Experiment No: 1

Title: To Prepare and Standardize 0.1N Perchloric acid. Reference: Practical Pharmaceutical Analysis by Dr. G. D. Devala Rao. Equipment Required: Volumetric Flask, Pipette, Conical flask, Measuring cylinder, and burette. Material required: Perchloric acid, Glacial acetic acid, acetic anhydride, Potassium hydrogen phthalate, Crystal violet indicator. Chemical reaction:
COOK HClO4 COOH COOH COOH KClO4

Theory: Titrimetric method employing non aqueous solvents are extensively used for assay of certain materials which cannot be easily titrated in aqueous solution. In non aqueous solvent such as glacial acid, weak organic bases and their salts can be titrated with perchloric acid solution. Procedure: Step-1 Preparation of 0.1 N Perchloric acid:a. In a 100ml volumetric flask mix 0.85 ml of perchloric acid with 50 ml of glacial acetic acid, 3.0 ml of acetic anhydride. b. Cool and add more glacial acetic acid to make 100 ml. Allow the solution to stand for 24 hours before use. Step-2 Standardisation of 0.1 N Perchloric acid:a. Pipette out 10 ml of 0.1N Potassium hydrogen Phthalate solution into a dry conical flask and add 2 to 3 drops of crystal violet indicator. b. Titrate the contents of the flask against perchloric acid until the colour changes to green. c. Repeat the titration for concordant values. Observation: Vol. of 0.1 N KHP (ml) Burette reading Vol. of HClO4 used (ml) Initial (ml) Final (ml) 10 0 X X 10 0 X X 10 0 x X

N1V1 = N2V2
Where, N1 = normality of KHP (0.1), V1 = volume of KHP (10 ml) N2 = normality of HClO4 (?), V2 = volume of HClO4 (X ml) N2 = (N1V1)/V2 = (0.1 * 10)/X =Y Result: Normality of given Perchloric acid is found to beY N

Experiment No: 2
Title: To find the strength of sodium acetate or potassium acetate using perchloric acid by non aqueous titration. Reference: Mendham, J.; Denny, R. C.; et al. Vogels Text Book of Quantitative Chemical Analysis, 6th edition; Person Education , 2008. Equipment Required: Volumetric Flask, Pipette, Conical flask, Measuring cylinder, and burette. Material required: Perchloric acid, acetic anhydride, acetic acid, sodium acetate, crystal violet, p-nitrobenzene, Potassium hydrogen phthalate. Theory: Titrimetric method employing non aqueous solvents are extensively used for assay of certain materials which cannot be easily titrated in aqueous solution. In non aqueous solvent such as glacial acid, weak organic bases and their salts can be titrated with perchloric acid solution. Chemical reaction:
COOK HClO4 COOH CH3COOH COOH COOH KClO4

CH3COONa

HClO4

NaClO4

Procedure: Step-1: Standardization of 0.1 N Perchloric acid:1. Take 2.1 ml of 70% perchloric acid, add 5.2 ml of acetic anhydride and dilute to 250 ml with acetic acid. 2. Weigh about 0.2 mg Potassium hydrogen phthalate and add 2.5 ml of acetic acid. 3. Warm & cool to room temp. 4. Add it p-nitrobenzene as indicator & titrate against perchloric acid and find out normality of perchloric acid. Step-2: Titration of sodium acetate with standard HClO4:1. Prepare 0.1 N sodium acetate soln. 2. Pipette out known amount of sol. in flask and add crystal violet as indicator. 3. Titrate it with perchloric acid & find the normality & then strength. Observation: Step 1: Titration between KHP vs HClO4 Vol. of 0.1 N KHP (ml) Burette reading Initial (ml) 10 0 10 0 10 0

Vol. of HClO4 used (ml) Final (ml) X X x X X X

N1V1 = N2V2
Where, N1 = normality of KHP( 0.1), V1 = volume of KHP (10 ml) N2 = normality of HClO4 (?), V2 = volume of HClO4 (X ml) N2 = (N1V1)/V2 = (0.1 * 10)/X =Y 5

