PRACTICAL LAB MANUAL - JRU

PRACTICAL LAB MANUAL

LAB MANUAL PHARMACEUTICALCHEMISTRY II

D.PHARMA 2nd YEAR

SL NO. LIST OF EXPERIMENT

1To perform the qualitative analysis of given sample of acetyl salicyclic acid(Aspirin)

2To perform the qualitative analysis of given sample of caffeine

3To perform the qualitative analysis of given sample of paracetamol

4To perform the qualitative analysis of given sample of acetazolamide

5To perform the qualitative analysis of given sample of chlorpromazine hydrochloride

6To perform the qualitative analysis of given sample of penicillin(Benzylpenicillin)

7To perform the qualitative analysis of given sample of Ampicillin

8To perform the qualitative analysis of given sample of sulphonamides

9To perform the qualitative analysis of given sample of barbiturate

10To perform the qualitative analysis of given sample of Carbohydrate

11To carry out the Identification test for the given sample of urea

12To carry out the Identification test for the given sample of alcohol(Ethanol)

13To carry out the Identification test for the given sample of aldehyde (Benzaldehyde)

14To carry out the Identification test for the given sample of ketone(Acetone)

15

To carry out the Identification test for the given sample of carboxylic acid(Benzoicacid)

16To carry out the Identification test for the given sample of phenol(Resorcinol)

17 To carry out the Identification test for the given sample of ester(ethyl aceatate)

18 To carry out the Identification test for the given sample of amines(Aniline)

19 To prepare acetanilide from aniline.

20 To prepare Aspirin from salicylic acid.,

21 To prepare picric acid from phenol

22 To prepare 2,4,6- Tribromophenol from Phenol.

EXPERIMENT NO.- 1

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample of acetylsalicyclic acid(Aspirin)

APPARATUS REQUIRED-

Test tube, test tube holder, test tube stand, mortar pestle, fractional weights.CHEMICALS REQUIRED-

Dilute sodium hydroxide, sodium nitrite solution, sulphuric acid and hydrochloric acid.

CHEMICAL TESTS FOR ASPIRIN-

SL NO. EXPERIMENT OBSERVATION INFERENCE

1 Boil about 0.5 g with 10 ml of 1Nsodium hydroxide solution forthree minutes, cool, and add 10 mlof dilute sulphuric acid. Filter,dissolve the precipitate in waterand to this solution add ferricchloride test solution.

2 Boil about 0.5 g with 10 ml of 1Nsodium hydroxide solution for 3minutes, cool, and add 10 ml ofdilute sulphuric acid.Filter and tothe filtrate add 3 ml of sulphuricacid, and warm.

RESULT:-

The given organic compound is found to be __________________

EXPERIMENT NO.- 2

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample of caffeine.

APPARATUS REQUIRED- Test tube, test tube holder, test tube stand, mortar pestle, fractionalweights.

CHEMICALS REQUIRED- Dilute hydrochloric acid, potassium chlorate, dilute ammonia solution,tannic acid solution, 0.1 N iodine, sodium hydroxide.

CHEMICAL TESTS FOR CAFFEINE-

SL NO. Experiment Observation Inference

1 Take 10 mg in a porcelain dish,add 1 ml of hydrochloric acid and0.1 g of potassium chlorate andevaporate to dryness on a waterbath. Expose the residue to thevapours of dilute ammoniasolution.

2 To a saturated solution add a fewdrops of tannic acid solution.

3 To 5 ml of saturated solution, add1.5 ml of 0.1 N iodine. Add a fewdrops of dilute Hcl. Add sodiumhydroxide to the precipitate.

RESULT- The give organic compound is found to be ______________

EXPERIMENT NO.- 3

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample of paracetamol

APPARATUS REQUIRED- Mortar and pestle, volumetric flask, UV Spectrophotometer.

CHEMICALS REQUIRED- Sodium hydroxide, paracetamol, distilled water.

PROCEDURE:-

By UV Spectrophotometry: The tablets were assayed spectrophotometrically using the followingprocedures. The average weight of tablet from each sample was determined by weighing tentablets and dividing the result by ten.

1. Two tablets were then crushed using a clean pestle and mortar (i.e. from each sample).2. For each sample, powder containing 0.05g (50mg) of paracetamol was accurately weighed and

transferred into different 100ml volumetric flasks. All the 7 samples were labelled using a penand a masking tape.

3. To Each volumetric flask, 50ml of 0.1 M NaOH and 100ml of distilled water were added, andsonicated for few minutes to dissolve the drug molecule. After sonicating, the volume wasmade to 100ml with distilled water.

4. The mixture in each flask was then mixed well and filtered through a filter paper into cleanbeakers.

5. From the filtrate, 10ml was taken using a pipette and transferred into a 100ml volumetric flask;distilled water was then added to make up the volume.

