LABORATORY MANUAL CHE102 CHEMISTRY LAB

LMCHE-102 Chemistry Lab

Some rules to be followed in the Lab: General Rules: 1. Entry without lab coat in chemistry lab is strictly prohibited.
2. Mobile phones should be switched off and kept in the bag during lab hours.

3. No group discussions are allowed in the lab. 4. Clean the apparatus as well slab after your experiment is finished. 5. Do not do any indiscipline activity in lab as you are under strict cc-TV surveillance. 6. Always bring the lab manuals with you. 7. Do not use laptop while performing the experiments. 8. Switch off electrical apparatus after their use. 9. Do not throw filter papers in sink dispose all solid waste in dustbin. 10. Liquid waste must be deposited in the waste containers designated for Chlorinated organic solvents, Organic solvents and aqueous waste. Precautionary Rules: 1. Never pipette out strong acids and bases with your mouth, it can be dangerous, therefore use measuring cylinders for such chemicals. 2. Never try to smell the chemicals as it can be dangerous for you. 3. Cap the bottles after taking chemical because uncovered bottles can be a source of harmful fumes. 4. In case of any accidental spill over of any chemical on you, report your teacher or lab technician immediately. 5. Report your lab technician if any breakage of glass apparatus takes place.

LMCHE-102 Chemistry Lab

TABLE OF CONTENTS S. No. Title of the Experiment Page No. With the help of complexometeric titration how you will 3-6 1 determine the hardness of given hard water sample by using EDTA. Provided standard hard water.(1ml of S.H.W.=1mg of CaCO3) Identification of elements present in given compound. 2 7-10 Determination of the dissociation constant of acetic acid using 11-14 3 pH-meter. Determination of Strength of hydrochloric acid 15-18 4 solution(approximately N/10) by titrating it against sodium hydroxide solution conductometrically To test the validity of Beer-Lamberts law using colorimeter and 19-23 5 to determine unknown concentration of solution Estimation of nickel in the given sample using dimethyl 24-26 6 glyoxime. Determination of the rate constant of hydrolysis in case of ethyl 27-30 7 acetate using an alkali. Determine the strength of given solution of ferrous ammonium 31-33 8 sulphate by titrating against potassium dichromate solution Separation of a mixture of organic compounds by thin layer 34-37 9 chromatography. Some important instruments in chemistry lab 10 38-40

LMCHE-102 Chemistry Lab

Experiment No. 1 1. Experiment: To determine the hardness of the given hard water sample by EDTA method. You are provided with Standard Hard Water (1 ml of SHW =1 mg of CaCO3). Equipment required: Burette, Burette stand, Titration flask, Pipette, Beakers, funnel etc. Materials required: Standard Hard water, Sample hard water, EDTA solution, Eriochrome black-T (EBT), Buffer Solution. 2. Learning Objectives: I) The purpose of this experiment is to determine the hardness of water by measuring the concentrations of calcium in water samples by titration. II) To gain knowledge about complexometric titration. III) To know the purpose of EDTA used IV) To know about use of buffer solution: The buffer being used has composition NH4Cl and NH4OH. Its pH is the order of 10.5. V) To know the purpose of indicator

Theory: Complexometric titrations are mainly used to determine the concentration of divalent cations such as calcium, magnesium, zinc, copper, lead etc. Hard water is the water that has high concentration of calcium and magnesium ions. The Ethylenediaminetetraacetic acid (EDTA) is the most commonly used complexant. EDTA is a chelating agent. It is able to form coordination complex with metal ions present in hard water. Although these reactions are easy to perform, it is necessary to maintain well defined pH. How does the indicator work? The indicator used here is organic molecule capable of forming a colored complex with the metal cations to be determined which is less stable than the complex formed by the cation with the corresponding titrant (EDTA). The indicator used is Eriochrome black-T. (i) When indicator is added to hard water it combines with free metal ions present in water giving metal indicator complex which is wine red in colour.
3 LMCHE-102 Chemistry Lab

HIn2- +

M2+

MIn-

H+

(M=Ca2+, Mg2+)

(Wine red) (ii) When EDTA solution is added to the titration flask it combines with the free metal ions giving metal EDTA complex, which is stable and colourless. H2Y2+ M2+ MY2+ 2H+

(Colourless) (iii) When all the free metal ions are exhausted, next drop of EDTA removes the metal ion engaged with indicator and the original blue colour of indicator is restored. H2Y2+ MIn MY2+ HIn2- + H+ (Blue)

3. Outline of the Procedure: (A) Standardization of EDTA solution: a. Fill the burette with EDTA solution. b. Pipette out 10 ml of standard hard water in the titration flask. Add to it 2-3 ml of buffer solution and two drops of Eriochrome Black-T indicator. A wine red color appears. c. Titrate this solution against EDTA solution taken in a burette till wine red color changes to blue. d. This is the end point. Recovered the volume of EDTA consumed as A ml. Repeated the procedure to get at least three concordant readings. Table1: Standardization of EDTA S. No. Burette readings Initial (R1) 1. 2. 3. 4. 5. 6. Final (R2) Volume of EDTA Consumed (R2 - R1) mL