Step 2: Titration between sodium acetate vs HClO4 Vol. of 0.1 N sodium Burette reading acetate (ml) Initial (ml) Final (ml) 10 0 a 10 0 a 10 0 a

Vol. of HClO4 used (ml) a a a

N1V1 = N2V2
Where, N1 = normality of sodium acetate (?), V1 = volume of sod. acetate (10 ml) N2 = normality of HClO4 (Y), V2 = volume of HClO4 (a ml) N2 = (N1V1)/V2 = (Y * a)/10 =b Step 3: strength of sodium acetate = normality * equivalent weight =b*q = M g/litre Result: Normality of given sodium acetate is found to beb N. Strength of sodium acetate is found to be.M g/lt . Caution: 1 All the apparatus should be clean and dry before being used. 2. Balance and weights should be calibrated.

Experiment No: 3
Title: To Prepare and Standardize N/10 HCl with NaOH pH metrically. Reference: Mendham, J.; Denny, R. C.; et al. Vogels Text Book of Quantitative Chemical Analysis, 6th edition; Person Education , 2008. Equipment Required: Volumetric Flask, Conical flask, Measuring cylinder, pH meter and burette. Material required: Buffer soln. of pH- 4 and pH- 7, N/2 oxalic acid soln., N/2 NaOH soln., N/10 HCl soln. Chemical reaction: COOH COONa 2NaOH H 2O COOH COONa

HCl

NaOH

NaCl

H 2O

Theory: pH meter is an electronic digital voltmeter, scaled to read pH directly. A glass rod has a asymmetry potential which makes it impossible to relate a measured electrode potential directly to the pH of the soln. and it necessary to calibrate the electrode. A pH meter therefore always includes a control so that with the electrode assembly placed in a buffer soln. of known pH, the scale reading of the instrument can be adjusted to the correct value. Procedure: Step-1:- Standardisation of NaOH: a. Pipette out 20 ml of oxalic acid in conical flask. b. Add 2-3 drops of indicator. c. Add NaOH soln. dropwise with constant shaking till the end point reached. d. Repeat the procedure to get three concordant readings. Step-2 Standardisation of N/2 HCl using pH meter with NaOH:a. Switch on the pH meter and wait for 10-15 minutes. b. Standardized the pH meter by immersing glass electrodes in standardized buffer soln. having pH- 4 and then in buffer soln having pH- 7. c. Adjust the value of pH meter exactly to the value of buffer used. Also adjust the temperature to normal room temperature. d. Take 50 ml HCl soln. and add 50 ml of water. Stirred the content thoroughly. e. Immersed the glass electrode in the beaker and clamped in it. f. Clamped the burette to the stand and add NaOH from burette to beaker a g. Note the pH value continued till we get a constant pH. h. Plot a graph between the pH and NaOH volume used.

Observation: Vol. of 0.1 N oxalic acid (ml) 10 10 10 Burette reading Initial (ml) Final (ml) 0 X 0 X 0 X Vol. of NaOH used (ml) X X X

N1V1 = N2V2
Where, N1 = normality of oxalic acid (0.1), N2 = normality of NaOH (?), V1 = volume of oxalic acid (10 ml) V2 = volume of NaOH (X ml)

N2 = (N1V1)/V2 = (0.1 * 10)/X =Y Vol. of NaOH used (ml) 10.0 10.5 11.0 11.5 12.0 12.5 13.0 pH A A A(end point) A B B B

N1V1 = N2V2
Where, N1 = normality of NaOH (Y), N2 = normality of HCl (?), V1 = volume of NaOH (11 ml) V2 = volume of HCl (50 ml)

N2 = (N1V1)/V2 = (Y * 11)/50 =Q Result: The normality of HCl is ..Q N

Experiment No: 4
Title: To Prepare and Standardize N/10 HCl and N/10 acetic acid with NaOH pH metrically. Reference: Mendham, J.; Denny, R. C.; et al. Vogels Text Book of Quantitative Chemical Analysis, 6th edition; Person Education , 2008. Equipment Required: Volumetric Flask, Conical flask, Measuring cylinder, pH meter and burette. Material required: Buffer soln. of pH- 4 and pH- 7, N/2 oxalic acid soln., 0.1 NaOH soln., N/10 HCl soln. and 0.1 N acetic acid. Chemical reaction:
COOH COOH 2NaOH COONa COONa

