LABORATORY MANUAL ON ENGINEERING CHEMistay 268 Q.7. How silica can be removed Saeed ‘i as. Si lowing methods : Ans. Silica can be removed by following j (9 By adding magnesia or dolomite lime to water. Magnesia acts as Mj magnesia by adsorption or by the formation of magnesium silicate. (49 By the use of ferrous sulphate or sodium aluminate as coagulants. They act as Fe(OH), or Alon, which enmesh finely suspended silica particles, (OH). Silica is removed jy the demineralised water through a strongly basic special anion exchanger, demineralised water ? ate é {vis the water which is completely free from soluble salts. It is obtained by ion exchange proce. How will you differentiate between organic and inorganic solids ? als (such as sand, gravel etc.) and salts, Ger solids represent the inorganic solids, Q.10. Explain the im Ans. (i) Knowledge Treatment units and for di (@ The quantity o sedimentation tank, resp; portance of examination of solids in water, of the organic or volatile fraction of solids in ‘necessary for the design of biological isposal of sewage by dilution, “ f inorganic and organic-settleable solids helps in esigning the grit chamber and ectively. BIO TECHNOLOGY o EXPERIMENT 22.8 ject To analyse the Antacid Tables Provided to you, Theory in the food Pipe (oesophagus) and th : e heat Sometimes the acidity leads te oss 8 BU heart attack to the patient, Dot ibe Di man aly Prseribe Digene ables or Digene "quid asa medicine for Acidity, The Digene tbls AR in addition to some vedas rea, Verna, soleette, Goa contains erythrosine and Poncesb- e Ci ilk i The alysis of Ami acd able, involves e stone one nc ation Of Total Alkatini , solatog aTP® eel determined by nn Ueto AL(OHD,, MgOn), solution (say HCI) ve indian Me oF he ag cator, Tae SNe cna 3h,0 MeOH), + 2146) Ian — Mec. + 2H,0 MOM), + Ho) Cac, + 2H,0 : y — Ho therefore, gives a feeling of pain 'g of Vomiting and flatulence to the 8 Of heart bum, which is called angina pectoris ie., mild and Ca(OH), : The amount only \eous solution against a standard:ENVIRONMENTAL CHEMISTRY AND BIOTECHNOLOGY 269 Ny ination of alkalinity due to milk of M: (i) Determinati of Magnesia only: The: 2 ir (etre titration of the known volume of the san ee eumoum of Mg can by estanated bo le solution against El i ic mplexor ples gainst EDTA using Eriochrome Black-T cotndicator, Let EBT be represented as Na;H,Y. I ionses in solution to give 2Na’ and a strong chelating fon as shown NasH,Y ——>» 2Na’ + H,Y> nen Eriochrome Black T is added to the sample solution at Wi Mg?" ions when ith AI, Ca’ and Mg he Ripe | BS HD? + Me” ——> MgD™ + Indicator Wine Red PH-10 is gives a wine red coloured complexes EDTA solution added from burette it combines with ionsto form respective EDTA complexes. HY? +a?” ——> CaEDTA Complex Y* + 2H" 2 w SHY + APt ——> Al,(EDTA), Complex Y* + 6H" ON end point when all free ‘metal ions are consumed, the excess EDTA abstracts metal ions from weak ee cator complex to form stronig metal - EDTA complex releasing indicatorin free form which shows a metal - colour change fa se Nes light ne ae Less stable. Winered...«, More stable Ble ‘To get alkalinity due to only Mg(OR),, the other ions AT or Ca™ are removed by precipitating them as Calcium oxalate Aluminium oxalate by adding calcium and aluminium precipitating buffer, [NH,Cl + NH,OH + (NH,), C04] 0" COO. _ 2 Ci | ca or Ca"* + C0, —>> CaC,0, coo" coo Cal. oxalate (white ppt) Similarly, AP" + C,0,7" ——> ALCO) aluminium oxalate (white ppt.) Precipitate of calcium oxalate and aluminium oxalate (white ppt.) is filtered and filtrate is treated with EDTA solution to give alkalinity due to Mg”* ions alone, ‘The alkalinity due to Ca” and Al™* can be calculated be subtracting alkalinity due to Mg** from the value of total alkalinity determined by first titration of HCI against the sample solution Chemical Required : N ; N (9 Standard HCI solution (3). (i Standard EDTA solution () ; (ii) Buffer solution of NH,Cl +NH,OH; (i») Buffer soluti - Fist eee) Buffer solution of NH,CI + NH,OH and (NH,) C,O, Indicator Phenolphthalein, End Point Disappearance of pi ¢ of pink col nd Titration ie hdicator Eriochy J evapgn "= Black (EBT), Colour. Shange from wine red to light blue.iS ‘TORY UAL 7 270 LABORAI MANUAL ON ENGINEERING CHEMistRy. Procedure Step 1: Preparation of the solution ‘Take the given two anti-acid ina pestle and morter and make a paste of it in 20 ml of distil it to.250 ml measuring flask. Give a number of washings to the pestle and morter and t completely into the measuring flask. Make up the volume upto the mark by adding distill flask and shake it vigorously. Step 2: Determination of total