Separation and Purication of the Components of an Analgesic Powder

Darvin Yi with contributions from Eric Welin and Christina Su October 24, 2011

Abstract Separation and purication is an important cornerstone of organic chemistry (and of chemistry in whole). In this experiment, we have separated and puried the contents of an analgesic powder: Goodys R Extra Strength Headache Powder. We have separated out this powders three active ingrediates: acetaminophen (4-acetamidophenol) (1), aspirin (acetylsalicyclic acid) (2), and caeine (3). After separating these components using their known solubility, acidity, and basicity properties, we analyzed each samples purity through melting point analysis, thin layer chromotagraphy (TLC), and a Ferric Chloride test to check for presence of salicylic acid in the aspirin sample. The melting point test revealed that the acetaminophen sample was slightly impure with a melting point of about 4 C lower than stated in the literature, the aspirin sample was relatively pure with a melting point equivalent to literature, and caffeine was quite impure with a melting point about 15 C lower than stated in the literature. The TLC showed a slight impurity in the acetaminophen sample. Finally, the Ferric Chloride test turned slightly purple on aspirin, showing some presence of salicylic acid. Thus, we have shown that the proven techniques for separation and purication of a solid sample can be applied to real life mixtures, such as an analgesic powder.

(a) Acetaminophen

(b) Aspirin

(c) Caeine

Figure 1:
powder.

The three components of the analgesic (a) Acetaminophen has a melting point of

169 170.5 C. It is also insoluble in methylene chloride. We should also note how acetaminophen is indeed soluble in ethanol (EtOH). (b) Aspirin has a melting point of 135 C. Aspirin will dissolve in methylene chloride. Because aspirin is a carboxylic acid, it will become a water-soluble salt in the presence of an aqueous base. (c) Caeine has a melting point of 235 C. Similar to aspirin, it will also dissolve in methylene chloride. Caffeine will become a water-soluble salt in the presence of an aqueous acid.

For the rst part of this experiment, we will utilize the idea of solubility in DCM. Thus, when the analgesic powder is dissolved in the DCM and ltered, we will be able to separate the acetaminophen and inactives, which are insoluble in DCM, from aspirin and caeine, both of which are soluble in DCM. After that part, we can separate acetaminophen from the inactives in a similar way using the knowledge that acetaminophen will dissolve in EtOH but the inactives will not. The second part of the experiment involves separating out aspirin from caeine. Aspirin is acidic, as it is a carboxylic acid. Caeine, however, is more basic, as it has some amine groups. The two amide functional groups of carbon on the right side of the Many chemical industries focus on the important role caeine molecule (as shown in gure 1c) are not as of separation and purication of materials. In this basic as the two amine groups on the left side of the experiment, we will focus on separating and purify- molecule. This is most likely caused by the polarity ing the three main components of Goodys R Extra of the carbon-oxygen doublebond being in such close Strength Headache Powder. The three main compo- proximity. Having such a polar bond near by will help nents of this analgesic powder are shown in gure (1). delocalize the lone pair of electrons on the nitrogen,
Department Lab

of Physics, Princeton University A.B. T.A. for CHM 303 Conductor of Partner Method A Experiment. Images taken from Miami Dade College Chemistry Site.

making the nitrogens of the amide group much less basic. The nitrogens on the amine group, however, have much more localized lone pairs. Thus, these two amine groups make the caeine molecule slightly basic. When an acid in an organic solvent is introduced to an aqueous base, it becomes deprotonated and forms an aqueous salt. Thus, as its solubility changes, the salt will now be in the aqueous layer of the whole solution. Similarly, when a base dissolved in an organic solvent is introduced to an aqueous acid, it becomes protonated and forms an aqueous salt. We can now see two dierent ways in which aspirin can be separated from caeine. In this experiment, we will focus on separating aspirin and caeine using an acid extraction (see gure (2)). An important point about aspirin is that it undergoes a hydrolysis reaction to form salicylic acid (gure 2). Thus, aspirin should have minimum contact with water throughout the experiment. To test for salicylic acid, a ferric chloride test can be performed. Salicylic acid will yield an intensely dark red, blue, purple, or green coloration in the ferric chloride test. This happens when the ferric chloride molecules react with the phenol (in this case, salicylic acid). This reaction will yield a complex of phenol and Fe(III), which is the source of the intense coloration. In the reaction of aspirin to salicylic acid, 1 mole of aspirin react with water to form 1 mole of salicylic acid. 1 mole of aspirin has a mass of 180.157 grams while 1 mole of 138.12 grams.

