EXPERIMENT No:4 Date: PAPER CHROMATOGRAPHY AIM To separate constituents present in a mixture containing lead and cadmium ions. ORY Paper chromatography is a type of chromatography in which a special adsorbent paper is used instead, Moisture adsorbed by this acts as a stationary phase and the solvent as a moving phase. The mixture to be separated or analyzed is put at one end of the paper strip as a small spot. The solvent moves up the paper due to capillary action and the components of the mixture rise up at different levels and thus get separated from one another. Lead and cadmium ions will move up at different rates and therefore get separated. Since the ions are colorless, this strip is sprayed with sodium sulphide solution so that lead ions form a black spot and cadmium ions form a yellow spot. Mobile phase will be a dilute solution of HCl. PROCEDURE ¢ Make a spot of mixture of soln. of lead nitrate and cadmium mixture in the center of the reference line and dry the spot. © Add dil, Soln, of HCI in a glass jar, and suspend chromatographic paper strip with the help of a glass rod in such a way that the spot does not directly dip into the solvent. © Keep the jar undisturbed and allow the mobile phase to move 10-12 cm, above the pencil line. ‘¢ Remove the paper strip and mark the solvent front with the pencil. ¢ Dry the strip and then spray sodium sulphide solution over it. # Identify the ion from its color and calculate Rf Value. RESULT The Rwalue of Pb*ion (black spot) is = ‘The R,value of Cd*ion (yellow spot) is =OBSERVATIONS SL | COLOUR OF | DISTANCE DISTANCE R-VALUE No. | SPOT TRAVELLED TRAVELLED BY THE BY THE COMPONE(cm) SOLVENT(cm) R,of Pb**= Distance moved by Pb*= Distance moved by the solvent R, of Cd*= Distance moved by Cd*= Distance moved by the solventEXPERIMENT N DATE: PREPARATION OF FERROUS AMMONIUM SULPHATE (MOHR’S SALT) AIM : To prepare ferrous ammonium sulphate crystals, APPARATUS: china dish, wire gauze , CHEMICALS REQUIRED : Ferrous sulphate Ammonium sulphate and dilute sulphuric acid THEORY : Mohr salt is prepared by mixing equimolar mixture of hydrated ferrous sulphate and ammonium sulphate in water containing small amounts of dil. Sulphuric acid FeSO..7H;0 + (NH,):SO, FeSO,.(NH,):SO,.6H,0 + H,0 PROCEDURE : Take 250 ml beaker and add 7g of FeSO, and 3.5 g of (NH,);SO,.Add 2 to 3 ml of dilute sulphuric acid to prevent hydrolysis of FeSOx. In another beaker boil about 25 ml of water and add the boiling hot water to the contents of the first beaker.stir it well with a glass rod until the salt is completely dissolved.Transfer the solution into the china dish and concentrate it to crystallization point.Place the china dish containing solution in a beaker full of cold water.On cooling crystals of mohr’s salt separates out. RESULT : Pale green crystals of Mohr salt are obtained. Mass of Mohr salt = gEXTNO:2 Date: PREPARATION OF STARCH SOL. AIM To prepare a lyophilic sol of starch MATERIALS REQUIRED + Soluble starch © Distilled water + 250 ml beaker + 50 ml beaker © Glass rod + Funnel + Filter-paper + Tripod stand © Wire-gauze + Bunsen burner THEORY Lyophilic means ‘liquid-loving’ or ‘solvent- attracting’. This means that in this colloidal solution there is a strong attraction between the dispersed phase and dispersion medium. Starch forms lyophilic sol when water is used as the dispersion medium. The formation of sol is accelerated by heating. Starch sol can be prepared by heating it and water at 100 °C. It is quite stable and is not affected by the presence of any electrolytic impurity. PROCEDURE