Classification Tests for Hydroxyl- and Carbonyl- Containing Compounds

Janelle Angela O. Siy, Mary Jane S. Sta. Ana, Samantha Hope A. Sy, Kevin G. Turingan, Fraz Joseph Theodore B. Yap, and James Christian J. Yuchongco Group 10 2B Medical Technology Organic Chemistry Laboratory

This very interesting experiment helped students identify the presence and structure of a hydroxyl, aldehyde, or ketone in an unknown compound using seven different tests. The first three tests done, solubility of alcohols in water, Lucas test and Chromic test helped identify the structure of alcohols, whether it is primary (if the alpha carbon atom where the OH group is attached is only attached to one alkyl group), secondary (if the alpha carbon atom is attached to two alkyl groups), or tertiary (if the alpha carbon atom is attached to three alkyl groups). The fourth test done was the 2, 4-Dinitrophenylhydrazone (2,4-DNP) test and this was used to identify the presence of carbonyl groups- aldehydes and ketones. More specifically, as stated in our Laboratory Manual, non-conjugated (aliphatic) carbonyl compounds usually yield yellow color in the reaction while conjugated (aromatic) ones yield orange red color. Hence, the 2, 4-DNP test is used to identify the presence of carbonyls in a compound and also distinguish its aromaticity. The fifth and sixth tests were the Fehlings and the Tollens Silver Mirror tests. Both of these were used to identify the presence of an aldehyde, thus distinguishing an aldehyde from a ketone. Only aldehydes yield a positive result in this test while ketones are non-reactive. Last but not the least, the Iodoform Test was used to see the presence of CH3CO group or methyl carbonyl group in a substance. The samples that were used are n-butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, acetaldehyde, n-butyraldehyde, benzaldehyde, acetone, acetophenone, and isopropyl alchohol.

ABSTRACT

INTRODUCTION

Figure 4- The structural formula of tert- butyl alcohol [1] The objectives of this experiment are to be able to Tert-butyl alcohol is a tertiary hydroxyl compound since distinguish whether a compound is hydroxyl- or carbonyl- the alpha carbon atom at which the hydroxyl group is containing, to be able to differentiate the three types of attached is surrounded by three alkyl groups, one methyl alcohols, aldehydes from ketones, and explain theon each of the three sides. mechanisms involved in the differentiating tests. The Hence, answering one of our objectives, the three types samples used were ethanol, n-butyl alcohol, sec-butylof alcohols are primary, secondary and tertiary alcohols, all alcohol, tert-butyl alcohol, benzyl alcohol, acetaldehyde, n- of which are part of the hydroxyl group. butyraldehyde, benzaldehyde, acetone, acetophenone, and isopropyl alcohol.

Figure 1- The structural formula of ethanol Ethanol is a primary two carbon hydroxyl compound.

Figure 5- The structural formula of benzyl alcohol Benzyl alcohol is an aromatic hydroxyl group.

Figure 2- The structural formula of n-butyl alcohol N-butyl alcohol is a primary hydroxyl compound since Figure 6- The structural formula of acetaldehyde Acetaldehyde, also called ethanol, is an aliphatic, nonthe hydroxyl (OH) group is attached to a primary alpha carbon atom. Primary alpha carbon atoms refer to the conjugated, carbonyl (C=O) compound, more specifically carbon atoms that are attached to only one alkyl group. In a aldehyde, since it is terminal. Aldehydes are reactive to oxidation. this case, it is attached to a propyl group.

Figure 3- The structural formula of sec-butyl alcohol Sec-butyl alcohol is similar to n-butyl alcohol except that Figure 7- The structural formula of n-butyraldehyde N-butyraldehyde, also called butanal, is a four carbon it is a secondary hydroxyl compound since the hydroxyl aliphatic, non-conjugated aldehyde. group (OH) is attached to a secondary alpha carbon atom. It can be seen in the figure that OH is attached to the second carbon atom, having an ethyl group on one side, and a methyl group on the other. Hence, it is surrounded by not one, but two alkyl groups.