6. From the resultant solution above (6), 10ml was taken with a pipette into a 100ml volumetricflask and 10ml of 0.1M NaOH was added, distilled water was then added and make up thevolume (5µg/ml).

7. The UV Spectrophotometer was put at zero by running a baseline (between 200-400nm) using0.1 M NaOH solution as blank. The absorbance of each sample was determined at 257nm, byputting small amount of the sample into a cuvette, and the cuvette was put into the machine.

8. The same procedure was repeated for the standard using 100mg of the powdered standard, andabsorbance determined, which was used to calculate the percentage content (in mg) ofparacetamol from each brand.

9. The concentration of each sample was also determined using Beer Lambert’s law according to

IP 8.

RESULT- The average weight of different brands of Paracetamol tablets were calculated andtabulated in

EXPERIMENT NO.- 4

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample ofacetazolamide.

APPARATUS REQUIRED- Test tube, test tube holder, test tube stand, mortar pestle, weights.

CHEMICALS REQUIRED- sodium hydroxide, zinc powder, hydrochloric acid, and coppersulphate.

CHEMICAL TESTS FOR ACETAZOLAMIDE-


1

2

RESULT- The given organic compound is found to be ____________

EXPERIMENT NO.- 5

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample ofchlorpromazine hydrochloride


CHEMICALS REQUIRED- Dilute Hydrochloride acid, potassium chlorate, dilute ammoniasolution, tannic acid solution, 0.1 N iodine, sodium hydroxide.

CHEMICAL TESTS FOR CHLORPROMAZINE HYDROCHLORIDE-


1 Dissolve 5 mg in 5 ml ofsulphuric acid.Divide in 2 portions.

Warm one portion.

Add 0.2 ml of 0.1 Npotassium dichromate toanother portion.

2 Dissolve 5 mg in 5 ml of waterand add one drop of ferricchloride test solution.

3 Determine the melting point.

4 Test for Chloride(a) Dissolve in 2 ml of water aquantity of the substance beingexamined equivalent to about 2mg of chloride ion. Acidify withdilute nitric acid and add 0.5 mlof silver nitrate solution.

(b) Introduce into a test tube aquantity of the substances beingexamined equivalent to about 1omg of chloride ion, add 0.2 g ofpotassium dichromate and 1 mlof sulphuric acid. Place a filterpaper strip moistened with 0.1 mlof diphenylcarbazide solutionover the opening of the test tube.

RESULT- The give organic compound is found to be_____________

EXPERIMENT NO.- 6

AIM OF THE EXPERIMENT-To perform the qualitative analysis of given sample of penicillin(Procaine penicillin)


CHEMICALS REQUIRED-Dilute hydrochloric acid, 0.1% w/v solution of sodium nitrite, b-naphthol solution containing 1 g ofsodium acetate.

CHEMICAL TESTS FOR PROCAINE PENICILLIN-

SL NO Experiment Observation Inference

1 Azo Dye Test: Dissolve 0.1 g in 2ml of dilute hydrochloric acid withthe aid of heat if necessary, cool inice, and add 4 ml of 0.1 % w/vsolution of sodium nitrite, and pourthe mixture in 2 ml of b-naphtholsolution containing 1 g of sdiumacetate.

RESULT- The give organic compound is found to be_______________

EXPERIMENT NO.- 7

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample of Ampicillin.

APPARATUS REQUIRED-Test tube, test tube holder, test tube stand, mortar pestle, fractionalweights.

CHEMICALS REQUIRED- Solution of Ninhydrin, potassium cupri-tartrate solution.

CHEMICAL TESTS FOR AMPICILLIN-


1 Place 0.1 ml of a 0.1 % w/vsolution of ninhydrin on a filterpaper, dry at 105 °C, superimpose0.1 ml of 0.1% of the substancebeing examined, heat for 5 minutesat 105°C, and allow it to cool.

2 Suspend 10 mg in 1 ml of water;add 2 ml of a mixture of 2 ml ofpotassium cupri-tartrarte solutionand 6 ml of water.

RESULT- The given organic compound is found to be____________

EXPERIMENT NO.- 8

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample ofsulphonamides(Phthalylsulphathiazole).

APPARATUS REQUIRED-Test tube, test tube holder, test tube stand, mortar pestle, fractionalweights.

CHEMICALS REQUIRED- Lead acetate paper, dilute sodium hydroxide, sodium nitrite solution,sulphuric acid, hydrochloric acid, resorcinol.

CHEMICAL TESTS FOR PHTHALYLSULPHATHIAZOLE-


1 Heat stronglyphthalylsulphathiazole and bring apaper dipped in lead acetate closeto the mouth of test tube.

2 Boil 2 g with 20 ml of sodiumhydroxide solution for 10 minutes,cool, add 15 ml of hydrochloricacid and slowly add sodiumbicarbonate solution until no morecarbondioxide is evolved.Recrystallize the precipitate fromwater. Dry at 105°C and take itsmelting point.

