Experiment 6: Colorimetric Assay of Aspartame

VIKTORIA FARIAN AND JORDAN FIEDOR

1 Seton Hill Drive
Greensburg, PA 15601

Abstract
We found the absorbance in Aspartame by using a spectrometer called, a Colorimeter. The
spectrometer shines light of four different wavelengths through the solution and then measures the
absorbance of light that was collected by the chemical solution. This allows scientists to determine
how light is absorbed by the aspartame that is inside of the solution. The aspartame was blue which
means it was absorbing red light. The two-average absorbance’s of the sweetener came out to be .247
and .226. 

Introduction then diluted with distilled water to the 10mL

In Experiment 6, a modified biuret
reagent will be used to determine the
concentration of Aspartame which is a
dipeptide derivative found in commercial
sweeteners. The complex of the aspartame
solution is a dark blue. Since the complex of
this solution is colored, we use the absorbance
of visible light to analyze the amount of
aspartame that is present in each available
sweetener during the experiment. To perform
the analysis of how much aspartame is present,
a spectrometer is used. The name of the
spectrometer is Colorimeter. This spectrometer
shines light of four wavelengths through the
sample. Then it measures the absorbance of
light by the solution. The Colorimeter will be
set at 635 nm which is the red wavelength
because the color of the solution is blue. The
amount of light that will be absorbed and the
concentration are proportion to each other and
that can be presented with Beer’s Law. Beer’s
Law is LOLOL where the absorbance is the
letter A, the molar absorptivity is the length in
centimeters which is the letter B, and the
concentration in mol/L is shown as the letter C.
Beer’s Law is used to find the unknown
solution concentrations in a standard solution.  Results
Experimental
For the concentration of 0.5 of
aspartame solution, 1mL of stock aspartame
was pipetted into a 10mL flask and then 2 mL
of biuret reagent was added. The flask was
mark. This step was repeated for the
concentrations of 1mg/mL, 1.5mg/mL,
2mg/mL, and 2.5mg/mL going up by 1mL of
aspartame stock solution each time, but keeping
the same amount of biuret reagent (2mL). Each
flask was then inverted to mix. Each mixture
was then transferred to a test tube and was left
undisturbed for 15 minutes. After 15 minutes,
the absorbance level was measured and
recorded for each concentration (3 trials for
each) using a Colorimeter (see Abstract for
usage). 0.4 grams of sweetener was then
measured and recorded straight into a
volumetric flask. 5mL of distilled water was
added to the flask in order to dissolve the
powder. 2mL of biuret reagent was added to
the mixture and then distilled water was added
up to the 10mL mark in order to dilute the
mixture. The flask was inverted for mixing,
transferred to a test tube, and set aside for 15
minutes, undisturbed. Absorbance of the
concentration of the sweetener was measured
using the Colorimeter and results were
recorded.
Table 1. Standard solution of absorbance
Standard Abs Abs Abs Average
Solution A1 A2 A3 Abs
0.5 0.103 0.104 0.099 0.102
1.0 0.192 0.186 0.172 0.183

1
 1.5 0.301 0.297 0.289 0.296
2.0 0.386 0.390 0.385 0.387 11.48 mg Aspartame X
=
2.5 0.534 0.530 0.527 0.530 400.1mg Sweetener 1,000 mg

Table 2: Absorbance level of sweetener made X = 28.69 mg

Sweetene Brand Mass Abs Abs Abs Avg
r Solution (g) Rdg Rdg Rdg Abs Class average of aspartame mg/packet = 33.4
A1 A2 A3 True Value of aspartame = 37.0 mg/packet
#1 Great .4001 .247 .247 .247 .247
Conclusion
Value
Overall, experiment 6 was a very easy
#2 Great .4001 .223 .227 .226 .225 to do with techniques and equipment that were
Value not confusing and very easy to use. Even
though the numbers were on the lower side of
the class average and the true value, we still
found success because we completed the
experiment with no issues. In the future for this
experiment different types of sweetener could
be used to see the different value of aspartame
in each sweetener packet.

Graph 1. Average absorbance of Aspartame

standard solution
References
Aspartame concentration in sweetener solution  University of Alabama Birmingham,
(#1) UAB Nutrition Trends. UAB School of
0.247 = 0.212x – 0.0184 Health Professions,
x = 1.25mg/mL https://www.uab.edu/shp/nutritiontrends
/recipes-food-facts/food-facts/artificial-
Aspartame concentration in sweetener sweeteners , November 22, 2017.
solution (#2)
0.245 = 0.212x – 0.0184
x = 1.148mg/mL

1.25 mg 10 mL
× =12.5 mg Aspartame
1 mL 1

1.148 mg 10 mL
× =11.48 mg Aspartame
1 mL 1

12.5 mg Aspartame X
=
400.1 mg Sweetener 1,000 mg

X = 31.24mg
2