2

SEPARATION AND IDENTIFICATION OF TWO ORGANIC COMPOUNDS

Compounds A & N (dissolved in CH2Cl2)
Extraction with aqueous NaOH

Organic layer containing N in CH2Cl2

Use simple distillation to remove CH2Cl2

Aqueous layer containing A as a sodium salt

Add excess HCl and use suction filtration to filter out A

Isolated N, to be purified

Isolated A, to be purified

This is a 6-hr experiment. Carry out the experiment carefully and seriously. You should take full responsibility of any loss (partial or total) of compounds during the experiment. NO extra sample will be provided.

PART 1: Separation of the Acidic and Neutral Compounds

(refer to videos on ICLM website- General Resources Lab Techniques) The unknown mixture, provided in a small plastic bag, contains an acidic (A) and a neutral (N) compound (abt 2.0 g each).

I. Separation by Solvent Extraction

I.1. As a preparation for the solvent extraction, check the separatory funnel. Fill in a few mL of de-ionized water. Check whether the stopcock leaks. Fit the plastic stopper to the funnel and invert the funnel. Check whether the stopper leaks. Change the funnel and/or stopper if necessary. Place the funnel on the retort

ring. I.2. Transfer the solid sample to a 250mL conical flask and dispense 70 mL of dichloromethane (CH2Cl2) into it to dissolve the sample. [Note the label on the dichloromethane bottle and make sure it is 70 mL.]

I.3. Transfer the solution to the separatory funnel (which should be on the retort ring) with the help of a glass funnel.

I.4.

Dispense 10 mL of CH2Cl2 to rinse the conical flask and transfer the washing to the separatory funnel. [Why rinse? How many mL of organic solution (CH2Cl2 solution) is there?]

I.5.

Add 40 mL of NaOH (2% w/v in water) to the CH2Cl2 solution. With the stopper tightly held, invert the funnel with the drain tube towards the fumehood. Shake the funnel vigorously and immediately release the pressure by opening the stopcock. Always release the built-up pressure in the separatory funnel after each vigorous shaking. Repeat about 3 times. [Incomplete extraction will result in difficulty in recrystallization.]

I.6.

Allow the two layers to separate and remove the stopper of the funnel. Drain the lower (organic) layer into a conical flask; pour out the upper (aqueous) layer into a beaker. [Why the organic layer is the lower layer?]

I.7.

Return the organic layer to the separatory funnel.

I.8. Extract the organic layer again with another fresh 40 mL of 2% NaOH. Combine the aqueous layer in the beaker and return the organic layer to the separatory funnel. I.9. Extract the organic layer a third time with 20 mL of de-ionized water. Drain the organic layer into a dry conical flask. Combine the aqueous layer in the beaker. [How many mL of organic layer is there in the conical flask? How many mL of aqueous layer is there in the beaker?]

II. Recovery of Neutral Compound by Simple Distillation

II.1. Dry the organic layer by addition of anhydrous Na2SO4 (one full spoonful). [Why should the organic layer be dry?]

II.2. Filter the dried organic layer to a dry round-bottom flask. (If the round-bottom flask is not dry, rinse with some acetone and discard the acetone into non chlorinated waste solvent bottle in fumehood B12, G7 or L7.)

II.3. Use simple distillation to remove CH2Cl2 and recover the crude product N. [Remember to put in 2pcs boiling chips.]

II.4. Recrystallization of the crude N after the Solubility Tests.

III.

Recovery of Acidic Compound by Acidification

III.1. Cool the aqueous layer in an ice-water bath and cautiously add 5 ml of concentrated HCl from a pre-adjusted dispenser. Using a glass rod, check if this mixture is acidic to Congo Red paper. If not, add a few more ml of concentrated HCl.

III.2. Recover the precipitated A by vacuum filtration and wash with cold de-ionized water.

III.3 Recrystallization of the crude A after the Solubility Tests.

IV.

Solubility Tests

Solubility test results are CRUCIAL for recrystallization on Week 2. Incorrect solubility test result may lead to no crystals or very low yield. IV.1. Add a small amount (match-head size, about 10 mg) of the compound into a test tube, followed by approximately 2 ml of solvent (de-ionized water or ethanol).

IV.2. Record the solubility at room temperature. [Sometimes the sample sticks to the test tube inner wall instead of dissolving. Excess sample may result in a saturated solution which can no longer dissolve any more sample.]

IV.3. Warm the test tube in a beaker of hot water and record the solubility.

For a successful recrystallization, the compound must be soluble in hot solvent, but relatively insoluble in cold solvent. [Why?]