3 Heat 0.1 g with 50 mg of resorcinoland 1 ml of sulphuric acid for about1 minute, cool, pour into water andmake alkaline with sodiumhydroxide solution.


EXPERIMENT NO.- 9

AIM OF THE EXPERIMENT- To perform the qualitative analysis of given sample ofbarbiturate(Thiopentone sodium)

APPARATUS REQUIRED-Test tube, test tube holder, test tube stand, fractional weights.

CHEMICALS REQUIRED- Dilute acetic acid, sodium nitroprusside, sodium hydroxide,dilutehydrochloric acid 15% w/v solution of potassium carbonate, potassium antimonate solution,magnesium uranyl acetate solution.

CHEMICAL TESTS FOR THIOPENTONE SODIUM-


1 Acidify 5 ml of 5.0% w/v solutionwith dilute acetic acid and filter.Wash and recrystallize. Wash theorecipitate with water. Dry. Takethe melting point.

2 Dissolve 1 mg of crystals obtainedin the above test and to it add 1 mlof 0.1 N NaOH. Add about 1 mg ofsodium nitroprusside and after 15minutes, add 1 ml of dilute HCl.

3 Evaporate the filtrate obtained inchemical test(1) and the residue leftbehind gives the tests for sodium.

Test for sodium(a) Dissolve 0.1 g of the substancesbeing examined in 2 ml of water.Add 2 ml of a 15% w/v solution ofpotassium carbonate, heat toboiling. Allow to cool in ice waterand if necessary rub the inside ofthe test tube with a glass rod.

(b) Acidify a solution of substancebeing examined with 1N acetic acidand add a large excess ofmagnesium uranyl acetate solution.


EXPERIMENT NO.- 10

AIM OF THE EXPERIMENT-To perform the qualitative analysis of given sample of Carbohydrate

APPARATUS REQUIRED- Beaker, glass rod, volumetric flask 100 ml and 500 ml

CHEMICALS REQUIRED- Sulphuric acid, naphthol, Fehling's solution

CHEMICAL TESTS FOR -Molisch’s Test:

This is a common test for all carbohydrates larger than tetroses. The test is on the basis that pentosesand hexoses are dehydrated by conc. Sulphuric acid to form furfural or hydroxymethylfurfural,respectively. These products condense with α-naphthol to form purple condensation product.

Furfural α-Naphthol

Fehling’s Test:

This forms the reduction test of carbohydrates. Fehling’s solution contains blue alkaline cuprichydroxide solution, heated with reducing sugars gets reduced to yellow or red cuprous oxide and isprecipitated. Hence, formation of the yellow or brownish-red colored precipitate helps in the detectionof reducing sugars in the test solution.

Benedict’s Test:

As in Fehling’s test, free aldehyde or keto group in the reducing sugars reduce cupric hydroxide inalkaline medium to red colored cuprous oxide. Depending on the concentration of sugars, yellow togreen color is developed . All monosaccharides are reducing sugars as they all have a free reactivecarbonyl group. Some disaccharides, like maltose, have exposed carbonyl groups and are alsoreducing sugars,but less reactive

RESULT- The given organic compound is found to be_____________

EXPERIMENT NO.-11

AIM OF THE EXPERIMENT- To carry out the Identification test for the given sample of urea.

APPARATUS REQUIRED- Ignition tube, pair of tongs, test tube, test tube holder, china dish,tripod stand, wire gauze, funnel and filter paper.x

CHEMICALS REQUIRED- Sodium metal, ferrous sulphate, Lead acetate, sodium nitroprusside,sulphuric acid, sodium hydroxide, ferric chloride, silver nitrate solution, carbon tetrachloride, chlorinewater, 1N hydroxylamine hydrochloride, 1N KOH solution.

ORGANOLEPTIC EVALUATION:-


1 Physical test

2 Colour

3 Odour

4 Ignition test: Take a small amountof sample in spatula and ignite itover flame.

5 Solubillity tests

Sample+cold water

Sample+ hot water

Sample+ cold dil. NaHCO3

Sample + cold dil. NaOH

Sample+ cold conc. H2SO4

Sample + hot conc. H2SO4

Sample+dil.HCl

Insoluble in water, acid and alkali

6 Tests for unsaturation(a) Test with bromine water:Dissolve 1 ml of sample in waterand add few drops of brominewater.

(b) Baeyer's test: Dissolve 1 ml ofsample in water and add 1 ml ofdilute Na2CO3 solution and a fewdrops of KmnO4 solution. Shakewell.

