Objective

 To study the type of detector system used in HPLC.

 To identify the present of caffeine in soft drink.
 To determine the amount of caffeine in a sample of beverages.
 To determine the concentration of caffeine in soft drink.
 To apply the principles of reverse phase high performance liquid chromatography.

Abstract

This experiment provides an introduction to the application of High Performance

Liquid Chromatography (HPLC) to the solution of complex analytical problems. Cola
type drinks are complex chemical systems that contain varying amounts of caffeine.
The amount of caffeine present in these soda drink can be determined by HPLC. In
this experiment, sample was prepared with standard caffeine samples of 5ppm, 15ppm,
25ppm, 50ppm, and 100ppm by diluting with distilled water. High performance liquid
chromatogaphy is technique for the separation of the components of a sample by
means of a high pressure pump forcing the eluent through a column. Polar, dispersive
or ionic forces working on the samples allow its components to either be retained in
the stationary phase or eluted out by the mobile phase thereby producing responses
with different retention times. The area under each response (peak) can be used to
obtain its concentration. In this experiment, reverse phase HPLC was used to
determine the caffeine content of cola products. External calibration was done to
generate the standard curve where the respective concentrations of caffeine in the
samples were intrapolated from. In this experiment, serial dilution also will be
prepared to be as standard caffeine and to determine if caffeine is present in the soda
sample by use retention time. The increase the concentration, the bigger the getting
area. This shows that the higher concentration of caffeine will make a bigger effect.
Procedure

2. prepare serial dilution for

1. calculate for prepare caffeine standard 5ppm,
caffeine standard 1000ppm 15ppm, 25ppm, 50ppm and
from the caffeine standard 100ppm and mark up with
deionized water.

4. degas the sample by

3. obtain the sample and soft
placing it in a vacuum flask
pump or water aspirator. drink.

5. leave it under vacuum 6.filter the degas soda

until no more bubbles through 45L filter syringe.