SPECTROSCOPIC DETERMINATION OF CAFFEINE AND BENZOIC ACID IN SOFT DRINKS

Caffeine is a bitter, white crystalline xanthine alkaloid that is a psychoactive stimulant. In humans, caffeine acts as a
central nervous system stimulant, temporarily warding off drowsiness and restoring alertness. It is a commonly
found in coffee, tea, and soft drinks. The concentration of caffeine in tea ranges from 2.5 to 4.5 % as contrasted to
an average concentration of about 1.5% in coffee. Excess of caffeine causes adverse effect on humans
cardiovascular system.
Benzoic acid is a white crystalline solid. Its salts are used as food preservatives. Food preservation is the prevention
of chemicals decomposition and the development of harmful bacterial in foods. High amount of benzoic acid is
harmful for liver and it disturbs carbohydrate metabolism which may lead to accumulation of fat causing obesity. It
is added as a sodium benzoate in soft drinks.
Like many organic molecules, both caffeine as well as benzoic acid absorbs radiation with a wavelength around 272
nm and 229 nm respectively. If a series of caffeine standards are analyzed in this region of absorption and a Beers
law is plot prepared, then the amount of caffeine in another substance can be determined.

Industrial Method
1. Extraction of caffeine
Take 150 ml soft drink sample into a separating funnel and add 2.0g of sodium carbonate in order to make the pH
basic. Add 50ml of methylene chloride and shake it vigorously for at least 5 minutes. Allow it to settle for about 5
10 minutes. Collect the lower organic layer into a conical flask. Again add 50ml sample of methylene chloride,
shake and then collect the organic layer in the conical flask. This solution can be use as such for UV measurement.
2. Extraction of Benzoic Acid
Take 150 ml soft drink sample into a separating funnel and add few drops of dilute HCl in order to make the pH
more acidic. Then extract it in a similar as performed for caffeine.
3. Prepare a standard solution of benzoic acid and caffeine having concentration of 5, 10, 15, 20 and 25 ppm
in methylene chloride.
4. UV Measurement
The absorbance of all standard and soft drinks samples can be recorded in UV spectrometer. And plot a Beers law
curve of absorbance vs concentration for the caffeine and benzoic acid standards. Using the graph, the concentration
of caffeine and benzoic acid in each soda can be estimated.
 Laboratory Method
All solutions will contain 0.010 M HCl, so the sodium benzoate is protonated to make benzoic acid. Caffeine has no
appreciable basicity, so it is neutral at pH 2. We restrict ourselves to non-diet soft drinks because the sugar substitute
aspartame in diet soda has some ultraviolet absorbance that slightly interferes in the present experiment. We also
avoid darkly colored drinks because the colorants have ultraviolet absorbance. One should be aware that the
assumption is being made that the unknown contains no other substances which absorb at this wavelength.
Preparation of 10ml Calibration Standards
1. Prepare a set of benzoic acid solutions containing 2, 4, 6, 8, and 10 mg/L in 0.010 M HCl.
2. Prepare caffeine standards containing 4, 8, 12, 16, & 20 mg/L in 0.010 M HCl.
Preparation of Soft Drink Solutions
1. Warm ~ 20 mL of soft drink in a beaker on a hot plate to expel CO2. (Do not boil!)
2. Filter the warm liquid through filter paper to remove any particles.
3. After cooling to room temperature, pipette 2.00 mL into a 50-mL volumetric flask.
4. Add 10.0 mL of 0.010 M HCl and dilute to the mark.
Determination of Caffeine and Benzoic Acid
1. Record the UV spectrum of each of the 10 standards with water in the reference cuvette.
2. Record the wavelength of peak absorbance for benzoic acid (229 nm) and the wavelength for the peak absorbance
of caffeine (272 nm).
3. Measure the ultraviolet absorption spectrum of the diluted soft drink samples.
Data Table:
Caffeine Standard Benzoic Acid Standard
Conc. Abs. Conc. Abs.

Brand Absorbance Concentration of Benzoic Acid Concentration of Caffeine