N.K.B.R. COLLEGE OF PHARMACY & RESEARCH CENTRE MEERUT (U.P.

LAB MANUAL Session 2010-2011

PHARMACEUTICAL ANALYSIS - I (PHARM-112P) Year: - 1ST Prepared By: AMIT KUMAR

Senior Lecturer

Semester: - I

Submitted to: Prof. R. Hemalatha Principal NKBR Col. of Pharmacy & Research Centre

LIST OF EXPERIMENTS (B.Pharm Ist Sem.)

1. To study the Glass Apparatus. 2. To study the operation of Analytical Balance. 3. To carry out the calibration of the Burette, Pipette & Volumetric Flask. 4.

To prepare 100 ml 0.1N NaOH and 0.1N HCl solution.

5. To prepare & standardization of 0.1N NaOH solution. 6. To carry out assay of Boric Acid. 7. To prepare & standardization of 0.1N AgNO3 solution. 8. To carry out assay of NaCl by Mohrs method. 9. To prepare & standardization of 0.1N KMnO4 solution. 10. To prepare & standardization of 0.1N NH4SCN solution. 11. To carry out assay of sodium chloride by Volhards method. 12. To carry out assay of KI by Potassium Iodate Titration. 13. To prepare & standardization of 0.05M KIO3. 14. To carry out assay of ferrous sulphate by Redox Method. 15. To prepare & standardization of 0.1N Ceric Ammonium Sulphate.

EXPERIMENT NO. 1 Aim: To study the Glass Apparatus. Requirements: 1. Apparatus Required: Beakers, volumetric flasks, conical flask, glass road, measuring cylinder, pipettes, test tube holders, funnel etc. Procedure: Study of Glass Apparatus of the following 1. BEAKER A beaker is a simple container for stirring, mixing and heating liquids commonly used in many laboratories. Beakers are generally cylindrical in shape, with a flat bottom and a lip for pouring. Many also have a small spout to aid pouring as shown in the picture. Beakers are available in a wide range of sizes, from one millilitre up to several litres. Shape Beakers are often graduated, that is, marked on the side with lines indicating the volume contained. For instance, a 250 mL beaker might be marked with lines to indicate 50, 100, 150, 200, and 250 mL of volume. These marks are not intended for obtaining a precise measurement of volume (a graduated cylinder would be a more appropriate instrument for such a task), but rather an estimation. The presence of a lip means that the beaker cannot have a lid. However, when in use, beakers may be covered by a watch glass to prevent contamination or loss of the contents, but allowing venting via the spout.

Function: The primary function of a beaker is to hold and work with liquids. If graduated, it can serve to make approximate measurements of liquid volume. The beaker is made of specialized glass so it can be heated and cooled without breaking. This type of glass makes the beaker brittle and it must be handled carefully. 2. PIPETTE Pipette is a laboratory instrument used to transport a measured volume of liquid.

Function: Pipettes are devices that allow the users to extract or deliver small amounts of a liquid. Pipettes come in a variety of designs with only two shown. Some are graduated to deliver exact quantities, but most allow for a "drop at a time" delivery.

3. STIRRER

MAGNETIC STIRRER A magnetic stirrer is a laboratory device consisting of either a rotating magnet or stationary electromagnets creating a rotating magnetic field. This device is used to cause a stir bar (aka a flea) immersed in a liquid to spin very quickly, agitating or mixing the liquid.

Function: The function of a stirrer is to agitate liquids for speeding up reactions or improving mixtures. The picture on the left is a magnetic stirrer used with hot plates. The picture on the right is a typical glass stirrer used in beakers or test tubes. 4. TEST TUBE A test tube, also known as a culture tube or sample tube, is a common piece of laboratory glassware consisting of a finger-like length of glass or clear plastic tubing, open at the top, usually with a rounded U-shaped bottom. A large test tube designed specifically for boiling liquids is called a boiling tube. Test tubes are available in a multitude of lengths and widths, typically from 10 to 20 mm wide and 50 to 200 mm long. The top often features a flared lip to aid pouring out the contents; some sources consider that the presence of a lip is what distinguishes a test tube from a culture tube. Some test tubes have a flat bottom; some are made so as to accept a ground glass stopper or a screw cap. They are often provided with a small ground glass or white glaze area near the top for labeling with a pencil.

Function: The function of a test tube is to hold a small experiment, which would be used to conduct an investigation. The test tube is made of specialized glass so it can be heated and cooled without breaking. This same type of glass makes the test tube brittle and it must be handled carefully. 5. TEST TUBE HOLDER A test tube holder is used in a laboratory and it is used to hold/ support test tubes. Although it is called a test tube holder it is also used to hold/ support piettes, etc.

It is a clamp that holds a test tube -- for instance to hold a test tube while heating it without getting burned. It is made from a thick piece of metal wire that is shaped into a spring-loaded clamp to securely hold a test tube.

Note the difference between a test tube clamp, and a test tube rack. A clamp is a way to hold a single test tube while heating the contents inside without actually touching the test tube itself. A rack is instead used to hold many test tubes at once in an upright position. See the Related Questions links to the left for more information on test tube racks. Function: The test tube holder obviously is designed to hold test tubes. However, it can be used to hold pipettes and stirring rods, etc. as well. 6. THERMOMETER Thermometer is a device that measures temperature or temperature gradient using a variety of different principles. A thermometer has two important elements: the temperature sensor (e.g. the bulb on a mercury thermometer) in which some physical change occurs with temperature, plus some means of converting this physical change into a value (e.g. the scale on a mercury thermometer). Thermometers increasingly use electronic means to provide a digital display or input to a computer.

