Laboratory Techniques for Separation of Mixtures

Example 1

Identify which separation method is most suited for the following mixtures:

A mixture of a
Separation A mixture of A mixture of A mixture of
solid dissolved in
methods: solids liquids solid and liquid
a liquid

Evaporation

Distillation

Filtration

Chromatography

Solution

A mixture of a
Separation A mixture of A mixture of A mixture of
solid dissolved in
methods: solids liquids solid and liquid
a liquid

Evaporation NO NO YES* YES**

Distillation NO YES* YES* NO

 Filtration NO NO NO YES

Chromatography YES* YES* YES* YES*

* Success depends on the physical properties of the components in the mixture.

** Would work but filtration is so much faster.

Test Yourself

What method of separation would be most effective on the following mixtures:

a) Sea water
b) Gold nuggets in water.
c) A solution of alcohol (liquid) and water.

Answers

a) evaporation or distillation (chromatography not effective here)

b) filtration

c) distillation

Key Concepts and Summary

Mixtures can be physically separated by using methods that use differences in

physical properties to separate the components of the mixture, such as
evaporation, distillation, filtration and chromatography. Which separation
method used when attempting to separate a mixture depends on what kind of
mixture it is (what states of matter are present) and on the physical properties of
the components.
Exercises

1. What method of separation would be most effective on the following mixtures:

a) Vinegar (a solution of acetic acid (liquid) in water)

b) Loose tea leaves in tea.
c) Copper sulfate (solid) in water.
 2. Identify what physical change occurs during the following separation processes.

a) Distillation of a solution comprising of 50:50 acetone and water

b) Filtration to remove tea leaves from tea.
c) Evaporation for water from a sugar solution to obtain sugar crystals.

d) Taking a sand and salt mixing, mixing it with water, followed by filtration to remove
the sand, then evaporating the salt solution to retrieve salt crystals.

3. Propose a method of separate the following complex mixtures:

a) A mixture of sand, sea water (water and salt)

b) A mixture of marbles, small gold nuggets, and sugar

Answers

1. a) distillation; b) filtration; c) evaporation

2. a) The lower boiling liquid (acetone) would undergo a phase change (evaporation)
upon heating, then once the gaseous acetone comes in contact with the condenser it would
under another phase change (condensation). b) No phase changes, this simply involves
physical removal of the leaves via filtration. c) Water would undergo a phase change
(evaporation) upon heating.

c) The salt would dissolve in the water, then during the evaporation step, water would
undergo a phase change (evaporation) upon heating.

3. a) Filtration to remove the sand, then evaporating the salt solution to retrieve salt
crystals.

b) Manual separation of the marbles (removing by using your fingers), dissolve the rest in
water, then filtration to remove the gold nuggets, and then evaporation of the water to
retrieve sugar crystals.

Glossary

chromatography: is a separation technique based on how the different components in a

mixture have different affinity for the stationary and mobile phase

distillation: is a separation technique used to separate components of a liquid mixture by a

process of heating and cooling

evaporation: is a separation method used to separate of a mixture of a liquid with a dissolved

solid, involving removal of a liquid by evaporating it and leaving behind a solid
filtration: is a separation technique used to separate the components of a mixture containing
an undissolved solid in a liquid by using a funnel lined with filter paper to retain the solids
while letting the liquid through.

Lassaigne Test
The sodium fusion test, or Lassaigne's test, is used in elemental analysis for the qualitative
determination of the presence of foreign elements, namely halogens, nitrogen, and sulfur, in
an organic compound. It was developed by J. L. Lassaigne.
The test involves heating the sample with sodium metal, "fusing" it with the sample. A variety of
techniques has been described. The "fused" sample is plunged into water, and the qualitative tests
are performed on the resultant solution for the respective possible constituents.
The halogens, nitrogen, and sulfur are covalently bonded to the organic compounds are
converted to various sodium salts formed during the fusion. Typically proposed reactions are:
Na + C + N → NaCN
Na + C + N + S → NaSCN
2Na + S → Na2S
Na + X → NaX
The fate of the hydrocarbon portion of the sample is disregarded.
The aqueous extract is called sodium fusion extract or Lassaigne's extract.
Test for nitrogen
The sodium fusion extract is made alkaline by adding NaOH. To this mixture, freshly
prepared FeSO4 solution is added and boiled for sometimes and then cooled. A few drops
of FeCl3 are added and Prussian blue (bluish green) color forms due to formation of ferric
ferrocyanide along with NaCl. This shows the presence of nitrogen in the organic compound.