LMCHE-102 Chemistry Lab

(B) Determination of Total Hardness: a. Pipette out 10 ml of sample hard water in the titration flask. Add to it 2-3 ml of buffer solution and two drops of Eriochrome Black-T indicator. b. A wine red colour appears. Titrate this solution against EDTA solution taken in a burette till wine red colour changes to blue. c. This is the end point. Recovered the volume of EDTA consumed as B ml. Repeated the procedure to get at least three concordant readings. Table 2: Determination of Total Hardness S. No. Burette readings Initial (R1) 1. 2. 3. 4. 5. 6. Calculations: (a) Standardisation of EDTA solution: 1 ml of standard hard water = 1 mg of CaCO3 10 ml of S.H.W. A ml of EDTA 1 ml of EDTA = 10 mg of CaCO3 = A ml of EDTA = 10 mg of CaCO3 = 10/A mg of CaCO3 Final (R2) Volume of EDTA Consumed (R2 - R1) mL

(b) Calculation of total hardness: 10 ml of sample hard water Now 1 ml of EDTA B ml of EDTA 10 ml of sample hard water 1 ml of hard water sample = B ml of EDTA = 10/A mg of CaCO3 = B 10/A mg of CaCO3 = B x 10/A mg of CaCO3. = B x 10/A x 1/10 mg of CaCO3.

1000 ml of hard water sample = B x 10/A x 1/10 x 1000 mg of CaCO3. Hence total hardness = 1000 x B/A ppm
5 LMCHE-102 Chemistry Lab

4. Required Results: Parameters: Volumetric analysis. Relationships: determine how much EDTA is consumed for 1 mL of Standard Hard Water, which is then, can be used for the calculation of unknown hardness. Graphs: NA Error Analysis: Ask for the actual hardness and calculate the % error.

Results: The hardness of given sample hard water is =.......... ppm. % error =%. 5. Cautions: a. The burette, pipette and conical flask should be washed properly and then rinsed with distilled water. b. Redistilled water should be employed for preparing the EDTA solution. c. The colour change near the end point is very slow and thus should be observed very carefully. 6. Learning outcomes: To be written by the students in 50-70 words.

LMCHE-102 Chemistry Lab

Experiment No. 2 1. Experiment: Identification of elements (Nitrogen, Sulphur and halogen) present in given organic compound. Equipment required: Test tube, China dish, fusion tubes, glass rod, Bunsen Burner, tripod stand, funnel, beaker, filter papers etc. Material Required: Sodium metal, Ferrous sulphate (freshly prepared), sodium hydroxide, dilute HCl/H2SO4, acetic acid, lead acetate solution, Sodium nitroprausside solution, Conc. HNO3, Tollens reagent, NH4OH 2. Learning objective: How to determine the presence of various elements (nitrogen, sulphur and halogen) other than carbon and hydrogen in any organic compound. Theory: The main constituents of organic compounds are carbon and hydrogen. Besides this Nitrogen, Sulphur and halogen may also present in organic compounds. Nitrogen, sulphur and halogens in any organic compound are detected by 'Lassaigne's test'. The organic compounds are covalent compounds. The fusion of organic compound with sodium metal convert covalently bonded Nitrogen, sulphur and halogens into ionic compounds (NaCN, Na 2S and NaX here X = halogen). The ionic compounds formed during the fusion are extracted in aqueous solution, and can be detected by simple chemical tests. The aqueous solution obtained by extracting the fused mass in water is called sodium extract or Lassaigne's extract. Chemical equations: Detection of Nitrogen 6NaCN + FeSO4 Na4[Fe(CN)6] + Na2SO4

(Sodium ferrocyanide) 3Na4[Fe(CN)6] + 4Fe3+ Fe4[Fe(CN)6] 3 + 12Na+ (Ferric ferrocyanide) (prussian blue/green)
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Detection of Sulphur Na2S + Pb(CH3COO)2 PbS + 2CH3COONa

(Black ppt) Na2S + Na2[Fe(CN)5NO] Na4[Fe(CN)5NOS]

(Sodium nitroprausside)

(Sodium thionitroprausside) (Purple)

Detection of Halogen NaCN + HNO3 Na2S + HNO3 NaX + AgNO3 NaNO3 + HCN (g) 2NaNO3 + H2S (g) AgX (ppt) 3. Outline of the Procedure: (A) Preparation of Lassaignes extract: a. In a dry fusion tube take a small piece of sodium metal, and heat the fusion tube in the flame so that the sodium metal is melted completely. b. To this heated tube add carefully a small amount of given compound, and heat the fusion tube again in the flame till it gets red hot. c. Pour the red hot fusion tube immediately into a china dish containing 20 mL of distilled water (care should be taken that china dish should not contain any impurity or chemical which could interfere in the detection) d. Repeat the procedure for at least three times with fusion tube (add the fusion tube to the same china dish having distilled water) e. Crush all the fusion tube in china dish with the help of a clean glass road and boil the solution in china dish to evaporate so that the volume is reduced nearly half. f. Filter the content through filer paper and collect the solution obtained in a clean beaker, the solution obtained will be Lassaignes extract. + NaNO3 (X = Cl, Br, I)

LMCHE-102 Chemistry Lab

(B) Test for nitrogen: a. In a clean test tube take about 2 mL of the Lassaignes extract and add a freshly prepared solution of ferrous sulphate, the dirty green precipitate of Fe(OH)2 will appear in the solution. b. To this solution add a small amount of dilute sodium hydroxide solution and heat it gently in the flame. c. Dirty green precipitate may disappear to this solution add a small amount of dilute HCl, if the solution turns out be prussian blue or prussian green, nitrogen is present.

(C) Test for Sulphur: a. In a clean test tube take about 2 mL of the Lassaignes extract and add one mL of acetic acid and then add few drops of lead acetate, if black coloured precipitates appear in the solution, sulphur is present in the compound. b. In a clean test tube take about 2 mL of the Lassaignes extract and add few drops of Sodium nitroprusside, if the purple or violet colour appears, it shows the presence of sulphur in the compound.