Function: The thermometer is an instrument for determining temperature. Most laboratory thermometers are calibrated in the SI scale (degrees Celsius). 7. WASH BOTTLE A wash bottle is a squeeze bottle with a nozzle, usually used to rinse various pieces of laboratory glassware, such as test tubes and round bottom flasks. Most wash bottles are made up of polyethylene, which is solvent-resistant petroleum based plastic. Most bottles contain an internal straw allowing upright use. Usually, wash bottles are filled with deionized water. Wash bottles may also be filled with detergent solutions and rinse solvents such as acetone and ethanol. In biological labs it is common to keep sodium hypochlorite solution in a wash bottle to conveniently disinfect unneeded cultures. When pressure is applied to the bottle, the liquid inside becomes pressurized and is forced out the nozzle into a narrow stream of liquid. As always the case when transferring reagents between containers, for safety always ensure the label on the wash bottle matches the contents of the bottle, and do not mix reagents or return them to the original container. Additionally, wash bottles are not appropriate for long term storage and must never be used for dangerous reagents. Volatile liquids such as acetone

or methanol require a bleed hole to reduce dribbling - often a vent is integrated into the top of the internal straw.

Function: Wash bottles usually contain distilled water and makes for a convenient method to wash out test tubes or pipettes. Wash bottles can also be used to dispense chemicals into test tubes or micro-chemistry depression plates. 13. FLASKS Laboratory flasks are vessels (containers) which fall into the category of laboratory equipment known as glassware. In laboratory and other scientific settings, they are usually referred to simply as flasks. Flasks come in a number of shapes and a wide range of sizes, but a common distinguishing aspect in their shapes is a wider vessel "body" and one (or sometimes more) narrower tubular sections at the top called necks which have an opening at the top. Laboratory flask sizes are specified by the volume they can hold, typically in metric units such as milliliters (mL or ml) or liters (L or l). Laboratory flasks have traditionally been made of glass, but can also be made of plastic. List of flasks There are several types of laboratory flasks, all of which have different functions within the laboratory. The most common types of flask are: Erlenmeyer Flask or Conical Flask.

Round-Bottom Flask a flask with a spherical body and one or more necks with ground glass joints.

Florence Flask a flask with a round body and one longer neck without a ground glass joint.

Buchner Flask or sidearm flask a thick-walled conical flask with a short hose-connection tube on the side of the neck

Volumetric Flask for preparing liquids with volumes of high precision. It is a flask with an approximately pear-shaped body and a long neck with a circumferential fill line.

Dewar Flask a flask with a hollow space in the wall to hold a vacuum.

Function: The function of a flask is to hold and store liquids. If graduated, it can serve to make approximate measurements of volume. One of the main advantages of a flask is the fact that the mixing of its contents can be done with little or no spillage. The flask is made of specialized glass so it can be heated and cooled without breaking. This same type of glass makes the flask brittle and it must be handled carefully. A stopper or cover allows for storage of liquids. Result: We study all glass apparatus. Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 28-54.

EXPERIMENT NO. 2 AIM: To study the operation of Analytical Balance. Requirements 1. Apparatus Required: Analytical Balances, weight box & fractional weight box etc. Procedure: Study of Analytical Balance An analytical balance is used to measure mass to a very high degree of precision and accuracy. The weighing pan(s) of a high precision (0.1 mg or better) analytical balance are inside a transparent enclosure with doors so that dust does not collect and so any air currents in the room do not affect the balance's operation. The use of a vented balance safety enclosure, which has uniquely designed acrylic airfoils, allows a smooth turbulence-free airflow that prevents balance fluctuation and the weighing of mass down to 1 g without fluctuations or loss of product. Also, the sample must be at room temperature to prevent natural convection from forming air currents inside the enclosure, affecting the weighing. Analytical precision is achieved by maintaining a constant load on the balance beam, by subtracting mass on the same side of the beam to which the sample is added. The final balance is achieved by using a small spring force rather than subtracting fixed weights. Weight of rider is 10 mg. Function: The function of a balance is to mass objects. [NOTE: If the device is measuring an object using springs against gravity, the devise is correctly referred to as a scale and its measurements are described as weight not mass.] A balance uses a comparison of a known substance or calibration with the unknown object to determine the unknown object's mass. Result: We study the operation of analytical balance. Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 29-30.