Test for halogens

The sodium fusion extract is boiled with concentrated HNO3 followed by the addition
of AgNO3 solution which yields a white (dissolves in ammonium hydroxide) (AgCl) or yellow (AgBr)
(sparingly dissolves in ammonium hydroxide) or yellow AgI precipitate (insoluble in ammonium
hydroxide) if halogen is present.

Test for phosphorus

Sodium peroxide is added to the compound to oxidise phosphorus to sodium phosphate. It is
boiled with concentrated HNO3 and then ammonium molybdate is added. A yellow precipitate
indicates the presence of phosphorus.
Test for sulphur
Lead acetate test
The sodium fusion extract is acidified with acetic acid and lead acetate is added to it. A black
precipitate of lead sulphide indicates the presence of sulfur.

Sodium nitroprusside test

Freshly prepared sodium nitroprusside solution is added to the sodium fusion extract, deep violet
colouration appears due to formation of sodium thionitroprusside.
The following are the tests carried to identify the presence of amino group present in the organic
compound.

1. Solubility test
2. Litmus test
3. Carbylamine test
4. Nitrous acid test
5. Azo-dye test
6. Hinsberg test

(a) Solubility Test:

Amines are organic compounds which are basic in nature, so they dissolve in mineral acids like
hydrochloric acid. But this test is not a confirmation test for amines.
The chemical reaction is given below.
C6H5NH2 + HCl → C6H5NH3+Cl–
Note: If it is soluble in mineral acid then it may be an amine.

(b) Litmus Test:

Amines are basic in nature. So it turns red litmus paper blue. This test is also not a confirmation
test for amines.
The chemical reaction is given below.
CH3-CH2-NH2 + H2O → CH3-CH2-NH3+ + OH–
Note: The change in the colour of red litmus paper shows the given organic compound is a base.
(may be amine)
(c) Carbylamine Test:
This test is also called isocyanide test, because when amines are treated with chloroform in the
presence of alkali, isocyanide is formed. Both aliphatic and aromatic amines gives positive for
this test. Secondary and tertiary amines do not give this test.
The chemical reaction is given below.
R-NH2 + 3KOH + CHCl3 → RNC(isocyanide) + 3KCl + 3H2O
Note: An unpleasant odor confirms the presence of primary amine.

(d) Nitrous Acid Test:

This test can be used to identify primary, secondary and tertiary amines. An aromatic primary
amine reacts with nitrous acid to form a diazonium salt and at higher temperatures the formed
diazonium compounds undergoes decomposition.
C6H5NH2 + HNO2 → C6H5-N≡N+Cl–(diazonium compound)
Primary aliphatic amines react with nitrous acid to give nitrogen gas, which is seen as bubbles.
R-NH2 + HONO → R-OH + H2O + N2↑
Secondary amines react with nitrous acid to form a yellow oily nitrosamine.
R2-NH + HONO → R2N-NO + H2O
Tertiary amines react with nitrous acid to form soluble nitrite salts.
R3N + HONO → R3NH+NO2–
Note: Nitrosamines are highly carcinogenic substances.

(e) Azo-Dye Test:

This test is performed in order to confirm the presence of aromatic amines. Aromatic amines
forms a diazonium salts when treated with HNO2. These diazo compounds couple
with phenols like β-naphthols forms an orange azo dye which is insoluble in water.
The chemical reaction is given below.
C6H5NH2 + HNO2 → C6H5-N≡N+Cl–

Note: The presence of red orange dye confirms the presence of aromatic amines.
Distinguishing Test for Primary, Secondary and Tertiary
Amines

(f) Hinsberg Test:

The reagent used in this test is benzene sulfonyl chloride. Amines reacts with benzene sulfonyl
chloride in the alkaline medium.
Primary amines reacts with benzene sulfonyl chloride to produce substituted sulfonamide which
contains an acidic hydrogen and dissolve in basic medium.

A secondary amine forms a substituted sulfonamide which is insoluble in alakali because it does
not have acidic hydrogen.

A tertiary amine forms a substituted sulfonamide which is insoluble in alkali because it does not
have an acidic hydrogen.

