Malaysian Institute of Chemical

& Bioengineering Technology

Food Technology Department Document SOP /TP/011 Version 01

Title Determination of Caffeine in Soft Drink by HPLC Date 17/03/08
Page 1 of 3
1. Introduction
Caffeine has a mildly addictive effect on the body. It is therefore interesting to know exactly how much
caffeine is in certain beverages. One way to analyze caffeine content in beverages is by using high
performance liquid chromatography (HPLC).

2. Scope
Prepare standard solution of known caffeine concentration and compared them to caffeinated sample (soft
drink)

3. Procedure
3.1 Equipment and Supplies:
Chemicals
• HPLC solvent /mobile phase
• Caffeine standard
• Sample (soft drink)

Materials
• HPLC UV detector (254nm)
• Column C18 Nova Pack
• Vacuum filter
• Syringe & micro syringe
• Micro filter paper
• Volumetric flasks/ small vial for filter samples

3.2 Preparation of caffeine standard (stock solution)

a. Calculate for prepare caffeine standard 1000ppm from the caffeine standard.
b. From 1000ppm, prepare serial dilution for caffeine standard: 5ppm, 15ppm, 25ppm, 50ppm
and 100ppm with ready filter de ionize water.

3.3 Sample preparation

a. Obtain sample, soft drink
b. Degas the sample by placing it in a vacuum flask and connecting the flask to vacuum pump
or water aspirator. Leave it under vacuum until no more bubbles appears in the soda samples.
c. Filter the degas soda through 45чL filter syringe.

3.4 Instrument set up

a. Refers SOP /OP/060 –HPLC operation

4. Precautions
• All sample must be filtered
• Never run the column dry, make sure there is enough solvent in the reservoir.

5. Distribution List
1. Original – Document controller
 Malaysian Institute of Chemical
& Bioengineering Technology

Food Technology Department Document SOP /TP/011 Version 01

Title Determination of Caffeine in Soft Drink by HPLC Date 17/03/08
Page 2 of 3
2. Copy - Analysis Laboratory Food Technology Department

6. Appendix
• F/SOP/TP/011

7. Document History
Document No & Description Description Version Date
Issued
Determination of Caffeine in Soft Drink by HPLC New document 01 170308

Document End