H2 O

CH3COOH

HCl

2NaOH

NaCl

H 2O

CH3COONa

Theory: pH meter is an electronic digital voltmeter, scaled to read pH directly. A glass rod has a asymmetry potential which makes it impossible to relate a measured electrode potential directly to the pH of the soln. and it necessary to calibrate the electrode. A pH meter therefore always includes a control so that with the electrode assembly placed in a buffer soln. of known pH, the scale reading of the instrument can be adjusted to the correct value. Procedure: Step-1:- Standardization of NaOH: a. Pipette out 20 ml of oxalic acid in conical flask. b. Add 2-3 drops of indicator. c. Add NaOH soln. dropwise with constant shaking till the end point reached. d. Repeat the procedure to get three concordant readings. Step-2 Standardization of mixture N/2 HCl and N/2 CH3COOH using pH meter with NaOH:a. Switch on the pH meter and wait for 10-15 minutes. b. Standardized the pH meter by immersing glass electrodes in standardized buffer soln. having pH- 4 and then in buffer soln having pH- 7. c. Adjust the value of pH meter exactly to the value of buffer used. Also adjust the teperature to normal room temperature. d. Take mixture of 25 ml HCl soln. and 25 ml of acetic acid then add 50 ml of water. Stirred the content thoroughly. e. Immersed the glass electrode in the beaker and clamped in it. f. Clamped the burette to the stand and add NaOH from burette to beaker a g. Note the pH value continued till we get a constant pH. h. Plot a graph between the pH and NaOH volume used. 9

Observation: Vol. of 0.1 N oxalic acid (ml) 10 10 10 Burette reading Initial (ml) Final (ml) 0 X 0 X 0 X Vol. of NaOH used (ml) X X X

N1V1 = N2V2
Where, N1 = normality of oxalic acid (0.1), N2 = normality of NaOH (?), V1 = volume of oxalic acid (10 ml) V2 = volume of NaOH (X ml)

N2 = (N1V1)/V2 = (0.1 * 10)/X =Y Vol. of NaOH used (ml) 10.0 11.0 12.0 13.0 14.0 15.0 16.0 pH of the solution A A A(end poin) A B B(end point) B

N1V1 = N2V2
Where, N1 = normality of NaOH (Y), N2 = normality of HCl (?), V1 = volume of NaOH (12 ml) V2 = volume of HCl (25 ml)

N2 = (N1V1)/V2 = (Y * 12)/25 =Q

N1V1 = N2V2
Where, N1 = normality of NaOH (Y), N2 = normality of CH3COOH (?), N2 = (N1V1)/V2 = (Y * 15)/25 =Z Result: The normality of HCl is ..Q N The normality of CH3COOH is ..Z N V1 = volume of NaOH (15ml) V2 = volume of CH3COOH (25 ml)

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Experiment No: 5
Title: To carry out the extraction of Acetaminophenol from paracetamol. Reference: Verma, R. M. Analytical Chemistry. CBS publisher and distribution, 3rd edition, 2000, page no. 279-281. Equipment Required: Separating funnel, Measuring cylinder, glass rod, filter paper and beaker. Material required: Paracetamol and chloroform. Theory: Solvent extraction of solid is widely performed to extract both the organic as well as inorganic substances. The process is done by simple mixing of solute and solvent and filtering and some time drying as well. Procedure: 1. Weigh about 250 mg of powdered paracetamol. 2. Put it in separating funnel and extract with 10 ml of chloroform for 30 minutes. 3. Repeat three times and filter the extract. 4. Allow it to dry by general heat. 5. Note the percentage yield and melting point of the substance. Observation: Theoretical yield= 250 mg Practical yield= X g % yield= (Practical yield /Theoretical yield) * 100 = (X/250) * 100 =Y% Result: Percentage yield of extracted product is Y%.