Note:

• Tertiary amines: Precipitate in the test tube but dissolves in concentrated HCl.
• Secondary amines: Precipitate in test tube but does not dissolve in concentrated
hydrochloric acid.
• Primary amines: No reaction or precipitate, on adding concentrated hydrochloric acid
insoluble material is seen.

Materials Required:
1. Aniline
2. Potassium hydroxide
3. Sodium nitrite solution
4. Chloroform
5. Dilute hydrochloric acid
 6. Concentrated hydrochloric acid
7. Sodium hydroxide solution
8. Benzene sulfonyl chloride
9. β-Naphthol
10. Test tubes
11. Test tube holder
12. Bunsen burner

Procedure:

(a) Solubility Test:

1. Take the given organic compound to be tested in a clean test tube.
2. Add 2 to 3 drops of concentrated hydrochloric acid.
3. Shake well the solution and observe the solubility.

(b) Litmus Test:

1. Take the moist red litmus paper.
2. Place one to two drops of given organic compound on it.
3. Observe the change in the colour of red litmus paper.

(c) Carbylamine Test:

1. Take potassium hydroxide solid in a clean test tube and add 2ml of ethanol to it.
2. Warm the test tube until the potassium hydroxide dissolves completely in it.
3. To this add a few drops of chloroform and shake well.
4. To the above mixture add a small amount of the given organic compound to be tested
and warm gently.
5. Observe the odour of the resulting mixture.

(d) Nitrous Acid Test:

1. Dissolve the given organic compound in concentrated hydrochloric acid and water in a
clean test tube.
2. Cool the solution in ice bath for 0-5oC.
3. To the cooled solution add sodium nitrite dropwise and mix well.
4. Test the solution for the presence of free nitrous acid.
5. Add excess of nitrous acid solution if there is an absence of nitrous acid.

(e) Azo-Dye Test:

1. Simultaneously take three test tubes and mark it as A, B and C.
2. In test tube A dissolve the given organic compound in 2ml of dilute hydrochloric acid.
3. In test tube B prepare aqueous solution of sodium nitrite solution.
4. In test tube C dissolve naphthol in dilute sodium hydroxide.
5. Place all the three test tubes in an ice bath.Now add the contents in test tube B to the
mixture in test tube A.
6. The resulting mixture is added to the contents in test tube C.
7. Stir the mixture with a glass rod and observe the changes in the test tube.
(f) Hinsberg Test:
1. Take the given organic compound to be tested in a clean test tube.
2. Add 2ml of sodium hydroxide solution and 1ml of benzene sulfonyl chloride to it.
3. Shake the mixture well and add water to dilute the solution.
4. Cool the mixture under tap water and observe the formation of precipitate.
5. If precipitate formed then treat it with concentrated hydrochloric acid.

Observations:

Solubility test If it is soluble in mineral acid then it may be an amine

Litmus test It turns red litmus paper blue

Carbylamine An unpleasant odor confirms the presence of primary amine.

test

Nitrous acid Appearance of bubbles shows the presence of primary amines, yellow oily liquid shows
test the presence of secondary amines and formation of nitrite salts confirms the presence of
tertiary amines.

Azo-dye test The presence of red-orange dye confirms the presence of aromatic amines.

Hinsberg test Primary amines: No reaction or precipitate, on adding concentrated hydrochloric acid
insoluble material is seen.
Secondary amines: Precipitate in test tube but does not dissolve in concentrated
hydrochloric acid.
Tertiary amines: Precipitate in the test tube but dissolves in concentrated HCl.
Laboratory Preparation of Nitrobenzene, purification,Uses
Table of Contents
• Principle involved in laboratory preparation of Nitrobenzene
• Procedure for the laboratory preparation of Nitrobenzene
• Purification of Nitrobenzene
• Uses of Nitrobenzene
• Preparation of nitrobenzene video
• References
Principle involved in laboratory preparation of Nitrobenzene
In the laboratory, nitrobenzene is prepared by heating benzene with a mixture of Conc.
HNO3 and Conc. H2SO4 at 600C. It is an electrophilic substitution reaction as it takes place by
the attack of electrophilic reagent nitronium ion NO2+ at the position of high electron density
in the ring.