Figure 8- The structural formula of benzaldehyde

ZnCl2 and HCl is the reagent. The alcohol is protonated by this mixture, and H2O group attached to carbon is Benzaldehyde, also known as phenol, is an aromatic, replaced by the nucleophile Cl-, which is present in excess. Tertiary alcohols react immediately with Lucas reagent as conjugated aldehyde compound. evidenced by turbidity owing to the low solubility of the organic chloride in the aqueous mixture. Secondary alcohols react within five or so minutes (depending on their solubility). Primary alcohols do not react appreciably with Lucas reagent at room temperature. Hence, the time taken Figure 9- The structural formula of Acetone for turbidity to appear is a measure of the reactivity of the Acetone, also known as propanone, is a carbonyl group, class of alcohol, and this time difference is used to more specifically, a ketone. Unlike aldehydes, it isnt differentiate between the three classes of alcohols: terminal. Most ketones dont react in an oxidation reaction. no visible reaction at room temperature and cloudy only on heating: primary, such as normal amyl alcohol (1Pentanol) solution turns cloudy in 35 minutes: secondary, such as sec-amyl alcohol (2-Pentanol) solution turns cloudy immediately, and/or phases separate: Figure 10- The structural formula of acetophenone tertiary, such as tert-amyl alcohol (2-Methyl-2-butanol) Acetophenone is an aromatic ketone compound.

Figure 12- the reaction in Lucas Test. Chromic Acid test is also known as Jones test. [3] The Jones oxidation is an organic reaction used to oxidize alcohols using chromic trioxide and acid in water. A primary alcohol is oxidized to an aldehyde or all the way to a carboxylic acid, while a secondary alcohol to a ketone. Figure 11- The structural formula of isopropyl alcohol The mechanism begins with the reaction of CrO3 with acid Isopropyl alcohol is a secondary three carbon hydroxyl (often H2SO4) to form chromic acid or dichromic acid in compound. more concentrated solutions. The alcohol oxidation then In short, ethanol, n-butyl alcohol, sec-butyl alcohol, tert- occurs with chromic acid which in turn gets reduced in the butyl alcohol, benzyl alcohol, and isopropyl alcohol are all process. Hence, this is used to check the presence of under the hydroxyl group. Acetaldehyde, n-butyraldehyde, primary and secondary alcohols, both of which would and benzaldehyde are all under the aldehydes group while indicate a positive result, which would be the formation of acetone and acetophenone are under the ketone group. All green color. aldehydes and ketones are under the carbonyl group. Benzaldehyde, benzyl alcohol and acetophenone are the only ones that are aromatic because they have a conjugated and cyclic structure, benzene. The first test was to identify the alcohols solubility in water. The fewer the carbon atoms are in a substance, and the lesser the branches, the more miscible it is in water. The trend of solubility from most soluble to least soluble is as follows: primary ROH>secondary ROH>tertiary ROH>aromatic ROH This test was used to identify whether it is a primary, secondary, tertiary hydroxyl compound or an aromatic Figure 13- reaction of (chromic acid test) Jones Test The 2, 4- Dinitrophenylhydrazone (2, 4-DNP) Test is alcohol. This was also used to compare the quantity of used to determine the presence of carbonyl groups such as carbons the different samples have in relation with each aldehydes and ketones in a substance. A positive result other. The Lucas test uses the Lucas reagent to identify would yield a yellow-orange precipitate, due to the whether an alcohol is primary, secondary or tertiary. [2]characteristic of the formed dinitrophenylhydrazone. This Lucas' reagent is a solution of anhydrous zinc chloride in also helps determine whether the carbonyl group is concentrated hydrochloric acid. This solution is used to aliphatic or aromatic. An aromatic substance would yield classify alcohols of low molecular weight. The reaction is a more of the red-orange color while aliphatic ones would substitution in which the chloride replaces a hydroxyl yield more of the yellow color. group. A positive test is indicated by a change from colorless to turbid, signaling formation of a chloroalkane. It is based on the difference in reactivity of the three classes of alcohols with hydrogen halides. The differing reactivity reflects the differing ease of formation of the corresponding carbocations. Tertiary carbocations are far Figure 14- reaction mechanism of 2, 4-DNP test more stable than secondary carbocations, and primary The Fehlings test is used to determine the presence of carbocations are the least stable. An equimolar mixture of an aldehyde in the solution and it doesnt react with ketone. The reagent of this test is cupric ion in an alkaline

