Patrick Smith 3/10/2011 Chem.

475 Excedrin UV-Vis analysis Introduction: In this lab we were tasked with finding the amount of aspirin, acetaminophen, and caffeine in Excedrin tabs, Mr. Goodys, and Anacin. The lab encompassed over 3 weeks of method development, and data gathering to reach the end result. As such, with the method of choice being that of double beam UV-Vis spectrometry, a definition of the methods must be discussed. First, it must be defined what a double beam UV-Vis spectrometer does. Unlike the single beam, the double beam has a much more complicated layout. It uses rotating mirrors to allow light to absorb both into and out of the blank. Along with this blank in one of the cells the next cell will contain the analyte of interest. This allows more statistical measurements and certainty to be taken, because at each wavelength the analyte is being measured against a blank. The other important aspect of what allows this spectrometer to work is its monochromater. Without one, it would be impossible to get any noticeable data. This piece is one of the keystones of a spectrometer as it allows only single wavelength through its selector from a polychromatic light source. This works by having the polychromatic light entering in through an entrance slit and then using a collimating mirror to focus the light onto a diffraction grating. This grating when set at a certain angle will disperse this polychromatic light and give off the desired wavelength to the focusing mirror which reforms the image that entered. This is then passed onto the exit slit, which is then emitted to our detector giving us our reading. Further filters can be added as well, to ensure that even after the monochromater has done its job no other wavelengths are allowed to reach the detector other then the desired one. This ability to select for only one wavelength at a time, and to increase our sensitivity by being able to measure against a blank at every said wavelength, was very valuable in our attempt to quantify the components of our 3 medicines. The reason being is that each individual compound would absorb at different wavelengths, and by using

contrived solutions and standards of the 3 medicines we would be able to find a way to identify through our linest algorithm the quantity of each. The linest program allowed us the ability to analyze, and find 2 of our 3 known compounds in our tablets, and then use the algorithm to find the remaining one. To do this we first had to spend the first week making up our known solutions of aspirin, acetaminophen, and caffeine in order to get baseline data for them. These extinction coefficients were essential as they allow us to derive single equations at each wavelength. These constants at each absorbance help us to identify, and prove that through Beers law of A=bCE that the only unknowns we have if we know the constants of extinction coefficients, are the concentrations of our compounds. With this idea in place it is then possible to analyze through the Linest function, and know the answer that is given is our concentration of the components we seek. Problems arose during this process though, as the results given indicated contradictory evidence of what theory should have given us. Most specifically the acetaminophen in theory was supposed to be the highest absorber above aspirin at around the 220-240-wavelength area. However, due to a dilution mistake on the acetaminophen, it appeared the exact opposite of this with aspiring far outstripping the acetaminophen in absorbance at this maximum thus contradicting the prediction. This problem was not noticed until the Excedrin tabs, Mr. Goodys, and the contrived solutions of known combined compounds were taken the second week. It illustrated the importance of the extinction coefficients because with them invalidated by this mistake the results in the linest function were disastrous. As a result of the misfortune however of this mistake we were better able to see that these constant values in beers law really are necessary to get statistically sound results and data. Thus, in the third week we proceeded with making a set of new contrived solutions with mixtures of different concentrations of the known compounds, and also mixtures of just pure compound as well. We then threw out the extinction coefficients for the previous weeks, and decided to use new ones to analyze our tablets. As well, it was decided that this time that along with Excedrin, Anacin would be used instead of Mr. Goodys. The reasoning was that Anacin only contained caffeine and aspirin, and thus with only two species in it, it would provide an excellent test of our method as there would be no overlapping interference from the two absorbances of aspirin and

acetaminophen. As well we decided to take other precautionary steps to ensure that the data had no chance of being skewed. .1 M HCl was blanked against this time rather then water before due to the fact that even though not much would be different in absorbing there could be a chance that it was noticeable enough to add error to that already present. As well for the Excedrin and Anacin solutions they were filtered through a .45micrommeter syringe filter to remove any of the filler-binding agents in the pills. As well they were diluted even more then the first time by a factor of 10 to ensure that it was as dilute as possible eliminating any binding agents whatsoever. Data and Results Figure 1: Absorbance data of pure compound solutions. Figure 2: Sample Spectra of Excedrin compounds. Figure 3: Sample Spectra of Anacin tabs. Figure 4: Extinction coefficient data from contrived solutions. Excedrin Data: Group 1 Aspirin Acetaminophen Caffeine Anacin Data: Std. Std. Std. Std. Group 1 Dev. Group 2 Dev. Group 3 Dev. Group 4 Dev. 0.644 0.581 0.744 0.725 Aspirin 358.8 369.1 408.9 398.52 0.638 0.573 0.737 0.72 Caffeine 30.2 33.6 36.2 33.29 Excedrin Group averages with propagated error: Drug Compound Acetaminophen Average 243 Std. Deviation 1.56 Std. Std. Std. Std. Dev. Group 2 Dev. Group 3 Dev. Group 4 Dev. 0.756 1.04 0.813 1.09 258.3 262 254.8 245 0.6301 0.864 0.676 0.908 221 259.5 243.9 251.2 0.751 1.029 0.805 1.08 56.1 61 57.5 55.3