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Experiment No: 6
Title: To determine the end point by Conductometry for Strong acid v/s Strong base. Reference: Kasture, A. V.; Wadodkar, S. G.; Mahadik, K. R.; Mole, H. N. Pharmaceutical Analysis. Nirali Publication, 6th edition, 2002, page no. 216-217. Equipment Required: Conductometer, Volumetric Flask, Pipette, Conical flask, burette and beaker. Material required: Sodium hydroxide, oxalic acid, Hydrochloric acid, and phenolphthalein. Theory: The determination of equivalence point of a titration by conductometric methodis based o the course of conductance varies in different manner before and after the equivalence point. This is because of electrical conductance of a soln. depending upon the number of ions present in soln. and their ionic mobilities. Thus on plotting the conductance against the volume of titrant added, we get two straight lines, the point of intercept of these lines give the equivalence point. Procedure: Step-1:- Standardisation of 0.1 N NaOH: a. Pipette out 20 ml of 0.1 N oxalic acid (Weigh 0.63 gm of oxalic acid and make the volume to 100 ml) in conical flask. b. Add 2-3 drops of indicator. c. Add 0.1 N NaOH soln. (dissolve 1 g of NaOH in 250 ml distilled water) dropwise with constant shaking until the solution becomes pink in colour. d. Note the end point. Calculate the normality of NaOH solution. e. Repeat the procedure to get three concordant readings. Step-2 Titration of HCl with NaOH:a. Prepare 0.1 N HCI. b. Set the conductometric apparatus & proceed the titration by taking 50 ml of 0.1N HCI in a beaker. c. Dip the conductometric cell in it. d. Add 0.1 N NaOH solution with constant stirring & take the reading. e. Repeat the above steps to take 8-10 readings when the conductivity starts increasing. f. Plot the readings on a graph to determine the end point. Observation: Vol. of 0.1 N oxalic acid (ml) 10 10 10 Burette reading Initial (ml) Final (ml) 0 X 0 X 0 X Vol. of NaOH used (ml) X X X

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N1V1 = N2V2
Where, N1 = normality of oxalic acid (0.1), N2 = normality of NaOH (?), V1 = volume of oxalic acid (10 ml) V2 = volume of NaOH (X ml)

N2 = (N1V1)/V2 = (0.1 * 10)/X =Y Vol. of NaOH used (ml) 2.0 4.0 6.0 16.0 Conductometric reading (-1) A A A A B B B

Volume of NaOH use at which end point is reached= M ml calculate from graph

N1V1 = N2V2
Where, N1 = normality of NaOH (Y), N2 = normality of HCl (?), V1 = volume of NaOH (M ml) V2 = volume of HCl (100 ml)

N2 = (N1V1)/V2 = (Y * M)/100 =Q Result: The normality of HCl is ..Q N

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Experiment No: 7
Title: To standardize the 0.1 N HCl and 0.1 N CH3COOH with NaOH by Conductometry. Reference: Kasture, A. V.; Wadodkar, S. G.; Mahadik, K. R.; Mole, H. N. Pharmaceutical Analysis. Nirali Publication, 6th edition, 2002, page no. 219-220. Equipment Required: Conductometer, Volumetric Flask, Pipette, Conical flask, burette, microbureette and beaker. Material required: Sodium hydroxide, oxalic acid, Hydrochloric acid, acetic acid and phenolphthalein. Theory: The determination of equivalence point of a titration by conductometric methods based o the course of conductance varies in different manner before and after the equivalence point. This is because of electrical conductance of a soln. depending upon the number of ions present in soln. and their ionic mobilities. Thus on plotting the conductance against the volume of titrant added, we get two straight lines, the point of intercept of these lines give the equivalence point. Procedure: Step-1:- Standardisation of 0.1 N NaOH: b. Pipette out 20 ml of 0.1 N oxalic acid (Weigh 0.63 gm of oxalic acid and make the volume to 100 ml) in conical flask. b. Add 2-3 drops of indicator. c. Add 0.1 N NaOH soln. (dissolve 1 g of NaOH in 250 ml distilled water) dropwise with constant shaking until the solution becomes pink in colour. d. Note the end point. Calculate the normality of NaOH solution. e. Repeat the procedure to get three concordant readings. Step-2 Titration of mixture of HCl and acetic acid with NaOH:a. Prepare 0.1 N HCI and 0.1 N acetic acid. Mix both the solution. b. Set the conductometric apparatus & proceed the titration by taking 50 ml of mixture in a beaker. c. Dip the conductometric cell in it. d. Add 0.1 N NaOH solution with constant stirring & take the reading. e. Repeat the above steps to take 8-10 readings when the conductivity starts increasing. f. Plot the readings on a graph to determine the end point. Observation: Vol. of 0.1 N oxalic acid (ml) 10 10 10 Burette reading Initial (ml) Final (ml) 0 X 0 X 0 X Vol. of NaOH used (ml) X X X