7 Nature: Dip a wet litmus paper inthe sample.

EXTRA ELEMENTAL DETECTION

Lassaigne's extract was prepared for the given sample.


1 Test for Nitrogen: To 1 ml of sodiumextract, add very little quantity of FeSO4crystals. If precipitate is not obtained, add adrop of sodium hydroxide. Heat thecontents to boiling and add dilute H2SO4.

2 Test for Sulphur(a) To 1 ml of sodium extract, add fewdrops of sodium nitroprusside solution.

(b) To 1 ml of sodium extract, add aceticacid followed by lead acetate solution

(c) Sodium plumbite test: To lead acetatesolution, add a drop of NaOH and observe.Then add excess of NaOH. Add sodiumextract.

3 Test for Nitrogen and Sulphur: To 1 mlof sodium extract, add ferric chloridesolution

4 Test for Halides(a) To 2 ml of sodium extract, add diluteHNO3. Boil the solution to expel H2S orHCN, if present, and then add few drops ofsilver nitrate solution.

(b) To 2 ml of sodium extract, add diluteHNO3. Add 1 ml of carbon tetrachlorideand a few drops of freshly preparedchlorine water.

TESTS FOR AMIDE


1 Reaction with NaOH solution:Take 0.2 g of the substance in aboiling tube and add 2 ml of 10%NaOH solution. Boil the contents.Bring a glass rod dipped in conc.Hcl near the mouth of the tube.

2 Hydroxamic Acid Test: In aboiling tube, take 0.1 g of thesubstance with 2 ml of 1Nhydroxylamine hydrochloride and2 ml of 1N KOH solution. Boilthe mixture for 5 minutes; cooland add 2-3 drops of ferricchloride.

3 Biuret test: Heat 0.1 g ofsubstance in a dry test tube, asublimate of biuret with theevolution of NH3 is formed onthe sides of test tube. Dissolvethis in 2 ml of distilled water; adda drop of very dilute CuSO4solution and 2 ml of NaOHsolution.

4 Urea nitrate tesrt: To 0.5 g ofurea, add 3-5 drops of conc.HNO3 and heat. Cool.

RESULT-The given organic compound has _____________ as an extra element and ____________________as the functional group.

EXPERIMENT NO.- 12

AIM OF THE EXPERIMENT-To carry out the Identification test for the given sample ofalcohol(Ethanol)

APPARATUS REQUIRED- Ignition tube, pair of tongs, test tube, test tube holder, china dish, tripodstand, wire gauze, funnel and filter paper.

CHEMICALS REQUIRED- Iodine solution, sodium bicarbonate, sodium hydroxide, conc.Sulphuric acid, hydrochloric acid, bromine water, potassium permanganate, sodium metal, ferroussulphate crystals, sodium nitroprusside, ferric chloride, nitric acid, silver nitrate solution, carbontetrachloride, chlorine water, ceric ammonium nitrate, salicylic acid, Lucas’ reagent, sodiumdichromate, 3,5-dinitrobenzoyl chloride, potassium dichromate, resorcinol.



1 Physical test

2 Colour

3 Odour

4 Ignition test: Take a small amount of samplein loop and ignite it over flame.

5 Solubillity tests

Sample+cold water

Sample+ hot water





Sample+dil.HCl


6 Tests for unsaturation(a) Test with bromine water: Dissolve 1 ml ofsample in water and add few drops of brominewater.

(b) Baeyer's test: Dissolve 1 ml of sample inwater and add 1 ml of dilute Na2CO3 solutionand a few drops of KmnO4 solution. Shakewell.

7 Nature: Dip a wet litmus paper in the sample.




1 Test for Nitrogen: To 1 ml ofsodium extract, add very littlequantity of FeSO4 crystals. Ifprecipitate is not obtained, add adrop of sodium hydroxide. Heatthe contents to boiling and adddilute H2SO4.

2 Test for Sulphur(a) To 1 ml of sodium extract, addfew drops of sodium nitroprussidesolution.

(b) To 1 ml of sodium extract, addacetic acid followed by leadacetate solution

(c) Sodium plumbite test: To leadacetate solution, add a drop ofNaOH and observe. Then addexcess of NaOH. Add sodiumextract.

3 Test for Nitrogen and Sulphur:To 1 ml of sodium extract, addferric chloride solution

4 Test for Halides(a) To 2 ml of sodium extract, adddilute HNO3. Boil the solution toexpel H2S or HCN, if present, andthen add few drops of silvernitrate solution.

(b) To 2 ml of sodium extract, adddilute HNO3. Add 1 ml of carbontetrachloride and a few drops offreshly prepared chlorine water.

FUNCTIONAL GROUP DETECTION

Test for AlcoholsSL NO. Experiment Observation Inference1 Cerric-ammonium nitrate Test: To

1 ml of the substance, add a fewdrops of ceric ammonium nitratereagent.

