Experiment 8B: Qualitative Analysis

Martiene Gabrielle G. Amper 1 and Thea Alissa D.Arceno2

1
College of Public Health, University of the Philippines Manila
Ermita, 1000 Manila
2
College of Public Health, University of the Philippines Manila
Ermita, 1000 Manila

Abstract

Qualitative analysis is an experiment that allows one to determine the functional

groups present in an unknown substance. This experiment aims to identify the unknown

through performing several tests which include Sodium Metal Test and Lucas Test for the

identification of alcohols; Ferric Chloride Test, Bromine in Water Test, and Millons Test for

the identification of phenols; 2,4-DNPH Test, Schiffs Test, Tollens Test, Iodoform Test,

Fehlings Test, Molisch Test, and Benedicts Test for the identification of aldehydes and

ketones and Hinsberg Test for the identification of amines. Also, in the test for carboxylic

acid and its derivatives, the formation of esters is made possible through the reaction of

carboxylic acid and alcohol while also included here are the hydrolysis of the acid

derivatives: benzamide, ester, and anhydride. Hyrdoxamic acid test was also performed.

There were two unknown samples given- a clear solution and a fine solid sample. Both

underwent series of tests in order to verify the results. After attaining positive results from

Millons test, Bromine in Water Test and Ferric Chloride test (for the solid sample) and

Hinsberg Test (for the liquid sample), and in consideration with the solubility test

performed, the unknowns were identified as Tyrosine (for the solid sample) and

Methylamine (for the liquid sample).

I. Introduction

Qualitative analysis is a branch of chemistry that aims to recognize the elements or group of

elements present in an unknown compound(Britannica, 1998). There are five areas of inquiry in an

organic unknown analysis: the physical properties, classification by solubility, spectroscopic analysis,

classification tests for functional groups, and synthesis of solid derivatives (Organic Qualitative Analysis).
 A functional group determines the chemistry of an organic compound. Atoms that are bonded in

specific ways consist these functional groups. They are also responsible for the chemical behavior of the

parent chain. The reactivity of these compounds will determine the chemical composition of a given

substance (Chang, 2010).

In performing this qualitative analysis functional groups namely alcohols, phenols, aldehydes,

ketones, amines, carboxylic acid and its derivatives were tested. The goal of this experiment is to

determine the functional group/s present in the given substances and to identify the specific unknowns.

Series of tests of the functional groups are needed to be performed in order to attain results which can be

use as basis of knowing ones compound. Avoid contaminating the reagents and the glasswares to

ensure a reliable data.

II. Methodology

E. Tests for alcohols and phenols

1. Reaction of alcohols (n-butyl alcohol, sec-butyl alcohol, and tert-butyl alcohol were used as the test

samples)

a. Reaction with sodium metal

In a 5-ml test tube, 20 drops of the alcohol was placed then followed by the addition of the Na

metal. The evolution of gas of the different alcohols were observed and compared.

b. Lucas test

20 drops of Lucas reagent and 10 drops of a given alcohol were placed in a 5 mL test tube. The

test tube was then shaken vigorously and a stopper was placed afterwards. The time the insoluble layer

appeared was recorded. Also, the rate of formation of a layer in the three samples were compared.

c. Reactions with Potassium Dichromate

In a 5ml test tube, 20 drops of a given alcohol was placed . 2M H 2SO4 was used to acidify the

solution and 5 drops of 3% K 2Cr2O7 solution were added. The respective reactions were observed and

noted.

2. Reactions of Phenol (phenol,a-naphthol, catecol and resorcinol were used as test samples)
a. Ferric Chloride Test

20 drops of 95% ethanol and 2 drops of a given phenol were placed in a 5 mL test tube. 5 drops

of 3% FeCl3 solution were added and mixed. The respective observations were recorded.

b. Bromine Water Test

In a 5 mL test tube, 20 drops 95% ethanol and 5 drops of a given phenol were placed. Bromine

water was then added dropwise. The test tube was shaken until the bromine color persisted.

c. Millons Test

5 drops of Millons reagent and 5 drops of a given phenol were placed in a 5 mL test tube and

then shaken. For 2 minutes, the solution was heated in a waterbath. The respective reactions were

observed and recorded.

