Gervais Manizabayo A00015909 CHEM 121 LAB REPORT Tuesday, 9th July 2013 GROUP MEMEBERS: Oscar Gumiriza,

Aimable Rukundo, Theoneste Manishimwe, Gervais Manizabayo

1. ABSTRACT In this experiment, a thin-layer chromatography (TLC) was used to determine the composition of various analgesics (Acetaminophen, caffeine, Aspirin and ibuprofen) and use the results to identify unknown compounds. In this experiment, two plates were prepared and then spotted. On the first plate, starting from left to right, solutions of acetaminophen, aspirin, caffeine, then aspirin were spotted. On the second plate, solutions to be identified noted labeled 1, 2, 3, were spotted in order to be identified. Both Plates were taken in the development chamber containing the development solvent. The development solvent was seen to rise gradually. The plates were removed from the development chamber, dried and placed under short wavelength UV lamp in order to mark all observable spots. . On the first plate, the distance travelled by Acetaminophen, Aspirin, Caffeine, Ibuprofen were found to be 6.25cm, 9.25cm, 5.9cm, and 10.25cm respectively. On the other hand, the distance travelled by the solution 1, 2, 3 were found to be 6.5 cm, 9.78cm, and 5.25cm respectively. According to the Rf Values calculated using the formula (Rf = found to be Acetaminophen, Ibuprofen, Caffeine respectively. the solutions 1, 2, 3 were

2. INRODUCTION Thin-layer chromatography is one of the easiest of the many chromatographic techniques. A thin layer of a suitable solid substance is coated on a sheet of glass or plastic. A very small sample of the mixture to be analyzed is spotted onto the sheet. By immersing one edge of the sheet in an appropriate liquid developing solvent, the solvent is drawn up the sheet by capillary action, and the compounds of interest are carried along at different rates, effectively separating the components. This is commonly called developing the plate. After the plate has been

developed, it is examined under ultraviolet (UV) light, which allows you to note the location of the spots. Experimental conditions in TLC include the solvent system used to dissolve the compounds, the absorbent coated onto the TLC plate, the thickness of the absorbent layer, and the relative amount of the material spotted onto the plate. Under an established set of experimental conditions, a given compound always travels a fixed distance relative to the distance the solvent front travels. This ratio of the distance the compound travels to the distance the solvent front travels is called the Rf value also called retardation factor or ratioto-front, and it is expressed as a decimal fraction.

3. EXPERIMENT SECTION Preparing the TLC Plates and iodine analysis. TLC plates were obtained from instructor. Using a pencil on the non-shiny side a horizontal line was drawn across the plate at about 1 cm from the bottom. In order to spot the respective substances, 5 spots were marked on the line for the respective substances (Acetaminophen, caffeine, Aspirin and ibuprofen), and the points were 1.8 cm apart along the line. These were the points at which the samples were spotted. Another plate (marked plate 2) was prepared in the same way but with 4 spots including 3 spots of three unknown solutions 1, 2, 3 and another spot for the reference solution. After performing the development and UV visualization, iodine analysis was conducted. At this stage, both plates were placed in a pre-heated beaker and the yellow spots appeared on the plates. After measuring the distance travelled by all solutions and solvents, here below is a table of value obtained. Known compounds, distance traveled by solvent: 14.75cm Acetaminophen Aspirin Caffeine Dist. Dist. Dist. Traveled=6.25cm Traveled=9.25cm Traveled= 5.9cm Rf= Rf= Rf= Ibuprofen dist. Traveled= 10.25cm Rf=

Unknown compounds, distance traveled by solvent: 14.25cm

Compound Compound 2, Compound 3, 1, dist. Dist. Dist. Traveled=6.5 Traveled=9.78cm Traveled=5.25cm Rf= Rf= Rf=

Results discussion

According to the table above, the Rf value for the Acetaminophen is closer to that of compound 1, so the compound 1 was seen to be Acetaminophen. In the same analysis, the table above shows that the compound 2 is Ibuprofen while the compound 3 is Caffeine. Among the compound identified, there were no aspirin, however; its Rf value is slightly different to that of the compound 2 as shown in the table above. 4. CONCLUSION With this lab, a thin-layer chromatography (TLC) was used to determine the composition of various over-the-counter analgesics, and how they will be identified using chromatography techniques. These chromatography techniques focused on of the fact that compounds of different components tend to move at different speeds along the flat surface from the paper used to differentiate them from each other. In this experiment, the RETARDATION FACTORS (Rf values) of known sample of Acetaminophen, caffeine, Aspirin and ibuprofen and those of unknown compounds were determined and then compared. In fact, compound with the same retardation factor were expected to have the same component, then expected to be the same. The experimental results showed that the Rf value of the unknown compound 1 (0.45) was seen to be very closer to that of Acetaminophen (0.423), and then the compound 1 was seen to be Acetaminophen. The same approach was used to identify compound 2 and 3 by comparing their retardation factors to the retardation factors of known compounds. The compounds 2 and 3 were found to be ibuprofen and caffeine respectively.

5. QUESTIONS

5.1. If the spots are made too large when preparing a TLC plate for development, there will be an overlapping of spots so that I may be very difficult distinguish and read the results. 5.2. If the spots are made too small when preparing a TLC plate for development, the spots become almost invisible, and this also can be an obstacle when reading the results. 5.3. The spots must be above the level of the development solvent in the developing chamber because the absorption of the solvent has to be equally distributed along the spots, and they have to wait for the solvent to go up (mobile phase). If they were lowered down, they will be dissolved into the solvent and the development wouldnt work. 4. If the spotting line and positions were marked on the plate with a ballpoint pen, the ink from the pen will stick to some of the solvent and will traveling itself through the paper making the results impossible to be read.

5.5. There are two methods than can be used to distinguish two spots that have the same Rf value but representing different compounds. The first method will be to make a different plate with three reference spots. Another one would be the use of Gas chromatography.

In gas chromatography, the mobile phase (or moving phase) is a carrier gas, usually an inert gas such as helium or an unreactive gas such as nitrogen. The stationary phase is a microscopic layer of liquid or polymer on an inert solid support, inside a piece of glass or metal tubing called a column (an homage to the fractionating column used in distillation). The instrument used to perform gas chromatography is called a gas chromatograph (or aerograph, gas separator). This would be very similar to the paper chromatography but instead of the solvent for the moving phase you will use the gas to get the results. 5.6. Acetaminophen is the most commonly used over-the-counter, non-narcotic analgesic. Acetaminophen is also a popular pain-reliever because it is both effective for mild to moderate pain relief and relatively inexpensive. Unlike acetaminophen, Aspirin can be stomach irritant. In fact, it can irritate any tissue it comes into contact with (thus chewing aspirin gum is not recommended because it can irritate the gums and mouth). In addition, for children, acetaminophen is preferred because the use of aspirin is associated with an increased risk of Reye's syndrome. Reye's syndrome affects all organs of the body but is most harmful to the brain and the liver. 6. REFERENCES Introduction to Organic Laboratory Techniques: A Microscale Approach, 4th Edition. Introduction to Organic Laboratory Techniques:. Google Books. N.p., n.d. Web. 17 Sept. 2012.Search Avantor Materials. Search Avantor Materials. N.p., n.d. Web. 17 Sept. http://goaskalice.columbia.edu/aspirin-vs-ibuprofen