is set up by having some adsorbent material placed on a plate. This is called the stationary phase. The stationary phase is placed in some solvent, called the mobile phase. Once small spots of the samples in question are placed on to the plate, the TLC can begin. As the mobile phase makes its way up the plate, it will dissolve the samples and carry it upwards. Thus, on a polar plate (such as silica gel), the substance that moved furthest can be identied as least polar and the substance that moved the least can be seen as most polar. This is because when the mobile phase takes up the dierent samples, the more polar sample will have a stronger interaction with the stationary phase, and thus, will be held back more. Quantitative analysis of this can be done by nding the retardation factor of the solutes (Rf ). The equation for the retardation factor is as follows: Length of Travel of the Mobile Phase Length of Travel of the Solute in Question

Rf =

Materials and Methods

Though there is only one way to separate and purify acetaminophen from the analgesic powder, there are two ways to separate aspirin and caeine. The rst method (A) separates aspirin and caeine by aqueous base extraction. The aspirin will become an aqueous salt after it becomes deprotonated by sodium hydroxide. The second method (B) separates aspirin and caeine by aqueous acid extraction. The caeine will become an aqueous salt after it becomes protonated by the hydrochloric acid. In this article, method B will be described. However, later on, results from another experiment modeling method A will also be shared. To summarize, the methods stated in the Organic Puzzle Book (11th ed.) pages 13-21 were used. The experiment detailed in this report will be method B, as described by the previously stated pages of the OPB. Preliminary Separation: The preliminary separation is the same for both method A and method B. 6.0mL of DCM was added to 1.006g of the analgesic powder in a 25mL conical ask. The ask was then heated in a 38 C hot water bath for 5 minutes. This marks the point where all the DCM soluble material (aspirin and caeine) should have been dissolved. The DCM insoluble material (acetaminophen and inactives) were then collected as solids by ltering the solution in a Hirsch funnel. 2.0mL of extra warm (heated in 38 C water bath as well) DCM was added to the conical ask to try to get as much of the residue into the Hirsch funnel as possible. Isolation and Purication of Acetaminophen:

Figure 2: This is the hydrolysis of aspirin yielding

salicylic acid. Salicylic acid has a melting point of 157 159 C. Testing for salicylic acid can be done with the ferric chloride test.

When a sample is pure, it will exhibit some characteristic melting point. However, when a sample is impure, the intermolecular forces between the crystals of the sample and the crystals of the impurities will break the highly organized structure of the original crystal lattice. Thus, because there is less order in the system as a whole, the melting point will drop. Thus, a melting point analysis of the sample will be an indicator of how pure the sample is. The lower the melting point is from the predetermined experimental melting point of a pure sample, the more impure it is. TLC is another way to check for impurities. TLC
This

gure was taken and modied from the Nottingham University Chemistry Bootcamp Site

Once the ltrate has been transfered to a small 25mL conical ask, 2.0mL of EtOH was added and this ask was heated in a 80. C hot water bath for 7 minutes. The contents of the ask were then ltered by Hirsch funnel. An extra 1.0mL of EtOH (heated by 80. water bath as well) was used to collect as much residue from the ask as possible. The ltrate was then dried until all the EtOH evaporated. The solid was then transfered to a medium test tube and dissolved by 1.0mL of water in a 80 C hot water bath. During the dissolving, another 1.0mL of water was added to help the process. Once all the crystals have dissolved, the solution was cooled at room temperature until recrystallization is complete. The crystals were collected by Hirsch funnel. Once the crystals dried, the melting point was found. For a breakdown of the Preliminary Separation, consult gure 3.

was then rapidly transfered to a small conical ask where crystals were collected via Hirsch funnel after cooling and chilling in an ice water bath. After the crystals have dried, the mass was recorded and the melting point was found. Isolation and Purication of Caeine (Method B): To the aqueous solution, 10% sodium hydroxide was added dropwise until the solution tested basic on litmus paper. 4.0mL of DCM was added. At this point, the caeine hydrochloride will have become deprotonated to reform caeine and will be dissolved in the DCM layer. The lower layer was removed to a clean medium test tube. Once 4.0mL of DCM was added again to the aqueous solution, the lower DCM layer was removed to the same test tube. 2.0mL of water was added to the test tube, and the DCM layer was extracted to a 25mL conical ask. The DCM layer was then dried over anhydrous sodium sulfate. Once the DCM solution was dried, the DCM was evaporated by rotary evaporator. The mass of the solid was recorded and the melting point was found. A summary of Isolation and Purication of Aspirin (Method B) and Isolation and Purication of Caeine (Method B) can be found in gure 4.