EXPERIMENT NO. 3 AIM: To carry out the calibration of the Burette, Pipette & Volumetric Flask. Requirements: 1. Apparatus required: Burette, Pipette & Beaker etc. 2. Chemicals required: Distilled water Procedure: Calibration of Burette: Clean the burette thoroughly. Weigh suitable beaker to nearest gm (W1). Fill the burette with distill water, sweep any air bubbles from the tip of the burette with drawn water until meniscus is at zero mark, record initial reading remove hanging drop if any. Run 10 ml of water from burette into previously weighed beaker. Record the weight of beaker (W2) and refill burette to zero reading. Run 20 ml of water into beaker and reweigh. Repeat the process for 30, 40 & 50 ml volume.
Apparent Volume (ml) Initial weight of burette (gm) W1 Final weight of burette (gm) W2 Actual Volume W2 - W1 / 1.0045 Correction / Tolerance AV - MV

10 20 30 40 50 Calibration of Pipette:

65.798 65.798 65.798 65.798 65.798

75.710 85.702 95.693 105.686 115.682

9.68 19.81 29.76 39.70 49.66

- 0.13 - 0.18 - 0.23 - 0.29 - 0.33

Clean & rinse the pipettes thoroughly and dry it. Weigh the suitable beaker to nearest gm (W1) Fill the pipette to zero mark level using distilled water. Remove any liquid on outside and release the pressure to allow liquid discharge into previously weighed beaker, allow the pipette to drain completely for 20-30 minutes. Check the weight of beaker. Note down the apparatus volume.
Apparent Volume Initial weight of Final weight of Actual Volume Correction /

(ml)

pipette (gm) W1

pipette (gm) W2

W2 - W1 / 1.0045

Tolerance AV - MV

65.798 65.798 65.798 65.798 65.798 65.798 65.798 65.798 65.798

66.299 66.258 66.288 67.213 67.211 67.216 75.317 75.219 75.389

0.49 0.45 0.48 1.40 1.40 1.41 9.47 9.37 9.54

- 0.50 - 0.54 - 0.51 - 0.59 - 0.6 - 0.58 - 0.52 - 0.62 - 0.45

2 10

Calibration of volumetric flask: Clean and rinse the volumetric flask with acetone and dry. Volumetric flask then weighed to rest gm and weight is recorded (W1) Volumetric flask then illed with distilled water at roo temperature until meniscus touches level mark. Vol. flask is weighed to nearest gm. The difference in two weights gives the actual weight of water from which actual volume is calculated. W2 W1 Actual volume = -------------1.0045 By measuring the tolerance value as per pharmacopia difference from the standard value measured.
Apparent Volume (ml) Initial weight of vol. flask (gm) W1 Final weight of vol. flask (gm) W2 Actual Volume W2 - W1 / 1.0045 Correction / Tolerance AV - MV

50

40.227 42.241 41.269 76.196 77.541 65.003

90.115 92.085 91.424 175.490 176.808 164.249

49.66 49.66 49.93 99.84 98.82 98.80

- 0.33 - 0.37 - 0.06 - 1.15 - 1.17 - 1.19

100

Result: We calibrate the following apparatus Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No EXPERIMENT NO. 4 AIM: To prepare 100 ml each 0.1N NaOH and 0.1N HCl Solution.

Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel & Pipette. 2. Chemical Required: NaOH Pelletes, Conc. HCl.

Procedure: Preparation 100 ml of 0.1N Sodium hydroxide solution (NaOH) Weigh accurately about 400 mg sodium hydroxide pellete in butter paper and transfer it into a 100 ml volumetric flask, dissolve in a little distilled water and finally diluted up to 100 ml with distilled water. Preparation of 0.1N Hydrochloride solution (HCl) Pipette out accurately about 0.85 ml hydrochloride into a 100 ml volumetric flask, dissolve in a little distilled water and finally diluted up to 100 ml with distilled water. Calculation For NaOH Normal solution 40 gm NaOH dissolve in 1000 ml distilled water gives 1N NaOH 4 gm NaOH dissolve in 100 ml distilled water gives 0.1N NaOH For HCl Normal solution 8.5 ml Conc. HCl dissolve in 1000 ml distilled water gives 1N HCl 0.85 Conc. HCl dissolve in 1000 ml distilled water gives 0.1N HCl Result: We prepare and submit 100 ml each 0.1N NaOH and 0.1N HCl Solution. Reference: IP 1996 Vol. II & A Text of Pharmaceutical analysis by Amit Kumar Page No 14.

EXPERIMENT NO. 5 AIM: To prepare & standardization of 0.1N NaOH solution. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette,

2. Chemical Required: NaOH Pelletes, Oxalic acid & Phnolphthalein as indicator. Principle: 2NaOH + C2H2O4.2H2O Sodium Oxalic acid Hydroxide C2O4Na2 + 2H20 Sodium Water oxalate
O C O C
C C O

Excess

NaOH +

HO

OH

HO

Phenolphthalein Colourless in acidic medium

Quinonoid form (Pink colour in alkaline medium)

O C O C

OH

Quinonoid form (Pink colour in alkaline medium) Procedure: Preparation 50 ml of 0.1N Sodium hydroxide solution (NaOH) Weigh accurately about 200 mg sodium hydroxide pellets in butter paper and transfer it into a 50 ml volumetric flask, dissolve in a little distilled water and finally diluted up to mark 50 ml with distilled water. Preparation 100 ml of 0.1N Oxalic acid solution Weigh accurately about 3.65 mg oxalic acid in butter paper and transfer it into a 50 ml volumetric flask, dissolve in a little distilled water and finally diluted up to mark 100 ml with distilled water.

