Analytical Chemistry Practical (Chem-2202P)

Experiment 1: Calibration of Glassware

Introduction

The fundamental goal of this experiment is to explore and understand the various techniques used for
measuring solutions with various types of volumetric glassware. Also, to understand the significance of precision
and accuracy, and how these concepts are affected by the types and choices of glassware used to measure specific
volumes of solutions. In order to calculate the value of uncertainty, the standard deviation and relative standard
deviation will be calculated and evaluated closely.

Experimental Procedure

Calibration of Pipets

- Weight a clean dry 50mL beaker five times and records each measurement precisely.
- Using the 1mL pipet, fill with water to the designated line and dispense water into the 50mL beaker and
weight five times. Record each measurement precisely. Repeat four more trials of this procedure for the
1mL pipet. (Make sure beaker is dried between each trial)
- Using the 2mL pipet, fill with water to the designated line and dispense water into the 50mL beaker and
weight five times. Record each measurement precisely. Repeat four more trials of this procedure for the
2mL pipet. (Make sure beaker is dried between each trial)
- Using the 5mL pipet, fill with water to the designated line and dispense into the 50mL beaker and weight
five times. Record each measurement precisely. Repeat four more trials of this procedure for the 5mL
pipet. (Make sure beaker is dried between each trial)
- Take the average of each individual trial for each pipet used. Then take the total average of all the
individual averages.
- Calculate the standard deviation and the relative standard deviation.

Calibration of Volumetric Glassware

- Weight a clean, dry 10mL volumetric flask five times. Record each measurement precisely.
- Weight a clean, dry 25mL volumetric flask five times. Record each measurement precisely.
- Weight a clean, dry 50mL volumetric flask five times. Record each measurement precisely.
- Fill a 10mL volumetric flask to the designated mark and weight the flask five times. Record each
measurement precisely. Repeat four more trials of this procedure for the 10mL volumetric flask.
- Fill a 25mL volumetric flask to the designated mark and weight the flask five times. Record each
measurement precisely. Repeat four more trials of this procedure for the 25mL volumetric flask.
- Fill a 50mL volumetric flask to the designated mark and weight the flask five times. Record each
measurement precisely. Repeat four more trials of this procedure for the 50mL volumetric flask.
- Take the average of each individual trial for each pipet used. Then take the total average of all the
individual averages.
- Calculate the standard deviation and the relative standard deviation.

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Data
Table 1. Dry weight of Volumetric Glassware in grams (g).
50mL Beaker 10mL Volumetric 25mL Volumetric 50mL Volumetric
Flask Flask Flask
32.0477g 8.986g 18.523g 35.966g
32.0482g 8.987g 18.521g 35.967g
32.0479g 8.987g 18.523g 35.966g
32.0480g 8.986g 18.524g 35.966g
32.0480g 8.986g 18.523g 35.965g

Table 2. Wet weights for the 1mL Pipet in grams (g). (fill the pipette and make it empty in beaker)
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
33.082g 33.113g 33.107g 33.070g 33.074g
33.083g 33.114g 33.106g 33.068g 33.074g
33.082g 33.112g 33.106g 33.068g 33.073g
33.081g 33.113g 33.105g 33.067g 33.073g
33.082g 33.111g 33.107g 33.067g 33.072g

Table 3. Wet weights of the 2mL Pipet in grams (g). (fill the pipette and make it empty in beaker)
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
34.079g 34.106g 34.074g 34.135g 34.084g
34.080g 34.107g 34.072g 34.134g 34.083g
34.079g 34.106g 34.073g 34.135g 34.084g
34.078g 34.105g 34.074g 34.134g 34.083g
34.078g 34.105g 34.073g 34.134g 34.085g

Table 4. Wet weights of the 5mL pipet in grams (g). (fill the pipette and make it empty in beaker)
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
36.972g 37.014g 36.944g 36.996g 36.959g
36.971g 37.013g 36.944g 36.995g 36.958g
36.972g 37.014g 36.945g 36.996g 36.959g
36.973g 37.013g 36.943g 36.997g 36.960g
36.972g 37.013g 36.943g 36.997g 36.958g

Table 5. Wet weights of the 50mL volumetric flask in grams (g).

Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
85.745g 85.753g 85.766g 85.794g 85.801g
85.743g 85.754g 85.764g 85.794g 85.800g
85.746g 85.756g 85.763g 85.794g 85.799g
85.743g 85.755g 85.764g 85.795g 85.799g
85.742g 85.755g 85.765g 85.793g 85.799g
Table 6. Wet weights of the 25mL volumetric flask in grams (g).
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
43.350g 43.293g 43.368g 43.343g 43.355g
43.351g 43.294g 43.368g 43.340g 43.354g
43.351g 43.293g 43.368g 43.341g 43.354g
43.350g 43.292g 43.367g 43.340g 43.353g
43.349g 43.294g 43.367g 43.339g 43.353g

Table 7. Wet weights of the 10mL volumetric flasks.

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Trial 1 Trial 2 Trial 3 Trial 4 Trial 5

18.922g 18.918g 18.901g 18.897g 18.818g
18.922g 18.916g 18.901g 18.897g 18.817g
18.923g 18.919g 18.899g 18.896g 18.818g
18.923g 18.918g 18.899g 18.899g 18.818g
18.922g 18.917g 18.900g 18.896g 18.817g

Calculations

To calculate the average weight of a trial. You add all the data points collected in the specific trial and then divide
that total number by the number of data points there are.

x1 + x 2 + x3 + x 4 + x 5
Mean Calculation : n
Example: Mean Calculation (Using the data for trial 1 of the 10mL pipet)

18.922 g+18.922 g +18.923 g+ 18.923 g+18.922 g

5
= 18.9224g

Standard Deviation Formula

2
√ x −avg ¿ +¿ ¿ ¿
1

Relative Standard Deviation Formula

standard deviation
x 100
total average
Example: Standard Deviation Calculation- (Using data for wet weight of 1mL Pipet)