alkalinity £e. Acid-Base Titration Pipt ou 20 lf his solution ito a conical ask, Add2-3 drops of phenolphhalein indicator. Tit with N/20 HCI solution taken in the burette slowly with constant shaking till the disappearance of pink at : ive. the end point, Repeat the same titration 3-4 times to get the concordant reading, um Step 3: Determination of Alkalinity due to Mg(OH),, ie, EDTA Titration Pipet out 100 ml ofthe sample solution into a250 ml beaker add 25 ml ofcaleiuman; buffer solution while constantly stiring the mixture witha glassrod, Allow the precipi 20 minutes, Filter the solution through dry filter paper (whatmann-42) ina dry beaker, and add 5 ml buffer solution and S drops of Eriochrome black-T. Indicator led vate. Tansy ransfer the Contents 4 water, Stopper ihe aluminium pespitat Hate to settle down for |s. Take 20 mlofthe filtrate Titrate it against x EDTA solution tll wine red colour changes to pure light blue colour, Observations Volume of ant-acid solution prepared = 250 ml Volune of sample solution taken in the titration flask = 20 mi for both the titration Table for acid-base titration as usual : Table for the complexometric (EDTA) titration as usual. Calculations Let the first titre value i.e. volume of x HCl used against 20 ml of the sample solution = v Let the second titre value ie. volume of x EDTA used against 20 ml ofthe sample solution= Applying | normality equation : | NV, = N,V, | HCI SampleSoln. | es = Nyx20 | Wino N, (Normality ofthe sample alkalies suchas [Mg(OH),, AIH), and Ca(OH) * 55,495 “a0 | We know strength (gL!) = Normality x Eq, wt. Strength w.rt, Mg (OH), = Normality x Eq, wt. of Mg(OH), oe ea9 pile © 99g * 29 8b" = X ‘Amount of Mg (OH), in the two tablets of Digene Sample =x g/L”! Amount of Mg(OH), in one tablets = > gt Similarly strength wart. ACOH); = Normality x Eq, wt of Al(OH), |a RONMENTAL CHEMISTRY. AND BIOTECHNOLOGY = Ty 28= Yo ; Yee a ‘Amount of AI(OH), in one tablet = > gL : similarly strength of Ca(OH),, if present) + Amount of Ca(OH), in one tablet =, 5 gL, ‘Amount of Mg(OH), present from the second titre value NyV, = N,V, EDTA’, Sample soln; 5078 = Noe N, (Normality of solution) w.r.t, Mg” ion= =3=1 a i = Th x29 a = vee The solution of EDTA taken for titration = 100 ml, rat So, amount of Me(OH), in one tablet = 23ye" = 125Y'eL" Alternative Method for total alkalinity. in Amtacia, | of pinkish colour, Repeat to get concordant reading. "Calculations = (50 ml-—ttre value) © NABV, 2 0.05% 365 x Vj Grams of HCI neutraised= 5s = apg = Normality x Eq, wt. of Ca(OH), Strength ofthe sample w.rt, Ma(OH), excluding other shalsNematiy xEq.wt. Total volume of the solution in which two tablets were dissolved Therefore amount of Me(OH); in total Volume ie. 250 ml. of solution = ~-B* Solution of Antacid tablets or syrup is prepared as given in the above ex; Using a pipette in a titration flask and add 50 ml of N/20 HCI to it. Swirl the tit | #dd2 drops of phenophthalein as indicator and titrate against N/20 NaOH Let the volume of N/20 HCI neutralised by the antacid tablet be V, ml. It can be ealenifed as is =" (Volume of N/20 HCl added ~ Volume of N20 NaOH used) 271 =25Y' gut ‘periment. Take 0 ml of the solution tration flask for five minutes: Now from the burrette till fe appearence— ore LABORATORY MANUAL ON ENGINEERING CHENistay whore N= nonmality of HCI and (0.05 N), E= Eq. wt. of HCI (36.5) and V, = Vol. Of HCI neutralised by anaes tables So, amount of HCI neutralised by 1.0 g of antacid 0.2%365xV 1 a 1000 w® where W is the weight of antacid tablets taken, Note: The value of Mg(OH), determined by Acid-base titration may not be exactly same as determined EDTA titration. It is because we are not certain about the exact nature of weak alkalies other than Mg(OH),.. In ther word, the numerical value of Y and Y! may not be the same. Precautions (9 The tablets should be powdered carefull into the measuring Mask. (9A number of washings of the complete transfer. ¥yand its paste with distilled water must be fully transtened Pestle and morte should be transferred tothe measuring ask to ensure (i) ‘The usual precautions during volumetric analysis should be followed. (©) The pH= 10 of the solution should be maintained by making use of basie buffer in case EDTA titration, (©) The turbidity inthe solution due to organic chemicals is the tablets may be removed by 5-10 ml of acetone or alcohol before making up the solution up to the mark, VIVA-VOCE Q.1. How can the analysis of Digene provided to you in solution form be undertaken ? Ans. The method described in the above expriment can be conveniently followed. Instead of two: tablets we should start with two spoons of the sample and note the volume of the two Spoons and also the total volume “of the Digene solution contained in the bottle. Q.2. Calculate the equivatent weight of Mg(OH),. f ‘ = Molecular weight 24 +2%17 Ans. Equivalent weight ofa base = Molecular weight _ 24 + 2x17 _ ns, Equivalent weight of a ba aaa 2 29 Acidity of the base in the total number of replacable OH “i ions in the base e.g. acidity of Mg(OH),istvo. Q.3, Calculate the equivalent weight of Al (OW);. Ans. Equivalent weight of Al(OH), = ee 27+3x17 _ 27451 EOE ae ANG, Q.4, Explain what do you understand by normality equation? 