Figure 3: This is a graphical description of the methods

of Preliminary Separation and Isolation and Purication of Acetaminophen.

Figure 4: This is a graphical description of the methods

of Isolation and Purication of Aspirin (Method B) and Isolation and Purication of Caeine (Method B).

Isolation and Purication of Aspirin (Method B): 3.0mL of 3M HCl was added to the DCM solution. Solution was mixed thoroughly via vigorous pipetting. After separation, the lower DCM layer was pipetted out into a clean medium test tube. 2.0mL of distilled water was added to the extracted DCM layer. After another thorough mixing, the lower DCM layer was pipetted out again. The residual aqueous layer was then combined with the previous residual layer to form the aqueous acidic solution. This will separate out the caeine from the aspirin, which is still in the DCM layer. The DCM solution will then be dried with anhydrous sodium sulfate. The DCM can be evaporated on a rotary evaporater. The solid was then heated in water in a 82 C hot water bath. It
This

TLC Analysis: The original analgesic powder, puried acetaminophen sample, puried aspirin sample, and puried caeine sample were dissolved in a 1:1 ratio of EtOH and DCM. To get everything to dissolve, the 1:1 solution of EtOH and DCM were slightly warmed in a 35 C water bath. The four solutions were then plated on a silicon gel plate via micropipette on the solute line. The plate was then placed in a 400mL beaker with a thin layer of 1015mL of 0.25% glacial acetic acid and ethyl acetate. Care was taken to make sure the mobile phase was under the solute line. After the TLC process was nished, the plate was taken out of the beaker and a line was drawn at the mobile phase front. After outlining

This gure was taken and modied from the Organic Puzzle Book, p.14 gure was taken and modied from the Organic Puzzle Book, p.15

each component of the substance with UV light analysis, the Rf value was computed. Iodine analysis was also conducted by Iodine analysis. Ferric Chloride Test: 3mg of aspirin was suspended in 1.0mL of distilled water in a small test tube. 2 drops of 2.5% aqueous ferric chloride were added. After mixing the contents of the test tube, the coloration was recorded.

the TLC plate was placed into the Iodine chamber, a yellow outline was formed on both acetaminophen spots formed in the original analgesic powder lane and the acetaminophen sample lane.

Ferric Chloride Test: Once ferric chloride was added to the suspended aspirin solution, the solution turned a very light purple. The solution was still clear; however, the color change was also indisputable. Thus, the color change signies that there Results was a reaction between the Fe(III) and a phenol, inRecorded Masses: The masses of each sample at the dicating that there was indeed presence of salicylic end of the experiments have been summarized in ta- acid. However, as the solution was still quite clear ble 1. The sample of aspirin could have been either and the purple was only a very light shade of purple, aspirin or salicylic acid (SA). Thus, on table 1, that there was only a limited presence of salicylic acid. sample has been labeled Aspirin / Salicylic Acid. Also listed in table 1 is the reported mass of each substance by the company that makes the analgesic powder. Table 1: Masses of Final Samples Sample Mass(mg) Correct(mg) Analgesic Powder 1006 Inactives 157 Acetaminophen 142 260 Aspirin / SA 420 520 Caeine 29 32.5 Melting Point Analysis: The results from the melting point analysis has been summarized in table 2. The two controls to determine the calibration on the melting point apparatus were benzoic acid and anthracene. The three values listed are the melting point values from this experiment (B)s samples, another experiment of method A (A)s samples, and the correct (C) value found in literature. All data in table 2 are raw measurements from the melting point apparatus. As we can see, the melting point of both benzoic acid and anthracene are quite close to the literature value. Thus, we can, for the most part, see how the rest of the melting point apparatuss readings will also be accurate. Thus, the corrected values are the raw values; the calibration measurements show no need for a correction. Table 2: Melting Point Results Sample B ( C) A ( C) C ( C) Benzoic Acid 122-4 122-3 Anthracene 215-7 215-7 Acetaminophen 165-6 170-2 169-71 Aspirin / SA 135-8 151-4 135 Caeine 220 205-7 235 Thin Layer Chromatography: The results of the Thin-Layer Chromatography can be seen in gure 5, a digital recreation of the TLC done in the experiment. As can be seen, the acetaminophen sample shows some sign of being contaminated by aspirin. Once 4