F. Tests for Aldehydes and Ketones (formaldehyde, acetaldehyde, acetone, benzaldehyde were used

as test samples)

a. 2,4-DNPH test

5 drops of 2,4-DNPH solution and 3 drops of an aldehyde/ketone were placed in a 5 mL test tube.

It was then heated in a water bath for 5 minutes and the respective reactions were observed and

recorded.

b. Bisulfite test

In a 5 mL test tube, 20 drops of methylamine and 5 drops of benzensufonyl chloride were placed.

It was then mixed and placed in an ice bath then the respective reactions were observed and recorded.

c. Schiffs test

20 drops of Schiffs reagent and 5 drops an aldehyde/ketone were placed in a 5 mL test tube. The

respective reactions were then observed and recorded.

d. Tollens test

In a 5 mL test tube, 20 drops Tollens reagent and 5 drops of an aldehyde/ketone were placed. It

was then heated in an ice bath and the respective changes were observed and recorded.

e. Iodoform test
 In a 5 mL test tube, 4 drops of an aldehyde/ketone, 10 drops distilled water, 5 drops 6M NaOH, 5

drops water, and 5 drops I2/KI solution were placed.The respective observations after 10 minutes were

recorded.

f. Fehlings test

20 drops of Fehlings reagent and 5 drops of an aldehyde/ketone were placed in a 5 mL test tube.

It was then heated in a water bath and the respective observations were observed.

G. Tests for the Molisch and Benedicts Tests (1% glucose, 1% sucrose, and 1%boiled starch were

used as test samples)

a. Molisch test

In a 5 mL test tube, 20 drops of Molisch reagent and 5 drops of a sugar were placed. One side of

the test tube was ran down with concentrated 2M H 2SO4. The test tube was then brought upright carefully.

The formation of a blue-violet ring was observed.

b. Benedicts test

20 drops of Benedicts reagent and 5 drops of sugar were added in a 5 mL test tube. It

was then heated in a water bath and the respective results were recorded.

H. Test for Amines

a. Hinsberg Test

Five different amines were used as test samples namely, methylamine, dimethylamine,

trimethylamine, aniline, and N-methylaniline. 20 drops of 10% NaOH, 5 drops of a given amine and 5

drops of benzene sulfonyl chloride were placed in a 5 mL test tube. It was then covered with a cork and

shaken for 5 minutes. The solution was tested with litmus. If the solution was not basic, 10% NaOH was

added dropwise until the red litmus paper turned blue. If there was a formation of precipitate, 40 drops of

water was added and the test tube was shaken. The respective observations were recorded.

I. Test for Carboxylic Acid and its Derivatives

1. Formation of Esters

a. Reaction of Carboxylic Acid and Alcohol

A pinch of salicylic acid and 20 drops of methanol were mixed in a 10 mL test tube. 5 drops of

concentrated H2SO4 was added then the mixture was heated in a water bath for 5 minutes. The

observations were recorded.

2. Hydrolysis of Acid Derivatives

a. Hydrolysis of a Benzamide

In a 5 mL test tube, a pinch of benzamide and 20 drops of 10% NaOH were added. A piece of

moist red litmus paper was placed over the mouth of the test tube during heating in a water bath. Any

change of color in the litmus paper and change in the odor of the mixture were noted.

b. Hydrolysis of an Ester

20 drops of ethylacetate was placed in a 5 mL test tube. 5 drops of 25% NaOH were added to the

solution. The test tube was loosely covered with a cork and was heated in a water bath for 15 minutes.

Using HCl, the solution was neutralized. Any change in the odor of the mixture was noted.

c. Hydrolysis of an Anhydride

20 drops of water and red and blue litmus papers were added to a 5 mL test tube. 20 drops of

acetic anhydride was then added. The test tube was then gently shaken and felt. The observations were

recorded.