9 Ans. Mathematically, N,V, = NV, itis clear from the equation that if N, =N,, V, must be equal 0 V2 ‘means that equi-normal solutions are equi-volumetric. Q.5. What are the constituents of anti-acid tablets? : Ye | Ans. Anti-acid tablets essentially contains milk of magnesia je, Mg(OH)>. It also contains Al(OH), 99 i b ic chemicals. ls small amount of Ca(OH),, which may or may not be Present, It also contians some organic che : composition varies from company to company, = 26 Q.6, Anti-acid tablets (Digene) or syrup are used ag a medicine for which disease ? eee: Ans. Anti acid tablets are used by the patients who suffer from acidity ie, the Son than required for digestion, Acidity errodes the musconal lining and ultimately lead to gastuenTaL CHEMISTAY AND BIOTECHNOLOGY ron nat arethe symptoms of Acidity? Qt es a feeling of pain in food pipe (oesopha; 273 es oesophagus) and the gas generated in the stomach the Ars omiting and flatulence to the patient. Acidity leads to hear burm which ieee tooth oa oe 20 called mild heart attack, Infact it has nothing to de wid herr aeige = Seo arene Xie, itis a disodium salt of ethylene diamine tetra carboxylic acid and can be written as HOOC-H;C., [etscoONa SN-cHy-CHyN Na*“00C-H30° ‘CH;-CooH c | 9.9. How is EDTA represented in aqueous soliitions? nit * “Ans, It is represented as Na,H,Y.. It ionises to give a strong shelating ion in solution. 5 Na,H3Y¥) ——>-2.Na" + H,Y7- = -Q.10. What name is given to the titration of EDTA with Mg?" and Ca” ions ?- Name the indicator Q. nplyed and the colour change at the end point. ‘Ans. The titration is referred to as complexomettic titration or complexometry.. The indieator is Eriochrome tukicT represented as HD*” and the colour change is from wine red to pure blue. "~ Q.11. How does the indicator works in EDTA titration ? | Ans. At pH= 10 it gives wine red coloured complexes with Ca”, Mg?" ions is solution. When EDTA added from burette, itcombine with free Ca”* and Mg” ions to form their respective complexes with EDTA. Atthe end point when all free metal ions are Consumed the excess of EDTA abstracts ions from weak metal. Q.12. Show the colour change iy EDTA titrations at the end point. ‘Ans. HD? + Mg"? > MgD~ + He 3 Indicator Wine Red qa HY? + Ca"? ——> Cav" +24" (EDTA) (CaEDTA Complex) Hay? + Mg’? > MgY? + 2H Mg-EDTA complex HY? + MgD" Mgy? + HD? + Ho Wine red complex More stable Blue with Indicator tess stable Q.13. Why pH = 10 is adjusted in EDTA Titration ? Ans. Athigher pH values, Ca(OH);, Mg(OH), and Al(OH), may get precipitated and the indicator changes iscolour. Mg-indicator complex becomes unstable and a sharp end point can not be obtain. Q.14. How the pH value is adjusted to 10? aes a Ans. The pH value is adjusted to about 10 by adding a buffer solution consisting NH,CI_ and NH,Ctin ‘molar ratio. Q.15. What is the effect on the end point with Eriochrome Black T indicator if the hard work sample does “Not contain Mg? 2 : Ans, Both Ca”* and Mg” react with EBT to form wine red complexes ata pH = 10. MP + HD > MDT} HT But the colour change from wine red to pure blue 'ed if not present in the sample solution, ‘not sharp with Ca-indicator complex. Mg™ have to be addLABORS TORY MANUAL ON ENGINEERING CHEMISTRY A EXPERIMENT 22,9 : Object determine the percentage composition of a mixture containing KNO, and KNo, ee _ To dete form by KMnO, method. ‘Theory ites react with oxidising agents such a acidified KMnO, solution i paso) "+ 5NOy 6H” ——> 2Mn’* + 3HO, + SNO, Itisnot possible to directly titrate nitrite (NO,) ion solution with acidifi in warm solution as Siven below: ied KMnO,, Durin, e (NO,) 8 acidificai vih 0, nitrous acids berated whichis volatile and unstable and canbe pana on tn vin Therefore, a measured volunie of standard KMNO, solution, acidified with dit H,SO, is treated withthe given mixture solution, added from the burette, until the pink colour of Permanganat NO,” ion gets oxidised to NO,” ion as per equation written above, N unreacted excess KMnO, left can be determined by titratin the