Figure 5: This is a digital recreation of the TLC

plate after the chromatography analysis. The spots on the plate were seen using a UV lamp. The Analgesic powder split up nicely into the three dierent bands of caeine, acetaminophen, and aspirin. The acetaminophen has a large spot at acetaminophen because too much acetaminophen solution was plated The Rf of acetaminophen can be calculated at 0.47. The acetaminophen also shows signs of being slightly contaminated with aspirin. The aspirin sample came out nicely without much contamination. The Rf of aspirin can be calculated at 0.67. The caeine solution came out nicely without much contamination. The Rf of caeine can be calculated at 0.12.

lane. However, the ferric chloride test does reveal that there was some salicylic acid production. The ferric Acetaminophen Purity: The acetaminophen pro- chloride test revealed a slight color change to a very duced in this experiment was relatively pure. This light purple. Because this purple was very light, we can be seen from the close melting point. Placing can understand how there was only a small amount the melting point of the experimental sample of ac- of salicylic acid present. etaminophen at 165.5 C and the correct literature Mass Analysis and % Recovery: From page 22 of value at 170 C, the experimental value has a 2.6% the Organic Puzzle Book (11th edition), we know that dierence from the accepted value. Because that is Goodys R Extra Strength Headache Powder has 530 a very small source of error, there was most likely mg of aspirin, 260 mg of acetaminophen, and 32.5 mg no explainable source of error. Slightly miscalibrated of caeine. Subtracting these values from our initial equipment is enough to cause error of this small mag- mass of the analgesic powder, we can nd that there nitude. A more telling factor, however, is the TLC are 193.5mg of inactives. However, as only 157mg analysis. In the TLC analysis, acetaminophen had were collected, the % recovery for the inactives can a Rf value of 0.47. This matches Rf value of ac- be seen to be 81%. Though this is not a great yield, etaminophen seen in the original analgesic. However, it is not enough to say there were any major experthere is also a small spot above the acetaminophen imental aws. The error, however, is not too small spot. This spot had an Rf value of 0.67. This Rf value to write o as equipment-related or stochastic errors. is equivalent to the Rf value of the aspirin in both The main source of error in the loss of the inactives the original analgesic sample and the aspirin sample. probably stems from when the solid residue from the Thus the contaminant is most likely aspirin. EtOH treatment ltration was transported onto the Caeine Purity: The melting point of caeine of to the cupcake cup. Most likely, all of the separated this experiments sample was 220 C. The accepted inactives were not salvaged and much was lost from value from literature is 235 C. Thus, this experimen- the Hirsch funnel to the cupcake cup. The % recovery tal melting point of the puried caeine has an er- of acetaminophen was 54.6. This recovery percentage ror percentage of 6.4. Again, this is such a small is quite low. This error is denitely not statistical, but deviation from the correct melting point that it is experimental. The error most likely originated from not enough to signify a major experimental error. trying to salvage the acetaminophen from the coniThough this is a large error, it is too large to ex- cal ask to the cupcake cup. A better experimental plain with a melting point apparatus miscalibration. model may be to preweigh the conical ask and then One of the main problems of caeine was the limited mass both the conical ask with the acetaminophen, yield. Thus, in the process of trying to scrape out as rather than to lose much of the product in the promuch caeine from the round bottom ask after the cess of transporting the acetaminophen from the ask rotavap as possible, multiple microspatulas were used. onto a cupcake cup. The % recovery of caeine is Many of these could have had some past residues on 89.2. This is quite a high % recovery and can still be them that were not fully cleaned o, leading to some explained through statistical anomaly. Finally, the contamination. Though slight traces of aspirin could % recovery of aspirin is 80.7. This is another low % have also been in the sample, this is unlikely due to recovery that can not be explained with stochasticthe fact that the TLC analysis shows no sign of as- ity. However, it can be explained by the dierence in pirin. There were no additional spots on the TLC mass of aspirin and salicylic acid. We found, through plate other than the caeine spot. The Rf value of the ferric chloride test, how there was indeed some this spot was 0.12, same as the Rf value of the caf- salicylic acid made in the process. From their given data, if all 520mg of supposed aspirin (2.9 moles) in feine spot of the original analgesic powder. Aspirin / Salicylic Acid : Though there was in- the analgesic powder hydrolyzed, a total of 400mg of deed some creation of salicylic acid, we can conclude salicylic acid would be formed. Thus, any product of that the aspirin sample was mostly pure. The melt- aspirin between 400mg and 520mg could be explained ing point of this aspirin sample was at 135 C, exactly by the production of salicylic acid. The lower % rethe same value as the accepted literature value of the covery was most likely due to the fact that some of melting point of aspirin. The TLC analysis showed the aspirin was converted to salicylic acid. the same conclusion of aspirins purity. TLC analysis of the aspirin sample showed only one spot with an Rf value of 0.67, same as the Rf value of aspirin from the original analgesic powder sample. Under both the UV illumination and the I2 , there were no dierences in the appearance of the aspirin spot from the aspirin lane from the aspirin spot from the analgesic powder 5 Salicylic Acid Production Though this experiments Method B results showed only a minor production of salicylic acid, a partner Method A reaction had a completely dark and opaque purple result from its ferric chloride test. The larger amount of salicylic acid produced from Method A experiments can be explained by the dierences in separation/purication

