Jess Turner Chem 219 page 1

Lab: FT-IR date: 5/5/2018

Chem 219: Determination of caffeine concentration in

beverages through FTIR spectroscopy

Jess Turner

Lab Partner: Dan Venezia

TA: Kevin Fischer

Date lab performed: 27/3/2018

Date report submitted: 10/4/2018

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ABSTRACT

The aim of the experiment was to determine the caffeine concentration in four drinks samples.

This was done through Fourier Transform Infrared (FT-IR) spectroscopy. A set of standard

solutions were made and an IR spectrum for each was taken. The absorption values obtained at

1655cm-1 were then used to create a calibration curve. Based on this curve the concentration of

caffeine in each sample could be calculated through the linear regression of the curve and the

relationship between absorption and concentration given in Beer’s Law. It was found that the

beverage with the lowest concentration was the decaffeinated coffee sample (0.1921ppm),

followed by the black tea (0.4745 ppm), Goldberg’s cold brew (1.231 ppm) and the Kazaav

espresso (2.723 ppm) respectively. These results are to be expected, particularly the

decaffeinated coffee giving the lowest concentration of caffeine. However, there was some

difficulty in producing an accurate calibration curve; one data point had to be removed due to

unknown sources of error, which may limit the accuracy of the curve itself. This will in turn

have had an effect of the calculated concentration values for the beverages. Despite this, the

order of caffeine concentrations would likely remain unchanged. In conclusion, to confirm the

concentration values the calibration curve should be repeated to obtain a better curve and in turn

more accurate concentration values.

INTRODUCTION

Caffeine is probably the most frequently ingested pharmacologically active substance

across the globe. Appearing in many foods and drinks such as chocolate, coffee and soft drinks

it has become increasingly important for the general public to understand the effects of the

chemical. High consumption of the substance has been linked to several health effects,

especially for pregnant women. (3) For this reason, it is important that we know how much
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caffeine is in the foods and drinks we consume so that we can monitor uptake and avoid potential

health risks.

The use of FTIR provides several benefits over alternative methods of analysis. Firstly,

the Multiplex or Fellgett advantage allows all source wavelengths to be measured simultaneously

as opposed to successively, allowing a complete spectrum to be collected much more rapidly

than for a standard dispersive spectrometer. Additionally, the throughput or Jacquinot advantage

allows more light to get through than a dispersive spectrometer which is limited by the slit width.

Combined these two advantages allow a signal to noise ratio to be obtained which is much

greater than for other devices.

Experimental approach was simple; a set of standard solutions were made and analyzed

to make a calibration curve. Analysis was also conducted on four drinks samples; Kazaav

Espresso, Goldberg’s cold brew, black tea and decaffeinated coffee, which were diluted in

chloroform before analysis. The concentration of caffeine in each of these would then be

calculated by using the linear regression of the calibration curve.

EXPERIMENTAL

Theory. Infrared (IR) spectroscopy is one of the most valuable analytical techniques

available to scientists today. (1) A spectrum is produced based on the absorbance of light by a

molecule. For a molecule to absorb IR radiation it must change its dipole moment upon

vibration; symmetrical molecules such as O2 and N2 do not absorb IR radiation. (2) However,

larger molecules such as caffeine are IR active and absorb well allowing concentrations to be

determined. IR radiation can be used for both qualitative and quantitative analysis through shift

and absorbance values respectively. For this experiment the instruments quantitative analysis

must be used in which Beer’s Law is utilized. It shows the linear relationship between
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absorbance and concentration, allowing the concentration of caffeine in each of the samples to be
still no "materials and
calculated based off the calibration curve and its linear regression. (2) safety considerations"?

Instrumentation. Several caffeine samples were provided to be tested, as well as

reagent grade chloroform for caffeine extraction and caffeine for standard solution preparation.

The spectrometer used for analysis was the Shimadzu IR Affinity-1 with a PIKE MIRacle,

single-reflection attenuated total reflectance (ATR) attachment.

Procedures (4). 0.228g of caffeine was weighed by difference and used to create a stock

solution in chloroform in a 250 mL volumetric flask. A 10.00 mL aliquot of this solution was

used to create a second stock solution in chloroform in a 100 mL volumetric flask. This was

then used to create four standard solutions with concentrations as shown below.

Table 1: Preparation and concentrations of standard caffeine solutions

Standard 1 2 3 4
mL of SS2 0.5 1.5 3 4
Concentration (ppm) 0.0456 0.1368 0.2736 0.3648

The liquid samples were then prepared for analysis. 3.00 mL of each sample and

3.00 mL chloroform were transferred by micropipette into a 30 mL separatory funnel. After

phase separation 1.00 mL of the chloroform layer was diluted in chloroform to give a final

volume of 10 mL in a volumetric flask. This solution was used for the IR analysis. The FT-IR

analysis was conducted using the Shimadzu IR Affinity-1 with a PIKE MIRacle, single-

reflection attenuated total reflectance (ATR) attachment at 4 cm resolution.

inverse cm (i.e.,
wavenumbers)
Data Analysis. The main aim of the experiment was to use the measured absorption of

each sample to calculate the concentration of the caffeine in each one. To successfully do this a

standard curve was created using the absorption value for each standard solution at 1655 cm-1.