following chemical equation. The following reaction t Nu au fe just disappears, The ‘ove. Nitrate (NO;”) ion remains unaffected, The ig it with standard Mohr’s salt Solution as shown by takes place during the titration, 2KMnO, * 3 H;S0, ——» K,S0, + 2MnSO, + 31H,0 + 50 [2 FeSO, + H,S0, +O ——> Fe,S0,), + H,01] x5 2KMnO, +10 FeSO, +8H,S0, ——> K. To get accurate results, during the addition of nitrite pipette containing nitrite solution should be below the s KMnO, solution can be carried out with a standard ‘ammonium sulphate solution. The ionic equations tal MnO, + 8H" + 5¢ ——4 Mn? 250, + 2MnSO, + 5 Fe,(SO,), + 81,0 to the excess acidified KMnO, solution, the tip ofthe surface of the liquid, The bac k titration of unreacted Mohr's salt. FeSO,. (NH,), SQ,. 6 H,O i ferrous ion may be written as under iking place during the titrati + 41,0 [Fe rea MnO, + 5Fe*+ 8H* —_, Chemicals Required ( Sample of KNO, and KNO, solid mixtyure (A mixture of 95% KNO, may be provided) (i) Standard (0.1 N) KMnO, solution, (iii) SO, solution 0.75'N In” + 4H,0 + 5 Fe (iv) Standard x Mohr’s salt solution for back titration. Indicator Self indicator: (KMn0,) End Point Disappearance of pink colour. ( Nitrite solution in the burette), Procedure Step 1: Preparation of sample solution z 5. Transfer the !.010 1.5 g of commercial sample of potassium nitrite on a previously mipiehed vere ss ing sk Contents to a washed beaker and dissolved it in cold water, Transfer the comers a “ater completely along with the washings. Make up the solution up to the mark with distil ie Step II: Titration of standard KMnO, solution against the prepared sample i ff 50, witha graduated Measure $0 ml, of standard 0.1 N KMnO, solution into a titration add 50 mlof ‘din Step. Tale cylinder and warm the content to 40°C. Fill up the burrete with KNO, solution prepar‘ 275 é. BIOTECHNOLOGY. s ee CHEMISTRY AND : gi ; eee Foe ica tem iS CITAgE slowly with constant stirring vi the} MnO so tion care sare one reguls can be cbiained by dipping the tip este under a feeder de ps decotourised, Bel ti 10, solution slowly, repeat the tit just aevotou joint add the KNO, 8 } “eho, san a ator nase fet concordant readings. Let the concordant volume of sample kK in on ee cae " sO oa which only NO,” ion reacts = V,) sit s Be are sample taken = Wg, z i ‘ : a eof 0-1 NKMn0, Solution taken = 50 ml ae Uk CANO shes seagate KNO, from the mixture solution used ='V; mi Ne of0.1Nsoltion of KMnO, reacted = (50 Vt? 3 raise that equi normal solution are equi voliimetii¢ ie.“ : NAVE SN Vere St Volume of0.1 N KMnO, solution taken = 50m as Total volurhe of the prepared sample solution =280 mi." { ; \olume of KNO, solution used against $0 ee of O.1 N KMnO, taken= V, ml Applying, N/V, "= N,V, (NOS) (KMNO,) Therefore, MixN, = 0.1x(50-V,) 01x G0-V) Ni(Normality wat: KNO, solution)= ———"7-——") i satin Strength = Normality xq, wt-of KNO, (85) 01 OSV, Peat oc 5 Si Wado, oe Xell ce 1000 i ! == 1000. Original strength of sample: = 250: 4w Percentage of KNO inthe sample = xi Ly Ferentage of KNOs inthe sample = (100A) ve ea } Precautions (Usually KMnO, i aways taken in the burette, but in this case NaNO. burette. 5 5 | Ui) Add about an e: burette, Gil) Avoid the use of a burette having a rubber tape. Read the Upper meniscus while taking burette reading with KMn0, titrations, In case a brown Precipitate appears durin; Reject the titration ; in that case. Shake the contents While preparing so! times, te 2 OF KNO, solution is taken in the qual volume of dil: H,SO, to the solution to be titrated before adding KMnO, from the 8 the titration, it shows insufficient addition of dil. H,SO,. of the titration after every small addition of KMNO, from the aa aie cf Iskion of KMnO, it shouldbe dissolved by the process of extraction i the tip of burette © WD: Nitetes arg casily oxidised to Nitrates by atmospheric oxygen: So itis Deter to keep : Aipping inthe solution in the titration fesk while delivery. ‘276 LABORATORY MANUAL ON ENGINEERING CHEMistRy VIVA-VOCE Q. 1. Suggest two methods for the estimation of NO, ions in solution. Ans. (i) Redox titration of NO,” ions with acidified KMnO solution, (ii) Nitrites can be titrated against Ce($O,), solution. The excess of Ce (SO,), can be known byt. with standard Mohr’s salt solution, aa wn by titration Q.2. Give ionic equation for reaction of nitrite ion with permanganate ion, 3 Ans, 2MnO, + SNO,” + 6H’ ——+ 2Mn™ + 5NO, +3 HO. d Q.3. Why direct titration of NO,” ion with acidified KMnO, is not possible? Ans. It is because during acidification with dil, HSO,, nitrous acid is formed, which being unstable and volatile may be paritally lost. and Q.4. Why