Camille Loscalzo Kyle Reichl 4/21/2011 Chem 213-103 Synthesis of 3-Carbethoxycoumarin Introduction In terms of organic synthesis, esterification and

condensations reactions make a powerful combination. Generally, esterification is one of the most significant and fundamental chemical reactions in organic chemistry.1 Types of esterification reactions are numerous, and include Fischer esterification, transesterification, and more. Condensation reactions just as abundantly contribute to the field of organic chemistry. In general, condensation reactions involve the combination of two larger molecules through the release of a small molecule. Biologically, condensation reactions are essential to the formation of macromolecules from monomers, such as the dehydration (through which water is the small molecule released) of glucose and fructose to produce sucrose, a disaccharide. Novel frameworks of polymers that are porous, crystalline, and composed of covalent bonds are being constructed via condensation reactions.2 In this experiment, a member of the coumarin family of heterocylic compounds is synthesized by a coupling of esterification and condensation reactions. Naturally occurring coumarins are responsible for the fragrant odor in a number of plants, including vanilla grass, sweet grass, and sweet clover. Both natural and synthetic coumarins demonstrate broad pharmaceutical activity, including anti-tumor, anti-viral, anti-inflammatory, anti-oxidant, antimicrobial and enzyme inhibition functionality.3 4-Hydroxycoumarins comprise an industrially standard category of coumarin derivatives, even so much that 4-hydroxycoumarins themselves are often loosely called coumarins. Coumarins, in this sense, are most renowned for their strong

anti-coagulate properties. As anticoagulants, coumarins serve as the active ingredient in drugs and rodenticides in which they function by inhibiting Vitamin K activity. Transesterification and aldol condensation reactions specifically are utilized for the formation of a number of coumarins. Transesterification involves the use of an ester as a starting product to synthesize another different ester. Aldol condensation involves the reaction of an enol or enolate ion with a carbonyl group to eventually produce a conjugated enolate. The process is described in detail in the following balanced chemical reaction and mechanism:

Figure 1: Transesterification and aldol condensation to synthesize 3-carbethoxycoumarin

The above reaction is initiated by the deprotonation of diethyl malonate by the weak base piperidine. The deprotonated diethyl malonate then attacks the carbonyl group of salicylaldehyde, displacing an electron pair in the double bond onto the oxygen molecule of the bond. The negatively charged oxygen molecule is then protonated by the acid ethanol. Aldol condensation is initiated as the hydroxyl group of salicylaldehyde-diethyl malonate intermediate attacks an intermolecular carboxyl group. This action once again displaces an electron pair in the double bond onto the oxygen molecule of the bond. When the electron pair reforms the carboxyl bond with carbon, the ether group is displaced as the conjugate base of ethanol. This final step of the reaction involves dehydration to 3-carbethoxycoumarin through the addition of heat. 3-Carbethoxycoumarin will be synthesized from salicyladehyde and diethyl malonate via a transesterification followed by an aldol condensation reaction. The reaction will be monitored by TLC, and 3-carbethoxycoumarin produced will be isolated by vacuum filtration. Characterization of the product will be evaluated by melting point and IR, 60 MHz 1H NMR, 400 MHz 1H NMR, and 400 MHz 13C NMR spectroscopy. Experimental 3-Carbethoxycoumarin. Ethanol (4 mL), piperidine (20 drops), and glacial acetic acid (4 drops) were added to a solution of salicylaldehyde (1.1 mL) and diethyl malonate (1.7 mL) in a 25-mL round-bottom flask. The flask was equipped with a water condenser mounted by a plastic drying tube filled with Drierite. The reaction mixture was refluxed and stirred for two hours and monitored by TLC (40% EtOAc/hexane). The solution was cooled to room temperature and placed in an ice bath. The resulting white solid was washed with cold ethanol (3 mL) and collected via vacuum filtration. The product was recrystallized (95% ethanol), and the crystals

were allowed to dry for five days. IR 984.6-1371.5, 1126.8, 1448.5-1482.7, 1560.2-1607.5, and 2917.9-3062.8 cm-1; 60 MHz 1H NMR 1.191-2.042 (t, 3H), 4.242-4.596 (m, 2H), 7.158-7.786 (m, 4H), and 8.534 (s, 1H) ppm; 400 MHz 1H NMR 1.4002-1.4358 (t, 3H), 4.3901-4.4436 (m, 2H), 7.3310-7.3762 (m, 2H), 7.6434-7.6626 (m, 2H) and 8.5501 (s, 1H) ppm; 400 MHz 13C NMR 14.2291, 61.9111, 76.9240-77.5623, 116.6594, 117.8088-118.0921, 124.9153, 129.5998, 134.4085, 148.6650, 155.0453, 156.7295, 162.9104 ppm. Results and Discussions 3-Carbethoxycourmain was synthesized by the reaction of a salicylaldehyde and diethyl malonate solution with ethanol, piperidine, and acetic acid under reflux conditions. The product was isolated via vacuum filtration, recrystallized, and then characterized by means of melting point and a number of different spectroscopy analyses. Salicyladehyde and diethyl malonate acted as the essential starting reagents for the synthesis of 3-carbethoxycoumarin. The functional groups and relative polarities of these molecules made then amendable to transesterification and aldol condensation reactions. Piperidine acted as a weak base to initiate transesterification by deprotonating one of the extremely acidic hydrogen molecules located on a carbon adjacent to carboxyl groups on either side. Ethanol was added to perform the protonation of the salicyladehyde-diethyl-malonate intermediate, following the initial carbonyl group attack. Acetic acid was added as an organic solvent for the reaction to proceed in, maintaining an appropriate concentration of free hydrogen ions. The reaction was monitored by TLC using 40% ethyl acetate in hexanes as the mobile phase specifically designed for the separation of salicylaldehyde and 3-carbethoxycoumarin. Salicylaldehyde has one carbonyl group to the two carbonyl groups of 3-carbethoxycoumarin. Therefore, salicylaldehyde is a less polar molecule than 3-carbethoxycoumarin and has a lower