√ 33.0820 g−33.0886 g ¿2 +¿ ¿
= 0.009844542

Example: Relative Standard Deviation Calculation- (Using data for wet weights of 1mL pipet)

0.009844542
x 100
33.0886
= 0.029752066 %

Averages, Standard Deviation, and Relative Standard Deviation of the Wet Weights of the
1mLStandard
Averages, Standard Deviation, and Relative Pipet Deviation of the Wet Weights of the
Trial 1 Trial2mL
2 Pipet Trial 3 Trial 4 Trial 5
Average 33.0820g
Trial 1 33.1126g
Trial 2 33.1062g
Trial 3 33.0732g
Trial 4 33.0690g
Trial 5
Total Average
Average 34.077g 34.1058g 33.0886
34.0732g 34.1344g 34.0838g
Standard Deviation
Total Average 0.009844542
34.09484
Relative Standard
Standard Deviation 0.012731737
Deviation
Relative Standard 0.029752%
Deviation 0.0373421%

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Averages, Standard Deviation, and Relative Standard Deviations of the Wet Weights of the
5mL Pipet
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
Average 36.972g 37.0134g 36.9438g 36.9962g 36.9588g
Total Average 36.97684
Standard 0.000787989
Deviation
Relative Standard
Deviation 0.002131%

Averages, Standard Deviation, and Relative Standard Deviation of the Wet Weights of the
10mL Volumetric Flask
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
Average 18.9224g 18.9176g 18.9000g 18.8970g 18.8176g
Total Average 18.89092
Standard Deviation 0.021213857
Relative Standard
Deviation 0.1122966%

Averages, Standard Deviation, and Relative Standard Deviation of the Wet Weights of the
25mL Volumetric Flasks
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
Average 43.3502g 43.2932g 43.3676g 43.3406g 43.3538g
Total Average 43.34108
Standard Deviation 0.014232459
Relative Standard
Deviation 0.0328383%

Averages, Standard Deviation, and Relative Standard Deviations of the Wet Weights of the
50mL Volumetric Flask
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5
Average 85.7438g 85.7546g 85.7644g 85.794g 85.7996g

Total Average 85.77128

Standard Deviation 0.024489416
Relative Standard 0.028552%
Deviation

Experiment 2: Construction of Calibration Curve

Introduction

In analytical chemistry, a calibration curve is a general method for determining the concentration of a substance in
an unknown sample by comparing the unknown to a set of standard samples of known concentration. A calibration
curve is one approach to the problem of instrument calibration; other approaches may mix the standard into the
unknown, giving an internal standard.

The calibration curve is a plot of how the instrumental response, the so-called analytical signal, changes with the
concentration of the analyte (the substance to be measured). The operator prepares a series of standards across a
range of concentrations near the expected concentration of analyte in the unknown. The concentrations of the

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standards must lie within the working range of the technique (instrumentation) they are using. Analyzing each of
these standards using the chosen technique will produce a series of measurements. For most analyses a plot of
instrument response vs. concentration will show a linear relationship. The operator can measure the response of
the unknown and, using the calibration curve, can interpolate to find the concentration of analyte.

The data - the concentrations of the analyte and the instrument response for each standard - can be fit to a
straight line, using linear regression analysis. This yields a model described by the equation y = mx + c, where y is
the instrument response, m represents the sensitivity, and c is a constant that describes the background. The
analyte concentration (x) of unknown samples may be calculated from this equation.

Many different variables can be used as the analytical signal. For instance, chromium (III) might be measured using
a chemiluminescence method, in an instrument that contains a photomultiplier tube (PMT) as the detector. The
detector converts the light produced by the sample into a voltage, which increases with intensity of light. The
amount of light measured is the analytical signal.

Most analytical techniques use a calibration curve. There are a number of advantages to this approach. First, the
calibration curve provides a reliable way to calculate the uncertainty of the concentration calculated from the
calibration curve (using the statistics of the least squares line fit to the data).

The chief disadvantages are (1) that the standards require a supply of the analyte material, preferably of high
purity and in known concentration, and (2) that the standards and the unknown are in the same matrix. Some
analytes - e.g., particular proteins - are extremely difficult to obtain pure in sufficient quantity. Other analytes are
often in complex matrices, e.g., heavy metals in pond water. In this case, the matrix may interfere with or
attenuate the signal of the analyte. Therefore a comparison between the standards (which contain no interfering
compounds) and the unknown is not possible. The method of standard addition is a way to handle such a situation.

Experimental Procedure

1. The first step in creating a graph using Microsoft Excel is entering the data. The data should be in two adjacent
columns with the x data in the left column. Labeled the columns but this is not necessary to create the graph.

Note that your absorbance data should include the origin {concentration = zero, absorbance = zero}:

2. Position the cursor on the first X value (i.e., at the top of the column containing the x values, or "Concentration”
values), hold down the left mouse button and drag the mouse cursor to the bottom Y value (i.e., at the bottom of
the column containing the y values, or "Absorbance" values). All of the X-Y values should now be highlighted (do
NOT highlight the labels!):

3. Click on the “Insert” tab at the top of the toolbar.

4. Under the “Chart” section, choose “Line”. At the bottom of this menu, choose “All Chart Types”.

5. Under “Chart Type” choose “XY (Scatter)”, and pick the first option, (without lines). (On older versions of Excel,
you will have to choose the “chart subtype” – choose the option that plots the points without lines).

6. After you click “OK”, a reduced version of your graph will appear; it should be a set of points that makes a
straight line with a positive slope.

7. If you wish, you may label your graph (i.e., put a title at the top of the graph). You may wish to label the y-axis
“absorbance” and the x-axis “concentration”. You may also modify the style of line, the legend, the data markers,
and other options if you wish, although this is not necessary.

8. To determine the slope and intercept of your line, move the mouse cursor to any data point and press the left
mouse button. All of the data points should now be highlighted. Now, while the mouse cursor is still on any one of

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the highlighted data points, press the right mouse button, and click on “Add Trendline” from the menu that
appears. 9. From within the "Trendline" window, select the type of “Trend / Regression Type” you want – for a
Beer’s Law plot the function should be “Linear”.