Nitrite solution is taken in the burette instead of KMnO, ? Ans. Itis to avoid the reaction of dil. H,SO, with nitrites directly, which may otherwis that may get decomposed. Nitrous acid does not react with KMnO, instantaneously, Q.5. Why the tip of the pipette containing nitrite solution should be dipped below thesurface of the liquid during titration ? produce HINO, and | Ans. It is to avoid the oxidation of nitrite to nitrate by atmospheric oxidation. More-over formation of nitrous acid is avoided, which may get decomposed. Q.6. What is end point of KMn0,j titration with nitrites, Why ? Ans. It is disappearance of pink colour and not appearance of pink colour. It is because KMnO, self indicator, but it is not taken in the burette. Q.7. Give the principle of estimation of nitrite ions with ceric sulphate solution. Ans. The sample solution containing NO,” ions is treated with excess of standard ceric sulphate and the excess of sténdard ceric sulphate can be determined with standard ferrous sulphate Mohr’s solution. Q'8..Give the reaction between nitrite and ceric ions in the form of an ionic equation. Ans. 2Ce* + NO, + H,O ——> 2Ce* + NO, +2H". Q.9. Write a chemical equation for the titration of ferrous ions with ceric ions equation also. Ans. Chemical equation : 2FeSO, + Ce(SO,), ——> 2Fe,(SO,), + Ce(SO,), Ferrous Ceric Ferrie Cerous sulphate sulphate sulphate sulphate lonic equation can be written as Fe™ ——> Fe™ + ¢ (Oxidation half) Ce + ——>+ Ce™ (Reduction half) overall ionic if equatio: Fe + Ce 5 Fe’ + or? Ferrous Cerie Ferrious _ Cerous jon ion ion ion i: Q. 10. Which of the two ceric sulphate solution or Mohr’s salt solution can be taken in the burette? 3 Ans. Either of two can be taken in the burette and the other in the titration flask during standardisation 4 step. Only the colour change at the end point would be reversed. Q. 11. Why not sample solution be taken in the titration flask but in the burette ? Ans. It is to avoid the oxidation of nitrite by atmospheric oxidation. More-over formati is avoided, which may get decomposed. ‘on of nitrous acid_AVIRONMENTAL CHEMISTRY AND BIOTECHNOLOGY. 277 12, Why the use ofa burette with rubber ta ins, When KMnO, comes in contact with o ager which aetas reducing agents, tn 1D is avoided in KMn0, titration ? ganic matter stich as rubber or paper ete, it attacks organic Q.13. Why warming is not necessary in the titration of KMnO, against Fe** ions 2 is because ferrous ion (Fe? id a : Q.14. Why heating is required in the titration of KMnO, with oxalic acid? ‘Ans. A number of bonds have to be broken in oxidisi slow. Inorder to make it rapid, warming is required, COOH 60-70°¢ ing oxalate ion into CO, and H,O and the reaction is Q.15. Why shaking is required after small addition of KMnO, titrations ? Ans. KMnO, should always be added little ata time with constant shaking, otherwise a brownish precipi- tate of MnO, is formed. Q.16. Why a brown colour appears during KMnO, titrations sometimes ? Ans. It shows that either H)SO, has not been added or has been added in insufficient quantity. In such case through away the solution and start afresh, Q.17. Why itis better to store KMnO, is coloured stoppered bottles 2 Ans, A solution of KMnO,, once properly prepared is quite stable as long as it is completely protected from dust, access of reducing gases and direct sunlight. It is therefore, better to preserve the solution in stoppered bottles of dark brown colour. EXPERIMENT 22.10 Object To determine the amount of acetic acid and present in a given sample of vinegar. Theory ‘Acetic acid (CH, COOH) is an essential component of Vinegar, which is used as preservation in pickles and other foodstuffs. It is a dilute acid and can be conveniently estimated by titration with a standard solution of strong base (NaOH). But NaOH is not a primary standard, its standard solution cannot be prepared directly. The solution prepared being of approximate normality has to be first standarised. For this purpose, oxalic acid being a primary standard is suitable. So, the estimation involed two steps. (0. Standardisation of approx. N/10 NaOH solution with N/10 oxalic acid salution, (ii) Estimation of acetic acid with standardised NaOH solution, Chemical Required : (/) Approx. N/10 NaOH solution, (i) N/10 oxalic acid solution, (iii) Phenolphthalein as indicator Chemical Equations: First Titration : COOH COONa 1 +2Na0H——> | +2H,0 COOH COONa Second Titration : HyC—COOH+ NaOH + H\C—COONa+H,0 Indicator Phenolphthalein for both the titrations. End point : Disappearance of furnish colour. Procedure : (i) Standard of NaOH provided Transfer 20 ml of approx. N/10 NaOH solution