Rf value. After the synthesis had proceeded for 20 minutes, TLC analysis indicated that the reaction mixture lacked any trace of salicylaldehyde (a higher spot with an Rf value of 0.306), but only contained 3-carbethoxycoumarin (a lower spot with an Rf value of 0.0918). The presence of both salicyladlehyde and 3-carbethoxycoumarin (a lower spot with an Rf value of 0.102) was detected in the starting mixture and the co-spotting lane during TLC analysis. Furthermore, the results of the 3-carbethoxycoumarin synthesis were exceptionally positive in terms of product yielded and product purity. The percent yield of 3carbethoxycoumarin was extremely high at 74.9%. The product unaccounted for in the yield may be accounted for in a number of ways. Transesterification and aldol condensation does not always induce a 100% conversion of products to reactants, as the calculation of theoretical yield assumes. Therefore, much of the product that is unaccounted for may never have been synthesized due to the limits of the mechanism and the quality of the experimental materials and starting compounds. In addition, there is always loss of product due to glasswear transfers and purification protocols. Purification was simple and efficient: the crude 3-carbethoxycoumarin was washed with cold ethanol and isolated via vacuum filtration, followed by recrystallization with 95% ethanol. Ethanol was utilized for purification because its presence throughout the synthesis did not interfere with the formation and stability of 3-carbethoxycoumarin. The purity of the product was verified by a number of analysis techniques. The accepted melting point of 3carbethoxycoumarin is 92-94C, and the experimentally determined melting point of the product was within this range (92C). An impure sample would have a depleted melting point, and the melting point range would be much more broad than that measured for this experiment.

Furthermore, the various forms of spectroscopy preformed on the product shows indicate a lack of serious contamination by any foreign molecules. Overwhelming evidence indicating that 3-carbethoxycoumarin was indeed synthesized by means of this experiment was found using IR, 60 MHz 1H NMR, 400 MHz 1H NMR, and 400 MHz 13C NMR spectroscopy. IR spectroscopy confirms the synthesis of 3-carbethoxycoumarin by peaks indicative of every functional group present in 3-carbethoxycoumarin. In addition, the spectrum is able to differentiate salicylaldehyde from 3-carbethoxycoumarin by the existence and absence of certain specific peaks. For instance, there were peaks in the 1470-1430, 13801370, and 1490-1440 cm-1 ranges, which are indicative of carbon hydrogen bonds in the umbrella and scissor bends that would appear in 3-carbethoxycoumarin but not salicylaldehyde (due to the ester functional group in the product). Each NMR analysis revealed concrete evidence that 3-carbethoxycoumarin was produced by the transesterification and aldol condensation reaction. The 60 MHz and 400 MHz 1H NMRs showed hydrogen peaks in all the correct positions. Furthermore, these spectra lack hydrogen peaks that would be shown if the sample was contaminated by salicylaldehyde, such as a single peak of an integral value of one hydrogen at 9.0-10.0 ppm (representative of a aromatic alcohol hydrogen). Both 1H NMR spectra also revealed characteristic hydrogen quartet peaks at around 3.5-4.8 ppm, each with an integration of 2 hydrogen molecules. These indicate a CH2 group split by a adjacent methyl group. Lastly, the 13C 400 MHz showed the correct number (12) chemically different carbon molecules. In conclusion, the synthesis of 3-carbethoxycoumarin from salicylaldehyde and diethyl malonate was highly successful. Not only have melting point and IR, 60 MHz 1H NMR, 400 MHz 1H NMR, and 400 MHz 13C NMR spectroscopy each individually confirmed that the

product synthesized is 3-carbethoxycoumarin, but furthermore, 3-carbehoxycoumarin was produced in high yield (74.9%) and high purity. References 1. Ishihara, K.; Ohara, S.; Yamamoto, H. Science, 2000, 290.5494, 1140. 2. Cote, A. P., et al. Science, 2005, 310.5751, 1166. 3. Riveiro, M. E.; De Kimpe, N.; Moglioni, A.; Vazquez, R.; Monczor, F.; Shayo, C.; Davio, C. Current Medicinal Chemistry, 2010, 17.13, 1325-1338. 4. Minard, B.; Bortiatynski, J.; Masters, K.; Halmi, T.O.; Williamson, K.L. Lab Guide for Chemistry 213, 2010-2011, p.p. 1-306.