M) (0.5 ml) = C2 (3 ml) ( 1M) (2 ml) = C2 (3 ml)

C2 = 0.17 M  C2 = 0.67 M 
Tube 3: Tube 6:

C1V1 = C2V2 C1V1 = C2V2

( 1M) (1 ml) = C2 (3 ml) ( 1M) (2.5 ml) = C2 (3 ml)

C2 = 0.33 M  C2 = 0.8M 
QUESTIONS: [11]
1. Measurement at a fixed wavelength is done to obtain quantitative information, such as concentration 
Absorbance measurement as a function of wavelength to provide qualitative information that assists in solving the identity
and structure of a pure substance.
 (2)

2. The equation of a graph cannot be used because this line assumes linearity for infinity and from the Beer-Lambert law, we
know that the graph plateaus at a value of 2.  (1)

3. Peptide bonds,  aromatic phenol groups, and prosthetic groups such as NAD, FAD FMN etc.  (3)

4. Manually measure the absorbance readings of the test tube which contains all molecules except the molecule under
investigation, then subtract the values when measuring the one containing the molecule under investigation.  (1)

5. a) quartz 
b) quartz / plastic / glass  (2)

1
6. When the reading is 2 or above, the spectrum has reached a plateau according to the Beer-Lambert law

If this happens, the solution needs to be diluted further so that we continue working in the linear region. The concentration
must be multiplied by the dilution factor that the correct concentration can be calculated.  (2)

CALCULATIONS: [10]
1. What is the micromolar concentration of a 100% NaCl solution (Mr=58.44) (3)

100% NaCl

100 g NaCl in 100 ml
m (NaCl) = C x V X Mr
100 g = C X (0.1 L) (58.44)

C = 17.11 M = 1.7 X 10-6 µM

2. How would you make 150 ml of a 65% Methanol solution? (3)

65 % = 65 ml (MeOH) in a 100 ml solution



97.5 ml (MeOH) in a 150 ml solution 
Add 97.5 ml of concentrated MeOH to 52.5 ml of dH2O to obtain 150 ml of a 65% MeOH solution. 
3. How would you make 0.5 L of 700 μM Tris-HCl, pH 6.5 (Mr=121.14g/mole) (4)


m (Tris) = C x V x Mr
= (700 x 10-6 M) (0.5 L) (121,14)
= 0.042 g = 42.4 mg 

Weigh out 42.4 mg Tris in some dH2O, adjust pH to 6.5 with HCI and make up to a final volume of 0.5 L (500 ml) with dH2O 

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