Cassandra Saikin

Quantitative Analysis Lab #9

May 2, 2008

Determination of Caffeine in Beverages by HPLC

Purpose: To determine the amount of caffeine (mg/serving) in various caffeinated beverages
and compare it to the label value.
Procedure: In order to create a calibration curve of standard caffeine concentrations, a solution
of 1,000 ppm concentration was made, and then diluted four different ways with HPLC water to
create four standard measurements 100 ppm, 50 ppm, 40 ppm, and 20 ppm. HPLC water was
used to correspond with the HPLC chromatograph. Everything went according to plan.
Data:
Sample
100 ppm (standard)
50 ppm (standard)
40 ppm (standard)
20 ppm (standard)
Diet Coke
Mt. Dew
Rockstar Energy
Caffeinated Tea

Height
656832
370760
282683
140016
110476
303842
886486
396251

Area
5737535
3167627
2587427
1256578
1017902
3144431
7635249
Not usable

Calculations:
Determining ppm of selected beverages
See attached Excel spreadsheet
Determining mg/mL of selected beverages
1000ppm = 1000 mg/L
Diet Coke
14.73867 ppm = 14.73867 mg/L
14.72867 mg/L x 1L/1000 mL = 14.72867 mg/L = .01474 mg/mL
1000 mL
Mt. Dew
44.09244 ppm = 44.09244 mg/L
44.09244 mg/L = .04409 mg/mL
1000 mL

Mg/Serving

48 mg/serving
54 mg/serving
80 mg/serving
20-40 mg/serving

Cassandra Saikin
Quantitative Analysis Lab #9
May 2, 2008

Rockstar Energy
132.5949 ppm = 132.5949 mg/L
132.5949 mg/L = .1326 mg/mL
1000 mL
Caffeinated Tea
58.12051 ppm = 58.12051 mg/L
58.12051 mg/L = .05812 mg/mL
1000 mL
Determining mg/serving
Serving Size for: Tea = 200 mL; Soda = 370 mL
mg/serving = mg/mL x serving size (mL)
Diet Coke
.01474 mg/mL x 370 mL = 5.454 mg/serving
Mt. Dew
.04409 mg/mL x 370 mL = 16.31 mg/serving
Rockstar Energy
.1326 mg/mL x 370 mL = 31.82 mg/serving
Caffeinated Tea
.05812 mg/mL x 2000 mL = 11.62 mg/serving
Percent Difference
Calculated value actual value x 100%
Actual value
Diet Coke
5.454 48.00 x 100 = 88.64%
48.00
Mt. Dew
16.31 54.00 x 100 = 69.80%
54.00
Rockstar Energy

Cassandra Saikin
Quantitative Analysis Lab #9
May 2, 2008

31.82 80.00 x 100 = 60.23%

80.00
Caffeinated Tea
11.62 20.00 x 100 = 41.90%
20.00
Results:
Sample

Ppm

mg/mL

mg/serving

Actual
mg/serving

% Difference

Diet Coke
Mt. Dew
Rockstar
Caf. Tea

14.73867
44.09244
132.5949
58.12051

.01474 mg/mL
.04409 mg/mL
.1326 mg/mL
.05812 mg/mL

5.454 mg/serv.
16.31 mg/serv.
31.82 mg/serv.
11.62 mg/serv

48 mg/serv.
54 mg/serv.
80 mg/serv.
20-40 mg/serv.

88.64%
69.80%
60.23%
41.90%

Discussion: The calculated mg/serving of Rockstar Energy contained the most caffeine of all the
beverages, while the label value also had the largest mg/serving. Even though the calculated
value was 60.23% different, it still was accurate in having the most amount of caffeine. The
amounts of caffeine in two of the four drinks were coinciding with the serving on the label in
comparison to each other. However, Diet Coke had the least calculated amount while tea should
have had the lowest according to the label. This error could be traced to the filtering process; the
soda could have been filtered more than the tea, or the tea could have been filtered less than the
soda. An error in filtering could also have attributed to the large percent differences between the
calculated values and the label values. There might have been too much filtering, which caused
a lot of the caffeine to be lost. Its very unlikely that the error is in the calibration curve, which
was used to determine the ppm (concentration) of caffeine in the beverages. The R 2 value is very
high (.994), which should give an accurate measurement. Overall, determining caffeine by
HPLC analysis is an okay method for determining relative amounts of caffeine per serving, but is
not very accurate in coming close to the actual mg/serving.