3. Hydroxamic Test for Acid Derivatives

The following were used as test samples for this test: ethylacetate, benzamide, acetic anhydride,

and benzoylchloride. In a 5 mL test tube, 5 drops of the sample and 20 drops of 7% methanolic

hyrdroxylamine hydrochloride were mixed. The solution was then added with 10% methanolic KOH until

the mixture turned basic after which 10 more drops were added. The solution was heated to boiling in a

water bath then was cooled slowly. 7% methanolic HCl was then added until the solution turned acidic

afterwhich, 10 drops of 3% FeCl3 were added. The respective observations were recorded.

III. Results and Discussion

 Series of tests were performed to determine what functional group/s are present in the given

unknowns. The results from the series of tests can be seen below:

Table 1.1 Results from reaction with Na metal

Sample Results

n-butyl alcohol Fast evolution of gas

(Primary Alcohol)

sec-butyl alcohol Moderate evolution of gas

(Secondary Alcohol)

Tert-butyl alcohol Slow evolution of gas

(Tertiary Alcohol)

Unknown 1 No evolution of gas

Unknown 2 No evolution of gas

The sodium test for alcohols is used to determine the presence of alcohols. The dissolution of

sodium metal with brisk effervescence indicates the presence of an OH group. (Ahluwalia & Dhingra,

2000). Since there was no evolution of gas when both unknown samples where tested, neither of the two

is an alcohol.

Table 1.2 Results from Lucas test

Sample Results

n-butyl alcohol No layer formation

(Primary Alcohol)

sec-butyl alcohol Moderate layer formation

(Secondary Alchol)

Tert-butyl alcohol Fast layer formation

(Tertiary Alcohol)

Unknown 1 No layer formation

Unknown 2 No layer formation

Lucas test is used for the distinction between primary, secondary and tertiary alcohols. In this

test, Zinc Chloride is used as a catalyst for the reaction to occur. The three alcohols can then be

differentiated by noting the time it took for the two distinct layers to separate. For tertiary alcohols,
separation of layers happens immediately; secondary alcohols within 5-10 minutes; primary alcohols have

no separation of layers. (Ahluwalia & Dhingra, 2000)

After performing this test, it was observed that both unknowns did not form any layer of

separation which may indicate that these two could be primary alcohols.

Table 1.3 Results from the reaction with K2Cr2O7

Sample Results

n-butyl alcohol Dark green solution

(Primary Alcohol)

sec-butyl alcohol Dark green solution

(Secondary Alchol)

Tert-butyl alcohol Yellow orange solution

(Tertiary Alcohol)

Unknown 1 Yellow orange solution

Unknown 2 Yellow orange solution

This test can be used to determine the presence of primary and secondary alcohols, as well as

aldehydes through the formation of a blue-green solution. Tertiary alcohols do not react with potassium

dichromate (VI), thus, the solution remains yellow orange in color. (Clark, Chem Guide, 2003)

The unknowns could possibly be tertiary alcohols since the resulting solution remained yellow

orange in color.

Table 2.1 Results from reaction of R-COOH with R-OH

Sample Results

Salicylic Acid Minty odor; clear solution

Unknown 1 No minty odor

Unknown 2 No minty odor

Esters are derivatives of carboxylic acids which are formed from the reaction of carboxylic acid

and alcohol. The process of forming an ester is formed is called esterification. An example of which is the

formation of methyl salicylate, a compound with minty odor, by heating salicylic acid and methanol, with

the help of H2SO4. (Fromm, 1997)

 After heating the two unknowns with methanol, it did not produce a solution with minty odor.

Therefore, the two unknowns could not be salicylic acid.

Table 2.2 Results from hydrolysis of Benzamide

Sample Results

Benzamide Pungent odor;

red litmus->blue

Unknown 1 No pungent odor;

No change in color of litmus

Unknown 2 Slightly pungent odor;

No change in color of litmus

Hydrolysis, spefically under alkaline conditions, involves the reaction between amides and

hydroxide ions, which in this case is NaOH. In the test performed where benzamide was hydrolyzed, a

pungent smelling, basic compound was formed, which is due to the formation of ammonia. (Clark, Chem

Guide, 2004)

The first unknown, could not be benzamide because the resulting solution did not have a pungent

odor and it did not become basic. The second unknown, although it did have a slightly pungent odor, it still

did not turn basic which means that ammonia was not formed. Hence, none of the unknowns are

benzamide.