Standardization: Now pipette out 10 ml oxalic acid in a conical flask, add 1 2 drops of phenolphthalein as indicator in to conical flask and titrated with 0.1N sodium hydroxide solution until pink colour appears. Calculation and Observation Table Contents of Conical flash 10 ml oxalic acid solution + 1-2 drops of Hph solution Burette Reading Volume of Titrant used 0.1NaOH Initial Final 0

Difference of first, second & three reading Average (Used 0.1NaOH) = ------------------------------------------------------3 Oxalic acid N1 X V1 0.1 X 10 = 0.1 x 10 N2 = -------------V2 Result: Normality of 0.1N NaOH solution was found to be-------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 14-15. NaOH N2 X V2 N2 X

EXPERIMENT NO. 6 AIM: To carry out assay of Boric Acid. Requirements:

1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette, 2. Chemical Required: Boric acid (sample), NaOH pellets, oxalic acid, Phenolphthalein as indicator etc. Principle: Principle (Assay): H3BO3 + NaOH Na2B4O7.10H20 Boric acid Sod.hydroxide Sodium borate
O C O C
C C O

Excess

NaOH +

HO

OH

HO

Phenolphthalein Colourless in acidic medium

Quinonoid form (Pink colour in alkaline medium)

O C O C

OH

Quinonoid form (Pink colour in alkaline medium) Principle (Standardization): 2NaOH + C2H2O4.2H2O Sodium Oxalic acid Hydroxide C2O4Na2 + 2H20 Sodium Water oxalate
O C O C
C C O

Excess

NaOH +

HO

OH

HO

Phenolphthalein Colourless in acidic medium

Quinonoid form (Pink colour in alkaline medium)

O C O C

OH

Quinonoid form (Pink colour in alkaline medium) Procedure: Assay of Boric acid: Weigh accurately about 2 gm, dissolve in a mixture of 50 ml of water and 100 ml of glycerin, previously neutralized to phenolphthalein solution. Titrate with 1M sodium hydroxide using phenolphthalein solution as indicator. Each ml of 1M sodium hydroxide is equivalent to 0.06183 gm oh H3BO3. Standardization Preparation 100 ml of 01M Sodium hydroxide solution (NaOH) Weigh accurately about 4 gm sodium hydroxide pellets in butter paper and transfer it into a 100 ml volumetric flask, dissolve in a little distilled water and finally diluted up to mark 100 ml with distilled water. Preparation 50 ml of 01M Oxalic acid solution Weigh accurately about 6.3 gm oxalic acid in butter paper and transfer it into a 50 ml volumetric flask, dissolve in a little distilled water and finally diluted up to mark 50 ml with distilled water. Standardization Procedure: Now pipette out 10 ml oxalic acid in a conical flask, add 1 2 drops of phenolphthalein as indicator in to conical flask and titrated with 01M sodium hydroxide solution until pink colour appears. Calculation and Observation Table Assay of Boric acid Contents of Conical flash 10 ml oxalic acid solution + 1-2 drops of Hph solution Burette Reading Initial Final 0 Volume of Titrant used 0.1NaOH

Find out the percentage purity of sample by applying following formula: Burette reading x Actual Normality x I.P. factor X 100 % Purity = -------------------------------------------------------------------Approximately Normality x Weight of sample (gm) Standardization of 0.1N NaOH solution Contents of Conical Burette Reading flash Initial Final 10 ml oxalic acid solution + 1-2 drops of Hph solution 0 Volume of Titrant used 0.1NaOH

Difference of first, second & three reading Average (Used 0.1NaOH) = ------------------------------------------------------3 Oxalic acid M 1 X V1 0.1 X 10 = NaOH M2 X V2 M2 X

0.1 x 10 M2 = -------------V2 Result: Percentage purity of boric acid was found to be ----- and Molarity of 01M NaOH solution was found to be-------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 1O9-110.

EXPERIMENT NO. 7 AIM: To prepare & standardization of 0.1N AgNO3 solution. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette, 2. Chemical Required: Silver Nitrate, NaCl, potassium chromate solution as Indicator etc.

Principle: AgN03 Silver Nitrate AgNO3 Silver Nitrate + NaCl sodium chloride + AgCl + silver chloride NaNO3 sodium nitrate + KNO3 potassium nitate

K2CrO4 potassium chromate

Ag2CrO4 silver chromate

Procedure: Preparation of 0.1N Silver Nitrate Solution: Weight accurately 16.9 gm of Silver Nitrate and transfer it in a 1 litre volumetric flask. Dissolve it in distilled water and make up the volume up to mark. Preperation of 0.1N NaCl solution. Weigh accurately 5.84gm of sodium chloride and dilute up to 1000 ml with distilled water. Standardization: Now pipetted out 10 ml of 0.1N NaCl solution in a conical flask and add 1, 2 drops of potassium chromate solution as indicator and titrated with 0.1N AgNO 3 until red colour precipitate appears. Calculation and observation table Standardization of 0.1N AgNO3 solution Contents of Conical Burette Reading flash Initial Final 10 ml 0.1N NaCl +1, 2 0 drops of potassium chromate as indicator Volume of Titrant used 0.1 AgNO3

Difference of first, second & three reading Average (Used 0.1 AgNO3) = ------------------------------------------------------3

NaCl N1 X V1 0.1 X 10

AgNO3 N2 X V2 N2 X

0.1 x 10 N2 = -------------V2 Result: Normality of 0.1N AgNO3 was found to be --------------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No169.

EXPERIMENT NO. 8 AIM: To carry out assay of NaCl by Mohrs method. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette, 2. Chemical Required: Silver Nitrate, NaCl, potassium chromate solution as Indicator etc.