(D) Test for Halogens: a. In a clean test tube take about 2 mL of the Lassaignes extract and add one mL Conc. HNO3 to this solution add 1 mL of tollens reagent the formation of precipitate indicates the presence of halogens in the given compound Identification of halogen b. If the precipitate formed are white in colour which are readily soluble in NH4OH, these indicates the presence of chloride in the given compound c. If the precipitate formed are pale yellow in colour which are partially soluble in NH4OH, these indicates the presence of bromine in the given compound d. If the precipitate formed are yellow in colour which are insoluble in NH 4OH, these indicates the presence of iodine in the given compound.

LMCHE-102 Chemistry Lab

4. Required Results: Parameters: Qualitative analysis of various elements present in the given organic compound Relationships To Be Determined: Which test is applicable to confirm the presence of an element Graphs/Plots: NA Error Analysis: NA Result: _____ elements are present in given organic compound.

5. Cautions: a. b. c. d. e. Sodium metal should be handled very carefully. China dish and test tubes should be properly cleaned. Take distilled water in china dish. Sodium metal should never be allowed to come in contact with water. On addition of FeSO4 a dirty green precipitate comes out which are not true indicator for the presence of nitrogen in the compound, these appear due to the formation of ferric hydroxide which is soluble in dilute sodium hydroxide.

6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Experiment No. 3 1. Experiment: Determination of the dissociation constant of acetic acid using pHmeter. Equipment required: pH meter, 100 mL beaker, pH electrode, burette, funnel etc. Material required: Acetic acid, sodium hydroxide, buffer solution of pH 4 and pH 7. 2. Learning objectives: I) II) III) Students will learn the basics of pH meter and how to use pH meter. To monitor the total pH of a solution and to determine equivalence point of titrations that involves ions. To calculate the dissociation constant of weak acid.

To set temperature pH reading

Glass electrode To set knob at pH Buffer Solution of pH 4 & 7 Fig: pH meter

Theory: The strength of an acid is experimentally measured by determining its equilibrium constant or dissociation constant (K). Since strong acids are strong electrolytes, they are ionized almost completely in aqueous solutions. It is not meaningful to study the ionic equilibrium of strong acids and calculate their equilibrium constants as the unionized form is present to such a small extent. Hence, the study of ionic equilibrium and calculation of K is applicable only to weak acids.
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e.g. Acetic acid ionizes feebly as, CH3COOH (aq) + H2O (l) K = [H3O+] [CH3COO-]/[ CH3COOH] pKa is a modern method of expressing acid strengths. pK a is determined by measuring the changes in pH of acid solution at different amounts of the base added. During the titration of an acid with a base, the pH of the solution rises gradually at first, then more rapidly and until at the equivalence point, there is a very sharp increase in pH for a very small quantity of added base. Once past the equivalence point, the pH increases only slightly on addition of excess base. The titration curve is obtained by plotting changes in pH at different amounts of the base added and the equivalence point is determined. 3. Outline of the Procedure: a. Switch on the pH meter after connecting the pH electrode to it. b. With the help of temperature knob shown in the Fig set the temperature to the room temperature. c. Make sure that pH knob is pointing towards pH as shown in Fig. d. Take buffer solutions of pH 4 and pH 7 which will be provided by the lab technician. e. Put the pH electrode in pH 7 solution and set the pH on screen to 7 with calibration knob present on the pH meter. f. Then wash the electrode and put it in pH 4 solution and set 4 on the screen. g. Your pH meter is now ready to take readings of unknown solutions. Dont touch any of the button now onwards till the end of the experiment. h. Pipette out 50 mL of the given weak acid into a 100 cm3 beaker. Immerse electrode assembly into the acid. Measure the pH of the acid. i. Fill a burette with the base (0.1 N sodium hydroxide).
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H3O+ (aq) + CH3COO- (aq)

j.

Add 0.5 mL of the base from burette to the acid. Stir the solution thoroughly and measure the pH after addition.

k. Continue adding 0.5 mL of base and noting down pH at each successive addition. l. When the pH begins to show a tendency to increase rapidly (e.g. from 6-9), add only small increments (say 0.2 mL) of the base and measure the pH after each addition. Continue till there is only a slight increase in pH on the addition of the base. m. Again add 1mL of base for 3-4 time more and note down the change in pH. n. Plot a graph of pH (ordinate) against the volume of sodium hydroxide added (abscissa). Determine the equivalence point and hence the pH at half equivalence point. This gives the pKa value of the acid. OBSERVATIONS AND CALCULATIONS: S. No Volume of NaOH added (ml.) 1. 2. 3. 4. 38. 39. 40. 0 0.5 1.0 1.5 =........................(on x-axis i.e. vol. of NaOH in mL) Observed pH

1) Equivalence point

2) Half equivalence point =............... (take half of the vol. of NaOH at Eq. Pt. in mL) 3) pH at half equivalence point =(on y-axis)

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LMCHE-102 Chemistry Lab

pKa of the given weak acid = pH at half equivalence point =.................... pKa = - log Ka Ka = Antilog10 (-pH at half equivalence point) 4. Required Results: Parameter used: pKa = - log Ka, hence the value of dissociation constant (Ka) can be calculated Relationships to be determined: Effect of addition of a base to an acid. Graph:

Equivalence point pH

Half equivalence point

Volume of alkali added (mL) With the help of plot, Equivalence point is calculated. Error Analysis: Ask for the original value from your instructor for % error calculations. 5. Cautions: 1. Handle the glass electrode very carefully. 2. Switch on the pH meter at least 10 minutes before the start of the measurements. 3. Stir the solution thoroughly before taking the reading. 4. Let the reading stabilize for tome time (15 seconds) before taking the reading. 6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Experiment No. 4 1. Experiment: To find the strength of hydrochloric acid solution (approximately N/100) by titrating it against sodium hydroxide solution conductometrically. Equipment required: Conductivity Bridge, conductivity cell, beaker, funnel, burette. Materials required: 0.1 N NaOH solution and approximately HCl solution. 2. Learning objectives: I) Students will learn the basics of Conductometery and how to use Conductometer. II) To monitor the total conductance of a solution and to determine the end points of titrations that involve ions. III) Students will get knowledge about conductometric titrations.
Value of Cell Constant

To set at the value of cell constant Conductivity Electrode

Conductivity Electrode

Range To set temperature

To set Cond. or Cell constant Fig.: Conductometer Theory: Conductometry can be used to detect the equivalence point (end point) of a titration. This method is based upon the measurement of conductance during the course of titration. The conductance varies differently before and after the equivalence point. This is due to the reason that electrical conductance of a solution depends upon the number of ions present and their ionic mobilities i.e. speeds. When conductance values are plotted against volume of titrant added, two straight lines are obtained; the point of intersection of the lines gives the end point. For studying HCl vs NaOH titration, a known volume of HCl is taken in a beaker and
15 LMCHE-102 Chemistry Lab

NaOH solution in the burette. The conductance of acid solution is noted initially as well as after successive additions of small amounts of NaOH solution. Conductance of acid solution in the beginning is very high due to presence of highly mobile H + ions. On adding NaOH solution, the H+ ions are replaced by slow moving Na ions, decreasing the conductance of solution. [H+ + Cl -] + [Na + + OH-] Na++ Cl- + H2O

When neutralization is complete, further addition of NaOH will cause the conductance to increase due to excess of highly mobile OH - ions. The conductance will thus be minimum at the equivalence point. Thus if conductance values are plotted against the volume of NaOH added, a curve of the type xyz is obtained. The point of intersection (i.e. point Y) corresponds to the end point. 3. Outline of the Procedure: a. Determine the cell constant of the given conductivity cell which is written on the neck of the cell. b. Connect the conductivity cell to the conductometer. c. Set the function switch to check position. Display must read 1.000. If it does not, set it with CAL control at the back panel. d. Put the Function Switch to Cell Constant and set the value of the cell constant determined in step-1 with the help of cell constant Knob shown in the Fig. e. Set the temperature control to the actual temperature of the solution under test. f. Rinse the conductivity cell with the solution whose conductivity is to be measured. g. Take 50 ml of the given HCl in a 100 ml beaker. h. Wash the conductivity cell with distilled water and then rinse it with it with the given HCl solution. Dip the cell in the solution taken in the beaker. i. Set the range with the help of range knob shown in the Fig to 20. j. Set the Function Switch to Conductivity and read the display. This will be the exact conductivity note it down. k. Take alkali (NaOH) in the burette and add 0.2 mL of it into the beaker containing HCl. Stir and determine the conductivity.
16 LMCHE-102 Chemistry Lab

l. Repeat the procedure of addition of 0.2 mL of NaOH and noting down the conductivity in the observation table. m. Take 25-30 readings in the ways. After each addition, stir the solution gently. n. Plot a graph between observed conductivity value along Y-axis against the volume of alkali added along x-axis. The point of intersection gives the amount of alkali required for neutralization of acid. OBSERVATIONS AND CALCULATIONS: Volume of HCl taken = 50 ml Normality of NaOH solution = 0.1 N OBSERVATION TABLE S. No Volume of NaOH added (ml.) 1. 2. 3. 4. 5 22. 23. 24. 0.2 0.4 0.6 0.8 1.0 Observed conductivity (mmoh/cm)

From graph the volume of NaOH used is (calculated by drawing perpendicular on Xaxis from the point of intersection) = A ml (also called as equivalence point). Applying normality equation N1V1 = (HCl) N2V2 (NaOH)

N1 x 50 = 0.1 x A N1 = a N
17 LMCHE-102 Chemistry Lab

We know strength in grams per litre = Normality x Eq. Wt. Therefore, strength of acid = a x 36.5 g/litre = Y g/litre. 4. Required Results: Parameters: Normality equation Relationship to be determined: Effect of adding a base to an acid on conductivity and strength of an acid Graph: When a graph is potted between volume of the alkali added and conductance then a V shaped graph is obtained. The point of intersection will give the end point.

X Conductance Z Y Equivalence point A ml Volume of NaOH added

Error Analysis: % error: Ask your instructor for the actual value and the calculate error in your result. Result: Strength of given HCl solution = ________ g/litre. 5. Cautions: a. The solution taken in the burette should be about ten times stronger than that taken in the beaker so that the volume change of latter solution is negligible on the addition of the former solution. b. After every addition of NaOH solution, the solution must be stirred thoroughly. 6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Experiment No. 5 1. Experiment: To test the validity of Beer-Lamberts law using colorimeter and to determine unknown concentration of solution. Equipment required: Colorimeter, test tubes, burette, pipette, beakers etc Material required: Distilled Water, 0.01 M Potassium permangnate. 2. Learning Objectives: I) Students will learn the basics of colorimetry and how to use colorimeters II) Students will gain practice in preparing solutions through dilution and in calculating solution concentrations III) Students will use algebraic representations to describe data IV) Students will learn how to use a calibration curve to determine the unknown concentration of a solution Filter (to set at )

To set O.D at zero

Fig: Photograph of Colorimeter Theory: When a monochromatic light of intensity I is incident on a transparent medium, a part of it is absorbed, a part, I, is reflected and the remaining part, I, is transmitted.