Table 2.3 Results from hydrolysis of Ester

Sample Results

Ethylacetate Sweet -> Sour Odor

Unknown 1 No sweet -> sour odor

Unknown 2 No sweet -> sour odor

Hydrolysis of ester is just the reverse reaction of esterification. It proceeds when an ester reacts

with an aqueous base, such as NaOH, to produce a carboxylic acid product and alcohol. In the test

performed, the ester, ethyl acetate, which is a sweet smelling compound was hydrolyzed to produce

acetic acid, a sour smelling compound, and ethanol. (Fromm, 1997)

 Since both of the unknowns did not form a sour smelling compound from a sweet smelling one, it

is evident that the two unknowns were not ethyl acetate.

Table 2.4 Results from Hydrolysis of Anhydride

Sample Results

Ethylacetate Turns blue litmus -> red

Unknown 1 No change in color of litmus

Unknown 2 No change in color of litmus

In this test of hydrolysis of anhydride, ethylacetate turned the blue litmus paper into red which

means that the solution was originally basic then it was turned into an acidic one. Both unknowns showed

negative results from this test.

Table 2.5 Results from Hydroxamic Acid Test for Derivatives

Sample Results

Ethyl acetate White ppt; magenta solution

Benzamide White ppt; brown solution

Acetic Anhydride White ppt; red-orange soln

Benzoylchloride White ppt; violet solution

Unknown 1 White ppt; yellow solution

Unknown 2 White ppt; pale yellow soln

The hydroxamic acid test, also known as Feigl test, is performed to convert ester into hydroxamic

acid by reaction with hydroxylamine. This hydroxamic acid formed is reacted with FeCl 3 to produce a

colored complex as shown in the reaction below:

Carboxylic acids do not give this test, but rather acid derivatives such as acid anhydrides and

chlorides. Most imides, some amides and nitriles also give a positive result to this test. Primary and
secondary nitro compounds also give a positive result due to its aci conformation. (Ahluwalia, V.K. &

Dhingra, Sunita, 2000)

After performing this test to the two unknowns, the results obtained were more clear, yellow-

colored solutions, and not colored complexes which means that none of the unknowns are acid

derivatives.

Table 3.1 Results from Ferric chloride test

Sample Results

Phenol Reddish brown solution

A-naphthol Dark blue solution

Catechol Navy blue solution

Resorcinol Purple solution

Unknown 1 Reddish brown solution

Unknown 2 Clear, yellow solution

This test is used to determine the presence of phenols and enols by reacting a simple with FeCl 3.

A positive result for this test is indicated by a dark colored solution, may it be blue, green, purple, or any

other dark color. (Ahluwalia, V.K. & Dhingra, Sunita, 2000) Considering this, it can be deduced from the

results that unknown 2 is not a phenol or an enol since it did not produce a dark colored solution. On the

other hand, there is a possibility that unknown 1 is a phenol since the color of its product matched that of

the resulting solution of phenol.

Table 3.2 Results from Bromine Water Test

Sample Results

Phenol White ppt

A-naphthol Cloudy, pomelo-colored solution

Catechol Brown solution

Resorcinol Clear, light pink solution

 Unknown 1 White ppt

Unknown 2 Clear Solution

The Bromine water test can also be used to detect the presence of phenol by adding bromine

water into the solution. The presence of a white precipitate would indicate the presence of phenol since

the reaction between the phenol and the bromine water would result to 2,4,6-tribromophenol which is a

white solid. (Clugston, Michael & Flemming, Rosalind, 2000) With this, it can be deduced that unknown 1

contains phenol since it had a white precipitate in the bromine water test while unknown 2 shows

otherwise, which means that it is not or it does not have phenol as one of its functional group.

Table 3.3 Results from Millons test

Sample Results

Phenol Brick red solution

A-naphthol Dark brown

Catechol Navy blue solution

Resorcinol Pomelo-colored solution

Unknown 1 Reddish brown solution

Unknown 2 Clear Solution

The Millons test is usually used to detect the presence of tyrosine which is the only amino acid

with a phenolic group. The positive result for this test is the formation of a red precipitate or red complex.