1.87 Aspirin Caffeine 255 1.85 56

Anacin Group averages with propagated error: Drug Std. Compound Average Deviation 1.35 Aspirin 384 1.34 Caffeine 33 Discussion of Results and Data: This data that was collected provided many conclusions that are valuable for understanding UV-Vis spectroscopic practices. As well, it also showed us how our method that was developed was verified. Finally, the data obtained was necessary in determining the reproducibility and accuracy of our results and the spectroscopic method. First, the most important thing that needed to be verified was that both our contrived solutions, and the pure compound solution data matched the same absorbance pathways. As seen in figure 1, the three absorbances of both Aspirin (green) Acetaminophen (red) and Caffeine (blue) follow a set pattern shown. Caffeine absorbs almost exclusively in the 275.0-295.0 nm range. No other species absorb at this wavelength so in the contrived solutions with caffeine by itself or with combinations of caffeine and other drugs the method put forth should in theory show the same absorbance peak for caffeine. The solutions without it should show no absorbance at all. Acetaminophen and aspirin both have their peaks at 235.0-245.0 nm with Acetaminophen showing the stronger absorbance. Again a similar peak should be shown in our contrived solution as predicted by our method set forth. Figure 4, which contains our contrived solutions shows that this prediction holds up with the given spectra following the same absorbance lines as in figure 1 with our pure solutions. This proof of what we had gathered in theory before conducting the experiment was very important because it showed that we could produce accurate extinction coefficients. This then showed that Beers Law truly is proven where the only variable

we have is the concentration of analyte, which then dictates our extinction coefficients and absorbances. It is this relationship that even makes it possible for us to identify different species in our two products. The spectroscopic relationships do not end there though. Figure 4 comes into play when trying to understand figures 2 and 3 of the Excedrin and Anacin data. The lines in figure 4 verify that with the contrived solutions when you combine solutions with acetaminophen and caffeine rather then getting 2 separate identifiable peaks you instead are given one peak that is the sum of the two. This is critical in identifying our unknown compounds as the spectral data in either of the figures 2 or 3 show only one absorbance line for each test done by a group. By knowing that these two peaks of acetaminophen and aspirin add together, it allows us to use our contrived solution data where we gathered extinction coefficients for when there was only one of the two species present and use it to identify either the acetaminophen or aspirin using the linest program. Along with one of these, the caffeine can easily be indentified because it absorbs at minimum absorbance of the other two. With two out of the three found we can then use the lines program to extrapolate what the third value must be in order to add up to the given absorbance. The overall data gathered in this lab was relatively accurate as shown in graphs on figures 2,3,4 visually, and by the standard deviations given by the linest program. In particular though there were a few problems that lead to some systematic error in our procedure. Some of these were fixed, but others were unable to be addressed in the span of the experiment. First, during the two weeks that we did our experiment we discovered that significant systematic error had been added in by using water as our blank rather then the .1 M HCL that we used as our solvent in solution preparation. It was believed that because HCL and water nearly absorb in the same way it would not have an impact, but the mistake highlighted the basic need of any uv-vis spectroscopic procedure, which is that you must blank against what your solvent is no matter what. This error was then rectified the third week and results became more streamlined between groups. Second, were the filler portions of the Excedrin and Anacin. This caused problems during the first two weeks as when making the dilute solutions the filler components could not be completely filtered out. As a result when put into the

spectrometer some of the light passing through the sample hit these fillers that were in the product solutions, and caused abnormal readings then what was expected, and increased systematic error. The third week this was rectified again by using a .15-micrometer filter rather then a .45-micrometer filter to allow only the soluble parts of the solutions to pass through. This again helped to lower our systematic error as the filler was almost completely removed. Third, there was the problem of a non-streamlined process in the solution making of both the contrived solutions, and the unknown samples. To change this, it was necessary to force each group to make the contrived solutions exactly the same way in the same procedure, and to make the dilution for the unknown solutions the same as well. By doing this there was a decreased chance of dilution error being present, and again systematic error was removed. Finally, the one main error that was unavoidable that was systematic of our method was that of Caffeine and its consistently low readings from what was expected in theory. This can easily be explained by the fact that acetaminophen, and aspirin are in much higher concentrations then caffeine in each pill. As a result, the caffeine becomes so dilute during the dilution of the unknown solution that the caffeine approaches the detection limit more so then the other two. This results in a little more uncertainty, but it is one that is hard to overcome as it is necessary to be so dilute in order to effectively read the absorbances of the acetaminophen and aspirin as to not over load the solution. This lab proves the value beyond measure spectroscopy in the modern world. In the realm of antibiotics and medicine it is vitally important to know that each pill does contain what its package says. This method we used allows us to verify it in a quantities method that is both reproducible, and prone to little error. When peoples lives are in the balance this is a must, and shows the true value of chemistry in our daily lives. Not only is this method valuable due to its sheer ease of use, and reproducibility, but it also is verifiable by other methods as well. This same experimental data could be confirmed by using Standard additions at each concentration to verify the same data we obtained using the double-beam spectrometer. However, while this is perhaps the best way to verify it, it shouldnt be used due to the amount of length in time that it would take to complete. Another method that could be used to verify this in a different practice is by using HPLC. By using a form of chromatography we can separate the three

components of the unknown solutions, and then quantify them using an appropriate method. This way to interchange chemical laboratory methods to obtain the same result truly shows why spectrometry is such a core backbone of chemistry.