10. At the bottom of the same menu, choose the option “Display Equation on Chart”.

11. Click “OK”. A line and an equation should appear on the graph, as shown below. Notice that this equation is in
the format {y = mx + b}, and numerical values are provided for the slope and intercept. The value of the intercept
should be close to zero, a small number, but it may not be exactly zero. In my example, the slope is 21033 and the
intercept is 0.0039: Using other software to plot calibration curve: If you don’t have MS Excel (or access to this
program), there are several free programs available that will serve the purpose.

Plot and print the calibration curve separately.

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Experiment 3
DETERMINATION OF ALKALINITY OF WATER
INTRODUCTION

Alkalinity is primarily a way of measuring the acid neutralizing capacity of water. In other words, it is the ability to
maintain a relatively constant pH of water. The possibility to maintain constant pH is due to the hydroxyl,
carbonate and bicarbonate ions present in water. The ability of natural water to act as a buffer is controlled in part
by the amount of calcium and carbonate ions in solution. Carbonate ion and calcium ion both come from calcium
carbonate or limestone. So water that comes in contact with limestone will contain high levels of both Ca++ and
CO32- ions and have elevated hardness and alkalinity.

ENVIRONMENTAL SIGNIFICANCE

Alkalinity is important for fish and aquatic life because it protects or buffers against rapid pH changes. Higher
alkalinity levels in surface waters will buffer acid rain and other acid wastes and prevent pH changes that are
harmful to aquatic life. Large amount of alkalinity imparts bitter taste in water.

The principal objection of alkaline water is the reactions that can occur between alkalinity and certain cations in
waters. The resultant precipitate can corrode pipes and other accessories of water distribution systems.

Wastewaters containing excess caustic (hydroxide) alkalinity are not to be discharged into natural water bodies or
sewers. Alkalinity as carbonate and bicarbonate of saline water is very important in tertiary recovery processes for
recovering petroleum. Alkaline water offers better wetting to the formation rock and improve oil release. As an
additional benefit, ions that provide alkalinity absorb on rock surfaces occupying adsorption sites and decrease the
loss of recovery chemical by adsorption. The alkalinity value is necessary in the calculation of carbonate scaling
tendencies of saline waters.

The alkalinity acts as a pH buffer in coagulation and lime-soda softening of water. In wastewater treatment,
alkalinity is an important parameter in determining the amenability of wastes to the treatment process and control
of processes such as anaerobic digestion, where bicarbonate alkalinity, total alkalinity, and any fraction
contributed by volatile acid salts become considerations.

PRINCIPLE

The alkalinity of water can be determined by titrating the water sample with Sulphuric acid of known values of pH,
volume and concentrations. Based on stoichiometry of the reaction and number of moles of Sulphuric acid needed
to reach the end point, the concentration of alkalinity in water is calculated.

When a water sample that has a pH of greater than 4.5 is titrated with acid to a pH 4.5 end point, all OH-, CO 32-,
and HCO3-1 will be neutralized. For the pH more than 8.3, add phenolphthalein indicator, the colour changes to pink
colour. This pink colour is due to presence of hydroxyl ions. If sulphuric acid is added to it, the pink colour
disappears i.e. OH- ions are neutralized.

Then add mixed indicator, the presence of CO32- and HCO3-1 ions in the solution changes the colour to blue. While
adding sulphuric acid, the color changes to red, this color change indicates that all the CO 32- and HCO3-1 ions has
been neutralized. This is the end point.

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MATERIALS REQUIRED

APPARATUS REQUIRED CHEMICALS REQUIRED

1. Burette with Burette stand 1. Standard sulphuric acid

2. Pipettes 2. Phenolphthalein
3. Pipette sucker 3. Mixed Indicator or Bromocresol Green
4. Conical flask (Erlenmeyer Flask) or Methyl Red
5. 250 mL Measuring cylinders 4. Ethyl alcohol
6. Volumetric flask 5. Distilled Water as Blank
7. Beakers

PROCEDURE

PREPARATION OF REAGENTS

Sulphuric Acid Solution (0.02N):

• Take approximately 500 mL of distilled water in a 1000 mL standard flask.

• Pipette 20 mL of concentrated 0.1 Normality Sulphuric acid and add slowly along the sides of the standard flask.

• Then make up the volume up to 1000 mL mark. Now the strength of this solution is 0.02 N.

Phenolphthalein Indicator Preparation: Weigh 1g of phenolphthalein and add to 100 mL of 95% ethyl alcohol or to
100 mL of distilled water. Use the readymade Phenolphthalein indicator available in lab.

Mixed Indicator Preparation: Dissolve 100 mg Bromocresol green and 20 mg of methyl red in 100 mL of 95% ethyl
alcohol or use 100 mL of distilled water. You may use methyl orange prepared solution in lab.

TESTING OF WATER SAMPLE

1. Rinse the burette with 0.02N Sulphuric acid and discard the solution.
2. Fill the burette with 0.02N sulphuric acid and adjust it to zero.
3. Fix the burette in the stand.
4. Using a measuring cylinder exactly measure 100 mL of sample and pour it into a 250 mL of conical flask.
5. Add few drops of phenolphthalein indicator to the contents of conical flask.
6. The colour of the solution will turn to pink. This colour change is due to alkalinity of OH -1 ions in the
sample.
7. Titrate it against 0.02N sulphuric acid till the pink color disappears. This indicates that all the hydroxyl
ions are removed from the water sample.
8. Note down the titter value (V1). The value of titration is 0.5mL .This value is used in calculating the
phenolphthalein alkalinity.
9. To the same solution in the conical flask add few drops of mixed indicator.
10. The colour of the solution turns to blue. This colour change is due to CO 32- and HCO3-1 ions in water
sample.
11. Continue the titration from the point where stopped for the phenolphthalein alkalinity. Titrate till the
solution becomes red. The entire volume (V2) of sulphuric acid is noted down and it is accountable in
calculating the total alkalinity.
12. Repeat the titration for concordant values.