If you find the solubility test results of one compound (either A or N or both), in a single solvent Z (ethanol or water) as:
Cold Solvent Z NS Hot S

NS: not soluble/poorly soluble

S: soluble

Carry out recrystallization by single solvent Z.

If your compound (either A or N or both) has the following solublities:

Cold Water Ethanol NS S Hot NS S

NS: not soluble/poorly soluble

S: soluble

Carry out recrystallization by solvent pair (ethanolwater mixture).

PART 2: Purification and Identification of the Acidic and Neutral Compounds Technique A: Recrystallization by Single Solvent
A.1. Use either 100% de-ionized water or 100% ethanol as the recrystallization solvent.

A.2. Place the crude compound in a 100-mL conical flask and add a small amount of the solvent (about 10 ml). Add 2 boiling chips and heat the mixture to a gentle boil on the hot plate. Then add hot boiling solvent until all the solid has just dissolved.

CAUTION: If your solvent is ethanol, take special care to control the heating rate so that the ethanol is not completely evaporated off.

A.3. If there is appearance of some insoluble oil at the bottom of the flask, do a hot gravity filtration. To do hot gravity filtration, the funnel and filter paper should be kept hot so that solute will not precipitate on cold filter paper. Therefore, a conical flask with ~5 mL of hot solvent will be heated on hot plate and the funnel is placed on the conical flask. When the solvent vapor rises, the funnel will be heated. Filter the hot solution as fast as possible to minimize precipitation of solute.

A.4. Set the solution aside to cool at room temperature for about 10 minutes and then cool in an ice-water bath (10 minutes) to allow complete crystallization.

Do not disturb or shake the solution during crystallization. Generally, large crystals are produced by allowing crystallization to proceed slowly and without disturbance. A.5. After crystallization is complete, filter the crystals by vacuum filtration and wash the crystals with cold solvent. Disconnect the vacuum for a moment and wash

the crystals with a few mL of cold solvent. Apply vacuum again to remove the solvent.

Technique B: Recrystallization by Solvent Pair

B.1. Use the solvent pair (mixture of ethanol and water) for recrystallization.

B.2. Place the crude compound in a 100-mL conical flask with about 10 mL of ethanol. Add 2 boiling chips and heat on a hot plate until the mixture begins to boil. While maintaining the mixture at a gentle boil, add ethanol until all the solid just dissolves.

CAUTION: Control the heating rate so that ethanol is not completely evaporated off.

B.3. Add hot boiling de-ionized water drop-wise until the solution becomes turbid, and just fails to clear up when it is swirled and boiled.

B.4. To the boiling mixture add ethanol drop-wise until the turbidity just disappears.

B.5. Set the solution aside to cool at room temperature for about 10 minutes and then cool in an ice-water bath (10 minutes) to allow complete crystallization.

B.6. Recover the crystals by vacuum filtration. Disconnect the vacuum for a moment and wash the crystals with a few mL of cold de-ionized water. Apply vacuum again to remove the solvent.

B.7. Dry both compounds under an infra-red lamp.

B.8. Determine the mass of the dried A and N compounds.

V.

Identification of Compounds

V.1. Determine the melting-point range of A and N.

V.2. As the melting range is totally unknown, a combination of a trial run and an accurate run will save a great deal of time. In the trial run, the rough melting point range is determined; in the accurate run, the melting point range is then accurately measured.

V.3. The parameters for the trial run are as follows:

Set point Gradient Max point

60C 20C/min 245C

Record the rough melting point range of both unknown. (The range should not be more than 20oC)

V.4.

The parameters of the accurate run are as follows:

Rough m.p. -5 oC 1 C/min Rough m.p. +5 oC

Set point Gradient Max point

Determine the accurate melting point range of both unknown. (The temperatures should have one decimal place as shown on the machine)

V.5.

Identify A and N by their melting-point range with reference to the table below.

List of Possible Compounds Present in the Unknown Mixture

Acidic Compound m-Toluic acid Benzoic acid Cinnamic acid o-Chlorobenzoic acid Adipic acid Salicylic acid Succinic acid m-Hydroxybenzoic acid Phthalic acid Diphenic acid p-Chlorobenzoic acid Neutral Compound Diphenyl Napthalene Benzil Phenanthrene Dibenzalacetone Acetanilide Benzamide Benzoin Benzenesulphonamide Benzanilide Anthracene Melting-point Range (C) 110111 121122 133134 140142 153154 158161 183185 200201 206208 227228 240243 Melting-point Range (C) 6971 8081 9495 99101 110112 113115 129130 135137 150152 160163 216217