solution. Aldehydes, when mixed with copper ion, would There was no need to prepare the Lucas reagent reduce the copper and create a mossy green solutionsince it is available in the lab. Three test tubes were which will eventually form a brick red precipitate of prepared and labeled as n-butyl chloride, sec-butyl cuprous oxide, Cu2O.

chloride, and tert-butyl chloride. One mL of Lucas reagent was put on each of the three test tubes, and then two to three drops of each sample were placed in their respective test tubes. The test tubes were shook for a few seconds. The three test tubes were Figure 15- reaction mechanism of Fehlings test then placed on the test tube rack for the samples to The Tollens Silver Mirror Test also has the same function settle in. The rate of formation of the cloudy as the Fehlings test. Its aim is to identify the presence of a carbonyl group and further differentiate them as either an suspension and the formation of two layers were aldehyde or a ketone. The reagent of this test consists of observed. 3. Chromic acid test (Jones Oxidation) silver ion from mixing silver oxide and ammonia. Aldehydes would reduce the reagent to metallic silver while For this test, seven compounds, as well as seven ketones wont have any reaction. The positive result must test tubes were used. The seven test tubes were look like a formation of a silver mirror. labeled accordingly as n-butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, n-butyraldehyde, benzaldehyde, acetone and acetophenone. Each contained one mL each of their corresponding samples. Two drops of 10% aqueous K2Cr2O7 solution and 5 drops of 6M H2SO4 were then added to each test tube. Color change and its rate were Figure 16- the reaction of Tollens test The last test, Iodoform test, is used to see the presence observed. 4. 2, 4- Dinitrophenylhydrazone test of CH3CO in aldehydes and ketones. A positive result would produce yellow precipitate, and if there is no CH3CO Five test tubes with their respective labels to their group in the sample being tested, then it will get acorresponding samples, 1 mL each, were utilized in different result. this test. The samples were acetone, acetaldehyde, n-butyraldehyde, benzaldehyde and acetophenone. Five drops of 95% ethanol was added to all five test tubes. These test tubes were shook thoroughly. Three drops of 2, 4-DNP were then added afterwards. The results exhibited were noted. When there was no reaction for some test tubes, they were subjected to 15 mins of settling at room Figure 17- the reaction of Iodoform test temperature. After that, the observations were EXPERIMENTAL recorded. A. Compounds Tested (Samples Used) 5. Fehlings Test The aliphatic hydroxyls: Ethanol, N-Butyl alcohol, sec-butyl alcohol, tert-butyl alcohol, isopropyl alcohol Since the Fehlings reagents were already available in the lab, there was no need to prepare it. However, The aromatic hydroxyl: benzyl alcohol The aliphatic aldehydes: acetaldehyde, n-there was a need for the students to mix equal amounts of Fehlings A and B. Six mL of Fehlings A butyraldehyde was put inside a small graduated cylinder, then it The aromatic aldehyde: benzaldehyde was mixed with 6 mL of Fehlings B solution. There The aliphatic ketone: acetone were five test tubes, with the labels acetaldehyde, The aromatic ketone: acetophenone n-butyraldehyde, acetone, benzaldehyde and B. Procedure acetophenone. The Fehlings reagent was navy blue 1. Solubility of Alcohols in H20 in color. One mL of Fehlings reagent was added to Five test tubes were labeled as ethanol, n-butyl each of the test tubes. Three drops of each sample alcohol, sec-butyl alcohol, tert-butyl alcohol, and were then dropped on their corresponding test tubes. benzyl alcohol. Using the Pasteur pipet, 10 drops of The tubes were placed in a water bath to boil for 10 the corresponding samples were added in each of the-15 minutes. After, the results were observed. five test tubes. One mL of water was added to each of the five test tubes and the test tubes were shaken. A fourth of an mL of water was added when cloudiness occurred. This was done repeatedly until the solution became homogenous. The amount of water that was needed to dissolve each compound Figure 18 Fehlings reagent (Fehlings A + B) was taken note of.