2 Sodium test: To 2 ml of theunknown, add a small piece ofsodium metal

3 Ester Formation:Heat about 0.5ml of the sample with 1 ml of theacetic acid and a few drops of conc.H2SO4 in a dry test tube for abouta minute. Cool and pour thecontents in a beaker containingwater; observe the odour.

4 Reaction with Acetyl chloride: To1 ml of the substance add 2-3 dropsof acetyl chloride. Bring a roddipped in NH4OH in contact withthe vapours evolved.

5 Lucas’ test: To 1 ml of alcohol in asmall test tube, add quickly 6 ml ofLucas’ reagent at 26-27℃, close thetube with cork, shake and allow itto stand. Observe the mixture.

6 Benzoylation: Take about 500 mgof powdered 3,5-dinitrobenzoylchloride in a dry test tube and add 2ml of the sample. Warm themixture in a beaker hot water untila clear solution is obtained. Coolthe tube.

7 Confirmatory TestIodoform test:- Take 5 ml of ethylalcohol and 2 ml of 10% solution ofsodium hydroxide. Add iodinesolution drop by drop until slightiodine colour persists. Keep thetube in warm water bath for 15minutes. Take out and cool

RESULT- The given organic compound has _______________ as an extra element and ----------------as the functional group.

EXPERIMENT NO.- 13

AIM OF THE EXPERIMENT- To carry out the Identification test for the given sample ofaldehyde(Formaldehyde)

APPARATUS REQUIRED- Ignition tube, pair of tongs, test tube, test tube holder, platinum loop,china dish, tripod stand, wire gauze, funnel and filter paper.

CHEMICALS REQUIRED- Sodium bicarbonate, sodium hydroxide, conc. Sulphuric acid,hydrochloric acid, bromine water, potassium permanganate, sodium metal, ferrous sulphate crystals,sodium nitroprusside, ferric chloride, nitric acid, silver nitrate solution, carbon tetrachloride, chlorinewater, 2,4-dinitrophenylhydrazine solution, sodium bisulphate, Schiff’s reagent, Fehling’s solution A,Fehling’s solution B, Benedict’s reagent, resorcinol.



1 Physical test

2 Colour

3 Odour

4 Ignition test: Take a small amount ofsample in loop and ignite it over flame.

5 Solubillity tests

Sample+cold water

Sample+ hot water





Sample+dil.HCl


6 Tests for unsaturation(a) Test with bromine water: Dissolve 1ml of sample in water and add fewdrops of bromine water.

(b) Baeyer's test: Dissolve 1 ml ofsample in water and add 1 ml of diluteNa2CO3 solution and a few drops ofKmnO4 solution. Shake well.

7 Nature: Dip a wet litmus paper in thesample.








3 Test for Nitrogen and Sulphur:To 1 ml of sodium extract, addferric chloride solution




Sl NO. Experiment Observation InferenceTest for carbonyl group

1 2,4-DinitrophenylhydrazineTest: To 2,4-dinitrophenylhydrazine reagentA(2 ml), add organic compoundwith vigorous shaking.

Test for Aldehydes2 Schiff’s test: To Schiff’s reagent,

add a few drops of organiccompound.

3 Tollen’s Test: To a small portionof organic compound, addTollen’s reagent and heat on awater bath for 5 minutes withoutdisturbing the test tube.

4 Fehling’s test: Mix 1 ml each ofFehling’s solution A and B in atest tube and add a small quantityof organic compound in presenceof sodium carbonate solution, Boilthe solution for 5 minutes on awater bath.

5 Benedict’s Test: To 4 ml ofBenedict’s reagent, add 3-4 dropsof unknown organic compound.Heat to boiling in a water bath.Confirmatory Test: To 2 ml ofaqueous solution of substance,add resorcinol and 2 ml of conc.Sulphuric acid.

RESULT- The given organic compound has ___________ as an extra element and_________________ as the functional group.______

EXPERIMENT NO.- 14

AIM OF THE EXPERIMENT- To carry out the Identification test for the given sample ofketone(Acetone)


CHEMICALS REQUIRED- Sodium bicarbonate, sodium hydroxide, conc. Sulphuric acid,hydrochloric acid, bromine water, potassium permanganate, sodium metal, ferrous sulphate crystals,sodium nitroprusside, ferric chloride, nitric acid, silver nitrate solution, carbon tetrachloride, chlorinewater, 2,4-dinitrophenylhydrazine solution, sodium bisulphite, ethanol, m-dinitrobenzene, potassiumiodide.