In case of aqueous solutions,

is negligible as compared to

and

Beers Lamberts Law: It states that the decrease in intensity of incident light with thickness of absorbing medium is directly proportional to the intensity of incident light and concentration of absorbing medium. (1)
19 LMCHE-102 Chemistry Lab

Where C is concentration of solute expressed in mole/litre, l is the length of the cell and is a constant characteristic of the solute called molar extinction coefficient or molar absorptivity. Further also called as optical density (OD) or absorbance (A). Since absorbance A of the medium is given by (2) From equation (1) and (2) A = Cl Transmittance, T of a solution is the ratio of transmitted by the solution. i.e., the fraction of incident light

A plot between absorbance and concentration is expected to the linear. Such a straight line plot, passing through the origin, shows that Beer- Lamberts law is obeyed. This plot, known as calibration curve can also be also employed in finding the concentration of a given solution. 3. Outline of the Procedure: a. Connect the instrument to the mains and put on the power switch. b. Adjust the wavelength knob to the 40 wavelength region on scale (approximately). c. Open the lid on the cell compartment and insert a cuvette containing the distilled water. Close the lid. d. Adjust the reading on the digital screen to zero optical density with the knob shown in the Fig. e. Remove the cuvette and close the lid tightly again. Empty the cuvette and rinse it with standard solution of KMnO4 (0.001M) [which will be provided to you by lab technician]. Fill the cuvette this solution and note the optical density. f. Change the wave length to the next high value using set wavelength knob every time and note down corresponding optical density. Make table with wavelength on LHS and OD at RHS.

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LMCHE-102 Chemistry Lab

Table 1: Optical density of KMnO4 solution with various wavelengths for the determination of max S. No. 1. 2. 3. 4. 5. 6. Wavelength () in nm O.D.

g. Plot a graph between wavelength on the x-axis and O.D. on the y-axis. Find the value of
max

( wavelength where O.D is maximum)

O.D

max

Wavelength (nm)

h. Now set the max value in the colorimeter with the help of the knob used for setting wavelength and place the cuvette containing Distilled water in the cell compartment. Set O.D to zero again. i. Prepare KMnO4 solution in water with composition 10%, 20%, 30%, 40%, ----------- 100%. 10% composition means 10ml of KMnO 4 and 90ml of water or 1ml of KMnO4 and 9ml of water. Table 2: Optical density of various compositions of KMnO 4 solutions at S. No. 1. 2. 3. 4. 5. 6. 7. 8. 9. Composition (%) 10 20 30 40 50 60 70 80 90 O.D.
max

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LMCHE-102 Chemistry Lab

j.

Note down the absorbance (OD) of series of solution of KMnO 4 prepared above (from 10% to 100%) by the method described above. Do not change wavelength now.

k. Plot a graph between O.D against composition. (If a straight line is obtained Lambert - Beers a law is verified) l. Now take a solution of a unknown concentration and note down optical density. Find out the concentration of the unknown solution from graph. Table 3: Optical density of unknown KMnO4 solution at Composition (%) Unknown O.D

max

4. Required Results: Report the results of unknown solution in gram/litre. Parameters:

max,

absorbance and concentration.

Relationships to be determined: Between concentration and absorbance. Graph: A plot between absorbance and concentration is expected to the linear. Such a straight line plot, passing through the origin, shows that Beer- Lamberts law is obeyed. This plot, known as calibration curve can also be also employed in finding the concentration of a given solution.

O.D

Composition of KMnO4 solution (%)

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LMCHE-102 Chemistry Lab

Error Analysis: To obtain the error bars. % error: Ask your instructor for the actual value and the calculate error in your result. 5. Cautions: a. Handle the glass cuvettes very carefully. b. Switch on the colorimeter at least 10 minutes before the start of the measurements. c. There should be no air drop outside the cuvette, and wipe off cuvette vary carefully with the help of soft tissue paper so that no water molecule is attached on outside and cuvette should not contain any finger impression. d. Let the reading stabilize for tome time (15 seconds) before taking the reading. 6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Experiment No. 6 1. Experiment: Estimation of nickel in the given sample using DMG. Equipments required: Beakers, Suction pump, sintered glass crucible, oven, glass rod etc. Chemical required: Nickel ammonium sulphate, Dimethyl glyoxime, concentrated ammonia, Ethanol, HCl etc. 2. Learning objectives: I) To know about gravimetric analysis technique. II) The students will learn how the transition complex formation is helpful to determine amount of particular ion in given salt. III) How to use sintered glass crucible and suction pump. Theory: Nickel dimethyl glyoxime is prepared by the action of alcoholic solution of dimethyl glyoxime on soluble nickel salts such as Nickel chloride or Nickel ammonium sulphate in presence of NH 4OH solution or alkaline medium. Dimethyl glyoxime is a chelating agent. It forms a coordination complex with Ni 2+ ions. The coordination number of the central metal atom is 4. The oxidation number of Ni is +2. The complex has square planar geometry. Chemical Reaction:

Red ppts
H O H3C
2

O N Ni N O H C C CH3 CH3

OH

H3C

C C

N N O

NiSO4
H3C N OH

2NH4OH
H3C

(NH4)2SO4

2H2O

3. Outline of the Procedure:

Procedure: Note: Apparatus should be cleaned properly prior to use, glass beakers shall be first washed with some organic solvent like acetone/methanol and subsequently rinsed with conc. HCl in order to remove traces of ammonia solution