2. Lucas Test

Since the explanation for the different tests and their balanced chemical equations were quite lengthy, they were indicated in the introduction instead. 1. Solubility of Alcohols in H20 The alcohol that was most soluble in water, needing the least amount of water to be miscible to it, was Figure 19 Researcher (Sy,ethanol. This is because it is a primary alcohol, and Samantha) with one of the samples reacting withmoreover, among all the alcohols tested, ethanol had Fehlings reagent. Two layers are seen here, greenthe least number of carbon atoms. Tert-butyl was on top, blue at the bottom. This was the transition of miscible in water however it required more water to do so. This is because tertiary hydroxyls have a color before it turned brick red. harder time bonding with water as compared to primary ones. Benzyl alcohol wasnt miscible in water at all because the hydroxyl group is attached to an aromatic compound, which is only sp2 hybridized, Figure 20 A closer look of the color of the which makes it even more immiscible. sample in figure 19 Table 1 Solubility of Alcohols in water
Alcohol Structural Formula Amount of H2O in mL needed to produce a homogenous dispersion 1 mL Solubility in water

RESULTS AND DISCUSSION

Figure 21 The samples

Ethanol

Most readily Miscible Miscible Miscible Least readily Miscible

Figure 22 heating the samples in a water bath on top of the hot plate Sec-

n-butyl alcohol

2mL 2.5mL 3mL

butyl alcohol 6. Tollens Test Tollens test was already present in the laboratory; Tertthat was why there was no need to prepare the butyl reagent. Five samples were used in this test, namely, alcohol

acetaldehyde, benzaldehyde, acetone, nBenzyl 3mL Immiscible butyraldehyde, and acetophenone. All had their alcohol own test tubes with the corresponding labels. Each of the five empty test tubes contained 1mL of fresh Tollens reagent. Two drops of each of the five samples were placed in their respective test tubes. 2. Lucas Test The mixture was shook and was allowed to stand for N-Butyl alcohol didnt have any reaction with this 10 minutes. When no reaction occurred for some of test because Lucas Test is non-reactive towards them, the tubes were placed in a warm water for 5 primary hydroxyls. The sec-butyl alcohol slowly minutes. The results were recorded. formed a cloudy mixture while the tert-butyl alcohol 7. Iodoform Test formed an immediate cloudy solution which later on Two drops of each sample, acetaldehyde,produced two distinct layers. This is because Lucas acetone, acetophenone, benzaldehyde, andtest gives positive result for tertiary hydroxyls. This isopropyl alcohol were placed in their own vials. is because the Lucas test needs a significantly stable Twenty drops of 10% KI (potassium iodide) solution carbocation. was placed in each tube. Twenty drops of fresh chlorine bleach (5% sodium hypochlorite) were placed while the tube was being slightly shaken. The formation of a yellow precipitate was watched out for.

Table 2 Reaction of Alcohols with Lucas Test being aromatic, yielded a clear yellow liquid with red Substance Structural Reaction with orange precipitate. formula Lucass test Table 4 Carbonyl groups reaction to 2, 4-DNP N-butyl alcohol No reaction test
Substance Sec-butyl alcohol Tert-butyl alcohol Cloudy (slow) Cloudy (immediate) 2 layers Acetaldehyde Structural formula 2, 4-DNP test result Yellow liquid with yellow ppt.