into a clean ‘itation ask. Add a drop of phenolphthalein indicator. The solution will tur pink in colour. Fil the burette278 LABORATORY MANUAL ON ENGINEERING CHEMISTRY. with N/10 oxalic acid solution and titrate. Add the acid’solution slowly with constant strrr disppearance of pink colur. Repeat the titration to’get concordant readings. (ii) Estimation of Acetic acid from the sample solution Fill the burette with standardised NaOH solu- tion. Transfer 20 ml of the sample solution into a washed titration flask. Add a drop of Phenolphathalein indicator. Tirate with NaOH solution from the burette till the appearance of light pink colouration, Repeat the experiment to get concordant readings. Ing until the Observations ‘Volume of the respective solution taken in the titration flask for each titration = 20 ml. Record the observation of the two titration in separate tables as usual. Let the volume of oxalic acid solution used in the first titration be x ml. Similarly, let the volume of standardised NaOH solution used be y ml. Calculation Applying normality equation: 1 x NiV)/=_NGV, Sor) 20=) 5 kxvorNy= STON WOH ovaicacia Exact Normality of NaOH = ——N 200 : uke Again, We have NAV) =. NyVy 0FN,*20= Sy Asaigeca "NOW N, lity wert. acetic acid = =~ jy s (Normality wrt. acetic acid =

HO — HC — COO Na+ 3H,0 1,C — COOH J,C— COONa Citric acid Sod. citrate COOH COONa +2,Naoly ——> [ +2110 COO Na Sod, Oxalate lender or in a mortar and mix thoroughly. Transfer (0 ml beaker and find the weight of the material lost by evaporation. Step 1: Preparation of Fruit Extract, Pulp the fruit ina bl. about 25 g of the pulped material into a previously weighed 25 transferred. Add about 100 ml of water and boil for about 30 minutes replacing the water I Cool, transfer to a 250 ml volumetric flask and make up to volume mark. Filter ifmecessary. ‘Step It: Estimation of citric acid (Acidity). Pipette out 50m of the sample prepared into a250 ml conical flask and dilute it with recently boiled distilled water. Add one or two drops of phenophthalein indicator and titate with standard alkali. Express the acidity in terms of grams of anhyydrous eitric acid per 100 g of fruit (percentage), ‘Step IIT: Standardisation of NaOH solution. Pipette out 20 ml of NaOH solution into a washed titration flask, Add a drop of Phenophthalein. The solution turns pink in colour. Fill the burette with N/10 oxalic acid. Titrate by constant slow addition of oxalic acid solution into the titration flask while stirring the flask until the disappearance of pink colour. Repeat the experiment fo get concordant reading, Calculation Wt. of the pulp transferred Vol. of the digest made upto = 250ml Vol. of the aliquot pipetted out = SOml Vol. of alkali titrated = Vil Novmality of the alkali = N (determined by standardisation) Vol. of | N alkali used = V*Nml I ml of | N alkali is eqivalent to 0.064 g of citric acid (Eq. wt of anthydrous citric acid is 64.04). 250x100 _ V XN 0.064 * “oy 72% i i ah a in fruit Percentage ofeitric aci Report the acidity ofthe fruit in terms of g of anhydrous eitric acid per 100g of fruit (ow)280 LABORATORY MANUAL ON ENGINEERING CHEMISTRY Note : Ifthe juice is coloured, dilute a small volume of the juice with large quantity of water. Then the colour becomes so pale that the colour change of the indicator daring titration can easily be observed. The value of (Normality) N can be calculated from step III as under : Let the volume of 0.1 N oxalic acid solution used against 20 ml of approx 0.1 NNaOQH =xml Applying normality equation N,V, = N,V, 2 orNxx=0.1 x 200rN= Result : Acidity of the fruit, ie., percentage of citric in the fruit = 2% EXPERIMENT 22.12 Object To determine the amount of reducing sugar in the given fruit juice sample. Theory The reducing sugars present in the fruit juice reduces the alkaline solution of cupric salt (Fehling’s solution) to red cuprous oxide. Methylene blue, a redox indicator is employed to detect the end point of the titration. The red colour of the cuprous oxide formed during the reaction will be marked by the intense blue colour of the indicator. But when all the cupric ions of the Fehling’s solution are reduced to cuprous by the sugar solution (fruit juice.), then the next few drops of sugar solution added reduce and thus decolourise the indicator so that the red colour of the cuprous oxide becomes visible which is taken as the end point of the titration. ‘Chemical Equation CuSO,+2NaOH > Cu (OH), +Na, SO, Cu(OH), > Cu0+H,0 2Cu0+R-CHO + Cu,0+R-COOH Reddish The reducing sugars will be often expressed in terms of glucose, since glucose is the most predominant reducing sugar present in the fruits, Reagents Required 1. Fehling’s solution A. Dissolve 69.28 g CuSO,. SH,O in water and make upto 1 litre, 2. Fehling’s solution B. Dissolve 346 g of Rochelle salt (pottassium Sodium tartarate, KNa C,H, 100g of NaOH in water and make upto litre. 