You disregarded my
previous comment: "
how was this
determined? It is critical
to your accuracy."
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From here the concentration of caffeine in each sample can then be determined using the

equation of linear regression from the curve and solving for x. (i.e. y=mx+c where y =absorption,

m =gradient, x =concentration and c =intercept.)

RESULTS

Table 2: Recorded absorptions from standard caffeine solutions at 1655 cm-1

Conc/ ppm Abs at 1655cm-1

0.0456 -0.002372
0.1368 0.1053
0.2736 0.004017
0.3648 0.004274

Table 3: Calculated concentrations of caffeine in each sample using the standard curve created
from the standard caffeine solutions.

Abs at 1655 cm-1 Conc / ppm (3 points) Conc / ppm (4 points)

Goldberg Cold Brew 0.02423 1.231 -12.16
Decaf 0.001165 0.1921 -1.483
Espresso Kazaav 0.05735 2.723 -27.50
Black Tea 0.007434 0.4745 -4.385
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Figures 1-4 and 7-10 can be found in an appendix at the end of the report.

Figure 5: Standard curve from all four absorptions obtained at 1655 cm-1

Figure 6: Standard curve from all three absorptions obtained at 1655 cm-1
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DISCUSSION

The aim of this experiment was to determine the caffeine concentrations in four different

beverage samples through Fourier Transform-Infrared (FTIR) spectroscopy. Beer’s Law

(A=bc) shows the linear relationship between absorbance and concentration and so this can be

used to obtain the concentration from the measured results. The results show that the beverage

with the highest caffeine content was the Espresso Kazaav at 2.723. The beverage with the

lowest concentration was the decaffeinated coffee. This is to be expected given the nature of the

drink.

An important aspect of this experiment was the use of a standard curve to determine the

concentrations. However, the results obtained for this part of the experiment were not as would

be expected. Given the relationship shown in Beer’s law, one would expect the standard solution

with highest concentration of caffeine to have the largest absorption value, but as shown in Table

2 this was not the case; the sample with the second lowest concentration had the highest

absorption value at 0.1053 ppm. This result is almost certainly an anomaly given the general

trend seen with the other data points and was therefore omitted from the calculation of the curve.

However, the cause of such a large error cannot easily be identified. One possible explanation is

an instrumental cause. When calculating the concentration, it is assumed that all other

parameters including the path length remain constant. In reality it is very unlikely that this is the

case; the path length of the FTIR instrument is very small and therefore keeping it constant is

challenging. Deviation away from a constant path length can result in errors of concentration

calculation using this method.

There are, of course, other possible sources of error, such as human and experimental

error, but it’s hard to see where such a large human error would have come from.
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APPENDIX
Figure 1: Recorded data for standard solution 1; concentration =0.0456 ppm

Figure 1.1: Recorded data for standard solution 1 between 0 and 500 cm-1.
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Figure 2: Recorded data for standard solution 2; concentration =0.1368 ppm

Figure 2.1: Recorded data for standard solution 2 between 399 and 600 cm-1.
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Figure 3: Recorded data for standard solution 3; concentration =0.2736 ppm

Figure 3.1: Recorded data for standard solution 3 between 399 and 600 cm-1.
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Figure 4: Recorded data for standard solution 4; concentration=0.3648 ppm

Figure 4.1: Recorded data for standard solution 4 between 399 and 600 cm-1.
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Figure 7: Absorption data for Goldberg’s Cold Brew sample

Figure 7.1: Recorded data for Goldberg’s Cold Brew between 399 and 600 cm-1.
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Figure 8: Absorption data for Decaffeinated Coffee sample

Figure 8.1: Recorded data for Decaffeinated Coffee between 399 and 600 cm-1.
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Figure 9: Absorption data for Espresso Kazaav sample

Figure 9.1: Recorded data for Kazaav Espresso between 399 and 600 cm-1.
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Figure 10: Absorption data for Black Tea sample

Figure 10.1: Recorded data for Black Tea between 399 and 600 cm-1.
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LITERATURE CITED

1. Stuart, B. (2005) Kirk-Othmer Encyclopedia of Chemical Technology Ed 1. Infrared

Spectroscopy, John Wiley and Sons.

2. Smith, B. (2011) Fundamentals of Fourier Transform Infrared Spectroscopy, Second

Edition, p5-6, Taylor and Francis Group.

3. Nawrot, P, Jordan, S, Eastwood, J et al. (2010) Effects of Caffeine on Human Health,

Food Additives and Contaminants Vol 20. p1-30

4. Petrucci, G. (Jan 2018) Determination of caffeine content in beverages using Fourier

Transform infra-red spectroscopy Laboratory Manual, p 6-7