Principle: Assay & Standardization AgN03 Silver Nitrate AgNO3 Silver Nitrate + NaCl sodium chloride + AgCl + silver chloride NaNO3 sodium nitrate + KNO3 potassium nitate

K2CrO4 potassium chromate

Ag2CrO4 silver chromate

Procedure: Assay procedure: Pipette out 25ml of NaCl solution in a 250ml conical flask and add 1, 2 drops of potassium chromate solution as indicator. Now titrated with 0.1N AgNO3 solution until changes the colour white precipitate to red colour precipitate. Each ml of 0.1M silver nitrate is equivalent to 0.005844 gm of NaCl. Standardization Preparation of 0.1N Silver Nitrate Solution: Weight accurately 16.9 gm of Silver Nitrate and transfer it in a 1 litre volumetric flask. Dissolve it in distilled water and make up the volume up to mark. Preperation of 0.1N NaCl solution. Weigh accurately 5.84gm of sodium chloride and dilute up to 1000 ml with distilled water. Standardization Procedure: Now pipetted out 10 ml of 0.1N NaCl solution in a conical flask and add 1, 2 drops of chromate solution as indicator and titrated with 0.1N AgNO3 until red colour precipitate appears. Calculation and observation table Assay of sodium chloride Contents of Conical Burette Reading flash Initial Final 25ml of NaCl solution + 0 1, 2 drops of chromate solution as indicator Volume of Titrant used 0.1 AgNO3

Find out the percentage purity of sample by applying following formula: Burette reading x Actual Normality x I.P. factor X 100 % purity = -------------------------------------------------------------------Approximately Normality x Weight of sample (gm) Standardization of 0.1N AgNO3 solution

Contents of Conical Burette Reading Volume of Titrant flash used 0.1 AgNO3 Initial Final 10 ml 0.1N NaCl +1, 2 0 drops of potassium chromate as indicator Difference of first, second & three reading Average (Used 0.1 AgNO3) = ------------------------------------------------------3 NaCl N1 X V1 0.1 X 10 = 0.1 x 10 N2 = -------------V2 Result: Percentage purity of sodium chloride was found to be ------- and normality of 0.1N AgNO3 was found to be -----------------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 168-169. AgNO3 N2 X V2 N2 X

EXPERIMENT NO. 9 AIM: To prepare & standardization of 0.1N KMnO4 solution. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette, 2. Chemical Required: Potassium permanganate, oxalic acid, sulphuric acid etc.

Principle: COOH 2KMnO4 + 5 .2H2O + 3 H2SO4 COOH 70 OC ----- 2 MnSO4 + K2SO4 + 10CO2 + 8H2O pink col.

Procedure: Preparation of 0.1N KMnO4 solution: Weigh 3.2 gm of KMnO4 and transfer it to 250 ml beaker containg cold water and stir vigorously with a glass rod to effect rapid dissolution. Add more DW to the beaker and repeat the above process till all potassium permanganate gets dissolved. Now transfer in a 1 litre volumetric flask and dilute up to 1 litre with distilled water. Note: Avoid heat in the preparation of KMnO4 solution because traces of grease or other contaminants on the glass vessels used can catalyze its decomposition. Preparation of 0.1Noxalic acid solution: Weigh accurately 6.302 gm of oxalic acid and transfer to a 1000 ml volumetric flask and diluted up to 1000 ml with distilled water. Standardization of 0.1N KMnO4 solution with 0.1Noxalic acid solution: Pipette out 10 ml 0.1N of oxalic acid solution in a 250 ml conical flask, add 10 ml of dil. H2SO4 and heated at 70 OC few minutes. Now titrate this solution with 0.1N KMnO4 solution until pink colour appears. Precautions: 1. Sufficient acid must be present, otherwise formation of a brown colour during titration may observe. 2. Similar brown coloration can also be observed by using too high a temperature or by using a dirty flask. Therefore, Or Or Or KMnO4 5e 158.0 g KMnO4 5000 ml N

31.60 g KMnO4 1000 ml N 3.2 g KMnO4 0.1 N KMnO4

Calculation and observation table Standardization of 0.1M Potassium Permanganate solution Contents of Conical flash Burette Reading Volume of Titrant used 0.1N KMnO4 Initial Final 10 ml 0.1N of oxalic acid solution 0 + 10 ml of dil. H2SO4

Difference of first, second & three reading Average (Used 0.1N KMnO4) = ------------------------------------------------------3 Oxalic acid M 1 X V1 0.1 X 10 = KMnO4 M2 X V2 N2 X

0.1 x 10 M2 = -------------V2

Result: Mormality of 0.1M Potassium Permanganate was found to be ---------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 152.

EXPERIMENT NO. 10 AIM: To prepare & standardization of 0.1N NH4SCN solution. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette, 2. Chemical Required: AgNO3, NH4SCN, Ferric alum etc.

Principle: AgNO3 + NH4SCN AgSCN + NH4NO3 Silver ammonium silver ammonium Nitrate thiocyanate thiocyanate nitrate NH4SCN + Fe+3 Fe+ [Ff+3 (SCN)6]-3 Ammonium ferric ferric ferrithiocyanate Thiocyanate alum (Red colour ppt.) Procedure: Preparation of 0.1M Ammonium Thiocyanate Solution: Weight accurately 8.5 gm of ammonium thiocyanate and transfer it in a 1 litre volumetric flask. Dissolve it in distilled water and make up the volume up to mark. Preparation of 0.1M Silver Nitrate Solution: Weight accurately 16.9 gm of Silver Nitrate and transfer it in a 1 litre volumetric flask. Dissolve it in distilled water and make up the volume up to mark. Standardization: Now pipette 25 ml of a standard 0.1M AgNO3 solution in a conical flask, dilute with 50 ml of DW,add to it 2 ml of nitric acid and 1,2 drops of ferric alum as indicator and titrate with 0.1M ammonium thiocyanate solution until red colour precipitate appears.