The reaction involving preparation of nitrobenzene by nitration of benzene can be represented

as:

Procedure for the laboratory preparation of Nitrobenzene

A mixture of 35 mL of Conc. HNO3 and 40 mL of Conc.H2SO4 are taken in a round bottom
flask. To this flask, 30 mL of benzene is added with constant shaking. Then the mixture is
heated (refluxed) in the water bath at 60oC for about one hour till the yellow oily layer of
nitrobenzene having a bitter almond odor appears on the surface. The flask is then cooled, and
the nitrobenzene layer is separated using a separating funnel.
 Laboratory preparation of Nitrobenzene
Purification of Nitrobenzene
The impure nitrobenzene is contaminated with acid impurities. Thus, to neutralize the excess
of acid, it is first treated with Na2CO3, and then with water several times. Now, It is dried with
anhydrous calcium chloride (CaCl2) to remove water content, and then finally distilled at about
2110C using an air condenser to get pure and dry nitrobenzene.
 Distillation of Nitrobenzene
Uses of Nitrobenzene
• For the preparation of aniline
• For the manufacture of shoe polishes, floor polishes, and so on.
• As an oxidizing agent in organic synthesis.
• Also used for scenting cheap soaps.
References
• Morrison, R. T., & Boyd, R. N., Organic chemistry, Allyn and Bacon, Inc. 1987.
• March, J., Advanced Organic Chemistry, Wiley Eastern Limited, 1986.
• Bahl, B.S., A., Advanced Organic Chemistry, S. Chand and company Ltd, New Delhi,
1992.
• Finar, I. L., Organic Chemistry, Vol. I and Vol. II, Prentice Hall, London, 1995.

Laboratory method of Preparation of Aniline

Laboratory method of preparation of aniline

Principle: In the laboratory, when nitrobenzene is heated with tin and HCl, aniline is formed.
The reaction of tin (Sn) and conc HCl produce nascent hydrogen and SnCl2 or SnCl4, the
nascent hydrogen reduces – NO2 group to – NH2 group.

Method of preparation:

(i) Aniline formation: Conversion of nitrobenzene into phenylammonium ions

A mixture of nitrobenzene, tin and hydrochloric acid is taken in a flask attached with a reflux
condenser and heated at 700C for half an hour, this produces aniline.

Nitrobenzene is reduced to phenylammonium ions using a mixture of tin and concentrated

hydrochloric acid. The mixture is heated under reflux in a boiling water bath for about half an
hour. Under the acidic conditions, rather than getting phenylamine directly, you instead get
phenylammonium ions formed. The lone pair on the nitrogen in the phenylamine picks up a
hydrogen ion from the acid.

(ii) Separation of aniline: The produced aniline further reacts with HCl forming anilinium
hydrochloride salt. When an alkali solution is added to this salt, aniline is separated and remains
floating forming a layer of a dark brown oil.

The division of the compounds in a combination should be as almost quantitative as probable

in order to give some idea of the genuine proportion of each element. Also, it is very significant
to carry out the division in such a method that each compound is obtained in pure shape.

(iii) Extraction of aniline: When the oil undergoes steam distillation aniline is distilled along
with water in the receiver flask. When the distilled liquid is saturated by common salt aniline
is separated in the form of oil. This oil is then separated by a separating funnel.

Nitrobenzene is obtained by evaporating ether. It is also shaken with ether unconnectedly and
ether is collected. Aniline is obtained by evaporating ether.
Fig: preparation of aniline

(iv) Purification: Aniline so found, then dissolved in ether and dried by solid caustic potash.
When it is refluxed at 1840C we get pure Aniline. After this separate aniline from water and
dry it with anhydrous potassium carbonate.

Method of decontamination of aniline is Steam sanitization: Aniline has a very high boiling
point; it is immiscible with water, unstable in steam and having high vapor pressure at the
boiling temperature of the water.

Another common way, Prepare aniline oxalate, recrystallize from 95% ethanol, stimulate free
base with potassium carbonate, dry and distill it under reduced pressure. More widespread
purifications involve preparation of derivatives, such as the double salt of aniline hydrochloride
and cuprous chloride or N-acetylaniline (114 °C) which can be recreated from water.

Aniline is used:

▪ For preparing dyes and dye intermediates.

▪ For produce of antioxidants and vulcanization accelerators in rubber production.
▪ For a mixture of drugs, particularly sulpha drugs.
▪ For making isocyanates required for polyurethane plastics used for insulation.