Weigh a small quantity (1 g) of soluble starch on an electronic balance. Transfer the weighed quantity of starch into a watch glass and add few drops of distilled water to make a paste. Take about 100 ml of distilled water in a 250 ml beaker and heat the beaker till the water starts to boil. Pour the starch paste slowly into the boiling water while stirring using a glass rod. Continue boiling for about 3 minutes and then allow the beaker to cool. PRECAUTIONS+ The apparatus used for preparing the sol should be properly cleaned. + Distilled water should be used for preparing sols in water. + Starch should be converted into a fine paste before adding to boiling water. + Starch paste should be added in a thin stream to boiling water. + Constant stirring of the contents is necessary during the preparation of the RESULT Starch sol is prepared.EXPERIMENT NO: 3 Date: PREPARATION OF FERRIC HYDROXIDE SOL AIM To prepare a lyophobic sol of ferric hydroxide MATERIALS REQUIRED + 2% solution of ferric chloride + Distilled water « 250 ml beaker + Dropper + Wire gauze + Tripod stand + Bunsen burner THEORY Lyophobie means ‘liquid-hating’. That means in these sols, there is little or no interaction between the dispersed phase and the dispersion .These sols are easily precipitated by the addition of small amounts of electrolyte, by heating or by shaking, therefore these sols are relatively less stable than lyophilic sols. They need stabilising agents for their preparation. Examples of lyophobic sols include sols of metals and their insoluble compounds like sulphides and oxides Ferric hydroxide forms lyophobic sols on treatment with water. Ferric hydroxide sol is prepared by the hydrolysis of ferric chloride with boiling distilled water. The reaction takes place is as follows. Ba FeCl,(aq) + 3H,0(l)-+Fe(OH),(S) + 3HCI(aq) Ferric clride Red sol PROCEDURE Take 50 mL of water in a beaker. Boil the water and add ferric chloride solution dropwise to the boiling water using a dropper. Continue heating until a deep red or brown solution of ferric hydroxide is obtained. Keep the contents of the beaker undisturbed for some time at room temperaturePRECAUTIONS + Add ferric chloride solution dropwise. + Heating is continued till the desired sol is obtained. + Hydrochloric acid formed as a result of hydrolysis of ferric chloride is removed by dialysis process otherwise it would destabilise the sol. RESULT Ferric hydroxide sol is prepared.EXPERIMENT NO: 5 Date: TEST FOR PROTEINS, FATS AND CARBOHYDRATI EXPERIMENT Test for Proteins To a little of the given milk, 2ml of NaOH solution is added followed by 1% CuSOs solution. Xanthoproteic Tes To a little of the given milk, conc. Nitric acid is added and heated, OBSERVATION Bluish violet colourisation. Yellow precipitate INFERENCE Presence of Proteins. Presence of Proteins. Test for Fats Solul T The solubility of given substance in water and chloroform is noted. -anslucent spot test Press a little of the substance in the folds of the filter paper. To a few drops of phenolphthalein, borax is added followed by small amount of given oil. Insoluble in water but soluble in chloroform. Translucent or greasy spot appears on unfolded paper Pink colour disappears. Presence of Fats. Presence of Fats. Presence of Fats.Test for Carbohydrates A little of the given substance is dissolved in water. It is then treated with | mL of Molisch’s reagent followed by a few drops of cone. sulphuric acid along the sides of the test tube. Tollen’s Test: To little of the given substance add Tollen’s reagent and