RECRYSTALLIZATION

Alava, P.J., Alcausin, D.A., Andal M.I., *Bagon, N.E., Barretto, D., Bautista, C. 2D-Pharmacy, Faculty of Pharmacy, University of Santo Tomas

Abstract Most organic compounds that are synthesized in laboratories or isolated from natural products often have been contaminated by impurities. These impurities can be removed by recrystallization, a method of purifying organic solids based on solubility. This method is the most preferred technique for purification of such solids. In this experiment, acetylation of aniline with acetic anhydride was done to synthesize crude acetanilide. Also, the crude acetanilide product was purified by recrystallization. This is done by dissolving them to be crystallized in a hot solvent and then cooled slowly in an ice bath. As it cools, a small, pure seed crystal of the compound forms initially. Additional molecules attach to this crystals layer by layer. The molecules in the crystal have a greater affinity for other molecules of the same kind than they do for any impurities present in the solution. In effect, the process of crystal formation removes one kind of molecule from the solution. The percentage yield calculated from the experiment is 66.96 %. This is the pure acetanilide obtained from aniline and acetic anhydride.

Introduction Generally, organic compounds isolated either from reaction mixtures in the laboratory or natural sources contain small amounts of impurities. In order for them to be used for the manufacture of certain product like a drug, a purification process is required. Various purification techniques are widely used in the industry. For liquids, distillation is most normally used depending on differences in boiling points of the two liquids. In extraction, two immiscible phases is used to separate the substance from one phase into the other based on differences in solubility. In sublimation, separation is on volatile solids which pass directly into vapor phase without passing through the liquid phase. For organic compounds which lack this property, recrystallization method can be used. Recrystallization is the primary method for purifying solid organic compounds. It involves dissolving the impure (crude) product in a minimal amount of hot solvent so that the solution is saturated with the product crystals, and then cooling the saturated solution so that the crystals come out of solution, leaving the impurities

dissolved in the solvent. In this method, the selection of the solvent is the most important aspect. The organic compound that was purified in this experiment is acetanilide. Acetanilide (C6H5NHCOCH3) is an odorless white crystalline solid that is slightly soluble and non-toxic. It is a useful precursor to many pharmaceuticals such as acetaminophen and penicillin. The objectives of this experiment are to (a) synthesize acetanilide by the acetylation of aniline and (b) purify crude acetanilide product by recrystallization. Materials and Methods The selection of recrystallization solvent is the first step performed to determine the best solvent that will yield pure acetanilide through the acetylation of aniline and acetic anhydride. In this, a corngrain amout of pure acetanilide is deposited into three different test tubes. Distilled water was added to the first test tube, 95% ethanol to the second, and hexane to the third. The test tubes were then shaken and placed in a warm water bath (37-40 oC) for 1-5 minutes. After heating, the test tubes were cooled and the solubility of the pure

acetanilide in the different solvents was noted. After the determination of the best solvent, the first crystallization on the experiment was performed. In this, aniline was mixed with distilled water in an Erlenmeyer flask. Then acetic anhydride is slowly added. The flask was cooled in an ice bath to hasten crystallization. When crystals were completely formed, the solution was filtered with the use of a wet filter paper. The crude acetanilide was then dried and weighed. When it was completely dried, distilled water which is the chosen recrystallization solvent was poured into the crude acetanilide. This was heated in a water bath until the solid dissolves. Since the solution was colored, activated charcoal was added to remove those colored impurities. While the solution is still hot, it was filtered immediately using a fluted filter paper. The receiver containing the filtrate was then placed in a beaker with tap water to cool. When the solution is done crystallizing, it was filtered again and the crystals were collected then washed with distilled water and dried. The yielded pure acetanilide was weighed. Results and Discussion Choosing the best recrystallization solvent to use is perhaps the most difficult step in recrystallization process. Recrystallizing solvent is a solvent that shows the desired solubility behaviour for the substance to be crystallized. This is necessary to purify the organic compound. The primary consideration when choosing a recrystallizing solvent is the extent to which the compound and impurities are soluble in the solvent at high and low temperatures. Ideally, the compound to be recrystallized should be insoluble at room temperature, very soluble while heating, and insoluble upon cooling. The unwanted impurities should be soluble at room temperature or insoluble during heating. Moreover, the boiling point of the recrystallization solvent should be lower than the melting point of the

compound to be recrystallized. If the solvent's boiling point is higher than the compound's melting point, the compound will oil out. Oiling out occurs when a compound is insoluble in a solution at a temperature above the compound's melting point. As a result, the compound is deposited as oil, and not as crystals. (Jeffers, 2004) Also, an abundant quantity of crystals must be produced as the solution cools to room temperature or below.