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LMCHE-102 Chemistry Lab

a. Dissolve 1.0 g of nickel ammonium sulphate or Nickel chloride in distilled water in a beaker and dilute it with 20 ml of distilled water. Add 1.0 ml of concentrated HCl. b. Dissolve 0.6 g of dimethyl glyoxime in 15 ml of ethyl alcohol in a separate conical flask. c. Add dimethyl glyoxime solution to nickel ammonium sulphate solution along with stirring. d. Heat the mixture solution to 60-70oC on water bath. e. Add 6N NH4OH solution (1:1 NH3) or Ammonia solution slowly with constant stirring till precipitation starts. Add excess of 6N NH 4OH solution (means a few drops more even after precipitation) f. Allow the reaction mixture to stand for about 20 minutes so that mixture to settle down. g. Separate the precipitate by filtration through a sintered glass crucible (ask for this from your lab technician) under suction and wash with cold water. h. Remove the brilliant red precipitate formed and dry in air oven. i. Note the colour and weight of the product formed. Observation Colour of the compound = Weight of the ppt = . Calculations (C4H8O2N2)2Ni 288.69 Ni 58.69

Hence, weight of Nickel = 0.2033 weight of the precipitate

4. Required Results: Parameters: Colour and weight Relationship: weight of the samples with the molecular weight of known molecules. Graphs: NA

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LMCHE-102 Chemistry Lab

Error Analysis: % error: Calculate the actual amount of Ni present in starting nickel ammonium sulphate taken for the preparation of complex and calculate the % error. Result: The Nickel content present in Ni(DMG)2 complex = . % error 5. Cautions: a. Apparatus should be cleaned properly. Rinse the apparatus with Conc. HCl then wash with water properly. b. The acid should be handled with care and dropper or a measuring cylinder should be used for addition of it. c. Ammonia solution should be added after heating the reaction mixture. 6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Experiment No. 7 1. Experiment: Determination of the rate constant of hydrolysis in case of ethyl acetate catalyzed by HCl. Equipment required: Six conical flasks, burette, pipette, Stop watch, Water bath etc Material required: 0.1 NaOH, ethyl acetate, 0.5 N HCl, Phenolphthalein. 2. Learning objectives: I) To make the concepts of chemical kinetics understandable. II) To know how to determine the rate constant of a reaction. III) To gain knowledge about chemical kinetics of pseudounimolecular reactions. IV) Student will learn how to find the order of reaction with the help of rate constant at different time intervals. V) To prove that order of reaction is an experimental concept. VI) To gain knowledge about acid base titration. VII) To know the effect of temperature on rate of reaction. Theory: The reaction is catalysed by H+ ions of an acid (HCl). This reaction is an example of psuedounimolecular reactions. Since water is present in large excess, its concentration is practically constant throughout the reaction. The concentration of HCl (catalyst) also remains constant. Therefore, the rate of reaction depends upon only on the concentration of ester. Rate = -dx/dt = k [CH3COOC2H5]. Hence reaction is of first order. During the hydrolysis of ester, acetic acid is produced. Therefore, the progress of reaction is followed by determining the amount of acetic acid formed at different time intervals.
H

CH3COOC2H5 + H2O

CH3COOH + C2H5OH

A definite quantity of the reaction mixture is withdrawn after different time intervals and is titrated against a standard solution of alkali. The amount of alkali used is equivalent to the total amount of HCl present initially and the amount of acetic acid formed. The volume of alkali used at the start of reaction is equivalent to amount of

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LMCHE-102 Chemistry Lab

HCl alone. Hence, the amount of acetic acid formed (x) after different intervals of time can be calculated. The amount of acetic acid formed at the end of reaction is equivalent of initial concentration of ester (a). Suppose the volumes of alkali used required for the reaction, at the start, after time t and the end of reaction are V0,Vt,V respectively, then initial concentration of ester (a) is proportional to V -V0. The concentration of ester after time t is V - Vt K = 2.303 t log V - V0 V- Vt

If the value of K comes out constant during different intervals of time, then order of reaction will first order.

3. Outline of the Procedure: a. Take 50ml of 0.5 N HCl in a clean dry 250mL conical flask and about 10mL of pure ethyl acetate in a test tube, cork both of them and place them in a thermostat or water bath at 45-50 C. (Take water in a big beaker and put conical flask and test tube in it). b. Keep the 0.5N HCl and ethyl acetate in the thermostat or water bath for about 10 min to allow them to acquire the temperature of the bath (40-45 oC). c. In the mean time, fit the burette properly and fill it with 0.1N NaOH solution. d. At about 9 minutes add 25mL of ice cold water in a separate conical flask. e. After 10 minutes pipette out 5mL of the ethyl acetate from the test tube and add it to the flask containing 50 mL of 0.5 N HCl. Start stop watch at this moment from zero again. f. Shake the contents for 2-3 seconds and immediately pipette out 5 mL of reaction mixture and transfer it at once to first conical flask containing ice cold water. g. Keep the flask containing reaction mixture in the water bath again, it should be noted that reaction mixture will be kept in the water bath at constant temperate throughout the experiment. h. Titrate the solution in conical flask containing 25 mL ice cold water and 5 mL of reaction mixture against 0.1 N NaOH taken in burette by using phenolphthalein as indicator. Appearance of pink colour is end point. The volume of 0.1 N NaOH

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LMCHE-102 Chemistry Lab

used against the withdrawn sample of the ester and dil. HCl mixture is taken as V0. i. After about 9 minutes add 25mL of ice cold water in a separate conical flask. j. Pipette out 5 mL of mixture and add it to the conical flask containing ice cold water after 10 min. Titrate it against 0.1 N NaOH . This gives V t after 10 min.

k. Repeat the above procedure after every 10 min for taking readings upto 80 minutes. l. Place the remaining reaction mixture in the separate water bath at 60-70oC for about half to one hour time. Pipette out 5 mL of mixture and titrate it against alkali solution. This gives V.

