127.1 Expt 4
127.1 Expt 4
127.1 Expt 4
Soft drinks
Non-alcoholic beverages typically containing carbonated water, caffeine, soduim benzoate, sweeteners, and flavoring agents soft in contrast to hard drinks (alcoholic beverages)
Caffeine
white crystalline xanthine alkaloid and psychoactive stimulant acts as a central nervous system (CNS) stimulant most widely consumed psychoactive substance contains diuretic properties
Benzoic Acid
colorless crystalline solid and the simplest aromatic carboxylic acid salts are used as a food preservative
Sodium benzoate : possible cause of DNA damage and hyperactivity Excessive amounts may affect liver and kidney
Beer-Lamberts Law
A = ebc
A = absorbance, e = molar absorptivity, b = pathlength (in cm), c = concentration in moles/liter The amount of light absorbed by a chemical is directly related to the concentration of the chemical in a solution.
Where
Multicomponent Analysis
The spectrum of a mixture of two compounds X and Y is just the sum of the spectra from the individual components X and Y. At any wavelength at the spectrum of the mixture,
Atot = AX + AY
Choosing two wavelengths 1 and 2 at which both compounds show significant absorbance,
,2
The four values can be determined from the slopes of four Beer's Law plots:
A
vs. c at and Y
and
Objectives
To be able to create or produce absorption spectra of soft drinks and standards using dual beam spectrophotometer To be able to determine the concentration of caffeine and benzoic acid in soft drinks using spectrophotometric methods
Calibration Curve
CALIBRATION CURVES
a
graph showing how the experimental observable (the absorbance in this case) varies with the concentration of known standard solution Beer-Lamberts Law, A = bc
From
In
the experiment, it was used to know the absorptivity of caffeine and benzoic acid
Calibration Curve
CALIBRATION CURVES
Table 1. Concentrations of benzoic acid standards and their corresponding UV-Vis absorbance at the given wavelength
Absorbance 230nm 0.223 0.369 0.560 0.742 0.932 272nm 0.020 0.033 0.052 0.069 0.086
Figure 1. Standard calibration curve for benzoic acid standard, showing absorbance readings from 350-210nm
Calibration Curve
230nm 272nm
Slope (a R2
,benz)
0.0934
0.0086
0.9958
0.9965
Figure 2. Absorbance curve showing application of Beers Law at the absorbance of benzoic acid at wavelengths 230nm and 272nm. Y-intercepts set at 0 for 0 absorbance at 0ppm benzoic acid
Calibration Curve
Table 2. Concentrations of caffeine standards and their corresponding UV-Vis absorbance at the given wavelength
Absorbance 230nm 0.200 0.409 0.613 0.824 1.032 272nm 0.367 0.751 1.125 1.503 1.881
Figure 3. Standard calibration curve for caffeine standard, showing absorbance readings from 350-210nm
Calibration Curve
230nm 272nm
Slope (a ,caff) R2
0.0514 0.9998
0.0939 0.9999
Figure 4. Absorbance curve showing application of Beers Law at the absorbance of caffeine at wavelengths 230nm and 272nm. Y-intercepts set at 0 for 0 absorbance at 0ppm caffeine
Calibration Curve
a230,benz = 0.0934 a272,benz = 0.0086
A230,benz = 0.0934bCbenz A272,benz = 0.0086bCbenz
Prepare two dilutions of each sample (2:100, df=50, 4:100, df=25) Add 10mL 0.1M HCl Each sample analyzed for absorbance at and
Sample Analysis
Figure 7. Absorbance readings for Mountain Dew sample at 2:100 and 4:100 dilutions from wavelengths 350nm210nm
Figure 8. Absorbance readings for 7up sample at 2:100 and 4:100 dilutions from wavelengths 350nm-210nm
Figure 9. Absorbance readings for Sprite sample at 2:100 and 4:100 dilutions from wavelengths 350nm-210nm
Sample Analysis
Table 3. Different soft drinks brands and their corresponding UV absorbance readings at given wavelengths. The concentrations of the analyzed sample and overall samples itself were determined along with percent deviation from average values obtained from literature. *Mountain Dew: 157.2ppm caffeine, sprite: 0ppm caffeine, 7Up: 0ppm caffeine 172ppm benzoic acid for all soft drinks
Brand
Dilution Factor 25
(nm)
Abs
Percent Deviation*
272 229
0.364 0.814 0.187 0.426 0.074 0.620 0.048 0.316 0.064 0.583 0.033 0.278
Mountain Dew 50
Caffeine: -48.45% Benzoic acid: +0.7558% Caffeine: -47.29% Benzoic acid: +6.105% Caffeine: +4.742% Benzoic acid: -5.058% Caffeine: +10.60% Benzoic acid: -5.058% Caffeine: +2.893% Benzoic acid: -10.17% Caffeine: +4.150% Benzoic acid: -14.82%
272 229
82.86
182.5
25 Sprite 50
4.742
163.3
10.60
163.3
25 7Up 50
2.893
154.5
4.150
146.5
Sample Analysis
272,caff=0.0939 1
Mountain Dew Dilution Factor=25 (4:100) A272= 272,caffBCcaff + 272,benzBCbenz A230= 230,caffBCcaff + 230,benzBCbenz
Ccaff,final = 25(3.242 ppm)= 81.04 ppm Cbenz,final = 25(6.931 ppm)= 173.3 ppm
Sources of Error
Use of slightly lower wavelength (229 or 229.5nm) instead of maximum (230nm) for sample analysis
Absorbance readings would be slightly lower than if they were measured at 230nm
Caffeine has greater at 230nm, so lower apparent absorbance would have caffeine contribute greater absorbance via higher concentration
known amounts of caffeine and benzoic acid and check if UV-Vis analysis could accurately return the data
References
European Parliament and Council Directive 95/2/EC (1995) on food additivies other than colours or sweeteners. (1995). Official Journal of the European Communities L61, 18.3.95, (pp. 1-40). Hensrud, D. (2011). Nutrition and healthy eating - Coffee and health: What does research say? Retrieved August 2011, from Mayo Clinic: http://www.mayoclinic.com/health/coffee-andhealth/AN01354 Howell, J. A. (1997). Ultraviolet and Visible Molecular Absorption Spectrometry. In F. Settle, Handbook of Instrumental Techniques for Analytical Chemistry (pp. 481-506). New Jersey: Prentice Hall. Lino, C., & Pena, A. (2010). Occurrence of caffeine, saccharin, benzoic acid and sorbic acid in soft drinks and nectars in Portugal and subsequent exposure assessment. Food Chemistry , 121 (2), 503-508. Wibbertmann, A., Kielhorn, J., Koennecker, G., Mangelsforf, I., & Melber, C. (2005). Concise International Chemical Assessment: Benzoic Acid and Sodium Benzoate. World Health Organization. Hanover: United Nations Environment Programme. Wilson, J. (2011). Caffeine Content of Popular Drinks. Retrieved August 2011, from http://wilstar.com/caffeine.htm Yucesoy, C. (2000). Determination of some parameters which affect accuracy and precision in UV-Vis spectrophotometry. J. Fac. Pharm., Ankara , 29 (2), 7-18.