3. Methylene blue indicator solution 1% in water. 4, Neutral lead acetate solution 45% in water. 5 Potassium oxalate solution 22% in water. Procedure Extract ‘Step J: Preparation of Fruit Juice Sample Pulp the fruit in a blender and filter through Whatman No. 40 filter paper. Weigh and transfer about25 g of the filtered juice into a 250 ml volumetric flask. Add about 100ml of water and neutralize with 1 N NaOH. Add 2 ml of lead accetate solution,shake and allow to stand for 10 minutes. Add 2 ml of Potassium oxalate solution to remove the exces of lead, make up the volume with water to 250 mark and filter, Step 2: Titration of Fehlinig solution against the prepared Juice Extract Pipette out 5 ml each of Fehling a solutions A and B into a250 ml conical flask. Add about 50 ml of distilled water and 2 or 3 glass beads. Boil and contents vigorously and while boiling add the clarified fruit juice taken ina burette, till the blue colour just disappears Then add 0.5 ml of methylene blue indicator and allowit to boil _ for one minute. While boiling, complete the titration as quickly as possible by adding 2 to 3 drops of sugat_ 4H1,O)andlONMENTAL CHEMISTRY AND BIOTECHNOLOGY oo gontion 0F 5 10 10. second intervals, until the indicator is completely decolourized and the brick red coour of vows oxide becomes dominant (Do not interrupt boing for more than a few seconds as the indicator oes al 'as free access.into the flask). Calculate the content of reducing, inde fe u jucose per 1-0 ice mi i uadetégs a of glucose Per 1-00 B ofa juice (percentage) 10 ml of Fehling solution (A + B)~ 0.05 got glucose. Caleulation envi! ‘wt. of the fruit juice taken for analysis = we. a Volume made upto = 250m! me of the clarified juice reacted with 10ml of Febling’s solution : (A+B)= Vml(Titre Value) ig's soln (A +B) = 0.05 gm of Glucose © (g of glucose/100 g of juice) 0.05 250 VW tthe content of reducing sugars in the fruit juice given for analysisas g of glucose per 100 gofthe juice. vou! lomlof Feb percentage of reducing sugars in the j X% Repor Note For accurate determination of reducing sugars, it may be necessary to standardise the Fehling’s solution prepared by titration with standard glucose solution and to calculate the glucose equivalent value of 10 ml. by the procedure identical to that following in the case of the test sample, rather than taking Fehling solution franted that 10 ml. Fehling’s solutioin is equivalent to 0.05 g of glucose, Result Percentage of Reducing sugar = X % EXPERIMENT 22.13 Ovject : To determine the amount of Vitamin C in the given sample of fruit sample ‘Theory Kecorbic acid reduces the oxidation - reduction indicator dye, 2 6- dichloro phenol indopheno} to ¢ col ero tian, At the same time ascorbic acid is oxidised to dehydro-ascorbic acid. The excess unreduced eis rose pink in acid solution and therefore attainment of this colour during titration will indicate the end point of the reaction. Reagents Required 1. Metaphosphoric acid (HPO, ), 3% in water. 2. Ascorbie acid standard solution: 0.100 & Phosphoric acid. One ml of this solution contains fresh, just before use. 3. Dye solution: Dissolve 0.25 g of sodium salt of 2,6 dichloro phenol indophenol in about 500 ml of water fentaining 0.210 g of NaFiCO, and .ditute to I litre with water, ‘Store the solution in refrigerator and standardise with freshly prepared standard solution of ascorbic acid, every time, just before use. Procedure eninge J : Standardisation of the dye flask and dilute it with an equal vo! Kared dye solution taken in a burett until alight but rose F £9" 10 seconds. Determine the dye factor ic, me of ser ee i metaphe.2 : Preparation of the sample ‘Transfer about 100 g of fur hosphorie acid. ‘Transfer the content into a one litre volumetric NOS met “Phosphoric acid. Filter or centrifuge if necessary: of ascorbic acid dissolved and made up to 1 lit in 3% meta- 0.1 mg. of ascorbic acid. This solution should be prepared corbie acid solution into a 250 ml jd solu rapidly with the te out 10 mot Sohoric ack solution. Titrate rapidly wit ume of 37% metaphesP Oot colour is obtained which persists at Pitiguivalent to Lamlofthe dye. siya ptendter and blend it with 3% wt atce up the votume with 3% Pipette out 10 ml of standard as:282 LABORATORY MANUAL ON ENGINEERING CHEMIstry ‘Step 3 : Assay of Extract : Pipette out 10 ml of the acid extract ofthe fruit into a 250 ml