Observations and Calculations: S. No Time (min) 0 10 20 30 40 50 60 Volume of V - V0 NaOH (ml) V0 V10 V20 V30 V40 V50 V60 V V - V10 V - V20 V - V30 V - V40 V - V50 V - V60 V - Vt log (V - Vt) log(V - V0)

1. 2. 3. 4. 5. 6. 7. 8.

Calculations: Calculation of K at time 10, 20, 30 and 40 min will be done by the following formula: K = 2.303 log V- V0 t V- Vt min -1

log(V- Vt ) = - Kt/2.303 +log (V- V0) is equation of straight line. 4. Required result: Rate constant of ethyl acetate at given temperature is ..........

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LMCHE-102 Chemistry Lab

Parameter used: Rate constant (K): The value of K comes out constant during different intervals of time, and then order of reaction will first order. Relationship to be determined: between K, t and volumes of NaOH used. Graph

log(V - Vt )

slope = - K/2.303. With the help of slope, the value of be calculated. t (min) rate constant can

Error Analysis: Scope of result: We study the kinetics of hydrolysis of esters; hydrolysis of ethyl acetate has a very rapid rate that could be carried in short time. 5. Cautions: a. Use the ice cold water only. b. Perform the titrations properly. c. Always take alkali in burette. d. Shake the reaction mixture properly at regular intervals. 6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Experiment No. 8 1. Experiment: Determine the strength of given solution of ferrous ammonium sulphate (Mohrs salt) by titrating against potassium dichromate solution. Equipment Required: Titration flask, 50 mL Burette, pipette, funnel etc. Material Required: Potassium dichromate K2Cr2O7 reagent grade, Ferrous ammonium sulphate, Sulphuric Acid (concentrated). 2. Learning Objectives: I) Student will learn how to calculate the exact normality of Ferrous ammonium sulphate (Mohrs salt) by titrating with potassium dichromate solution. II) To gain knowledge about redox titration. Theory: This experiment is an example of redox titration. The loss of electrons is oxidation; the gain of electrons is reduction. Reduction/oxidation (redox) processes occur when electrons are transferred from a donor species (the reducing agent 2FeSO4(NH4)2SO4) to another acceptor species (the oxidizing agent K2Cr2O7). During titration of Fe2+ ions against K2Cr2O7, ferrous ions are oxidized to ferric ions by potassium dichromate in acidic solution. The completion of oxidation reaction is marked by the appearance of green colour by oxidized ferroin indicator. Reactions: The reaction between Potassium dichromate and Mohrs salt can be represented as: Chemical Reactions K2Cr2O7 + 4H2SO4 K2SO4 + Cr2(SO4)3 + 4H2O + 3[O] 2FeSO4 + H2SO4 + [O] Fe2(SO4)3 + H2O K2Cr2O7 + 6FeSO4 + 7H2SO4 K2SO4 + Cr2(SO4)3 + 3Fe2(SO4)3 + 7H2O Ionic Equations Cr2O72 + 14H+ + 6e 2Cr3+ + 7H2O Fe2+ Fe3+ + e6

Cr2O72 + 6Fe2+ + 14H+ Fe3+ + 2Cr3+ + 7H2O

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3. Outline of the Procedure: Titration of K2Cr2O7 soln. with Mohrs salt solution a. Pipette out 10 mL of Mohrs salt solution in the conical flask. b. Add approximately 4 mL of Conc. H2SO4 to the same flask. c. Add 2-4 drops of the Ferroin indicator. d. Titrate the reaction mixture with potassium dichromate solution taken in burette till a colour change from Wine red to green is obtained. e. Repeat the titration for three concordant readings.

Observations and Calculations Volume of Mohrs salt solution used in each titration = 10 mL Indicator used = Ferroin indicator Color change at end point = Wine red - green Equivalent weight of Mohrs salt Table. S. No. Burette readings Initial 1. 2. 3. 4. 5. 6. Final Volume of K2Cr2O7 Consumed (R2 - R1) mL = 392

Thus applying the normality relation NMohr VMohr = Thus NMohr

NdichrVdichr

= NdichrVdichr/VMohr

Thus strength (g/L) of Mohrs salt solution = Normality x Eq. Wt 4. Required Results: Strength of given Ferrous ammonium sulphate solution= g/litre. Parameters used: Volumetric parameters, Normality equation
32 LMCHE-102 Chemistry Lab

Relationship to be determined: determine how much amount of oxidizing agent required for Fe2+ ions solution that can be used to calculate the concentration of the latter. Plot/Graph: NA Error Analysis: Ask for the actual normality of Fe2+ ions solution and calculate the % error. Result: Strength of given Ferrous ammonium sulphate solution= g/litre. 5. Cautions: a. Always take Potassium dichromate solution in burette. b. Potassium dichromate acts as oxidizing agent in acidic medium. Therefore always add dil. H2SO4 in the reducing agent. c. Read the upper meniscus while taking burette readings because K 2Cr2O7 is d. coloured.

