METHODOLOGY
A. Preparation of Mobile Phase
A 600 mL of HPLC grade methanol was
diluted to a 400 mL distilled water into
a 1000 mL volumetric flask then was
swirled gently. After diluting the
methanol it was filtered through a
vacuum filtration setup.
B. Preparation of Caffeine
Standard
In the preparation of standard with the
concentration of 900 ppm (parts per
million), 0.025 grams of caffeine was
diluted in a 25mL volumetric flask
using the filtered methanol mobile
phase. The volumetric was then
covered with parafilm and then swirled
and labeled properly.
After labeling the volumetric flask, 3
ml of the sample were drawn into
needles syringe and then was filtered
using PTFE nylon syringe filter into a
clean dry vial. It was then covered and
labeled properly.
C. Preparation of the Commercial
Sample
In the preparation of the commercial
sample, 10 mL of energy drink sample
was diluted into a 50 mL volumetric
Flask using the 60 % methanol mobile
phase. The volumetric flask was then
covered with parafilm and was mixed
and swirled.
After mixing, the PTFE nylon syringe
filter was washed using 1 mL of the
commercial sample by injecting the
sample out. The washing was discard
and then 3 ml of the commercial
sample was drawn again and then
filtered using the PTFE into a clean dry
vial. It was then covered and labeled
properly.
D. Data Acquisition
The data obtained from the HPLC
analysis of commercial sample and
caffeine standard was then graphed.
Using the graph the concentration of
then commercial sample was
obtained.