heat in a water bath. Fehling’s solution test: To a little of the given substance, fehling’ and ‘B’ are added in equal amounts and heated in a water bath. Test for Starch Toa little of the given solution, add iodine solution. solution ‘A’ Violet colouration at the junction of the two liquids. Appearance of silver lining along the sides of the test tube. Reddish precipitate is obtained. Blue black colour. Presence of carbohydrates. Presence of reducing sugar Presence of reducing sugar. Presence of Starch.EXPERIMENT NO: DATE: QUANTITATIVE ANALYSIS: REDOX TITRATIONS PERMAN (OMETRIC TITRATIONS. TITRATION-1 AIM: To prepare M/20 Mohrs salt solution [FeS0..(NH,);S0,6H,O] and to estimate the strength of the given KMnO, solution by titrating it against standard Mohrs salt solution PRINCIPLE: KMn0O,acts as an oxidising agent. It oxidizes Fe"to Feions and the oxidising agent gets reduced. IONIC EQUATION: MnO; + 8H'+ Se” Mn?" + 41,0 Sx[Fe*Fe+ le] MnO.+ 5Fe*+ 8H’ ~Mn’+ SFe*+4H,O REQUIREMENTS: Burette, pipette, conical flask,KMnO,solution,Mohrs salt,4N H,SO, KMn0, acts as a self indicator END POINT: Colourless tums pink PROCEDURE: 1. Preparation of standard Mohr salt solution (a) Weigh the given Mohrss salt (b)Transfer the weighed Mohrs salt into 100m! standard flask through the funnel. Add 10ml of H,SO. (c) Make up the solution up to 100ml with H,O. (d)The last few drops may be added using a dropper. 2.Titration of Mohr's salt si ition with KMnO, solution (a)Wash the apparatus with water (b)Rinse the burette with KMnO, solution and fill it up to the ‘0” mark (©) Rinse the pipette with the prepared Mohr's salt solution, Pipette out 20ml Mohr's salt solution into a clean flask. (d)Add 4N H,SO, into it and titrate it against the KMnO, solution taken in a burette (e )Repeat the experiment to get concordant valueOBSERVATION ARTICLE WEIGHED WEIGHT (g) Weight of bottle + salt Weight of empty weighing bottle Weight of salt Mohr salt x KMnO, SLNo Volume Burette (ml) Volume of of Mobr's | Initial Final Reading | KMnO, salt Reading (mL) (mL) Calculation RESULT Molarity of KMnO. solution= M Strength of KMnO,solution= Molarity of KMnO,x 158 x 158 = g/mLEXPERIMENT NO: DATE: VOLUMETRIC ANALYSIS REDOX TITRATION PERMANGANOMETRIC TITRATION TITRATION - 2 To prepare M/20 of oxalic acid to estimate the molarity and strength of given KMnO, solution. [MnO,-+ 8 H'+ 5 ee Mn*+ 4 1,0] x2 [C,02-7 2CO,+2e]x5 02+ 2 MnO;+ 16 H-> 10 CO,+ 2 Mn+ 8H REQUIREMENT! Burette, pipette, conical flasks, KMnO, solution, oxalic acid crystals and 4N HSO, END POINT: Colourless to pink. NDICATOR : KMn0, act as a self indicator PROCEDURE: 1. PREPARATION OF STANDARD SO! 1. Weigh the given oxalic acid crystals TION OF OXALIC ACID 2. Transfer the weighted oxalic acid crystals into 100m! standard flask through a funnel. 3. Now, make the solution up to the 100mL mark. The last few drops of water should be added using a dropper. ION OF OXALIC ACID SOLUTION WITH KMnO, SOLUTIO! 2. 1. Wash the apparatus with water. Now rinse the burette with KMnO, solution. fill it with KMnO, solution up to the 0 mark, Rinse the pipette with oxalic acid solution. Pipette out 20ml of oxalic acid solution to a clean conical flask and heat it to 60-70°C, after adding 20ml of 4N H,SO,, Titrate it against the KMnO, solution taken in a burette2. End point is the appearance of pale pink colour, Repeat the experiment to get concordant values and record the observation. OBSERVATION ARTICLE WEIGHED WEIGHT (g) Weight of bottle + salt Weight of empty weighing bottle Weight of oxalic acid Oxalic acid x