Taylor White

Experiment A5 – IR spectroscopy of solvents and flu remedies.

Taylor White

Abstract:
This experiment was used to determine the volumetric fractions of two unknown samples of ortho-
xylene: para-xylene mixtures. These were done through the use of an attenuated total reflection
probe within an infrared spectrometer which was able to detect the degree of which these mixtures
absorbed infrared light at different wavelengths for ortho and para xylene. This experiment concluded
that the unknown mixtures came in ratios of 3-para: 1-ortho xylene for the first unknown and 1-para:
4-ortho xylene for the second unknown compound. This experiment also used a qualitative method
to determine the type of flue remedy used within three solid samples from different manufacturers.
These qualitative tests determined that both the Beecham and Tesco samples contained primarily
ibuprofen, whereas the Lemsip sample contained primarily acetaminophen (paracetamol) within its
sample.

Introduction:
IR spectroscopy is a technique that can be used for analysing a range of different samples which are
prepared in different ways depending on the state of the sample and the exact method of IR used.
Solid samples can be pressed into a disc when grinded together with KBr in this purely qualitative
analytical method. The aim of using this method for this experiment was to obtain results which
identified the different solid samples of flu remedies coming from different manufacturers. Liquid
samples can be readily added dropwise to the lens of an attenuated total reflection (ATR) probe. The
aim of using this method was to attain semi-quantitative results which could be used to determine
the volume fraction of para-xylene in the two unknown samples.

Experimental:
The procedure described in the manual was followed for this experiment. This consisted of using the
an ATR probe to analyse droplets of liquid samples of para-xylene and ortho-xylene in various different
fractional mixtures and these results plot the calibration graph that is used to calculate the volume
fraction of the unknown samples.

Reagents used: ortho-xylene, para-xylene and ortho/para mixtures of xylene in ratios 1:1, 1:2, 1:4,
2:1, 4:1 and two samples of unknown ortho/para ratios.

Excess of KBr powder, Lemsip’s, Tesco’s and Beecham’s flue remedies.

Results:
The first part of this experiment included using the ATR probe to test solvents that included different
isomeric variations of xylene, which were the ortho and para isomers. Using the samples with the
known volume fractions it became possible to plot a calibration curve that would be used to calculate
the volume fraction for the unknown samples.

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Table 1: The results obtained from the first test of every known compound and the two unknowns:

Run 1: Wavenumber Absorbance

Sample Volume Para Ortho Para Ortho AO/AP
Fraction (p)
Ortho xylene 0.00 n/a 738.77 n/a 0.629 0.000
Para xylene 1.00 792.75 n/a 0.501 n/a 0.000
1-para : 1-ortho 0.50 794.37 740.94 0.283 0.398 1.406
1-para : 2-ortho 0.33 794.94 740.16 0.196 0.49 2.500
1-para : 4-ortho 0.20 795.36 739.60 0.132 0.556 4.212
2-para : 1-ortho 0.67 793.82 741.67 0.357 0.301 0.843
4-para : 1-ortho 0.80 793.37 742.31 0.418 0.199 0.476
Unknown 1 x 793.51 794.07 0.396 0.235 0.593
Unknown 2 x 795.22 739.76 0.154 0.532 3.455

Table 2: Results of the second tests done on each individual sample for this experiment:

Run 2: Wavenumber Absorbance

Sample Volume Para Ortho Para Ortho AO/AP
Fraction (p)
Ortho xylene 0.00 n/a 738.78 n/a 0.641 0.000
Para xylene 1.00 792.74 n/a 0.500 n/a 0.000
1-para : 1-ortho 0.50 794.37 740.93 0.282 0.400 1.418
1-para : 2-ortho 0.33 794.94 740.15 0.197 0.491 2.492
1-para : 4-ortho 0.20 795.37 739.61 0.130 0.556 4.277
2-para : 1-ortho 0.67 793.83 741.67 0.346 0.297 0.858
4-para : 1-ortho 0.80 793.38 742.25 0.413 0.200 0.484
Unknown 1 x 793.53 742.03 0.395 0.236 0.597
Unknown 2 x 795.24 739.77 0.146 0.530 3.630

These tables of results also show the calculated values for the absorbance for both the ortho and para
samples divided against each other which is the value that will be plotted onto the calibration graph.

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4.500

4.000 y = -5.9253x + 4.8594

3.500 R² = 0.9068

3.000

2.500
AO/AP

2.000

1.500

1.000

0.500

0.000
0.00 0.20 0.40 0.60 0.80 1.00
Volume Fraction of Para Xylene.

Figure 1: Graph of the volume fraction relative to para-xylene plotted against

AO/AP.