Discussion

methods. In method A, aspirin and caeine in DCM were extracted with aqueous sodium hydroxide, resulting in sodium acetylsalycylate in water. Aspirin was later extracted by HCl addition to become aspirin and water. However, in method B, aspirin and caffeine in DCM were extracted with aqueous hydrochloric acid, resulting in aspirin in DCM. Thus, because aspirin was extracted to an aqueous solution, there was a higher chance that it would enter into the hydrolysis step. However, in the method B methods, the only real interaction aspirin could have had with water was during the addition and mixing of hydrochloric acid. Thus, it makes sense that much less salicylic acid was produced in method B than in method A. There will probably be a dierence in the production of salicylic acid depending on which chemical (HCl or NaOH) comes in contact with the aspirin. The two products of aspirin hydrolysis are salicylic acid (a monohydroxybenzoic acid) and acetic acid, both acids. Thus, in an acid environment, this reaction that produces acidic compounds would not be as favorable in an environment with a higher pH. Thus, NaOH will most likely drive this hydrolysis reaction forward in comparison to HCl. To test this, we can add aspirin straight into a concentrated NaOH sample and an equally concentrated HCl sample. We should nd that the NaOH environment will produce more salicylic acid. Smell of Vinegar : It is quite understandable how the smell of vinegar can be present when an old bottle of aspirin is opened. Vinegar is essentially acetic acid; thus, acetic acid is the main source of the vinegar smell. We also know that the products of the hydrolysis of aspirin is salicylic acid and acetic acid. Thus, when aspirin has been kept for a very long time in its bottle, some of it could have been exposed to moisture within the bottle. Thus, with ample amounts of time, the stochastically insignicant chance of a hydrolysis reaction in a new bottle can be magnied to something nontrivial, leading to production of both salicylic acid and acetic acid. This is most likely the source of the smell of vinegar within an old bottle of aspirin. Acetaminophen Soluble in DCM : If acetaminophen were soluble in DCM, we could not use the same experimental procedures. This would mostly come in during the fact that acetaminophen would not be ltered out from the initial treatment with warm DCM (as it would be soluble in DCM). We know that acetaminophen is a phenol, a class of

organic molecules that is slightly acidic. Thus, when treated with NaOH, it would also form an aqueous salt, and join sodium acetylsalicylate in the aqueous solution. Thus, when HCl is added, the aqueous solution will not only have water and aspirin, but it will also contain acetaminophen. Thus, we must devise of an alternate experimental pathway that can let us separate acetaminophen and aspirin. We know that aspirin is a carboxylic acid. In general, carboxylic acids are much stronger acids than phenols. Thus, we can extract out aspirin rst from the massive DCM solution via weak base extraction. There are many possible weak bases that can be used, including, but not exclusively, alanine, ammonia, methylamine, pyridine, and acetylacetone. The same general procedures can be followed after that point to recrystallize aspirin. After that, we can extract the acetaminophen with the sodium hydroxide solution (a strong base). The same procedures can be followed after that point to recrystallize acetaminophen. A ow chart representation of this separation schematic can be seen in gure 6.

Figure 6: This is a graphical description of the methods of purication and separation of the components of the analgesic powder if acetaminophen were soluble in DCM.

Conclusion: As we have shown, we can use acid/base extraction methods to separate out the components of an analgesic powder. The acetaminophen, aspirin, and caeine were all relatively pure; however, the aspirin did have some traces of salicylic acid, as shown by the ferric chloride test. By utilizing known properties of substances, we can creative methods that will give us the ability to separate and purify each individual substance within a mixture.

Honor Code
I pledge my honor that this paper represents my own work in accordance with university regulations. Darvin Yi