Calculation and observation table Standardization of 0.1M Ammonium Thiocyanate Solution Contents of Conical Burette Reading Volume of Titrant flash used 0.1 NH4SCN Initial Final 25 ml 0.1M AgNO3 +1, 0 2 drops of ferric alum as indicator Difference of first, second & three reading Average (Used 0.1 NH4SCN) = ------------------------------------------------------3 AgNO3 M 1 X V1 0.1 X 10 NH4SCN M2 X V2 M2 X

0.1 x 10 M2 = -------------V2

Result: Normality of 0.1M ammonium thiocyanate was found to be ----------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 170-171.

EXPERIMENT NO. 11 AIM: To carry out assay of sodium chloride by Volhards method. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette, 2. Chemical Required: Sodium chloride, silver chloride, ammonium-

thiocyanate, ferric alum etc. Principle: Principle (Assay): NaCl + AgNO3 AgNO3 + NaNO3 AgNO3 + NH4SCN AgSCN NH4SCN + Fe+3

+ NH4NO3

Fe+ [Ff+3 (SCN)6]-3 Red colour ppt.

Principle (Standardization): AgNO3 + NH4SCN NH4SCN + Fe+3

AgSCN + NH4NO3

Fe+ [Ff+3 (SCN)6]-3 Red colour ppt.

Assay procedure: Weight accurately 0.1gm NaCl and dissolve in 50 ml of water in a glass-stoppered flask.Add 50 ml of 0.1M silver nitrate, 5 ml of 2M nitric acid and 2 ml of dibutyl phthalate, shake well and titrate with 0.1M ammonium thiocyanate using 1,2 drops of ferric alum as indicator,until the colour becames reddish yello.Each ml of 0.1M silver nitrate is equivalent to 0.005844 gm of NaCl. Standardization Preparation of 0.1M Ammonium Thiocyanate Solution: Weight accurately 8.5 gm of ammonium thiocyanate and transfer it in a 1 litre volumetric flask. Dissolve it in distilled water and make up the volume up to mark. Preparation of 0.1M Silver Nitrate Solution: Weight accurately 16.9 gm of Silver Nitrate and transfer it in a 1 litre volumetric flask. Dissolve it in distilled water and make up the volume up to mark. Standardization Procedure: Now pipette 25 ml of a standard 0.1M AgNO3 solution in a conical flask, dilute with 50 ml of DW,add to it 2 ml of nitric acid and 1,2 drops of ferric alum as indicator and titrate with 0.1M ammonium thiocyanate solution until red colour precipitate appears. Calculation and observation

Assay of Sodium Chloride Contents of Conical flash 0.1gm NaCl + 50 ml of water + 50 ml of 0.1M silver nitrate + 5 ml of 2M nitric acid + 2 ml of dibutyl phthalate + 1,2

Burette Reading Initial Final 0

Volume of Titrant used 0.1 NH4SCN

drops of ferric alum as indicator Find out the percentage purity of sample by applying following formula: Burette reading x Actual Normality x I.P. factor X 100 % purity = -------------------------------------------------------------------Approximately Normality x Weight of sample (gm) Standardization of 0.1M Ammonium Thiocyanate solution Contents of Conical flash 25 ml of a standard 0.1M AgNO3 solution + 50 ml of DW + 2 ml of nitric acid + 1,2 drops of ferric alum as indicator Burette Reading Initial Final 0 Volume of Titrant used 0.1M NH4SCN

Difference of first, second & three reading Average (Used 0.1M NH4SCN) = ------------------------------------------------------3 AgNO3 M 1 X V1 0.1 X 10 = NH4SCN M2 X V2 M2 X

0.1 x 10 M2 = -------------V2

Result: Percentage purity of Sodium Chloride was found to be--------- and mormality of 0.1M ammonium thiosulphate was found to be -------------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 168-169.

EXPERIMENT NO. 12 AIM: To carry out assay of KI by Potassium Iodate Titration. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette,

2. Chemical Required: Potassium iodate, pot. Iodide, sodium thiocyanate etc. Principle(Assay): KIO3 + KI + HCl Pot. Pot. iodate iodide Principle (Standardization): KIO3 Pot. iodate + KI + HCl Pot. iodide I2 + KCl + H2O ICl + KCl + H2O

I2 + Na2S2O3 NaI + Na2S4O6 Sodium Sod.tetrathinate Thiosulphate Assay of Potassium Iodide: Weigh accurately about 0.35 g of KI, dissolve in about 10 ml of water, add 35 ml dil. HCl and add 5 ml of chloroform. Titrated with 0.05 M KIO3 solution until purple colour of iodine disappears from chloroform. Each ml of 0.05 M KIO3 if equivalent to 0.0166 g of KI. Preparation of solutions: Prepration of 0.05N KIO3 Solution: Weigh accurately 10.7 gm KIO3 and diluted up to 1000 ml with distilled water. Preparation of 0.1N Na2S2O3: Weigh accurately 25 gm sodium thiosulphate (Na2S2O3) and dissolve in small quantity of water and diluted up to 1000 ml with distilled water. Standardization of 0.05N KIO3: To 25 ml of KIO3 solution, add 2 gm of KI and add 10 ml of dil. HCl and add 1,2 drops of starch solution as a indicator and titrated with 0.05 N sodium thiosulphate (Na2S2O3) solution until blue colour appears. Calculation and observation Table