Table 1 presents the solubility of pure acetanilide in water, 95% ethanol, and hexane. In all system, hexane is insoluble whereas 95% ethanol is soluble. This shows that recrystallization will not occur if these two solvents were used. Also, extra precaution must be taken because both hexane and 95% ethanol are highly volatile and flammable. Conversely, water exhibits different result. It shows the desired solubility behaviour of an ideal recrystallization solvent. In water, pure acetanilide is insoluble at room temperature, soluble during heating, and insoluble upon cooling. In acetylation of aniline with acetic anhydride, the produced acetanilide exhibits an exothermic reaction. Exothermic reaction is the liberation of heat from the system. In cooling, the temperature is lowered, thus, the formation of crystals can be observed. The use of ice bath speeds up the crystallization. Table 2. The actual yield of crude and pure acetanilide, and its percentage yield. Weight of the crude acetanilide Weight of the pure acetanilide Percentage yield 1.70 g 1.50 g 50.51%

As shown in Table 2 on the previous page, the weight of crude acetanilide is 1.70 g. The obtained weight was affected by the impurities in the acetylation of aniline with acetic anhydride. During recrystallization process, the solution was colored due to high molecular weight, colored impurities present. This was removed with the use of activated charcoal, often called as Norit or activated carbon. It adsorbs the impurities because it has a large surface area and it can remove impurities more effectively. Only small amount of it was added because too much activated charcoal will result in excessive loss of the desired product. When the acetanilide was dissolved, the solution was filtered immediately while hot. In this case, filtering is the most effective way to remove the solid impurities especially because decolorization was applied. However, during filtration, some part of the solution was lost and was not subjected to filtration that resulted to reduction of the yielded pure acetanilide. Furthermore, washing of the crystals with distilled water was done to help reduce the amount of the solution that remains in the crystals. After the filtration and washing, drying of the acetanilide crystals was done by pressing-in of crystals with filter paper. Some crystals were remaining attached to some part of the filter paper and were not weighed, thus, reducing the weight of pure acetanilide to 1.50 g. To calculate the percentage yield, the limiting reagent and theoretical yield should be identified. Limiting reagent is the reactant that is used up first in a reaction. Theoretical yield is the maximum yield of pure crystals that could be obtained by cooling or evaporating a given solution. This refers to the quantity of pure crystals deposited from the solution. Below is the reaction of aniline and acetic anhydride.

Table 3. Amount anhydride in grams. Aniline 2 mL C6H5NH2 Acetic anhydride 3 mL (CH3CO)2O

of aniline and acetic 1.0217 g x (in 1 mL) 1.082 g x (in 1 mL) 2.0434 g 3.2460 g

Table 3 shows the computation to get the amount of aniline and acetic anhydride in grams from millilitres. This was used to calculate for the limiting reagent as shown in the next page.

Thus, the limiting reagent is aniline because it needs 2.96 g C6H5NH2 to react with 3 mL (CH3CO)2O. From this, we can solve for the % yield using the formula:

In this experiment, actual yield is the weight of the pure acetanilide which is 1.50 g and theoretical yield is the weight of the limiting reagent aniline which is 2.24 g. Substituting these values give:

The percentage yield calculated is 66.96 %. This is the pure acetanilide obtained from the 2 mL aniline and 3 mL acetic anhydride. References http://science.pc.athabascau.ca/labinfo.nsf/ 0/a39dc67b2da8ed33872570cd005ebbc1? OpenDocument http://www.wiredchemist.com/chemistry/inst ructional/laboratory-tutorials/recrystallization http://math.mercyhurst.edu/~cpharr/Courses /OChem2/OChemLab2/OCIIExp7SynthesisofAcetanilideCarboxylicAcidDerivatives.pdf http://www.erowid.org/archive/rhodium/che mistry/equipment/recrystallization.html http://www.chem.ualberta.ca/~orglabs/Tech niques%20Extra%20Info/Recrystallization.ht ml#theory http://www.foothill.edu/psme/armstrong/recr ystallization.shtml http://www.chm.uri.edu/bdeboef/chm292/rex tallization.pdf