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N1V1 = N2V2
Where, N1 = normality of oxalic acid (0.1), N2 = normality of NaOH (?), V1 = volume of oxalic acid (10 ml) V2 = volume of NaOH (X ml)

N2 = (N1V1)/V2 = (0.1 * 10)/X =Y Vol. of NaOH used (ml) 2.0 4.0 6.0 16.0 Conductometric reading (-1) A A A A B B B

Volume of NaOH use at which end point is reached= M ml calculate from graph

N1V1 = N2V2
Where, N1 = normality of NaOH (Y), N2 = normality of HCl (?), V1 = volume of NaOH (M ml) V2 = volume of HCl (50 ml)

N2 = (N1V1)/V2 = (Y * M)/50 =Q

N1V1 = N2V2
Volume of NaOH use at which end point is reached= N ml calculate from graph Where, N1 = normality of NaOH (Y), V1 = volume of NaOH (N ml) N2 = normality of CH3COOH (?), V2 = volume of CH3COOH (50 ml) N2 = (N1V1)/V2 = (Y * N)/50 =T Result: The normality of HCl is ..Q N The normality of CH3COOH is ..T N

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Experiment No:8
Title: Packing of column in column chromatography. Reference: Verma, R. M. Analytical Chemistry. CBS publisher and distribution, 3rd edition, 2000, page no. 285-288. Equipment Required: Column. Material required: Toluene, Silica gel, mesh no. 100, 200, column. Theory: Chromatography is a separative process which is very useful for separating molecular mixtures. In this technique advantage is takes of the differential adsorption of different components of a mixture by an adsorbent. For example, suppose a petroleum ether extract of green leaves is allowed to flow down through a glass tube filled with CaCO3 powder. The ether extract is a mixture of different components derived from green leaves such as green pigments, yellow pigments (xanthophylls) and another yellow substance (carotene). The green pigments are more strongly adsorbed and so are more firmly held by the powder consequently they are not able to more much down the column and remain near the top of the column. Yellow pigments are less strongly adsorbed so they move down the column. The adsorption column along with its coloured bands containing separated material is called a chromatogram.

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Experiment No:9
Title: To perform the TLC of given sample. Reference: Verma, R. M. Analytical Chemistry. CBS publisher and distribution, 3rd edition, 2000, page no. 285-288. Equipment Required: Glass slide, TLC jar, beaker and glass rod. Material required: Toluene, pet ether, methyl acetate, Silica gel, mesh no. 100, 200, ethanol, alcohol. Theory: Chromatography is a separative process which is very useful for separating molecular mixtures. In this technique advantage is taken of the differential adsorption of different components of a mixture by an adsorbent. In this technique an adsorbent coated on a glass plate, acts as the stationary phase. The mobile phase percolates through the adsorbent and carries along with it various components of a mixture which move with different speed and thus get separated. Calculation: Rf = distance travel by solute/ distance travel by solvent

Result: the Rf valute of given sample is..