6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Experiment No. 9 1. Experiment: Separation of amino acids by thin layer chromatography. Equipment required: Glass plates, beaker, Glass rod, Material required: Silica gel (for TLC), Glycine, Leucine, acetic acid, n-butanol, water, alcoholic solution of ninhydrin 2. Learning Objectives: I) II) III) Students will learn the basis of TLC and how to prepare the TLC plates. To calculate retention factor (Rf). Students will learn how to separate the mixture of organic compounds on the basis of Rf value Theory: Thin Layer Chromatography (TLC) is used extensively for qualitative analysis (tentative identification of mixture of two or more organic compounds).

Chromatography is based on the general principle of distributing the components of a mixture of organic compounds between two phases - a stationary phase and a mobile phase. In thin layer chromatography the stationary phase is a solid supported on a glass plate, while the mobile phase is a liquid (solvent). As the stationary phase is a solid, the basis of separation of different components is adsorption. Hence, the chromatography can be defined as the technique for the separation of a mixture of compounds where the separation is brought about by the differential movement of the individual compounds through a porous medium under the influence of a moving solvent. Thin layer chromatography (TLC) is type of adsorption chromatography, which involves separation of the components of a mixture over a thin layer of an adsorbent. A thin layer (about 0.2 mm thick) of an adsorbent (Silica gel or alumina) is spread over a glass plate of suitable size. The plate is known as thin layer chromatography (TLC) plate. In this technique a small amount of the material (to be separated), dissolved in an appropriate solvent is is applied as a small spot near one edge (about one cm above) of the TLC plate covered with thin layer of adsorbent. After the sample has been deposited on the adsorbent, the coated plate is placed in a beaker containing small amount of solvent in such a manner that level of solvent must be lower than the marked spot. As the solvent in a beaker moves up
34 LMCHE-102 Chemistry Lab

through the adsorbent by capillary action, the various components of mixture move up along with the solvent to different distances depending on this degree of adsorption. This results in the separation of various components of mixture. The relative adsorption of each component of the mixture is expressed in terms of its retention factor ie., Rf Value.

Retention Factor: The movement of any substance relative to the solvent front is called retention factor. It is constant for a given chromatographic system and characteristic of a substance. When the solvent front has almost reached the top of the adsorbent layer or three-fourth of it, the plate is removed from the beaker, dried and examined. TLC involves the following steps: (a) Preparation of a thin layer plate (b) Application of the materials to be separated on the plate (c) Development of the chromatogram plate in a solvent (d) Visualization or Location of components (e) Calculation of Rf values. 3. Outline of the Procedure: a. Take small amount of silica gel in a beaker and dissolve it in distilled water with constant stirring by a glass rod. Continue to stir until a uniform paste free from air bubbles is formed. Add some more water to obtain slurry of suitable consistency. (OR this slurry may be provided to you by the lab technician). b. Mark the base line on the glass plate about 1 cm from the bottom edge of the glass plate. (Mark only at edges with pencil). This line is just to take an idea that where 1 cm distance lies from the bottom where sample mixture is to be applied. c. Pour the slurry on to the clean and dry plate and prepare a uniform thin layer by glass rod.
35 LMCHE-102 Chemistry Lab

d. Allow the layer to dry for 5-10 minutes and then heat the plates in an electric oven at 100-120oC for about 20 min. e. Prepare solution by mixing two or more different organic compounds. (Given to you by lab technician). f. At base line and apply small sample of the mixture with the help of thin capillary tube in the centre. Take care the spots must be as small as possible. g. Allow the spot to dry. Place the glass plate in a beaker containing solvent to a depth of about 1 cm or less than that and allow the solvent to flow up until it nearly reaches the top of the plate. (take due care that spot must not touch the solvent) h. Remove the plate from the beaker, mark the position of the solvent front (on edge of the plate with pencil) and allow the solvent to evaporate. i. Spray with alcoholic ninhydrin and dry TLC plate in the oven for 2 min. j. Spots will develop on the plate. Take measurements of the distance moved by solvents and each component from the 1 cm mark. k. Calculate the Rf values of the components in the mixture. OBSERVATION: Distance moved by first component Glycine (b1) = cm Distance moved by second component Leucine (b 2) = cm Distance moved by solvent (a) = cm CALCULATIONS:

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LMCHE-102 Chemistry Lab

4. Required Results: Parameter used: Retention Factor (Rf): The movement of any substance. Relationship: determination of relative Rf value of mixture of compounds Plot/Graph: Draw slide with observed lengths of solvent and two components

b2 b1

Error Analysis: NA Result: Rf value for first component (Glycine) = .. Rf value for second component (Leucine) = Scope of Result: Thin Layer Chromatography (TLC) is used extensively for qualitative analysis and tentative identification of mixture of two or more organic compounds. TLC is a useful screening technique in clinical chemistry; for example, it can be used to detect the presence of drugs in urine. 5. Cautions: a. Make the slurry very carefully; it should not be very thick or very thin. b. Always prepare fresh silica slurry. c. Make sure that sample dot is always outside the solvent layer. d. Spots of mixture must be as small as possible. e. Dry TLC plates carefully. 6. Learning outcomes: To be written by the students in 50-70 words.

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LMCHE-102 Chemistry Lab

Some Common Apparatus used in Chemistry lab:

Apparatus for TLC

TLC plate in solvent in development phase

TLC plate in solvent in development Ninhydrin Spray apparatus phase

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LMCHE-102 Chemistry Lab

Burette with stand burette

How to read the

Graduated Pipette

Pipette with filler

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LMCHE-102 Chemistry Lab

Conical Flasks

Measuring Cylinders

Droppers

Beakers

Spatula

Water bath

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LMCHE-102 Chemistry Lab