1 Physical test

2 Colour

3 Odour

4 Ignition test: Take a small amount of sample onloop and ignite it over flame.

5 Solubillity tests

Sample+cold water

Sample+ hot water



Sample + hot dil. NaOH



Sample+dil.HCl


6 Tests for unsaturation(a) Test with bromine water: Dissolve 1 ml ofsample in water and add few drops of brominewater.

(b) Baeyer's test: Dissolve 1 ml of sample inwater and add 1 ml of dilute Na2CO3 solutionand a few drops of KmnO4 solution. Shakewell.

7 Nature: Dip a wet litmus paper in the sample.




1 Test for Nitrogen: To 1 ml ofLassaigne’s extract, add very littlequantity of FeSO4 crystals. Ifprecipitate is not obtained, add adrop of sodium hydroxide. Heatthe contents to boiling and adddilute H2SO4.

2 Test for Sulphur(a) To 1 ml of Lassaigne’s extract,add few drops of sodiumnitroprusside solution.



3 Test for Nitrogen and Sulphur:To 1 ml of sodium extract, addferric chloride solution.



FUNCTIONAL GROUP DETECTIONSL NO. Experiment Observation Inference

Test for carbonyl Group1 2,4-Dinitrophenylhydrazine

Test: To 2 ml of 2,4-Dinitrophenylhydrazine reagent,add organic compound withvigorous shaking.Tests for Aldehydes

2 Sodium Nitroprusside Test: To0.5 ml organic compound, add

about 1 ml of sodiumnitroprusside followed by additionof excess of sodium hydroxidesolution.

3 m-Dinitrobenzene Test: To 1 mlof the organic compoundinethanol, add 0.1 g of m-Dinitrobenzene followed byexcess of NaOH solution.

4 Iodoform Test: Take about 0.5ml of the substance and dissolve itin 2 ml of water(or methylalcohol). Add 1 ml of 10%sodium hydroxide and a solutionof iodine in potassium iodidepersists. Allow it to stand for 15minutes.

5 Schiff’s test: Add about 1 ml ofSchiff’s reagent to about 1 ml ofacetone. Keep it for few minutesand observe.Confirmatory Test

(a) Add solution ofbenzidinehydrochloride to thesolution of sample.

Add Nesseler’s reagentin very dilute aqueoussolution of substance.

Add nesseler’s reagentto conc aqueoussolution of substance.

(b) To 2 ml of aqueous solution ofsubstances, add 20% potassiumhydroxide solution and boil for aminute.

RESULT- The given organic compound has ⸑ ⸑ ⸑ ⸑ ⸑ ⸑

As an extra element and --------------__as the functional group.

EXPERIMENT NO.- 15

AIM OF THE EXPERIMENT- To carry out the Identification test for the given sample ofcarboxylic acid(Benzoic acid)


CHEMICALS REQUIRED- 1N Sodium hydroxide, ferric chloride test solution, methyl red.

CHEMICAL TESTS FOR BENZOIC ACID -

SL NO. Experiment Observation Inference1 Warm gently0.2 g with 20 ml of

water and add 1 ml of 1N NaOHand filter. To the filtrate add ferricchloride test solutions.

2 To few ml of the substance addmethyl red solution.

RESULT-

The given organic compound is found to be _________________.

EXPERIMENT NO.- 16

AIM OF THE EXPERIMENT- To carry out the Identification test for the given sample of phenol(Resorcinol)


CHEMICALS REQUIRED- Sodium bicarbonate, sodium hydroxide, conc. Sulphuric acid,hydrochloric acid, bromine water, potassium permanganate, sodium metal, ferrous sulphate crystals,sodium nitroprusside, ferric chloride, nitric acid, silver nitrate solution, carbon tetrachloride, chlorinewater, phthalic anhydride, sodium nitrite.



1 Physical test

2 Colour

3 Odour

4 Ignition test: Take a small amountof sample on loop and ignite it overflame.

5 Solubillity tests

Sample+cold water

Sample+ hot water






Sample+dil.HCl
















SL NO. Experiment Observation Inference1 Ferric chloride Test: To a 1 ml of

aqueous suspension of sample,add few drops of 5% ferricchloride solution.

2 Liebermann’s Nitroso Test:Dissolve an organic compound

(0.1 g) in 1 ml of conc H2SO4 inadry test tube and add a fewcrystals of sodium nitrite. Dilutethe resultant solution in water.Add sodium hydroxide to thissolution.

3 Phthalein Test: To 0.2 mg of theorganic compound in a dry testtube add 1 ml of phthalicanhydride followed by 0.5 ml ofconc. Sulphuric acid. Heat andcool. And then add excess ofsodium hydroxide.

4 Azo Dye Test: To 0.5 ml ofaniline add conc. Hydrochloricacid. Cool the solution to 0-5℃followed by organic compound insodium hydroxide (0-5℃)

5 Bromination: Dissolve 0.2 g ofsample in water. Add brominewater slowly and then in excess.

6 Confirmatory Test: To smallamount of phenol, add hot fumingnitric acid. Heat and then pour thecontents in water.

RESULT- The given organic compound has ________________as an extra element and__________________as the functional group.