Assay of Potassium Iodide Contents of Conical flash

Burette Reading Initial Final 0

Volume of Titrant used 0.05 M KIO3

Find out the percentage purity of sample by applying following formula: Burette reading x Actual Normality x I.P. factor X 100 % purity = -------------------------------------------------------------------Approximately Normality x Weight of sample (gm)

Standardization of 0.1M Potassium Iodate solution Contents of Conical flash 25 ml of KIO3 solution + 2 gm of KI + 10 ml of dil. HCl + 1,2 drops of starch solution as a indicator Burette Reading Initial Final 0 Volume of Titrant used 0.1N Na2S2O3

Difference of first, second & three reading Average (Used 0.1N Na2S2O3) = ------------------------------------------------------3 KIO3 N1 X V1 0.1 X 10 Na2S2O3 N2 X V2 N2 X

= 0.1 x 10 N2 = -------------V2

Result: Normality of 0.05N Potassium Iodate was found to be --Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 149-150.

EXPERIMENT NO. 13 AIM: To prepare & standardization of 0.05N KIO3 solution. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder, funnel, pipette,

2. Chemical Required: Potassium iodate, pot. Iodide, sodium thiocyanate etc. Principle: KIO3 Pot. iodate + KI + HCl Pot. iodide I2 + KCl + H2

I2 + Na2S2O3 NaI + Na2S4O6 Sodium Sod.tetrathinate Thiosulphate Procedure: Prepration of 0.05N KIO3 Solution: Weigh accurately 10.7 gm KIO3 and diluted up to 1000 ml with distilled water. Preparation of 0.1N Na2S2O3: Weigh accurately 25 gm sodium thiosulphate (Na2S2O3) and dissolve in small quantity of water and diluted up to 1000 ml with distilled water. Standardization of 0.05N KIO3: To 25 ml of KIO3 solution, add 2 gm of KI and add 10 ml of dil. HCl and add 1,2 drops of starch solution as a indicator and titrated with 0.05 N sodium thiosulphate (Na2S2O3) solution until blue colour appears. Calculation and observation Table Standardization of 0.1M Potassium Iodate solution Contents of Conical flash 25 ml of KIO3 solution + 2 gm of KI + 10 ml of dil. HCl + 1,2 drops of starch solution as a indicator Burette Reading Initial Final 0 Volume of Titrant used 0.1N Na2S2O3

Difference of first, second & three reading Average (Used 0.1N Na2S2O3) = ------------------------------------------------------3

KIO3 N1 X V1 0.1 X 10

= 0.1 x 10

Na2S2O3 N2 X V2 N2 X

N2 = -------------V2

Result: Normality of 0.05N Potassium Iodate was found to be --Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 149-150.

EXPERIMENT NO. 14 AIM: To carry out assay of Ferrous Sulphate by Redox Method. Requirements (i) Apparatus Required: Conical flask, Beaker, Measuring cylinder,

funnel, pipette, (ii) Chemical Required: Cerric ammonium sulphate, ferrous sulphate, NaOH, sulphuric acid, osmic acid, arsenic trioxide etc. Principle Principle (Assay): Fe2+ Fe3++ eCe4+ + e- Ce3+
-----------------------------------------------

Fe2+ + Ce4+ Fe3+ + Ce3+

-------------------------------------------------

The excess drops of cerric ammonium sulphate react with ferroin indicator gives end point (blue-green colour).

Fe+2
N

Fe+3
N

Ce4+ Cerric

Red colour (1, 10 Phenanthrolineferrous sulfate) (Ferroin indicator)

Blue-Green colour (1, 10 Phenanthrolineferric sulfate)

+ Ce+3 Cerrous

Principal (Standardization): Arsenic trioxide is the best primary standard for the standardization of cerric ammonium sulphate. NaOH is added to dissolve the Arsenic trioxide which is dissolve due to formation of sodium arsenite. As2O3 + 6NaOH 2Na3AsO3 + 3H2O Arsenic trioxide sodium arsenite Dilute H2SO4 is added to neutralixe the excess of NaOH. Cerric ammonium sulphate oxidizes the sodium arsenite into Sodium arsenate. 2 Ce(SO4)2 + Na3AsO3 +H2O Sodium arsenite Ce2(SO4)3 + Na3AsO4 + H2SO4 sodium arsenate

The trace of osmic acid is added to speed up the reaction between cerric ion and arsenate ion. At the end point all the arsenite ion are consumed. A slight excess of cerric ammonium sulphate react with ferroin indicator to give blue green colour at end point.