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Experiment No:10
Title: To perform the paper chromatography of given sample. Reference: Verma, R. M. Analytical Chemistry. CBS publisher and distribution, 3rd edition, 2000, page no. 289-291. Equipment Required: Paper, beaker and whatmans filter paper. Material required: Toluene, pet ether, methyl acetate, ethanol, alcohol. Theory: Chromatography is a separative process which is very useful for separating molecular mixtures. In this technique advantage is taken of the differential adsorption of different components of a mixture by an adsorbent. For example, suppose the separation of a mixture of amino acids by mean of a strip of a filter paper. A drop of a soln. containing the mixture of amino acids is placed at one end of the strip, is allowed to dry and its position on the paper is marked with pencil. Equal volume of phenol and water are shaken in a separating funnel. The upper layer I consist of water which is used as stationary phase while the lower layer II consist of phenol and this is used as mobile phase. The strip is hanged in a gas jar. The mixture of layer I is placed at the bottom of the jar so that the air inside become saturated with it. The layer II acts as the mobile phase is now placed in jar and closed it. The developers move into the paper by capillary action. As the developer flows down, the different amino acid present in the mixture, also moves down with different characteristics rates. When the developer reaches almost to the lower end of the strip the gas jar is opened and paper strip is removed. Since amino acids are colorless the strip is sprayed with 0. 1% sol. Of nyn hydrin and warmed when different spots of purple colou are observed. Calculation: Rf = distance travel by solute/ distance travel by solvent

Result: the Rf valute of given sample is.. 18

Experiment No:11
Title: To perform the TLC of given sample in the solvent mixture of methanol and toluene. Reference: Verma, R. M. Analytical Chemistry. CBS publisher and distribution, 3rd edition, 2000, page no. 285-291. Equipment Required: Beaker, glass rod and TLC jar. Material required: Toluene, Silica gel, slide, mesh no. 100, 200, and methanol. Theory: Chromatography is a separative process which is very useful for separating molecular mixtures. In this technique advantage is taken of the differential adsorption of different components of a mixture by an adsorbent. In this technique an adsorbent coated on a glass plate, acts as the stationary phase. The mobile phase percolates through the adsorbent and carries along with it various components of a mixture which move with different speed and thus get separated. Procedure: Step 1: Preparation of glass slide: a. Prepare silca gel by adding silica gel in water. b. Put slowly on glass slide to form uniform layer. c. Keep it in oven to dry it. Step 2: Preparation of saturated solvent soln. a. Take the solvent methanol and toluene in ratio 5:5 in a beaker b. Saturated the jar by covering with aluminum foil. Step 3: Perform the TLC. a. Place a one drop min ink and one drop red ink and one drop of extraction to slide with the help of capillary. b. Keep the slide in solvent soln. for 5 min. in a beaker and cover it with aluminum foil to make it saturated. c. Note the Rf Calculation: Rf = distance travel by solute/ distance travel by solvent

Result: the Rf valute of given sample is..

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Experiment No: 12
Title: To Prepare and Standardize 0.05 M EDTA Solution. Reference: Kasture, A. V.; Wadodkar, S. G.; Mahadik, K. R.; Mole, H. N. Pharmaceutical Analysis. Nirali Publication, 6th edition, 2002, page no. 219-220. Equipment Required: Volumetric Flask, Pipette, Conical flask, burette and beaker. Material required: EDTA, Calcium chloride, pH-10 Buffer solution, Solochrome Black-T (SBT). Theory: EDTA is very unselective reagent because it complexes with numerous doubly, triply and quadruply charged cations. In strongly acidic soln. the dye tends to polymerise to a redbrown product, and consequently the indicator is rearly applied in titrations of soln. more acidic than pH 6.5. Procedure: a. Pipette out 10 ml of 0.05 M of Calcium chloride solution into a conical flask. b. Then add 5 ml of pH 10 buffer solution. c. Add 2 or 3 drops of SBT indicator d. Titrate against EDTA solution. e. Repeat the titration for concordant values. Observation: Vol. of 0.05 M calcium chloride (ml) 10 10 10 Burette reading Initial (ml) Final (ml) 0 X 0 X 0 X Vol. of EDTA soln. used (ml) X X X

N1V1 = N2V2
Where, N1 = molarity of calcium chloride (0.05), N2 = molarity of EDTA soln (?), N2 = (N1V1)/V2 = (0.05 * 10)/X =Y Result: The exact molarity of given solution of EDTA is found to be......Y M... V1 = volume of calcium chloride (10 ml) V2 = volume of EDTA soln (X ml)

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