Spectrophotometry Lab Answer Sheet
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Spectrophotometry Lab
Question 1
a.
Chlorophyll reflects green light (540
-
560 nm) and absorbs other wavelengths. What biologically important
molecules other than chlorophyll absorb and reflect certain colors?
Nitrogenous base cytosine, adenine, thymine, uracil, and guanine.
b.
ls the absorbance of light critical to these molecules' functions or just a consequence of their molecular
structure? Explain your answer.
Each molecule has a unique structure, which makes them absorb and/or reflect certain wavelengths of
light.
Question
2
Why is it important to clean the sample tubes?
For the light to pass
absorb and not reflect on any foreign substance on the sample tube, it must be clean.
Wavelength
(nm)
Absorbance
350
0.41
380
0.64
410
0.65
440
0.35
470
0.32
500
0.46
530
0.78
560
1.34
590
2.01
630
2.25
660
0.82
690
0.14
Peak Wavelength: 630
Concentration
Standards (mg/mL)
Absorbance
1
0.06
10
0.39
20
0.85
30
1.32
40
1.76
50
2.25
Unknown
Absorbance
Concentration
A
1.55
35 mg/mL
B
1.87
42 mg/mL
C
1.47
32 mg/mL
D
1.16
27 mg/mL
Paste your standard curve here:
0
10
20
30
40
50
60
0
0.5
1
1.5
2
2.5
f(x) = 0.05 x − 0.03
R² = 1
Absorption: Standard Curve
Question 3
a.
What wavelength is the peak absorbance of Bromophenol Blue?
630
b.
Why is it important to recalibrate with your blank sample often?
In order to get an accurate reading. Just in case you used a different substance previously, you do not
want it affecting your current experiment.
c.
Would you expect a curve of the same shape for another molecule such as chlorophyll? Why or why
not?
It would be a slightly different curve, because every molecule has a unique pattern that allows it to
reflect and absorb certain light.
Question4
Do the plotted data points of your standard curve lie on a straight line?
Yes
Chlorophyll Absorption Spectra
Wavelength
(nm)
Absorbance
350
0.56
380
0.81
410
1.17
440
0.77
470
0.38
500
0.18
530
0.16
560
0.16
590
0.19
630
0.24
660
0.76
690
0.18
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Arial
10pt
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面
田旺 田
QUESTION 20
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is the only common disaccharide that is not a reducing sugar.
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