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CALCULATION

Phenolphthalein Alkalinity / Partial Alkalinity / P Alkalinity (mg/L)

= volume of H2SO4 (V1)x normality x 50 x1000 / volume of sample taken

Total Alkalinity (mg/L)= volume of H2SO4 (V2) x normality x 50 x 1000 / volume of sample taken

INFERENCE

Alkalinity is a measure of the capacity of water to neutralize acids. The predominant chemical system present in
natural waters is one where carbonates, bicarbonates and hydroxides are present. The bicarbonate ion is usually
prevalent. However, the ratio of these ions is a function of pH, mineral composition, temperature and ionic
strength. Water may have a low alkalinity rating but a relatively high pH or vice versa, so alkalinity alone is not of
major importance as a measure of water quality. Alkalinity is not considered detrimental to humans but is
generally associated with high pH values, hardness and excess dissolved solids. High alkalinity waters may also
have a distinctly flat, unpleasant taste.

Alkalinity should not exceed 200 mg/L for potable water. For the fresh water alkalinity ranges between 20 – 100
mg/L. If alkalinity of tested sample is within the limits specified in the standards the water sample is fit for drinking.

PRECAUTIONS

1. Do not keep the indicator solution open since it contains the alcohol which tends to evaporate.

2. The mixed indicator solution is containing dye in it; care should be taken so that it is not spilled to your
skin.

3. If it spills on your skin, the scar will remain at least for two to three days.

REFERENCES

(1) APHA Standard Methods for the Examination of Water and Wastewater - 20 th Edition. Method 2320.

(2) Methods for Chemical Analysis of Water and Wastes, EPA-600/4-79-020, USEPA, Method 310.1.

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Experiment 4
DETERMINATION OF CHLORIDE CONTENTS IN WATER
Purpose: This STM is to explain step by step method for estimation of Chloride in given sample of water.

Scope: Method is applicable to drinking and surface waters, domestic and industrial wastes, and saline waters.

Principal: This is a titrimetric method. An indicator, potassium chromate, is added to the sample, which loosely
bind up a few chloride ions. Silver nitrate solution is then titrated into the sample producing fine white precipitates
of silver chloride. When all the free chloride ions are complexed in this reaction the silver nitrate takes the bound
chloride ions from the indicators, producing the blood red precipitate, silver promote making the end of the
titration

Formula mg Cl-1/L = (A-B) x N x35450

Ml SAMPLE
A= ml titrated for sample, B= ml titrated for blank, N= normality of titrant

A standard of 500 mg/l chloride ions is run along with the sample analysis and standardized your solution.

Apparatus: Burette (25 ml), Erlenmeyer Flasks (250 ml), Volumetric Flasks (1000 & 100 ml)

Reagents:

Potassium chromate as an indicator: Dissolve 50 gm potassium chromate in a little distilled water, add silver nitrate
solution a drop at it ion until a definite a red precipitate is formed. Let stand 12 hrs, filter, diluted to 1 ltr.

AgNO3, 0.0141 M (The titrant): Dissolve 2.395 AgNO3 in distilled water and dilute to 1000ml water, Standardizes
against NaCl

Standard Sodium Chloride: NaCl (0.0141 M): Dissolve 824mg NaCl (Dried at 140C0) in distilled water & dilute to
1000ml. Concentration of this solution is 500mg/l

Note: Silver nitrate stains skin so avoid contact

Procedure:

 Measure 100ml sample into an Erlenmeyer flask.

 Treat the NaCl solution as if it were a sample, and run this through the procedure also. The solution will
turn milky white & then paled pink.
 End titration at this point.
 Also run the distill water blank.
 Add 1 ml potassium chromate (K2CrO4) indicator and mix.
 Titrated with AgNO3 titrant until red precipitate diffuses throughout the solution.
 End titration and record ml used.
 Calculate first for the titrant normality using ml titrated for the NaCl standards. It is N.
 When calculating mg Cl-/L for the samples use the titrant normality calculated above.

Note: The silver chloride Precipitate form in water sample of low chloride content is barely visible. It may not be
noticed.

Reference:

Drinking water Chemistry by Barbara A. Hauser (Lewis Publishers).

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Experiment 5
DETERMINE HARDNESS OF GIVEN WATER SAMPLE
Principle: Water hardness is traditional measure of the capacity of water to precipitate soap. Hardness is defined
as the sum of the concentrations of calcium and magnesium ions dissolved in water, both expressed as CaCO 3 in
mg/L. Total hardness is simple titration at pH 10.0 + 0.1, which makes use of an organic adsorption indicator to
define the endpoint. The indicator, Eriochrome Black T (viscous dark blue colour) is added to the sample. It loosely
binds up a few calcium and magnesium ions, creating a complex that exhibit a wine red color. The sample is
titrated against standard disodium salt of EDTA (titrant). EDTA ties up free calcium and magnesium ions. When
there are no more free ions left titrant combines with the ions which are loosely bound to the indicator. Indicator
then turns back to blue color and end point is reached.

The degree of hardness of drinking water has been classified in terms of the CaCO 3 equivalent
concentration as Soft (0-60 mg/L), medium (60-120 mg/L), hard (120-180 mg/L) and very hard (> 180
mg/L).
Scope: Method is applicable to drinking and surface waters and domestic and industrial wastes. To avoid large
titration volumes, use aliquot containing <25 mg CaCO 3. Generally water with total hardness over 200 mg/L as
CaCO3 would be considered as hard H2O.

Reagents:

 Buffer Solutions: Dissolve 16.9 g NH4Cl in 143 ml NH4OH (NH3 solution 33 %), add 1.25 g Mg EDTA (1.179 g
Na2EDTA.2H2O and 0.780 g MgSO4.7 H2O or 0.644 g MgCl2 .6H2O dissolved in 50 ml H2O may be substituted for
1.25 g Mg-EDTA), and dilute to 250 ml with H2O.