3. Chromic acid test (Jones Oxidation) The samples that yield a positive result for this test means that they are primary or secondary alcohols. Benzaldehyde Aldehydes also have a positive result with this test, ketone doesnt. N-butyl alcohol, sec-butyl alcohol, Nbutyraldehyde and benzaldehyde all fit in the category; therefore, they all got a positive result Acetone exhibited by the appearance of green color from the oxidized Chromium ion. Tert-butyl alcohol, being a tertiary hydroxyl group, wasnt reactive with this test. Acetone and acetophenone, both ketones, were acetophenone not reactive with this as well. Table 3 The reaction of aldehydes, ketones and alcohols to Chromic acid test (Jones test)
Substance n-butyl alcohol Sec-butyl alcohol Tert-butyl alcohol nbutyraldehyde Benzaldehyde Structural formula With chromic acid Immediate shift from orange to blue green Immediate shift from orange to blue green No color change

nbutyraldehyde

Yellow liquid with yellow ppt. Yellow turbid immiscible liquid Yellow liquid with yellow ppt.

Clear yellow liquid with red orange ppt.

5. Fehlings Test Acetaldehyde, n-butyraldehyde, and benzaldehyde all got a result of brick red or bright red precipitate with this test because this test aims to identify aldehydes, and all these three are aldehydes, thus the positive result. Acetone and acetophenone on the other hand had no reaction at all with this test since they are ketones. Table 5 Fehlings Test Reactions
substance Acetaldehyde Structural formula Reaction to Fehlings test Brick red ppt.

Orange to brown to mossy green Orange to green

nbutyraldehyde Acetone No color change Benzaldehyde Acetophenone No color change Acetone

Bright red ppt.

Brick red ppt.

None

4. 2, 4- Dinitrophenylhydrazone test acetophenone Acetaldehyde, n-butyraldehyde and acetone all got yellow precipitate because they are consists of carbonyl groups and they are aliphatic. Benzaldehyde, although aromatic in itself, just yielded a yellow turbid immiscible liquid and didnt 6. Tollens Test exhibit orange like precipitate. This is just a form of laboratory error. Acetophenone on the other hand, it

None

Acetaldehyde, N-butyraldehyde, and benzaldehydeC-OH group of the isopropyl alcohol would turn into got a positive silver mirror in this test because this CH3CO upon subjecting it to iodoform test. test is similar to Fehlings test, its positive result is Benzaldehyde got a different result because it from aldehydes, and all the three mentioned above doesnt contain any CH3CO group. are aldehydes. Acetone and acetophenone didnt get Table 7 Iodoform Tests Results Substance Structural formula Iodoform any reaction with this test since they are ketones. test result Table 6 Tollens Test Reactions
substance Acetaldehyde Structural formula Reaction to Fehlings test Silver mirror Acetaldehyd e Yellow ppt.

Benzaldehyd e nbutyraldehyde Benzaldehyde Silver mirror Acetone Silver mirror Acetopheno ne

Red immiscible droplets Yellow ppt.

Acetone

None

Yellow ppt.

acetophenone

None

Isopropyl alcohol

Yellow ppt.

7. Iodoform Test Acetaldehyde, acetone, acetophenone, and isopropylREFERENCES alcohol all formed yellow precipitate because this test[1] Bayquen, A.V., Cruz, C.T., de Guia, R.M., Lampa, aims to identify the presence of a methyl carbonylF.F., Pena, G.T., Sarile, A.S., Torres, P.C. (2009). group (CH3CO) in a compound. A positive result of Laboratory Manual in Organic Chemistry. Manila, this would indeed incur yellow precipitate. Seeing thePhilippines: C&E Publishing, Inc. structural formulas, all the ones mentioned above [2] Lucas Reagent contain CH3CO group, except for isopropyl alcohol. http://en.wikipedia.org/wiki/Lucas%27_reagent The reason why isopropyl alcohol got a positive [3]Jones Oxidation http://www.nameresult, as explained by Maam Daya, was because thereaction.com/jones-oxidatio