EXPERIMENT NO.- 17

AIM OF THE EXPERIMENT- To carry out the Identification test for the given sample of ester(ethyl aceatate)

APPARATUS REQUIRED- Ignition tube, pair of tongs, test tube, test tube holder, china dish, tripodstand, wire gauze, funnel and filter paper.

CHEMICALS REQUIRED- Sodium bicarbonate, sodium hydroxide, conc. Sulphuric acid,hydrochloric acid, bromine water, potassium permanganate, sodium metal, ferrous sulphate crystals,sodium nitroprusside, ferric chloride, nitric acid, silver nitrate solution, carbon tetrachloride, chlorinewater, phenolphthalein, hydroxylamine hydrochloride.



1 Physical test

2 Colour

3 Odour

4 Ignition test: Take a small amountof sample on spatula and ignite itover flame.

5 Solubillity tests

Sample+cold water

Sample+ hot water






Sample+dil.HCl








1 Test for Nitrogen: To 1 ml ofLassaigne’s extract, add very littlequantity of FeSO4 crystals. Ifprecipitate is not obtained, add adrop of sodium hydroxide. Heatthe contents to boiling and adddilute H2SO4.

2 Test for Sulphur(a) To 1 ml of Lassaigne’s extract,add few drops of sodiumnitroprusside solution.







Test for Esters

SL NO. Experiment Observation Inference1 Phenopthalien test: To an organic

compound (100 mg) add dilutesodium hydroxide solution (3 ml)and phenolphthalein solution (2drops) heat.

2 Hydroxamic acid: To 1 ml organiccompound, add 0.2 g ofhydroxylamine hydrochloride andabout 5 ml of sodium hydroxideto make the solution alkaline. Boilgently for 1-2 minutes. Cool andacidify with dilute hydrochloricacid. Add 2-3 drops of ferricchloride solution.

RESULT- The given organic compound has ________________ as an extra element and________________ as the functional group..

EXPERIMENT NO.- 18

AIM OF THE EXPERIMENT- To carry out the Identification test for the given sample ofAniline.


CHEMICALS REQUIRED- Sodium bicarbonate, sodium hydroxide, conc. Sulphuric acid,hydrochloric acid, chloroform, sodium nitrite, β-naphthol, benzoyl chloride, p-toluene sulphonylchloride or benzene sulphonyl chloride.



1 Physical test

2 Colour

3 Odour

4 Ignition test: Take a small amountof sample on loop and ignite it overflame.

5 Solubillity tests

Sample+cold water

Sample+ hot water






Sample+dil.HCl








1 Test for Nitrogen: To 1 ml of Lassaigne’sextract, add very little quantity of FeSO4crystals. If precipitate is not obtained, add adrop of sodium hydroxide. Heat the contentsto boiling and add dilute H2SO4.

2 Test for Sulphur(a) To 1 ml of Lassaigne’s extract, add fewdrops of sodium nitroprusside solution.

(b) To 1 ml of sodium extract, add aceticacid followed by lead acetate solution

(c) Sodium plumbite test: To lead acetatesolution, add a drop of NaOH and observe.Then add excess of NaOH. Add sodiumextract.

3 Test for Nitrogen and Sulphur: To 1 ml ofsodium extract, add ferric chloride solution.

4 Test for Halides(a) To 2 ml of sodium extract, add diluteHNO3. Boil the solution to expel H2S orHCN, if present, and then add few drops ofsilver nitrate solution.

(b) To 2 ml of sodium extract, add diluteHNO3. Add 1 ml of carbon tetrachlorideand a few drops of freshly prepared chlorinewater.


Test for amines

SL NO Experiment Observation Inference1 Hinsberg Test; To 0.1 ml of substance,

add 0.2 g of benzenensulphonylchloride and 5 ml 10% sodiumhydroxide solution with vigorousshaking. Observe the formation ofprecipitate of benzene sulphonylderivative. If no reaction occurs, heatthe contents on awater bath for 2-3minutes and then cool in ice and acidifywith HCl.

2 Carbylamine Test: To 0.2 ml of thesubstance, add 1 ml of chloroform and2 ml of sodium hydroxide solution.Heat the contents.

Azo Dye test: Take 3 test tubes. In thefirst test tubes, dissolve 0.5 ml of thesubstance in 3 ml of conc HCl and 5 mlof water. In the second test tube, take 2ml of 10% solution of sodium nitrite. Inthe third test tube, dissolve 0.1 g of β-naphthol in 2 ml of 10% NaOH. Coolthe 3 test tubes in ice cold water at0.5℃. add sodium nitrite solution to 1st

test tube followed by addition of β-naphthol.