 Indicator: Mix a few crystals (0.5 g) of Erichrom Black T and 100 g NaCl.

 EDTA standard solution: - 0.01 M: Weight 3.723 g Na 2EDTA 2H2O and dilute to 1 L with H2O. Standardize
against Ca standard solution as in 4th step of method. Store in polyethylene bottle and re-standardize
periodically.

 Calcium Standard Solutions: 1.000 mg CaCO 3/ml. Weight 1.000 g CaCO3 (primary standard or special reagent
low in heavy metals, alkalis and Mg) into 500 ml Erlenmeyer flask. Place funnel in neck and add, little at a time,
HCl (1+1) until all CaCO3 has dissolved. Add 200 ml H2O and boil for few minutes to expel CO2, cool, add few
drops of methyl red indicator, and adjust to intermediate orange with 3N NH 4OH or HCl (1+1) as required.
Transfer quantitatively to 1 L volumetric flask and dilute to volume.

 Standardization of EDTA: Dilute 25 ml standard calcium solution to 50 ml with distilled water. Add 1 ml buffer
then 1-2 drops of indicator and mix. Solution will turn red. Titrate it with EDTA till blue endpoint. Record ml
used as (a/b), where, a= calcium standard solution and b= mg of CaCO 3 equivalent to 1 ml EDTA

Procedure:

 Take 25 ml of sample and add 1-2 ml buffer solution

 Add few drops of indicator (solution will turn red).
 Titrate with EDTA standard solution slowly till blue endpoint.

Calculations:
Hardness (EDTA) as mg CaCO3/L = A x B x 1000
ml of sample

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Where

A = ml EDTA standard solution

B = a/b mg CaCO3 equivalent to 1.00 ml EDTA titrant

Precautions:

 Add NH3 buffer prior to titration.

 Use mask before adding NH3 buffer.
 Discard buffer solution (NH3 solution) after one month.

References:

 Official Method of Analysis of AOAC. Vol. 1, part 1 973.52 Hardness of water. First Action 1973. p, 323
 Barbora A Houser Drinking water Chemistry, Laboratory Manual Lewis Publishers. Total Hardness Sg.
 Standard Method for the examination of water and waste water 20th edition by Lenores Clesen. Arnol E
Greanberg, Andrew D Eaton, P. 2-3a

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Experiment 6
Gravimetric determination of nickel in a given sample
Objectives:

 To precipitate nickel from the solution by adding dimethyl glyoxime.

 Mass of nickel calculated from the mass of precipitate.

Theory:

Gravimetric analysis is one of the most accurate analytical methods available. It is concerned with the
determination of a substance by the process of weighing. The element or radical to be determined is converted
into a stable compound of definite composition and the mass of the compound is determined accurately. From
this, the mass of element or radical is calculated.

The gravimetric analysis involves a) precipitation b) filtration c) washing of the precipitate and d) drying, ignition
and weighing of the precipitate.

Following are the four fundamental types of gravimetric analysis:

1. Physical gravimetry

2. Thermogravimetry

3. Precipitative gravimetric analysis

4. Electrodeposition. These differ in the preparation of the sample before weighing of the analyte.

Physical gravimetry: Physical gravimetry involves the physical separation and classification of matter in
environmental samples based on volatility and particle size (e.g., total suspended solids). It is the most common
type used in environmental engineering.

Thermogravimetry: In this method the samples are heated and the changes in sample mass are recorded. Volatile
solid analysis is an important example for this type of gravimetric analysis.

Precipitative gravimetry: The chemical precipitation of an analyte occurs in the precipitative gravimetry. The most
important application of this technique in the environmental field is the analysis of sulphite.

Electrodeposition: It involves the electrochemical reduction of metal ions at a cathode and simultaneous
deposition of the ions on the cathode.

The steps commonly followed in gravimetric analysis are;

(1) Preparation of a solution containing a known weight of the sample.

(2) Separation of the desired constituent.
(3) Weighing the isolated constituent.
(4) Computation of the amount of the particular constituent in the sample from the observed weight of
the isolated substance.

Pecipitative Gravimetric Analysis:

Precipitative gravimetric analysis requires that the substance to be weighed be readily removed by filtration. In
order for a non-filterable precipitate to form, it must be supersaturated with respect to its solubility product
constant. However, if it is too far above the saturation limit, crystal nucleation may occur at a rate faster than
crystal growth (the addition of molecules to a crystal nucleus, eventually forming a non-filterable crystal). When

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this occurs, numerous tiny micro-crystals are formed rather than a few large ones. In the extreme case, micro-
crystals may behave as colloids and pass through a fibrous filter. To avoid this, precipitating solutions may be
heated. Because the solubility of most salts increases with increasing temperature, this treatment will lower the
relative degree of super saturation and slow the rate of nucleation. Also, one might add the precipitant slowly with
rapid mixing to avoid the occurrence of locally high concentrations.

Precipitative gravimetry is often practiced at high ionic strengths. This is to reduce the electric double layer
thickness (salting-out effect) of the slowly forming crystals. When this occurs, electrostatic repulsion between the
crystal and its precipitating molecules is reduced. Crystal growth can then occur more rapidly. It is very important
that the precipitate be pure and has the correct stoichiometry.

Co-precipitation:

For certain gravimetric analysis some other substances besides the desired substances also get precipitated in
small amounts, the phenomenon is called co-precipitation. It occurs when an unwanted ion or molecule becomes
trapped in the precipitate. This may be due to inclusion or occlusion. Inclusion is the term used for a single
substitution in the crystal lattice by an ion of similar size. Occlusion refers to the physical trapping of a large pocket
of impurities within the crystal. One technique for minimizing these problems is to remove the mother liquor, re-
dissolve the precipitate and then re-precipitate. The second time the mother liquor will contain fewer unwanted
ions capable of co-precipitation.

