Experiment 8

Experiment 8

Data Reduction and Analysis

Spectrophotometry

Determination of Chlorophyll-a Concentration in Spinach

Name

Lab Partner

Section

TA Name

Score

The following should be attached to this form:

Spreadsheet tables followed by
Plot of absorbance vs. wavelength followed by
Plot of absorbance vs. concentration of chlorophyll-a.
Copy of data sheet behind plots.

Experimental Data:
max Chlorophyll-a __________
Calibration Curve Data:
Solution

Concentration Chlorophyll-a

Absorbance, max

Stock solution
Dilution 1 (50% concentration)
Dilution 2 (25% concentration)
Dilution 3 (12.5% concentration)
Calibration curve trendline equation _______________________________
R2 value of trendline _________________________

Chlorophyll Extract:
Mass spinach __________________________________________
Total volume spinach extract _______________________________
Absorbance spinach extract, max ___________________________
Absorbance spinach extract, 750 nm ________________________
Corrected absorbance ____________________________________

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Spectrophotometry

Determine the concentration of chlorophyll-a in spinach.

a. Using the absorbance of the spinach extract and the equation of the trendline, determine the concentration
of the extract solution. Report the concentration in moles/L (M).
General Equation:

Calculation:

Concentration of chlorophyll-a in extract _________________

b. Calculate the number of grams of chlorophyll-a in the 25.00 mL spinach extract solution.
General Equation:

Calculation:

c. Calculate the concentration of the chlorophyll-a in spinach (mg chlorophyll-a/g spinach).

General Equation:

Calculation:

132

Experiment 8

Experiment 8

Data Reduction and Analysis

Spectrophotometry

Determination of Chlorophyll-a Concentration in Spinach

Name
Section

Lab Partner
TA Name

Score

Questions
1. Why was it important to force the trendline in the plot of absorbance vs. concentration through the
origin? Would adding the point (0, 0) to your data set have had the same effect? Why or why not?

2. Acetone is a good extraction solvent for plant pigments as it breaks down cell walls and is miscible
with the pigments. Can you be sure that you only extracted chlorophyll-a from your spinach leaf?
If not, what else might have been extracted?

3. Using the techniques from this experiment, how would you determine the answers to the above
questions in lab?

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Spectrophotometry

4. Identify possible sources of error in your experiment. Do not simply claim human error. Cite specific
examples from the design or your implementation of the experiment.

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