4 Benzoyl Chloride test: Suspend 0.5 mlof the substance in 10 ml of 5% sodiumhydroxide solution in a boiling tube.Add 1 ml of benzoyl chloride withconstant shaking. Shake vigorously for5-10 min until the odour of benzoylchloride has disappeared.

5 Confirmatory tests(a) Sample+ bleaching powder(b) Dissolve adrop of

compound in 2 ml of etherand dilute it with water. Adddilute solution of bleachingpowder and shake.

(c) Dissolve a small amount ofpotassium dichromate inconc. Sulphuric acid and tothis add a drop of thesubstance.

RESULT: The give organic compound has _____________as an extra element and _______as thefunctional group.

EXPERIMENT NO.- 19

AIM OF THE EXPERIMENT- To prepare Acetanilide from Aniline.

APPARATUS REQUIRED- 250 ml conical flask, 500 ml Beaker, filter paper, funnel, stirrer.

CHEMICALS REQUIRED-Aniline- 10 mlAcetic anhydride- 10 mlGlacial acetic acid- 10 ml

PROCEDURE

Take 10 ml Aniline, 10 ml acetic anhydride and 10 ml glacial acetic acid in a 250 ml conical flask.Shake and warm the contents for half an hour. Cool and pour the contents in ice cold water taken in500 ml beaker with stirring. Filter off the crude acetanilide, wash with cold water and recrystallizefrom boiling water along with small quantity of methylated spirit.

RESULT: The practical yield of acetanilide is ________________%.

EXPERIMENT NO.- 20

AIM OF THE EXPERIMENT- To prepare Aspirin (Acetyl Salicylic acid) from Salicylic acid.

APPARATUS REQUIRED- 250 ml conical flask, 500 ml Beaker, filter paper, funnel, stirrer.

CHEMICALS REQUIRED-Salicylic acid- 5 gAcetic anhydride- 10 mlConc. Sulphuric acid- 1-2 mlAlcohol- 30 ml

PROCEDURE

Take 5 g salicylic acid, 10 ml of acetic anhydride and 1-2 ml of conc sulphuric acid in 250 ml conicalflask. Shake and warm the contents for half an hour at 50-60℃. Cool and pour the contents in 100 mlcold water taken in a 500 ml beaker with stirring and filter off the crude aspirin. For recrystallization,dissolve the product in alcohol(30 ml) and pour the solution into warm water(75 ml). if the solidseparates, warm the mixture to dissolve the solid. The clear solution is allowed to cool slowly to getneedle shaped crystals of aspirin.

RESULT: The practical yield of aspirin is ________________%.

EXPERIMENT NO.- 21

AIM OF THE EXPERIMENT- To prepare Picric acid (2,4,6-Trinitrophenol) from phenol.

APPARATUS REQUIRED- Flat bottomed flask, 100 ml Beaker, filter paper, funnel, stirrer.

CHEMICALS REQUIRED-Phenol- 6 gConc. Sulphuric acid- 7.5 mlConc. Nitric acid- 22.5 ml

PROCEDURE

Take phenol(6 g) and conc. Sulphuric acid(7.5 ml) in a dry flat bottomed flask and heat the mixtureon a boiling water bath for 30 min. with frequent shaking to complete the formation of o and p-phenolsulphonic acid. Cool the flask in an ice bath and add conc. Nitric acid(22.5 ml) drop wise withconstant shaking. Allow the reaction mixture to stand for sometime and when evolution of red fumesstops, heat the flask on a water bath for one and half an hours with occasional shaking. Add cold wate(75 ml), mix well chill thoroughly in ice water. Filter the yellow coloured crude product using suctionpump. Wash with water to eliminate all inorganic acid and drain. Recrystallize from dilute alcohol(alcohol:water; 1:2), about 70 ml of mixed solvent is required. Picric acid is obtained as pale yellowcrystals.

RESULT: The practical yield of Picric acid is ________________%.

EXPERIMENT NO.- 22

AIM OF THE EXPERIMENT- To prepare 2,4,6-Tribromophenol from phenol.

APPARATUS REQUIRED- Flat bottomed flask, 100 ml Beaker, filter paper, funnel, stirrer, 250 mlconical flask.

CHEMICALS REQUIRED-Phenol- 5 gBromine – 9 ml

PROCEDURE

Dissolve phenol(5 g) in 100ml of water and add aqueous bromine solution(9 ml dissolved in about100 ml of water) gradually with continuous shaking. Pour the reaction mixtureinto ice cold water .Filter the crude product, wash with water and recrystallize with dilute alcohol to obtain colorlesscrystalline material.

RESULT: The %practical yield of 2,4,6-Tribromophenol is ___________

PRACTICAL LAB MANUAL - JRU

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