Conditions of Precipitation:

1. Precipitation should be carried out in dilute solutions.

2. The reagents should be mixed slowly and with constant stirring. This will assist the growth of large
crystals.
3. Precipitation is effected in hot solutions, provided the solubility and stability of the precipitate permit
it. Either one or both of the solutions should be heated to just below the boiling point. High
temperature assists (a) coagulation and increase (b) velocity of crystallization.
4. Crystalline precipitates should be digested for as long as possible, preferably overnight.

Precipitating Reagents:

Ideally a gravimetric precipitating agent should react specifically or at least selectively with the analyte. Specific
reagents which are rare, react only with a single chemical species. Selective reagents which are more common,
react with a limited number of species. In addition to specificity and selectivity, the ideal precipitating reagent
would react with analyte to give a product that is;

a) Easily filtered and washed free of contaminants;

b) Of sufficiently low solubility that no significant loss of the analyte occurs during filtration and washing;
c) Unreactive with constituents of atmosphere;
d) Of known chemical composition after it is dried or, if necessary, ignited.

The Gravimetric Estimation of Nickel:

The nickel is precipitated as nickel dimethyl glyoxime by adding alcoholic solution of dimethyl glyoxime
C4H6(NOH)2 and then adding a slight excess of aqueous ammonia solution.

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When the pH is buffered in the range of 5 to 9, the formation of the red chelate occurs quantitatively in a solution.
The chelation reaction occurs due to donation of the electron pairs on the four nitrogen atoms, not by electrons on
the oxygen atoms. The reaction is performed in a solution buffered by either an ammonia or citrate buffer to
prevent the pH of the solution from falling below 5. If the pH does become too low the equilibrium of the above
reaction favors the formation of the nickel (II) ion, causing the dissolution of Ni (DMG) 2 back into the mother
liquor.

A slight excess of the reagent has no action on the precipitate, but a large excess should be avoided because of the
possible precipitation of the reagent itself. The precipitate is soluble in the free mineral acids. It is therefore crucial
to avoid the addition of too large and excess of the reagent because it may crystallize out with the chelate. It is also
important to know that the complex itself is slightly soluble to some extent in alcoholic solutions. By adding small
amount of chelating agents will minimize the errors from these sources. The amount of the reagent added is also
governed by the presence of other metals such as cobalt, which form soluble complexes with the reagent. If a high
quantity of these ions is present, a greater amount of DMG must be added. The nickel dimethylglyoximate is a very
bulky precipitate. Therefore, the sample weight used in the analysis must be carefully controlled to allow more
convenient handling of the precipitate during the transfer to the filtering crucible. The compactness of the
precipitate is improved by adjusting the pH to 3 or 4, followed by the addition of ammonia solution.

A slow increase in the concentration of ammonia in the solution causes a slight increase in the pH gradually and
results in the precipitation of the complex. The result is the formation of a denser precipitate. Once the filtrate has
been collected and dried, the nickel content of the solution is calculated stoichiometrically from the weight of the
precipitate.

The structure of DMG & the complex with nickel ions is given below;

DMG

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Procedure:

 The given nickel solution is made up to 100mL in a standard measuring flask.

 20 mL of solution is pipetted into a 250 mL beaker.

 About 5 mL 1:1 HCl is added and diluted to 150 mL.

 The solution is heated to 70-80°C. 25 mL of 1% dimethyl glyoxime in alcohol is added, immediately

followed by dilute ammonia solution drop wise until it strongly smells of ammonia.

 The solution containing the precipitate is heated in a water bath for 30 minutes.

 The precipitate is allowed to stand for an hour.

 Filter the solution through a previously weighed sintered glass crucible.

 The precipitate is washed with cold water to free chloride.

 The crucible is placed in a dry 100 mL beaker and heated in the air oven at 110-120°C for 1 hour.

 It is cooled in a desiccator and weighed. Repeat drying until constant weight is obtained.

Calculation:

Mass of sintered glass crucible = a g.

  Mass of sintered glass crucible + nickel complex = b g.

  Mass of dimethyl glyoxime nickel complex = (b-a) g.

  288.69 of nickel complex contain 58.69 g of nickel.

Mass of nickel in (b-a) g of complex  .

Therefore, Mass of nickel in the whole of the given solution 

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Experiment 7
REDOX TITRATION WITH IODINE (Analysis of H2O2 and Vitamin C)

Introduction

The shelf life of pharmaceutical products must always be tested. Hydrogen peroxide used in disinfectants is slowly
reduced to water; it also photodegrades. Vitamin C is inherently unstable, being slowly oxidized by air. The ability
to accurately and reliably determine the concentration of the active ingredients in various formulations is an
important one. Both H2O2 and Vitamin C analyses through potentiometric titration of aqueous iodine with sodium
thiosulfate using a platinum ring indicator electrode following the progress of the titration curve. Alternatively they
are analysed by iodometric titration.

Background: Iodine is a moderately weak oxidizing agent; it is reduced to form the iodide anion, as follows:

I2(aq) + 2e- 2I-.(aq) E° = 0.621 V

The above redox reaction is completely reversible, and so the iodide anion is a moderately weak reducing agent
that will react with oxidizing analytes to produce iodine. In iodometric titrations, the analyte (an oxidizing agent)
reacts with an unmeasured excess of iodide to produce iodine: A red + 2I- Aox + I2

The iodine produced in this reaction is stoichiometrically related to the amount of analyte originally present in the
sample. The iodine may then be titrated to determine the analyte concentration in the sample. The nearly
universal titrant for iodine is thiosulfate; they react quantitatively as follows:

2S2O3 -2 + I2 S4O6 -2 + 2I-

So we see that reactions involving iodine can be used for the analysis of moderately strong reducing agents (by
reacting with I2 in iodimetry) or moderately strong oxidizing agents (through reaction with excess iodide in
iodometry).

