Jonathan Saperstein, CHEM 213B Synthetic #3 FFR Synthesis of Benzocaine via Fischer Esterification Introduction Benzocaine is an organic compound

that has played a major role in the field of medicine for more than 100 years. It is used as a local anesthetic and can be found in many ointments designed to relieve pain.1 For example, benzocaine is an active ingredient in many topical medications that treat for sunburns, toothaches, and hemorrhoids. It works as a numbing agent by blocking signals between nerves on areas where it is applied.2 Benzocaine is an ester, which is a large class of organic molecules. Esters serve many purposes. Some give off very potent fragrances (such as those of fruits) and can be found naturally in plants, while others make up polyesters, materials that give rise to fabric and plastics.3 Esterification reactions are very useful to organic chemistry, as there are many different ways in which to make esters. The classic method of forming an ester is through the Fischer esterification (which is what is used in this experiment). In this reaction, a carboxylic acid is treated with an alcohol under acidic conditions.4 Esters can also be made by reacting alcohols with acyl chlorides or acid anhydrides.5 Once an ester is created, it can be turned into another ester by means of transesterification, which is catalyzed by either an acid or base and involves treating an ester with an alcohol to produce a new ester and alcohol products. Finally, esters can be turned back into their alcohol and carboxylic acid constituents using an acid (reverse of the Fischer esterification) or a base (saponification).6

Figure 1. Electron-pushing mechanism for the synthesis of benzocaine from p-aminobenzoic acid. The formation of benzocaine in this reaction involves the Fischer esterification, in which ethanol and sulfuric acid were used to convert a carboxylic acid to an ester. First the carbonyl oxygen is protonated by sulfuric acid. One of the electron pairs of the carbonyl bond moves up to the oxygen, creating a positive charge at the carbon. The oxygen of ethanol then attacks the carbonyl carbon through nucleophilic addition. Next, ethanol deprotonates the newly added ethanol substituent. One of the hydroxyl groups then receives a proton from a protonated ethanol ion in solution. A lone pair of the oxygen of the second hydroxyl group moves to form a double bond with the central carbon while kicking off a water molecule. Finally, after an ethanol molecule deprotonates the positively charged carbonyl oxygen to restore a neutral charge, benzocaine is formed. The purpose of this experiment is to make benzocaine from p-aminobenzoic acid by converting the carboxylic acid on the p-aminobenzoic acid into an ester using absolute ethanol and sulfuric acid in the Fischer esterification. After the reaction, the product is purified using sodium bicarbonate and characterized using melting point, IR, 60 MHz 1H NMR, and GC/GC-MS.

Experimental Benzocaine.5 p-aminobenzoic acid (0.123 g, 0.90 mmol), absolute ethanol (1.5 mL, 25.69 mmol), and 3 boiling chips were added to a reaction tube, and the reaction mixture was heated until the solids dissolved (~5 minutes). After being cooled on ice, 18 M sulfuric acid (0.20 mL, 3.75 mmol) was added drop-wise (forming a precipitate), and the mixture refluxed for 45 minutes. The solution was then cooled to room temperature and was transferred to a 10-ml Erlenmeyer flask with water (3 mL). Next, sodium bicarbonate (3 mL, 78.56 mmol) was added dropwise to form white precipitate. Vacuum filtration and water (3 x 1 mL) isolated and rinsed the white product crystals (0.068 g, 45.90%); melting point range 85-88 oC; 1H-NMR (60 MHz, DMSO) 7.71 and 7.56 (d, 2H), 6.63 and 6.48 (d, 2H), 5.91 (s, 2H), 4.35, 4.25, 4.14, and 4.05 (q, 2H), 1.35, 1.26, and 1.15 (t, 3H); IR (ATR) max 3339.3, 2983.3, 1680.1, 1511.4, 1442.3, and1272.5; GC (dichloromethane, 1.0 uL) 40 oC to 200 oC, 8 oC/minute, product retention time 20.30 minutes, % composition 100%; GC-MS peaks at 119.95 and 165.00 g/mol.

Results and Discussion The synthesis of benzocaine involved the formation of an aromatic ester from a carboxylic acid by means of the Fischer esterification. This synthesis relied on the use of absolute ethanol and 18 M sulfuric acid to form the ester from p-aminobenzoic acid. After this reactant was added, the solution was heated and refluxed for 45 minutes (during which the solids dissolved). After the solution finished refluxing, sodium bicarbonate was added to neutralize the ester and make it insoluble in water. White benzocaine product precipitated out of the solution as a result. The crystals were collected using a Hirsch funnel and vacuum filtration and were rinsed