Fe+2
N

Fe+3
N

Ce4+ Cerric

Red colour (1, 10 Phenanthrolineferrous sulfate) (Ferroin indicator)

Blue-Green colour (1, 10 Phenanthrolineferric sulfate)

+ Ce+3 Cerrous

Procedure: Assay of ferrous sulphate: Weigh accurately 0.3 gm ferrous sulphate and dissolved in 30 ml water and add 15 ml 1M H2SO4 and then 1, 2 drops of ferroin solution as indicator. Now titrated with 0.1M cerric ammonium sulphate till blue green colour appears. Each ml of 0.1M cerric ammonium sulphate is equivalent to 0.01519 gm of FeSO4. Preparation of solutions Preparation of 0.1 M cerric ammonium sulphate solution Dissolve 65 gm. of cerric ammonium sulphate with the gentle heat, in a mixture of 30 ml of H2SO4 and 500 ml of water. Cool, filter the solution, if turbid, and dilute to 1000 ml with water. Standardization Standardization of 0.1M cerric ammonium sulphate with Arsenic trioxide Weigh accurately about 0.2 gm arsenic trioxide, previously dried at 105 0C for 1 hour, and transfer to a 500 ml conical flask. Add 25 ml NaOH solution then add 100 ml water, mix well, now add 30 ml of dil. H2SO4 and 0.15 ml osmic acid and 1, 2 drops of ferroin sulphate indicator, now titrated with 0.1N cerric ammonium sulphate until red colour changes to blue green colour. Calculation and observation table

Standardization of 0.1M Ammonium Thiocyanate Solution Contents of Conical flash Burette Reading Volume of Titrant used 0.1N Ce4 Initial Final 0.2 gm arsenic trioxide + 25 0 ml NaOH solution + 100 ml

water + 30 ml of dil. H2SO4 + 0.15 ml osmic acid + 1, 2 drops of ferroin sulphate indicator Now find out the normality by applying the following formula. N Weight of arsenic trioxide X 1000 = -----------------------------------------Burette reading X Eq.wt.

Assay of sodium chloride Contents of Conical flash 0.3 gm ferrous sulphate + 30 ml water + 15 ml 1M H2SO4 + 1, 2 drops of ferroin solution as indicator

Burette Reading Initial Final 0

Volume of Titrant used 0.1N Ce4

Find out the percentage purity of sample by applying following formula:

Burette reading x Actual Normality x I.P. factor X 100 % purity = -------------------------------------------------------------------Approximately Normality x Weight of sample (gm)

Result: Percentage purity of ferrous sulphate was found to be--------- and normality of 0.1N ceric ammonium sulphate was found to be -------------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 147-149.

EXPERIMENT NO. 15 AIM: To prepare & standardization of 0.1N Ceric Ammonium Sulphate. Requirements 1. Apparatus Required: Conical flask, Beaker, Measuring cylinder,

funnel, pipette, 2. Chemical Required: Cerric ammonium sulphate, arsenic trioxide, sodium hydroxide, osmic acid etc. Principle: Arsenic trioxide is the best primary standard for the standardization of cerric ammonium sulphate. NaOH is added to dissolve the Arsenic trioxide which is dissolve due to formation of sodium arsenite. As2O3 + 6NaOH 2Na3AsO3 + 3H2O Arsenic trioxide sodium arsenite Dilute H2SO4 is added to neutralixe the excess of NaOH. Cerric ammonium sulphate oxidizes the sodium arsenite into Sodium arsenate. 2 Ce(SO4)2 + Na3AsO3 +H2O Sodium arsenite Ce2(SO4)3 + Na3AsO4 + H2SO4 sodium arsenate

The trace of osmic acid is added to speed up the reaction between cerric ion and arsenate ion. At the end point all the arsenite ion are consumed. A slight excess of cerric ammonium sulphate react with ferroin indicator to give blue green colour at end point.
N

Fe+2
N

Fe+3
N

Ce4+ Cerric

Red colour (1, 10 Phenanthrolineferrous sulfate) (Ferroin indicator)

Blue-Green colour (1, 10 Phenanthrolineferric sulfate)

+ Ce+3 Cerrous

Procedure: Preparation of 0.1 M cerric ammonium sulphate solution Dissolve 65 gm. of cerric ammonium sulphate with the gentle heat, in a mixture of 30 ml of H2SO4 and 500 ml of water. Cool, filter the solution, if turbid, and dilute to 1000 ml with water. Standardization Standardization of 0.1M cerric ammonium sulphate with Arsenic trioxide Weigh accurately about 0.2 gm arsenic trioxide, previously dried at 105 0C for 1 hour, and transfer to a 500 ml conical flask. Add 25 ml NaOH solution then add 100 ml water, mix well, now add 30 ml of dil. H2SO4 and 0.15 ml osmic acid and 1, 2 drops of ferroin sulphate

indicator, now titrated with 0.1N cerric ammonium sulphate until red colour changes to blue green colour. Calculation and observation table Standardization of 0.1M Ammonium Thiocyanate Solution Contents of Conical flash Burette Reading Volume of Titrant used 0.1N Ce4 Initial Final 0.2 gm arsenic trioxide + 25 0 ml NaOH solution + 100 ml water + 30 ml of dil. H2SO4 + 0.15 ml osmic acid + 1, 2 drops of ferroin sulphate indicator Now find out the normality by applying the following formula. N Weight of arsenic trioxide X 1000 = -----------------------------------------Burette reading X Eq.wt.

Result: Normality of 0.1N Cerric Ammonium Sulphate was found to be ---------Reference: A Text of Pharmaceutical analysis by Amit Kumar Page No 148-149.