Solid iodine is not very soluble in water. As a result, iodine solutions are usually prepared by dissolving the solid in
the presence of potassium iodide. Iodine reacts with iodide to produce the triiodide ion:

I2(aq) + I- l3 - K = 710

Due to this reaction, solid iodine is soluble in solutions of iodide salts. However, most of the dissolved iodine is
actually present as triiodide, not as iodine. If a solution prepared by dissolving primary standard potassium iodate
is mixed with a slight excess of iodide, and then acidified with sulfuric acid, a standard iodine solution is prepared
by this .reverse disproportionation reaction.

IO3- + 5I-. + 6H+ 3I2 + 3H2O

The solution that results can be used to standardize sodium thiosulfate titrant, or to generate a known quantity of
iodine reagent in situ.

Analysis of Ascorbic Acid by Iodimetry

Ascorbic acid (vitamin C) is sometimes called an .anti-oxidant. (i.e. a reducing agent) by pharmacists and food
nutritionists. Iodine rapidly oxidizes ascorbic acid, C6H8O6, to produce dehydroascorbic acid, C6H6O6:

C6H8O6 + I2 C6H6O6 + 2I-. + 2H+

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Ascorbic acid is readily water-soluble, and direct iodimetric titration is a standard method for the analysis of
vitamin C in a variety of citrus fruits and in vitamin tablets. In our experiment, we actually perform an iodimetric
back-titration: we generate a measured excess of iodine in the sample solution and then titrate the unreacted
iodine with sodium thiosulfate. This back titration is still an iodimetric titration, since it is based on the reaction of
analyte with aqueous iodine.

Structures of Ascorbic acid: ascorbic acid, C6H8O6, to produce dehydroascorbic acid, C6H6O6

Analysis of Hydrogen Peroxide by Iodometry : Solutions of hydrogen peroxide are widely sold as disinfectants.
Since hydrogen peroxide is an oxidizing agent, it will react with iodide in a redox reaction:

H2O2 + 2H+ + 2I-. I2 + 2H2O

The reaction is not instantaneous, but it is fast enough to form the basis of a practical analysis by iodometric
titration. The sample is added to an acidic solution containing an unmeasured excess of iodide. After the reaction is
complete, the iodine produced by the above reaction is titrated with sodium thiosulfate.

Materials Required: 1M H2SO4,.solid potassium iodide, a standard solution (approx 0.01M) of potassium bi-iodate,
KH(IO3)2, 0.1M sodium thiosulfate, Na2S2O3, as titrant

Procedure

Standardization of Sodium Thiosulfate: The standard potassium bi-iodate solution is acidified in the presence of
excess iodide to produce a standard solution of iodine.

1. Add approximately 30 mL of deioinized water to three clean sample cups

2. Add approximately 5 mL of 1M H2SO4 to each cup.

3. Add approximately 0.4g of KI into one sample cup and swirl until dissolved.

4. Accurately pipette 5.00 mL of standard bi-iodate solution into the sample cup. The formation of iodine
should be apparent. Titrate immediately with thiosulfate.

5. Repeat steps 3-4 for at least two other sample cups.

Analysis of Hydrogen Peroxide:

An unmeasured excess of iodide is mixed with the sample under acidic conditions. The iodine produced by the
oxidation of iodide by the analyte is titrated with thiosulfate. The reaction between iodide and analyte needs
about 15 minutes to be complete.

1. Add approximately 40 mL of 1M H2SO4 to at least 3 samples.

2. Add approximately 0.6g of KI to each sample cup and swirl to dissolve.

3. Accurately pipette 0.40 mL of sample solution into each cup, swirl well, and cover with a watch glass.
Make sure the reactants are well mixed, because they can form two distinct layers, slowing the reaction.
Allow 15 minutes and then titrate each solution with thiosulfate to note end point.

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Analysis of Ascorbic Acid in Vitamin C Tablets:

A measured excess of iodine is generated in situ by the acidification of standard bi-iodate in the presence of excess
iodide. The unreacted excess is determined by thiosulfate titration.

In this procedure, you will grind vitamin C tablets before weighing and dissolving the sample. This action greatly
hastens the oxidation of ascorbic acid. In addition, the back-titration should be performed as soon as possible due
to the instability of iodine in acidic solutions. Thus, each analysis should be performed in its entirety before
proceeding to the next sample.

1. Add approximately 30 mL of deioinized water and 5 mL of 1M H 2SO4 to a sample.

2. Add approximately 0.7g of KI to the cup and swirl to dissolve.

3. Randomly sample a single vitamin C tablet, weigh it accurately in a weighing boat, and grind it using the
mortar and pestle. Once ground, use weighing paper to rapidly but accurately weigh 0.09-0.10g of the
powder. Pour the sample into the solution and swirl. Note that the binder in the tablet will not dissolve in
water. Be sure that all of the powder ends up in the solution.
4. Accurately pipette 10.00 mL of standard bi-iodate into the solution, swirl briefly and titrate immediately
with thiosulfate.

5. Clean the mortar and pestle and the spatula thoroughly and repeat steps 1-4 for at least two more
samples, using two new tablets.

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Experiment 8
Determining Solubility Product and effect of common ion
When a chemical species is classified as “insoluble”, this does not mean that none of the compound dissolves in
the given solvent or solution system. In reality, a measurable level of material does go into solution, but it is
sometimes considered negligible relative to the total amount of the chemical. Perhaps a better name for such salts
is “sparingly soluble.” The dissolving of a solid monovalent-monovalent salt, represented as MX, in an appropriate
solvent is represented by the general equation:

MX(s) M+ (aq) + X- (aq).

where the subscripts “(s)” and “(aq)” represent the solid and aqueous solution physical states, respectively. For a
set of given conditions, the precipitate has a definite solubility (or maximum amount that will dissolve) expressed
in units of grams/liter or moles/liter.

An equilibrium constant expression can be written for the above reaction, as: Keq = [M+] [X-]/[MX]. The
“concentration” of any solid material, such as MX, is proportional to its density and is constant. Thus, the term
[MX] is usually combined into the Keq value, giving: Keq [MX] = [M+] [X-] = Ksp, where “Ksp is called the “solubility
product” constant. Note: Square brackets indicate saturation or equilibrium molarities.