(Kumar, 2016) Since it was said that this test is usually used to detect the presence of tyrosine, it may be

deduced that unknown 1, having a deep red resulting solution which is a positive result for Millons test is

tyrosine.

Table 4 Results of the Hinsberg Test

Methylamine Precipitate formation

Dimethylamine No precipitation, clear pale yellow solution
Trimethylamine No precipitation, clear pale yellow solution
Aniline 2 layers, up-brown and bottom-cloudy
N-methylanine Brown precipitate, brown solution
Unknown 1 (solid) Clear solution
Unknown 2 (liquid) Clear, pale yellow, little formation of precipitate
 This table showed the results for the test of amines which is the Hinsberg test. The unknown 2

(liquid) showed promising results from the said test, unlike the unknown 1 (solid) which showed negative

results. Trimming down the choices, methylamine and trimethylamine were the two compounds which

showed similar results from the unknown sample. Though during the experimentation, there were some

technicalities that happened. Theoretically, the standard for trimethylamine should not show any evidence

of precipitation but due to contamination of reagents, the standard created formed some precipitate.

Another problem encountered during the experimentation was the lack of supply of the reagent,

specifically the methylamine. Due to this, the solubility test performed in experiment 7 was used as basis

of comparison. The ethylamine in the list portrays similar properties of that of the methylamine.

Table 5 shows the results of the solubility test performed. After series of analysis and comparison

of data, the unknown 2 (liquid) was recognized to be Methylamine.

Table 5 Results of Solubility Test

Sample Water 5% NaOH 5% HCl 5% Ether Conc. 5%H3PO4

NaHCO3 H2SO4
Ethylamine +++ +++ +++ ++ X ++ ++
Unknown 2 +++ +++ +++ +++ X ++ ++

The Hinsberg test is done to determine the presence of amines in a substance. This test is best

for distinguishing primary, secondary and tertiary amines. After addition of diluted HCl, a primary amine

will form a soluble sulfonamide salt which will eventually precipitate. A secondary amine in the same

reaction will directly form an insoluble sulfonamide. A tertiary amine will not react with the sulfonamide but

is insoluble. A soluble ammonium salt will be the result of the conversion of an insoluble amine after

adding a dilute acid. In this way the reaction can distinguish between the three types of amines (Curtin,

Fuson, Shriner, 1956). The rate at which the sulfonyl chloride reagent is hydrolyzed to its sulfonate salt

without the presence of amines, is more timely because of the heterogeneous nature of this system. The

amine dissolves and reacts immediately (if it is 1 or 2), with the neutralization by the base of the

resulting HCl. The sulfonamide derivative from 2-amines is usually an insoluble solid. However, the

sulfonamide derivative from 1-amines is acidic and dissolves in the aqueous base. Acidification of this

solution then precipitates the sulfonamide of the 1-amine (Chemistry of Amines). In the case of the given
unknown it was known to be the first degree amine which is the methylamine. The reaction involved in the

Hinsberg test of methylamine is shown below:

Retrieved from: https://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/amine1.htm

Methylamine is a colorless gas or liquid which resembles the punget odor of ammonia(Pub Chem , 2004).

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Chang, R. (2010). Chemistry, 10th edition. New York, NY: McGraw-Hill.

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Clark, J. (2003). Chem Guide . Retrieved May 9, 2017, from Chem Guide:

http://www.chemguide.co.uk/organicprops/alcohols/oxidation.html

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http://www.chemguide.co.uk/organicprops/alcohols/oxidation.html

Clark, J. (2004). Chem Guide. Retrieved May 10, 2017, from Chem Guide:

http://www.chemguide.co.uk/organicprops/amides/hydrolysis.html

Clugston, Michal & Flemming, Rosalind. (2000). Advanced Chemistry. Oxford: Oxford University

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 Fromm, J. (1997). Retrieved May 10, 2017, from

http://www.3rd1000.com/chem301/chem301v.htm

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