The graph was then used to determine the volumetric fractions of the two unknown samples that
were tested by taking the equation of the graph that was given by excel and inputting the obtained
and averaged value for the AO/AP in as the y value. Table 3 shows how the equation of the line
produced from the graph in figure 1 was rearranged to be able to determine the ortho/para xylene
fractions in the unknown samples by inserting the obtained values

Table 3: Worked answers for the volume fractions (x) of the unknown samples:

Equation of line Y= -5.9253x + 4.8594

Rearranged for x X= y - 4.8594/-5.9253
Unknown 1 X= 0.72
Unknown 2 X= 0.22

These results show that the ortho and para compositions of the unknowns samples are as follows:

Unknown 1: 3-para: 1-ortho

Unknown 2: 1-para: 4-ortho

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The second part of this experiment involved compressing solid samples into a disk to also be analysed
by IR spectroscopy to obtain qualitative results. Three samples of cold and flu remedies were analysed
by IR and the spectrums that were produced are included in the appendix section. The three samples
that were analysed were from different manufacturers which included Beecham, Tesco and Lemsip.
There are three pages within the appendix section which show the results that were obtained by
running them through the IR spectrometer. The results were them compared against a stored library
of commonly known compounds to which they were identified according to the closest chemical
match that could be made. There were two samples identified as being ibuprofen which were
‘Beecham’s’ and ‘Tesco’s’ samples. The third sample was then identified as being acetaminophen
(paracetamol) which was the sample from ‘Lemsip’.

Answers to “Requirements for full write-up”

There is a wide range of available sample holders for IR absorption spectroscopy of which the materials
that are used for them allow the infrared beams of light to pass through them while absorbing only a
minimum amount of the light, or letting the light pass with maximum transmission. The commonly
used examples of these sample holders include potassium bromide, tetrachloroethylene and sodium
chloride.

The attenuated total reflection (ATR) technique uses a beam of infrared light to pass through the
crystal just under the lens of the ATR probe, and then reflects the light once or more times off of the
surface which is in contact with the sample and then generates an evanescent wave which then
extends into the sample. The beam of light is then detected as it leaves the crystal and for this
technique to work the ATR crystal that is being used must have a higher refractive index than the
sample. There are a whole host of advantages to using ATR, one of the largest of these are that the
need for sample preparation is greatly reduced due to its ability to directly measure both solid and
liquid samples without any complex preparations. This also leads on to another advantage which is
how straightforward it is to operate, the steps for using this involve: cleaning the crystal, measuring
the background with the ATR, making sure when the sample is placed onto the crystal there is good
contact and finally measuring it. This procedure being so short compared to other analytical
techniques reduces the possibility of errors caused by the operators.

Systematic errors are those with apply to that particular experimental system and are caused by
defects within the actual experiment or by the processes with which it is carried out. These can be
difficult to detect and cannot effectively be statistically analysed. An example for a systematic error
for this experiment could have been that the samples of known fractions of ortho and para xylene
could have been inaccurate in their volumetric fractions. This would have impacted the calibration
graph that was used to calculate the ortho/para xylene fractions of the two unknown samples and
possibly yielded results that were not truly representative of the experiment. Random errors will
always occur within any experiment and will be able to produce anomalous results. They won’t include
any recognisable pattern because otherwise that would be a non-random and systematic error. The
chances of encountering a random error will increase with each successive measurement made due
to not always being able to reproduce the same measurement exactly multiple times in a row. An
example of how a random error could have occurred in this experiment would be that the ATR crystal
might not have been properly cleaned and wiped of the previous sample before a new one was added.
That could have possibly led to the contamination of the spectrum of the new one produced.

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This graph in excel generated an equation for the linear regression line but to calculate it in person to
check for its accuracy the following equation can be used:

𝑌 = 𝑏𝑋 + 𝑐 Equation: 1

This is the equation for a straight line where the following represent:
∑ {( ̅ )( )}
𝑏= ∑( ̅)
Equation: 2

𝑐 = 𝑦 − 𝑏𝑥̅ Equation: 3

These two equations for b and c calculate the gradient of the line and its Y intercept respectively. Using
the values from Table 4, it was possible to work out that b=-5.92527 and c=4.859445. These two
numbers are exactly what was obtained from the graph on Figure 1, which shows this.

Table 4: The averages of the values obtained in Table 1 and Table 2, alongside
calculated values to be used for the linear regression and correlation coefficient.

The calibration coefficient was also given on the graph from Figure 1, but this can be calculated from
the results obtained to compare it to what was stated on the graph.

∑ {( ̅ )( )}
r= / Equation: 4
{[∑ ( ̅ ) ][∑ ( ) ]}

Once again the values from Table 4, will be used but into Equation 4, this time they would
be used to obtain a value for r and then r2.

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Table 5. The values calculated from Table 4, were used here to solve for r in Equation 4.

With r2 calculated from this table it has turned out to be exactly the same as the value that was
obtained from the excel generated graph in Figure 1. The r2 value being 0.9068 shows that there is a
strongly negative correlation between the volume fraction of the samples analysed and the
absorbance values produced through the IR spectra.

Conclusion:
In conclusion, this experiment was designed to obtain the volume fractions of both ortho and para
xylene from two samples of unknown volume compositions using this semi-quantitative technique.
For the first unknown compound, the ratio was found to be 3-para: 1-ortho xylene and the second
unknown was determined to be 1-para: 4-ortho xylene. These were calculated using the calibration
graph that was produced through excel in Figure 1, and shown how the equation of the line was
rearranged in Table 3. The qualitative part of this experiment had determined that from the 3 solid
samples provided, ‘Beecham’s’ and ‘Tesco’s’ samples were both ibuprofen whereas the technique had
determined that ‘Lemsip’s’ sample was acetaminophen (paracetamol) and these IR spectrums can be
found in the appendix.

Appendix:
Page 7: IR spectrum of ‘Beecham’ sample.

Page 8: IR spectrum of ‘Tesco’ sample.

Page 9: IR spectrum of ‘Lemsip’ sample.