KMnO, SLNo Volume Burette (ml) Volume of oxalic of acid Initial Final Reading | KMnO, (mL) Reading (mL) CALCULATIONS: Molarity of given KMnO, solution = M Strength of given KMnO, solution = Molarity x Molecular mass of KMnO,= x 158 =g/L RESULT: Molarity of given KMnO,solution= — M Strength of given KMnO,solution= g/L ORGANIC ANALYSIS GENERAL RE. “TIONS OF FUNCTIONAL GROUPS1.TEST FOR UNSATURATION To a little of the Pink colour is discharged. organic compound Baeyer’s reagent is added. Presence of unsaturation. So ¢ Ye = C+ 2KMn0, +4 1,0 —> Pink of purple Io 3- - i -+2Mn0, + 20H OH OH Colourless 2, TEST FOR ALCOHOLS Toa little of the Brisk effervescence due organic compound to liberation of hydrogen in a test tube a small gas. piece of metallic sodium is added. Presence of hydroxyl group. 2ROH + 2Na —> 2RONa + Hz Esterification: A fruity smell due to To a little of the the formation of ester. organic compound acetic acid is added followed by 2 drops of conc. H2SOa, Warm the Presence of hydroxyl group. contents in a water bath and pour it into cold water. CHsCOOH + ROH ~ CHscoor + H20 3. TEST FOR CARBOXYLIC ACID To a little of the Effervescence due to Presence oforganic compound saturated solution of sodium bicarbonate is added. formation of COz. RCOOH + NaHCOs > RCOONa + H20 + COz carboxylic acid. Esterification: To a little of the organic compound Ethyl alcohol and 2 drops of cone. H2SO« are added. Warm the contents in a water bath and pour it into cold water. A fruity smell due to formation of ester. RCOOH + C2HsOH > RCOOCiis + H20 Presence of carboxylic acid. 4. TEST FOR PHENOLS To a little of the organic compound neutral ferric chloride is added and is diluted with water, Violet colourisation is developed. Presence of phenols. 6CsHsOH + FeCle > [(CeHs0),Fe] + 3HCI + 3H* Liebermann’s Nitroso reactiot Take a little of the sample, add NaNO, followed by concentrated H,SO,. ‘Then dilute with water. Add NaOH. Appearance of a deep green or blue colour which changes to red on dilution with water and restores original green or blue colour with NaOH. Presence of phenols. 5. TEST FOR CARBONYL COMPOUNDSTo a little of the organic compound 2,4-dinitro phenyl hydrazine reagent is added. Orange yellow crystals appear due to the formation of phenyl hydrazone. Presence of carbonyl compounds, no _ | 2. 4 Dinitro pheny! hydrarones (Orange of yellow ppe ) Silver Mirror/Tollen’s Appearance of silver Presence of aldehyde Reagent lining along the sides of group. the test tube. To a little of the organic compound Tollen’s reagent(Ammoniacal silver nitrate) is added, warmed over a water bath for § minutes. RCHO + 2[Ag(NHs)2]" + 30H” > 2Ag + 2H20 + 4NHs + RCOO™ Fehling’s Solution Test: To a little of the organic compound equal amounts of Fehling’s reagent A and B are added and warmed in water bath. Red precipitate is formed. OHO Cv" +SOH' RCO” + CuO ad Presence of aldehyde group. Add a small amount Red colourisation. Presence of ketones.of organic compound to Sodium nitroprusside solution followed by NaOH solution in drops, CHsCOCHs + OH” ~ CHsCOCH2” + HO [Fe(CN)sNO}*+ CHsCOCH2" > [Fe(CN)sNOCHsCOCH2]* 6.TEST FOR AMINES To a little of the organic compound dil. HCI is added and shaken well. Organic compound is Presence of amines. soluble. Dye Tes RNH2 + HCl > RNH.cI- Orange red colouration. _Presence of amines Dil. HCI is added to sodium nitrite crystals in a test tube. This solution is then treated with organic compound in n another test tube. B-naphthol is dissolved in sodium hydroxide solution. The two solutions are mixed together