Thin Layer of Chromatographic Analysis of Analgesic Tablets Danielle Karol H. Reyes, Karlo Jonathan A. Salem, Maria Daniela P.

Santos and Michael Ruben L. Tiu Department of Sports Science, College of Rehabilitation Sciences, University of Sto. Tomas, Manila, Philippines Abstract: In this experiment, Thin Layer Chromatography (TLC) was used to identify the active compound of an analgesic tablet. Through the use mortar and pestle, several over-the-counter drugs were crushed and solvent using Methanal and Toluene was created as the eluent. By immersing the one edge of the stationary phase in the mobile phase, the solvent went up by capillary action. The compounds were carried at different rates by separating the components. The TLC plate was examined under UV light to locate the dark spots of each components, which allowed the computation of the retardation factor (RF) values.

Introduction: Over-the-counter analgesics typically contain one or more of the following active ingredients: acetaminophen, aspirin, caffeine, ibuprofen or naproxen. chromatography (TLC) offers a simple method of analysis for these products. Chromatography is the collective term for a set of laboratory techniques for the separation of mixtures. It is widely used to describe a family of closely related separation methods. There are many types separation methods, but chromatography compared to other physical and chemical methods of separation is both a stationary and mobile phase; the sample is to be separated a number of times with these phases. The sample is carried through the system via the mobile phase, and the interactions that happen in the process are due to the differences in the physical and chemical properties. These differing similarities manage the rate at which the individual Thin layer

components of the sample pass over the stationary phase under the influence of the mobile phase. Thin layer chromatography (TLC) is a widely employed laboratory technique and is similar to paper chromatography. However, instead of using a stationary phase of paper, it involves a stationary phase of a thin layer of adsorbent like silica gel, alumina, or cellulose on a flat, inert substrate. Compared to paper, it has the advantage of faster runs, better separations, and the choice between different adsorbents. For even better resolution and to allow for quantification, high-performance TLC can be used. It is one type of chromatography where the stationary phase is a thin layer of adsorbent particles attached to the solid plate. A small amount of sample is applied (spotted) near the bottom of the plate, and the plate is placed in the mobile phase. This solvent is drawn up by capillary action to a predetermined height. Each component, being different in chemical and physical composition, will interact with the stationary phase at a different time (retention time), thereby creating the individual bands on the plate. The retention time or retention factor (Rf) is used to characterize and compare components of various samples. Results and Discussion: Table 1: Rf value of analgesics Analgesic Acetaminophen Aspirin Caffeine Sample Ibuprofen Mefenamic Acid Phenacetin Rf value 0.808 0.914 0.531 0.702 0.914 0.957 0.851

() ()

Figure 1: Rf value equation Calculations:

. = . = 3.7976 = 0.808 4.7

4.2958 = 0.914 4.7 2.4957 = 0.531 4.7

. = . =

3.2994 = 0.702 4.7 4.2958 = 0.914 4.7 4.4979 = 0.957 4.7

. =

. = . =

3.9997 = 0.851 4.7

Figure 2: Chromatogram of analgesics

The spots obtained under the UV light determines the Rf value of the 7 analgesic solutions. The RF value of each spot can be determined by dividing the distance the product traveled by the distance the solvent front traveled using the initial spotting site as reference. These values depend on the solvent used and the type of TLC plate and are not physical constants. The separation of compounds is based on the competition of the solute and the mobile phase for binding places on the stationary phase. The compounds present in the stationary phase has different polarity values. The less polar compound moves higher up the plate. The order of strength/weakness depends on the coating (stationary phase) of the TLC plate. The results show that Mefenamic acid has the highest polarity and the Caffeine has the lowest polarity while Aspirin and Ibuprofen has the same polarity value. Experimental: In performing a Thin Layer Chromatography analysis, the experiment requires the use of mortar and pestle, tablet, TLC plate, 250 mL beaker, watch glass, funnel, test tube and graduated cylinder. The experiment is divided into five procedures. The first step is the preparation of the sample, stationary phase (SP) and mobile phase (MP). To prepare the sample, the tablet was pulverize using a mortar and pestle. The analgesic powder transferred into a test tube and 2mL of Methanal and Toluene was added. After it is allowed to settle, the mixture was decanted to get the sample. The SP was a plastic back coated with a thin film of silica gel and the MP was a solution of ethyl acetate, methanol and acetic acid. After the preparation of the sample, SP and MP, the sample is applied to the SP. There should be a drawing on the SP indicating that it is the solvent front and 1cm at the bottom that is called the origin. Seven spots should be marked evenly in pencil along the origin. The next step was to immerse the SP in the MP. The spots on the SP should be above the level of the MP. The MP should also be allowed to move to the solvent front for the development of the chromatogram. After which, the SP was viewed under UV light in the UV viewing chamber to locate the dark spots and should be traced using a pencil.

Setup 1: Development of Chromatogram

References: http://en.wikipedia.org/wiki/Thin_layer_chromatography http://www.scribd.com/doc/47193409/1a-TLC-Analysis-of-Analgesic-Drugs http://www.scribd.com/doc/55852880/TLC-Analysis-of-Analgesic-Drugs http://www.scribd.com/doc/55852880/TLC-Analysis-of-Analgesic-Drugs