with cold water three time. This made the crystals very free of impurities. After these steps, the benzocaine product was then analyzed and identified. First the product of the reaction was analyzed by its melting point. A Mel Temp apparatus was used to find the melting temperature of the product. The crystals started melting at 85 oC and finished melting at 88 oC. The known melting range for benzocaine is 88-90 oC.8 The fact that the recorded melting range was so small (only 3 oC) and that the accepted values fell just above this range suggests that a relatively pure benzocaine product was synthesized from the reaction. Further evaluation of the product was needed to confirm its identification. The final product of the reaction was then analyzed through spectroscopic methods, the first being 60 MHz 1H NMR. On the NMR (Figure 2), a doublet peak at 7.71 and 7.56 ppm that has an integration value of 2.20 correlates to the two aryl hydrogens that are closest to the amine group. The next doublet peak 6.63 and 6.48 ppm has an integration value of 2.00 and corresponds to the two aryl hydrogens closer to the ester. The two hydrogens of the amine group are represented by a singlet at 5.91 ppm that has an integration value of 1.70. The quartet that comes right after, at 4.35, 4.25, 4.14, and 4.05 ppm and with an integration value of 1.89, represents the two hydrogens bonded to the carbon that adjacent adjacent to the ester. These peaks are very important because they signal that the ester product was actually synthesized and that the reaction was successful. Lastly, there is a triplet at 1.35, 1.26, and 1.15 ppm that corresponds to the hydrogens of the methyl group at the end of the ester. It has an integration value of 3.37 (three hydrogen atoms) as expected and the chemical shift falls right within the expected range of such protons. There are three peaks representing impurities on Figure 2, one at 3.32 ppm that indicates water, one at 2.50 ppm that indicates DMSO (the solvent used for the NMR), and one at 2.09 ppm that indicates acetone. DMSO can pull water molecules out of the air very quickly,

which is why there is such a high water peak on the NMR.9The acetone peak is present because the NMR tube was washed with acetone and it must not have all been rinsed away. Apart from these peaks, there are no unaccounted-for impurities. The next analysis that was performed to identify the reaction product was IR. The IR of the reaction product (Figure 3) shows a peak at 3339.0 cm-1 that corresponds to an N-H bond (primary amine). Also in Figure 3 is a peak at 2983.3 cm-1, which indicates a C-H bond of an aryl hydrogen. At 1680.1 cm-1 there is a sharp peak that corresponds to the carbonyl of an ester. This is the most important peak of the entire figure because it shows that the carboxylic acid of p-aminobenzoic acid was converted into the ester of benzocaine. The peak at 1511.4 cm-1 correlates to a benzene ring. Next, the peak at 1442.3 cm-1 corresponds to a methyl group (the one at the end of the ester). Finally, at 1272.5 cm-1 there is a peak that indicates a C-O ester bond, another confirmation that the reaction was successful. All of these peaks in the IR spectrum serve as evidence for the creation of the desired product, benzocaine. Figure 3 shows a very pure product with no evidence of starting material present. The last spectral analysis that was performed on the product was GC and GC-MS. The GC (Figure 4) shows the solvent (dichloromethane) peaks having retention times of 1.11 minutes and a product peak having a retention time of 20.30 minutes. The percent composition of the product peak is 100% because there is only one peak. Further analysis with GC-MS confirms the presence of benzocaine. Benzocaine has a molar mass of 165.19 g/mole and the GC-MS (Figure 5) shows a molecular ion peak at 165.00 g/mol. There is also another large peak at about 120 g/mol, which correlates to a loss of a CH3CH2O-group from the molecule. The benzocaine formed from the reaction was quite pure as evidenced by the spectral analyses. The only impurities seen were the slightly depressed melting point and those on the

NMR and were caused by the use of DMSO as a solvent. Moreover, the percent yield for the reaction was quite good at 45.90%. Loss of product could be attributed to the use of many pieces of glassware and loss during vacuum filtration (product stuck to filtration paper).

Conclusion Benzocaine is an important organic compound that has been used as a topical anesthetic for a very long time. The goal of this lab was to synthesize this product by means of the Fischer esterification, reacting p-aminobenzoic acid and ethanol under acidic conditions, and it did just that. After benzocaine was synthesized, it was isolated using saturated sodium bicarbonate and purified using vacuum filtration and water. The experiment was successful in that benzocaine was identified as the product and found to be quite pure. This was evidenced by a melting point range, recorded as 85-88 oC, that is only slightly lower than the literature range of 88-90 oC.8 IR and 60 MHz 1H NMR analyses were also taken, and they showed no starting material peaks. Moreover, a percent yield of product of over 45% demonstrated further experimental success.

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<http://www.sigmaaldrich.com/catalog/product/sigma/e1501?lang=en&region=US>. 9. McLain, S. E.; Soper, A. K.; Luzar, A. The Journal of Chemical Physics. 2007, 127, 174-186.