The ideal or thermodynamic solubility product expression is written in terms of the “activities” or “effective
concentrations” of all species, rather than in actual mass/volume or mole/volume concentrations. In solutions of
low concentrations, the activity coefficients are near unity, and the simplification of terms is appropriate for
equilibrium systems for most reactions.

For a given chemical species and solvent system, the main factor which affects the value of K sp is the temperature.
Most often, an increase in the temperature causes an increase in the solubility and value. However, there are some
exceptions, as in the case of a salt which dissolves with a loss of energy (i.e., an exothermic dissolution). For a given
temperature and set of experimental conditions, there are often kinetic limitations as to how fast crystalline solids
form. Thus, sufficient time should be allowed for the system to come to a true state of equilibrium.

The amount of a slightly soluble salt that dissolves does not depend on the amount of the solid in equilibrium with
the solution, so long as there is enough to saturate the solution. A non-symmetric salt, such as lead iodide or PbI2,
would have a reaction as:

PbI2(s) Pb2+ (aq) + 2I-(aq), with Ksp = [Pb2+][I-]2

As with any equilibrium constant, the K sp value holds under all conditions at the specified temperature. If there is
an excess of one ion over the other, the concentration of the second is suppressed (due to the common-ion effect),
and the solubility of the precipitate is decreased. Because the solubility product value always holds constant,
precipitation will not occur unless the product of [M +] and [X-] exceeds the value of Ksp. If the product of the ion
concentrations is just equal to Ksp, all the M+ and X- would remain in the solution (i.e., no precipitate or solid forms).

In this experiment, the relative solubility (and an approximate value of the K sp) of lead iodide will be determined by
direct observation. The procedure calls for the mixing of two standard solutions (one of a soluble lead salt, and a
second of a soluble iodide salt) in different proportions and allowing time for the resulting mixture to come to
equilibrium. In some of the mixtures, the solubility product constant for lead iodide will be exceeded, and
precipitation of PbI2 crystals will occur. In other mixtures, the final concentrations of lead and iodide ions will be
such that precipitation does not occur.

A simple product function “Q,” called the Ion Product will be defined so that:

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Q = {lead ion} x {iodide ion}2 = {Pb2+}{I-}2,

where curly brackets indicate prepared molarities before precipitation occurs (if it does occur). An experimental
value of “Q” will be calculated for each mixture, based on the total amount of lead ion and iodide ion added in
each solution. Clearly, the formation of solid crystals of PbI 2 will occur when the value of “Q” exceeds the solubility
product value of lead iodide (i.e., when Q > Ksp).

Similarly, precipitation will not occur when the value of K sp > Q. Thus, in this experimental procedure, if some
sample mixtures form PbI2 crystals and other solutions do not, the value of the solubility product constant lies
between Q values with precipitates and Q values without precipitates. The most precise value for K sp is given as:

Qmin with PbI2 > Ksp > Qmax without PbI2

The value for Ksp is determined with an uncertainty level equal to the difference of the two Q values. In order for a
more precise determination of K sp, the solution mixtures are designed such that the various Q values are rather
close together.

Caution: lead salts are toxic, so be careful handling the solutions, and make sure to wash your hands after you
complete the experimental work.

Preparation of lead iodide mixtures: Equimolar standard solutions of lead and iodide compounds are provided on
the reagent shelf. A 1.00 x 10─2 M Pb2+ solution is prepared by dissolving 3.312 grams of Pb (NO3)2 in 1.00 liter of
deionized water, while a solution of 1.00 x 10-2 M I- is prepared by dissolving 1.660 grams of KI in 1.00 liter of
deionized water.

Each student or group of students should assemble a set of nine clean, dry test-tubes. A number of burette stations
will be set up in the laboratory. Each station will consist of three delivery burets, one burette for each of the stock
solutions and a third for deionized water. Following the specifications listed in Table I, prepare a mixture series of
the lead and iodide stock solutions. To avoid local, temporary supersaturation, always mix the water first.

Note that the total volume of each mixture solution is 10.0 mL. Be sure that the solutions are mixed thoroughly,
and do not get the test tubes out of order. An easy check is to observe that the Q values progressively decrease
from #1 to #9 samples.

After preparing the solutions, allow each mixture to set for at least 30 minutes before checking for precipitation.
During this equilibration period, calculate the theoretical or maximum concentration of each ion in the mixture,
using the equation:

[Ion]mixture = [Ion]standard x mL's ion in solution mL's total mixture

Use these concentration values to calculate the “Q” value for each mixture, and record the data in Table I.
Colorless clarity (clearness) of solution indicates no precipitate. Golden cloudiness (lack of clarity) indicates
formation of a precipitate.

After the equilibration period is completed record under the “PbI2?” column whether or not a precipitate has
formed in each test tube. If all goes well, the mixtures of higher Q values will contain shiny, golden crystals of lead
iodide, while the tubes of lower Q values will have no solid.

Table I

# mL Pb2+ mL I- mL H20 [Pb2+] [I-] Q PbI2

1 4.00 5.00 1.00

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2 3.00 5.00 2.00

3 2.00 5.00 3.00
4 1.00 5.00 4.00
5 2.50 2.00 5.50
6 1.20 2.50 6.30
7 1.25 2.00 6.75
8 2.50 1.00 6.50
9 1.00 1.00 8.00

Data treatment and report: A single-page report will be submitted for this experiment. Under a full heading,
recreate the completed data entries of Table I. Be sure to indicate which test tubes contain precipitates of lead
iodide. Identify the smallest value mixture containing a precipitate and the largest Q value test tube which has no
solid crystals. The Ksp for lead iodide should lie within these two Q values, so report the experimental value of Ksp
within this range, as:

Qmin with PbI2 > Ksp > Qmax without PbI2

Actual Ksp for lead iodide is ________________

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