conical lask and dilute it with an equal volume of 3% meta phosphoric acid. Titrate with standard dye solution toa light rose ping end point which should persist at least for 10 seconds. Titrate rapidly and make a preliminary determination of the titre. In the next determination add most of the dye required and then titrate accurately. Express the ascorbic” acid content as mig of acid per 100 g of fruit, 3 Calculations > @) Standardisation of dye solution Volume of dye solution reacted with 10 ml of std ascorbic acid solution = V ml (Titre value) 10ml ascorbic acid solution = Img of ascorbic acid Therefore | ml of dye is equivalent to + mg of ascorbic acid = Dye factor (6) Aseorbic acid content of fruit Wu. of the fruit taken = |W g Volume made upto = 100m! Volume of the dye reacted with 10 ml of fruit extract = Tmi (Titre value) T x1000 x100 a Miligram of ascorbic acid in 100 g of fruit vxloxw. 28 Report the ascorbic acid content of the fruit as mg/100 g of fruit. Note: 1. It is difficult to see the end point when the fruit juice is highly coloured. In such case, add 1 ml of. chloroform to the reaction mixture (taken in a boiling tube) and the end pointis often when a rose pink colour is seen in the organic phase. The test and blank are treated the same way and vitamin C content calculated as previously. 2.2% oxalic acid can be used in place of 3% metaphosphoric acid. : Result; ] Amount of Vitamin = Zmg/100g of fruit a EXPERIMENT 22.14 : Object : To determine the amount of strach in a plant material sample. ; Theory 4 Strach is first made available from sugars present in the plant material by treatment with ethanol. Itis then hydrolysed to glucose by digestion with dilute HCI. The reducing sugar in the hydrolysate is then determined i : by titration with Fehling’s solution, ; (C,H,,0,),+nH,0 > n0,H,,0, 162 g of starch on hydrolysis yields 180 g of glucose. Therefore 1 g of gcse obtained on hydrolysis is equal to 0.9 g of starch. Procedure Step! : Liberation of Slarch Weigh and transfer 2 to 5 g of the powdered plant material into a 100 ml centrifuge tube. Wet it with a few drops of water, Add about 100 ml of 95% ethanol and stir the content with a glass rod for 30 minutes at inteNVIRONMENTAL CHEMISTRY AND BIOTECHNOLOGY sa arifyge ill the precipitate setles a the bottom, Filler and wash the residue about 50% ethanol until the cer ives no test for sur ‘test for sugar : Toa few ml ofthe filtrate in a small narrow test tube, add 2 drops of 10% aleoholic solution ofalpha naphthol. Add | ml of conc.H,,SO, slowly down the side of the test tube so as to form a layer beneath fhe aqueous solution. Ifsugars are presenta red ring will appear within a few seconds atthe junction ofthe two layers ‘ : ‘Transfer the residue to a250 ml conical flask with about 100 ml of water. Add about 20 mi of cone. HCI, Jace asmall funnel at the mouth of flask to prevent evaporation and digest the content on a hot plate for about sehour, Cool, make the content alkaline to litmus with sodium hydroxide, filter ifnecessary and make up toa Sefinite volume (250 ml) with water, Step 2 Titration of above extract with Fehling solution. For details refer to experiment 20.12 Calculation : For details refer to experiment 22.12 Wt. of the plant material taken = W g The volume of the solution made = 250ml Vol. of solution reacted with 10 ml of Fehling’s solution = A (ml) Titre value 10 ml of Fehling’s solution is equivalent to 0.05 g of glucose 1 g of glucose obtained on hydrolysis = 0.9 g of starch. 0.05 x 250 AxW ‘% of Strach in the plant materia *100*0.9=Z% Report the % of starch in the sample on moisture free basis. Note The removal of sugars prior to the acid hydrolysis of starch described in para I of the procedure can be avoided in the case of samples containing only negligible amount of sugars or when the total sugar content of the sample is known. Result Percentage of strach in the plant material = 2% AlternateMethod Procedure To | gfresh sample ground ina mortar with pestle, add 20 ml of 2N HCI and heat on a water bath for 2 hours. Cool, neutralise and make up to 100 ml. Thisis titrated while boiling against 10 ml of 0.0SN K,Fe(CN), to which a of 2.5N NaOH is added using methylene blue as indicator. A blank titration is run with 0.05 N glucose solution, | Caleulation I ml of 0.05N glucose = 9.01 mg glucose Letthe blank titre value be *X’ ml, then X mlof0.05 N glucose= X x 9.01 mg glucose To reduce 10 ml of 0.05N K,Fe(CN),= X 9.01 mg glucose (X x 9.01)% 100 x 100 x19 Titre value x W 1000 x20 W= Weight of sample in gram Percent starch in the unknown solution = 19 739 7 Factor for conversion of sugar to strach