In the Laboratory
Determination of Phosphorus in Cola Drinks
Diego Lozano-Calero and Pilar Martín-Palomeque
Servicio de Bioquímica, Hospital La Paz, Paseo de la Castellana, 261, 28046 Madrid, Spain
Silvia Madueño-Loriguillo
Departamento de Química Inorgánica, Universidad de Málaga, Campus de Teatinos, 29071 Málaga, Spain
Laboratory experiments can improve student inter- ter. Place 39 mL of the ammonium molybdate solution,
est in science. However, the contrary effect could occur 60 mL of the ascorbic acid solution and 125 mL of di-
if they are not well designed and seem tedious, too labo- luted sulfuric acid into a 250-mL volumetric flask. Fill
rious, and disconnected from daily life. to mark and mix.
Cola beverages are one of the most widely consumed
drinks and are most popular among students. Much at-
tention is being paid to possible consequences of exces-
sive consumption for human health. Intensive efforts are
being made to assess the erosive potential for teeth be-
cause of the beverages’ acidity (1, 2); adverse effects sec-
ondary to high caffeine intake (e.g., hypertension, aller-
gic reactions, gastrointestinal disturbances) (3–5); and
adverse effects on calcium metabolism due to their high
phosphoric acid content, which combined with low di-
etary calcium intake could increase the risk of suffering
from bone diseases (6–9).
We propose here the quantification of the phospho-
rus content in this kind of drinks by a different proce-
dure from that previously described by Murphy in this
Journal (10). We think this laboratory experiment will
seem very interesting to students.
Theory
The method consists of the formation of a complex
of phosphate ion with a molybdate compound in acid so- Figure 1. Plot of absorbance vs. concentration of the standard so-
lution: lutions.
7 H3PO4 + 12 (NH4)6Mo7O24 ?4H2O → Table 1. Sample and Calibration Curve Preparation
+ { Calibration Curve Sample
7 (NH4)3[PO 4(MoO3)12 ] + 51 NH4 + 51 OH + 33 H2O
Volume from working standard
0 0.5 1 1.5 2 2.5
The colorless hexavalent molybdenum phosphate solution (mL)
Volume from diluted sample
complex is reduced to a blue pentavalent form by ascor- 0.25
(mL)
bic acid in acid medium. The intensity of the blue color
Reducing solution (mL) 2 2 2 2 2 2 2
is measured spectrophotometrically at 830 nm (11).
Obviously, the sample cannot be colored; therefore Deionized water (mL) 3 2.5 2 1.5 1 0.5 2.75
we used a clear cola, Crystal Pepsi®, acquired in the Final volume (mL) 5 5 5 5 5 5 5
market. Final concentration expressed as
0 0.4 0.8 1.2 1.6 2
P2O5 (ppm)
Experimental Procedure
Preparation of Solutions Table 2. Absorbance Obtained for Sample and Standards
Stock Standard Solution. Weigh out 1.9412 g of dry
Solution A830
KH2PO4, reagent grade, and dissolve in deionized water
in a 1-L volumetric flask. Dilute to mark. The phospho- Standards, ppm
rus content of this solution expressed as P2O5 is 1000 0.4 0.255
ppm.
0.8 0.588
Working Standard Solution. From the stock stan-
dard solution place 2.5 mL into a 50-mL volumetric flask 1.2 0.866
and dilute to mark. From there deliver 4 mL into a 50- 1.6 1.151
mL volumetric flask and fill to mark. The final result- 2.0 1.436
ing solution is 4 ppm P2O5.
Sample, no.
Reducing Solution. Dissolve 1.00 g of ammonium
molybdate in 50 mL of deionized water and 1.76 g of 1 0.565
ascorbic acid in 100 mL of deionized water. Dilute 17 mL 2 0.571
of concentrated sulfuric acid in 200 mL of deionized wa- 3 0.563
Vol. 73 No. 12 December 1996 • Journal of Chemical Education 1173
In the Laboratory
Sample Preparation and Measurement mation from P2O5 to P content, the phosphorus present
in the sample was calculated by the following equation:
Let the sample stand at atmospheric pressure for
24 h before analysis in order to eliminate gases. Then ppm of P in sample =
transfer 2.5 mL of the sample into a 50-mL volumetric 20 × 20 × X ppm P2O5 × (62 mg P/142 mg P2O5)
flask. Fill to the mark with deionized water.
Dispense the volume of each solution into test tubes We found that Crystal Pepsi® contains an average
as indicated in Table 1. Warm the tubes for 45 min at 50 142 ppm phosphorus.
°C in a water bath. Read the absorbance against a blank
at 830 nm. Make at least three determinations of the
Literature Cited
sample.
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Clin. Immunol. 1993, 3, 160–162.
Our results are shown in Table 2. The linear cali- 4. Brikett, N. J.; Logan, A. G. J. Hypertens. 1988, 6(4 Suppl.), S620–S622.
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bration curve was obtained for the range 0.4–2 ppm of 6. Wyshak, G.; Frisch, R. E. J. Adolescent Health 1994, 15, 210–216.
P2O5 and was fitted by least squares (y = 0.72x – 0.02, 7. Hernandez, A; Stampfer, M. J.; Ravnikar, V. A.; Willet, W. C.; Schiff, I.; Francis,
r = .999; see Fig. 1). The concentration of phosphorus M.; Longscope, C.; Mckinlay, S. M. Epidemiology 1993, 42, 128–134.
8. Smith, S.; Swain, J.; Brown, E. M.; Wyshak, G.; Albright, T.; Ravnikar, V. A.;
present in the test solution was calculated by the equa- Schiff, I. Arch. Intern. Med. 1989, 149, 2517–2519.
tion of the calibration curve. Taking into account the di- 9. Spencer, H.; Kramer, L.; Osis, D. J. Nutr. 1988, 118, 657–660.
10. Murphy, J. J. Chem. Educ. 1983, 60, 420–421.
lutions made on the original sample and the transfor- 11. Charlot, G. Chemie Analytique Quantitative, Masson: Paris, 1974; p 478.
1174 Journal of Chemical Education • Vol. 73 No. 12 December 1996
