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SPECTROPHOTOMETRY
Experiment 3
Ebru AKHARMAN, Gebze Technical University, Turkey
AIM:
While materials absorb a portion of light in our environment, they reflect a portion of light.
There is a diferent place of this situation in biochemical experiments. If the concentration of
used solution or material is not know, the concentration of this solution or material should be
determined by colorimetry / spectrophotometry.
These two apparatus are used for calculating absorbance. Absorbance is linear proportion with
concentration. Thus, if absorbance is known, concentrations can calculated. The concentration of
methyl red will calculated in this experiment. Also, this experiment is a sample for using
colorimetry / spectrophotometry.
RESULTS:
=
avr.-
= = REP1andREp2 blank
REP1 REP2 avr. avr.
A: Absorbance A 0,529 0,461 0,495 0,463
: Molar extinction B 0,234 0,242 0,238 0,206
coefficient
C 0,153 0,157 0,155 0,123
: Lenght of the light path
: Concentration D 0,266 0,141 0,204 0,172
E 0,258 0,053 0,156 0,124
F 0,054 0,047 0,051 0,019
There is a linear relationship between the
Blank G 0,026 0,038 0,032 0
absorbance and the concentration of a
sample. unknown H 0,35 0,367 0,359 0,327
0,327
X= 93,43
0,0035
DISCUSSION: