Lab Manuel for Analytical Chemistry and Environmental Science

1. Experiment Name: Spectrophotometric determination of Cr (VI) from supplied sample

solution
There is increasing awareness of the presence of hexavalent chromium in the environment and the
effects on human health. Chromium is a grey, hard metal and is one of the most widely distributed
heavy metal in the Earth crust. It is found throughout the environment in three major oxidation
states which are Cr(0), Cr(III) and Cr(VI). The most stable form is Cr(III), occurs naturally, while
Cr(VI) and Cr(0) are generally introduced into the environment by various industrial processes and
anthropogenic activities.
How much and what types of environment requires for the conversion of trivalent chromium to
hexavalent form depend on range of factors, including the quality of used reagents, temperature
control, and the circumstances within the tanning process. What is more, Hexavalent chromium in
leather samples may come about due to mistakes in local production or can emerge later in the
manufactured leather. Chinese researchers have shown that leather containing high proportions of
unsaturated fatty acids results in the formation of chromium (VI). Irregularities in the tanning
process and certain additives, combined with the presence of chromium (III) compounds, can result
in the formation of chromium (VI) compounds. Meanwhile, scientists are targeting chrome tanning
agents, which are provided with additives that counteract the emergence of chromium (VI).
However, these materials are expensive, and the consumer should not assume that all imported
goods are safe. In such cases, the customer can only rely on responsible manufacturers who undergo
voluntary inspections of their batches and can prove it by having their goods tested by reputable
independent testing institutes. A study in 2011 found that the use of heat and adhesives in shoe
production promotes the formation of chromium VI. Leathers with a higher proportion of tannin
chromium (III) are more prone to the formation of chromium (VI) than leather with lower chromium
(III) content. The most widely used quantitative method for hexavalent chromium is a
diphenycarbazide absorption method using a UV-Visible spectrophotometer.

For any given compound, the amount of light absorbed depends upon the concentration, the path
length, the wavelength and the solvent. Absorbance is related to the concentration according to
the Beer-Lambert law: A= εcl, where ε is the extinction coefficient (M-1 cm-1), l is the solution path
length (cm) and c is the concentration (molL-1). Not all substances obey the linear Beer-Lambert
law over all concentration ranges. Therefore, you will construct a calibration curve that will provide
the relationship between concentration and absorbance under the conditions used for the analysis.
Procedure: Measure 0.0286 g K2Cr2O7 solid using electric balance and transfer into 100 mL
volumetric flask which makes 100 ppm solution of Cr(VI). For the preparation of 1 ppm, 2.5 ppm
5 ppm, 7.5 ppm, and 10 ppm Cr (VI) standard solutions in five different 100 mL volumetric flask
take 1.0, 2.5, 5.0, 7.5, and 10.0 mL from 100 ppm K2Cr2O7 solution using burette or graduated
pipette. Then add 2.0 mL of 1, 5-diphenylcarbazide solution (1 g 1,5-diphenylcarbazide was
dissolved in 100 mL acetone with 1 drop acetic acid) and 2.0 mL ortho-phosphoric acid solution
(5:3 ratio of ortho-phosphoric acid and distilled water) into each of the volumetric flask.
Afterwards, these volumetric flasks are made up to the mark by adding distilled water, shake the
solutions for well mixing, and kept them 5-7 minutes to be stabilized. After that, take the absorbance
of each solution by UV-visible spectrophotometer at absorption maximum 540 nm after baseline
correction. After recording the absorbance, in the graph paper make an absorbance versus
concentration calibration curve from the experimental data. Take the concentration of unknown
solution and from this absorbance of unknown solution, calculate the concentration of Cr(VI)
solution.
2. Experiment Name: Spectrophotometric determination of Fe (II) in supplied sample solution
UV-visible absorption spectrophotometry provides a convenient method of determination of
concentration of any substance which can be treated to form a coloured solution in which the colour
intensity is proportional to the concentration of the substance. This experiment is based on the
determination involving the formation of a complex species that absorbs in the visible region.
In the determination of Fe (II) in aqueous solutions, a tricyclic nitrogen heterocyclic compound, 1,
10-phenanthroline (C12H8N2, ortho-phenanthroline) is used as the ligand that reacts with metals
such as iron, nickel, ruthenium, and silver to form strongly coloured complexes. With ferrous ions
(Fe2+), it reacts in a ratio of 1:3 to form an orange red coloured complex [(C12H8N2)3Fe]2+ in aqueous
medium.
The molar absorption coefficient (ε) of the ferrous complex, [(C12H8N2)3Fe]2+ so obtained is 11,100
dm3 mol-1cm-1 at the wavelength of maximum absorbance, λmax = 508 nm. The large value is
indicative of strong absorption by the complex and forms the basis of the quantitative determination
of iron (II). The colour intensity is not affected by change of pH over the range 2-9 and is also stable
for a long time. However, a pH of about 4.5 is ordinarily recommended to prevent precipitation of
iron salts. Further, the cations like Ag+, Bi3+, Cu2+, Ni2+,Co2+ and anions such as perchlorate,
cyanide, molybedate and tungstate interfere significantly in this determination, therefore, these
must be absent in the analyte solution. As we are attempting to determine the concentration of iron
(II) ions in the analyte sample, it must be free from any iron (III) ions which may be present due to
the partial oxidation of the ferrous ions. This is achieved by adding a reducing agent before the
coloured complex is formed. Ferric ion (Fe3+) is reduced to ferrous state (Fe2+) by hydroxylamine
before complexation as per the following equation.
4Fe3+ + 2NH2OH = Fe2+ + N2O + 4H+ + H2O

Preparation of Reagents Solution

i) Standard ferrous solution (10 ppm (10 mg/dm3)); prepared as follows. Weigh 0.0702 g of
analytical grade ferrous ammonium sulphate hexahydrate [Fe(NH4)2(SO4)2.6H2O]. Quantitatively
transfer the weighed sample to a volumetric flask of 1 dm3 capacity and add sufficient distilled
water to dissolve it. Add 2.5 mL of concentrated sulphuric acid and make up the solution to the
mark.
ii) 1, 10-phenanthroline; prepared by dissolving 0.1 g of the reagent in 100 mL of distilled water.
The reagent can be stored in a bottle.
iii) Hydroxylamine hydrochloride; prepared by dissolving 10 g of hydroxylamine hydrochloride in
100 mL of distilled water.
iv) Sodium acetate (0.1 M); prepared by dissolving 10 g of sodium acetate in 100 mL of water.
Procedure: Take five 100.0 mL volumetric flask and pipette out 1.0, 5.0, 10.0, 15.0 and 25.0 mL
of the 10 ppm standard supplied iron solution. Take 100.0 mL of distilled water in another flask.
This is the blank solution. Add 1 mL hydroxylamine hydrochloride and 8 mL sodium acetate in all
of them except the blank. This will reduce all iron in the unknown sample to iron (II). Then add 5.0
mL 1,10-phenanthroline monohydrate. The solutions should turn red. Dilute upto the mark. After
that, take the absorbance of each solution by UV-visible spectrophotometer at absorption maximum
508 nm after baseline correction. Make a plot of absorbance at Y-axis vs. concentration of the
standard solutions at X-axis to get the calibration curve. (The linear region of the curve obeys Beer-
Lambert’s law and is used for the estimation of unknown samples.) Find the absorbance of the
unknown solution and use it to find the concentration of iron in the sample.
3.Name of the experiment: Determination of chloride content in supplied solution by Volhard’s
method.
Chloride, in the form of chloride (Cl–) ion, is one of the major inorganic anions in water and
wastewater. The salty taste produced by chloride concentrations is variable and dependent on the
chemical composition of water. Some waters containing 250 mg Cl–/L may have a detectable salty
taste if the cation is sodium. On the other hand, the typical salty taste may be absent in waters
containing as much as 1000 mg/L when the predominant cations are calcium and magnesium. The
chloride concentration is higher in wastewater than in raw water because sodium chloride (NaCl)
is a common article of diet and passes unchanged through the digestive system. Along the sea coast,
chloride may be present in high concentrations because of leakage of salt water into the sewerage
system. It also may be increased by industrial processes. High chloride content may harm metallic
pipes and structures, as well as growing plants.
In tanning process, to preserve the raw hides and skins at the primary stage, we apply huge amount
of salts in it, before the tanning operation we wash these raw hides and soak in water. That means
salt is dissolved in water. So it is necessary to maintain the content of chlorides. In Volhard method
a back titration with CNS- is done to determine the concentration of the chloride ions in a solution.
Before the titration an excess volume of AgNO3 solution is added to the solution containing Cl
forming a precipitate of AgCl.
Cl-(aq ) + AgNO3(aq) = AgCl ; Indicator Fe3+ is added after that solution is titrated with KCN
solution . Ag +(aq ) + AgNO3(aq) = AgCl
Ag +( aq ) + SCN–(aq ) = AgSCN (s )
Fe3+( aq ) + SCN–(aq ) = [ FeSCN ] 2+(aq)
Standardization of thiocyanate solution: Pipette out 10.0 mL of the standard silver nitrate (you
have prepared) solution into a conical flask. Add 5 mL of 6 M HNO3 and 1 mL of ferric alum
indicator. Now, titrate the solution with the supplied thioyanate solution from a burette until a faint
brown colour appears which no longer disappears upon shaking. It is essential to shake vigorously
during titration. Calculate the strength of supplied thiocyanate solution.
Procedure: Take 10.0 mL soaking liquor in a conical flask. Then add 3-4 mL of concentrate HNO3
to it using measuring cylinder and add 25.0 mL of 0.1 AgNO3 solutions to it by pipette. Add the
1mL of Nitrobenzene and 1 mL of ferric alum (indicator) to it. Then heat it upto 1.0 hours and then
cooled down. Titrate the residual AgNO3 solution with 0.1 M of SCN- and the end point is indicated
by persistent brown colour.

4.Experiment Name: Determination of total dissolve solid (TDS) in supplied sample

TDS refers to materials that we completely dissolve in water. It is defined as reduce upon
evaporation of no filterable sample on a filter paper. A well sample is filtered through a standard
glass filter and the filtrate is evaporated to a weighted and dried to constant weight at 179-181
degree Celsius. The increase in dish weight represents the total dissolve solids.

Procedure:
1. To measure dissolve solids take a clean dish which has been washed and dried in air oven in180
degree Celsius for an hour.
2. Then the empty evaporated dish is weighted.
3. The sample is mixed well and poured into a funnel with filter paper.
4. Then placed it in the hot air oven and care should be taken to prevent splattering of sample.
5. Then the sample is dried to get constant glass
6. The containing is cooled in a desiccator.
7. The dish is weighted as soon as it has cooled to avoid absorption of moisture due to hydroscopes
nature.
8. The weight with residue is noted.

Calculation: Weight of the clean evaporating dish W1=

Weight of the dish and residue W2=
Weight of the residue W= W2-W1

5.Experiment Name: Determination of chromium oxide from supplied leather sample

To determine chromium oxide in leather sample iodometry is used in which process an oxidizing
agent reacts with iodine salt to free iodine which is titrated with standard thiosulphate solution to
determine the quantity of freed iodine. In this process concentrated nitric acid is used to breakdown the
bond between collagen and chromium ion followed by perchloric-sulphuric acid mixture is added for
oxidation of chromium from trivalent to hexavalent form. Produced hexavalent chromium is reduced by
an acid solution of potassium iodide and iodine is liberated. The liberated iodine (I2 or I3-) reacts
with thiosulphate solution rapidly and stoichiometrically. Thus the equation stands at :
2𝐶𝑟 +6 + 6𝐼 − → 2𝐶𝑟 +3 + 3𝐼2 ----------- (1)
 3𝐼2 + 6𝑆2 𝑂3−2 → 3𝑆4 𝑂6_2 + 6𝐼 − ----------- (2)
∴ 6 𝑚𝑜𝑙 𝑆2 𝑂3−2 ≡ 2𝐶𝑟 +6
1
∴ 1 𝑚𝑜𝑙 𝑆2 𝑂3−3 ≡ 𝐶𝑟 +6
3
1
∴ 1000𝑚𝑙 1𝑀 𝑆2 𝑂3−2 ≡ × 53.0𝑔 𝐶𝑟 +6
3
1 53
∴ 1𝑚𝑙 0.1𝑀 𝑆2 𝑂3−2 ≡ × 1000 × 0.1𝑔 𝐶𝑟 +6
3

= 0.00253g 𝐶𝑟2 𝑂3
Preparation of sample: For dry sample: Reduce the leather to small pieces by stamping or
cutting. Sparing the mill and yielding a more uniform ground sample.
For wet sample: Dry the sample at a temperature not exceeding 50 º C. Condition in an atmosphere
of temperature 20 ± 2º C and relative humidity 65 ± 2 % for 24 hours before grinding. Grind the
whole sample and mix the leather powder and store in a clean, dry, airtight container.
1. Accurately weigh a mass of ground leather into a conical flask in the following way:
Type of leather Mass of leather
Full chrome 1.0 g
Good semi-chrome 2.0 g
Leather of low chromium content 2.0-5.0 g
2. Add 20 mL conc. HNO3 to it. Mixture is allowed to stand for 10 minutes.
3. Then add 20 mL of H2SO4 and HClO4 mixture and a few anti-bumping granules. Heat the
conical flask until the mixture begins to turn orange.
4. Then allow the mixture to cool for 15 minutes and dilute to 100 mL using distilled water.
5. Boiled the mixture for 10 minutes to eliminate chlorine and then allow it to cool.
6. Add 20 mL KI solution to the cooled mixture and left in the dark for 10 minutes.
7. Finally, titrate the solution with 0.1N Na2S2O3 using starch indicator. The titration will be
continued until the color turns into bottle green.
Data Record during Titration:

SL. Initial Reading Final Reading in Volume in mL. of Average

No. in mL. mL. Titrant. Volume in mL.
01 0 X X A
02 X Y Y-X
03 Y Z Z-X

6. Name of the experiment: Determination of chromium oxide from supplied leather liquor sample
The chromium liquor is one of the most used liquor in leather industry, especially in chrome tanned
leather. Chromium sulphate is the chrome liquor which is used profoundly in the tanning process.
Cr+3 is used around the world for tanning and is not harmful for environment . But Cr+6 is harmful
for health and sometimes exists in leather liquor. It can come about due to mistakes in production
or can emerge later in manufacturing process.
To determine chromium oxide in liquor iodometry is used in which process an oxidizing agent
reacts with iodine salt to free iodine which is titrated with standard thiosulphate solution to
determine the quantity of freed iodine. Thus the equation stands at :
2𝐶𝑟 +6 + 6𝐼 − → 2𝐶𝑟 +3 + 3𝐼2 ----------- (1)

3𝐼2 + 6𝑆2 𝑂3−2 → 3𝑆4 𝑂6_2 + 6𝐼 − ----------- (2)

∴ 6 𝑚𝑜𝑙 𝑆2 𝑂3−2 ≡ 2𝐶𝑟 +6
1
∴ 1 𝑚𝑜𝑙 𝑆2 𝑂3−3 ≡ 𝐶𝑟 +6
3
1
∴ 1000𝑚𝑙 1𝑀 𝑆2 𝑂3−2 ≡ × 53.0𝑔 𝐶𝑟 +6
3
1 53
∴ 1𝑚𝑙 0.1𝑀 𝑆2 𝑂3−2 ≡ × 1000 × 0.1𝑔 𝐶𝑟 +6
3

= 0.00253g 𝐶𝑟2 𝑂3
To determine the chromium content from liquor by 0.1M 𝑁𝑎2 𝑆2 𝑂3 it has to be titrated to collect
the required volume data.
Procedure: Take 25.0 mL supplied chromic liquor in a conical flask. Add 20 mL conc. HNO3 to
it. Mixture is allowed to stand for 10 minutes. Then add 20 mL of H2SO4 and HClO4 mixture and a
few anti-bumping granules. Heat the conical flask until the mixture begins to turn orange. Then
allow the mixture to cool for 15 minutes and dilute to 100 mL using distilled water. Boiled the
mixture for 10 minutes to eliminate chlorine and then allow it to cool. Add 20 mL KI solution to
the cooled mixture and left in the dark for 10 minutes. Finally titrate the solution with 0.1N Na2S2O3
using starch indicator. The titration will be continued until the color turns into bottle green.

7.Experiment Name: Determination of dissolved oxygen (DO) in supplied water sample.

Dissolved Oxygen (DO) levels in natural and wastewaters are dependent on the physical,
chemical and biochemical activities prevailing in the water body. The analysis of DO is a key
test in water pollution control activities and waste treatment process control.Dissolved oxygen
(DO) levels in environmental water depend on the physiochemical and biochemical activities in
water body and it is an important useful in pollution and waste treatment process control. Two
methods are commonly used to determine DO concentration: (i) The Iodometric method which
is a titration-based method and depends on oxidizing property of DO and (ii) The membrane
electrode procedure, which works based on the rate of diffusion of molecular oxygen across a
membrane.
It is based on oxidation of potassium iodide. The liberated iodine is titrated against standard
hypo solution using starch as a final indicator. Since oxygen in water is in molecular state and
not capable to react with KI, an oxygen carrier manganese hydroxide is used to bring about the
reaction between KI and O2 .Manganous hydroxide is produced by the action of potassium
hydroxide and Manganese Sulphate.
Chemical reaction: 2KOH + MnSO4 → Mn(OH)2 + K 2 SO4
2Mn(OH)2 + O2 → 2MnO (OH)2
2MnO(OH)2 + 2H2SO4 +4I- → 2MnSO4 +4H2 O +2I2
2I2 + 4Na2 S2 O3 → 4NaI + 2Na2 S4 O6
Sodium tetrathionate
 Starch + I2 → Starch iodide complex
(Blue in color)
Thus, 4 mol Na2 S2 O3 = 1 mol O2

The Azide Modification with the Winkler Method for DO Determination

This modification is used because of presence of nitrite ions. This occurs in effluents from
wastewater treatment plants employing biological processes, in river water and in incubated
BOD samples. It does not oxidize Mn2+ but doses oxidize I- to I2 under acidic conditions.
When the reduced form of nitrite (N2O2) is oxidized by oxygen, it is converted to NO2
- again, establishing the cycle again that can result in erroneous results, far in excess of amounts
that would be expected.
2NO2- +2I- +4H+ = I2 + N2O2 + 2H2O………(a)
N2O2 + 0.5 O2 +H2O = 2NO2 - +2H+ ………..(b)
When interference from nitrites is present, it is impossible to obtain a permanent end point. As
soon as the blue color of the starch indicator has been discharged, the nitrites formed by the
reaction (b) reacts with more iodide ions to produce I2 and the blue color of the starch indicator
will return. The nitrite interference is easily overcome with use of sodium azide (NaN3), which is
incorporated in the alkali-KI reagent. When sulfuric acid is added, following reactions happen:
NaN3 + H+ = HN3 +Na+ …………. (c)
HN3 +NO2- + H+= N2 +N2O+H2O ………..(d)

Required Chemicals: MnSO4 , KOH, H2SO4, Na2 S2 O3 , Starch and NaN3

Manganous sulphate solution: Dissolve 100 g manganous sulphate (MnSO4 . 4H2O) in 200 mL
distilled water.
Alkaline‐iodide‐azide solution: Dissolve 100 g sodium hydroxide in 100 mL distilled
water. Allow to stand for some days, during which any carbonate present sinks to the
bottom. Siphon off all of the clear liquid, add 30 g potassium iodide and 2 g sodium azide and
make up to 200 mL with distilled water. Store in plastic container.
Sodium thiosulphate (0.0125 mol/L): Dilute 125 mL 0.1 mol dm‐3 sodium thiosulphate to 1 L
with distilled water.
Potassium iodide solution (0.025 mol/L) Dry analytical grade potassium iodate at 120o
C. Dissolve 5.35 g in distilled water and dilute to exactly 1 L. This solution is stable for long
periods if stored in a glass stopper bottle.
Potassium iodide solution (10% w/ v) Dissolve 10 g KI in 100 cm3 distilled water.
Procedure: Measure the volume of water to fill up-to the brim of the supplied bottle. This is the
volume of the bottle. Fill the bottle with sample water by 6.0 mL less than its total volume. Add 2.0
mL of MnSO4 solution and 2.0 mL of alkali-iodide-azide reagent which go well below the surface
of the liquid. Mix by inverting the bottle several times. Add 2.0 mL of conc. H2SO4. Allow the acid
to run-down the neck of the bottle. Stopper the bottle and mix the content completely. The bottle is
kept for 10 minutes and the contests of the bottle are transferred into a conical flask. Rinse the bottle
with distilled water. The-I2 released is titrated with 0.025 M sodium thiosulfate until pale straw
color appease; add 1-2 drops of starch indicator. Titrate to the transition phase from blue to
colorless.
Standardization of Sodium thiosulphate: Prepare 100.0 mL 0.01M K2Cr2O7 (mol. wt. of
K2Cr2O7 = 294.18) solution by weighing analytical grade potassium dichromate in a volumetric
flask.
Take about 50 mL distilled water in a conical flask, add 1-2 g pure sodium bicarbonate to it, and
shake the conical flask well to dissolve sodium bicarbonate. Add 10-12 mL 10% iodate free KI
solution and 3-4 mL of conc. HCl. Pipette out 10.0 mL prepared K2Cr2O7 solution into this conical
flask. Then, cover with a watch glass, and let stand 5 minutes in your closed locker. Afterwards,
titrate with your thiosulfate solution until the brown iodide color becomes noticeably lighter.
Immediately add 1-2 mL starch indicator and continue to titrate until the deep blue color abruptly
changes to the pale blue-grey color produced by the Cr3+ present in the solution. From the known
weights of dichromate and the titration volume calculate the molarity of your thiosulfate solution.

8. Experiment Name: Determination of Chemical Oxygen Demand (COD) in wastewater sample.

The Chemical Oxygen Demand (COD) is a parameter that measures the amount of substances
dissolved or suspended in a liquid sample that can be oxidized by a strong chemical oxidant. It is
used to measure the degree of contamination and it is expressed in milligrams per liter of oxygen
(mg O2/L). It is an applicable method in inland waters (rivers, lakes or aquifers), sewage, rainwater
or water from any other source that may contain an appreciable amount of organic matter. This test
is useful for monitoring and control of wastewater treatment plants. It does not apply, however, to
drinking water, as there is a low content of oxidizable matter and the accuracy of the method would
not be appropriate. In this case the method of oxidizability with potassium permanganate is used.

The most general COD determination uses potassium dichromate in excess in an acidic medium
with the aid of silver sulfate (Ag2SO4) as a catalyst, and mercuric sulfate (HgSO4) added to remove
interference of chlorides. Dichromate oxidizes organic and inorganic matter in the sample, and it is
reduced from Cr+ 6 to Cr+ 3. The test is performed at 150 °C under total reflux for 1-2 hours. After
digestion, excess potassium dichromate is titrated with the Mohr salt using ferroin as indicator. The
solution color changes from green to red. The amount of oxidizable matter measured of oxygen
equivalent of proportioned to the potassium-di-chromate consumed.

Preparation of reagent solutions

1. Sulfuric acid, containing silver sulfate: Dissolve 10g silver sulfate, Ag2SO4, in 35mL water
under additional of portions of 965 mL sulfuric acid;
2. Potassium dichromate solution (K2Cr2O7:0.02 mol/L), containing mercury sulfate:
Dissolve 80g mercury sulfate (HgSO4) in 800 mL water and 100 mL sulfuric acid (5.1). Add
5.884g of K2Cr2O7 (dried for 2 hrs at 1050C) and bring to volume (1000ml) with water;
3. Ammonium iron (II) sulfate solution [(NH4)2 Fe(SO4)2.6H2O] = 0.120 mol/L: Dissolve
47.1g Ammonium iron (II) sulfate hexahydrate Cryst. Pure (NH4)2Fe(SO4)2.6H2O in water.
Add 20ml sulfuric acid (5.1) and after cooling to room temperature, dilute with water to 1000
mL
4. Ferroin and diphenylamine indicator solution.

Standardization of FAS: Prepare 100.0 mL 0.02M K2Cr2O7 (molecular weight of

K2Cr2O7 = 294.18 g/mol) solution by weighing analytical grade potassium dichromate in a
volumetric flask. Pipette out 10.0 mL of the supplied FAS solution into a conical flask. Add
approximately 25 mL of distilled water, 15 mL of H3PO4 _ H2SO4 mixture and a few drops of
ferroin indicator solution in it. Titrate the solution slowly with standard (0.02 M) dichromate
solution while stirring continuously until the solution becomes green colour. Carry out three such
titrations.

Preparation of sample: Pipette out 1.0 mL effluent sample in a 100 mL volumetric flask and
make up to level with distilled water.
Note: In case of lime liquor, take 0.5 mL effluent sample and make up to level with distilled
water.
Description of the procedure
Pipette 20.0 mL blank into a reaction flask and pipette out 20.0 mL of the sample for analysis into
another reaction flask and add boiling aids (use tweezers) in every reaction flask. Add 10.0 mL of
the mercury-containing potassium dichromate solution and mix thoroughly. Add cautiously and
slowly, by swirling, 30 mL silver sulfate containing sulfuric acid. During addition, cool the reaction
flask under running water or in ice bath in order to avoid local overheating and thus excluding as
far as possible and losses of volatile substance. Attach the reflux condenser and bring the reaction
mixture to boiling within 10 minutes, continue with gentle boiling for another 110 minutes. Keep
the temperature of the reaction mixture at (1483) oC and cool the reaction mixture to a temperature
of about 60 oC. Rinse the reflux condenser with water and contents of the flask are thus diluted to
about 100 mL. Lower the temperature of the contents of the flask to room temperature and add 2
drops of the ferroin indicator solution to the contents of the flask. Titrate the unreacted potassium
dichromate with ammonium Fe(II) sulfate until the color changes from blue-green to red-brown.

Calculation: The chemical oxygen demand of sample is calculated according to the following
equation:
C×F
COD (mg/L) =-------------------- (VB-VE) × D
VP
C = Concentration of the ammonium iron (II) sulfate solution
in mol/L, (here C= 0.12 mol/L)
F = Equivalent factor (here: F = 8000 mg/mol)
VB = Volume of the ammonium Iron (II) sulfate solution
consumed in the blank determination in mL
VE = Volume of the ammonium iron (II) sulfate solution consumed
during sample titration in mL
VP = Volume of the sample, (here VP= 20.0 mL)
 D = Dilution factor (The times of dilution)

9.Experiment Name: Determination of SO42- in supplied sample by gravimetric method.

Basics: Gravimetric analysis is based on conversion of ions/elements/radicals into a stable and pure
compound through precipitation adding suitable precipitating agent. The chemical that is added to
cause the precipitation is called the precipitant or precipitating agent. The solid precipitate can be
separated from the liquid components using filtration, and the mass of the solid can be used along
with the balanced chemical equation to calculate the amount or concentration of the desired
compounds in solution. In precipitation gravimetric method a number of steps involved like
reparation of the solution, precipitation, digestion of the precipitate, washing and filtering the
precipitate, drying and ignition, weighing and calculation.
Gravimetric method gives the most accurate results and is the recommended procedure for sulphate
concentrations above 10 mg/mL. The sulphate ions in the sample are precipitated by the addition of barium
chloride solution to water sample acidified with hydrochloric acid and kept near the boiling point.
SO42-+Ba2+=BaSO4
In highly alkaline water maintained near the boiling temperature, BaCO3 may get precipitated and
eliminate this, the sample is acidified. To precipitate sulphate ions as completely as possible, excess
barium chloride is used. The precipitate of BaSO4 is highly insoluble and hence there is considerable
tendency for most of the precipitate to form in colloidal condition which cannot be removed by
ordinary filtration procedures. To facilitate the conversion of colloidal form to crystalline form, the
samples at temperatures near the boiling point for a few hours are digested.
Procedure: Take 10.0 mL of supplied sulfate solution in a beaker. Add 0.5 mL conc. HCl and
dilute the solution to about 200 mL with distilled water. Heat the solution for boiling and add drop-
wise about 5% Barium chloride solution. Stir the solution constantly during addition. Allow the
precipitate to settle and test for complete precipitation (To do this add a drop of BaC1 2 solution
carefully down the side of the precipitate and note whether any turbidity appears). If the
precipitation is incomplete add a further portion of barium chloride solution (about 4 mL) with
constant stirring. Now keep the covered solution hot (a low flame heating or steam bath heating)
but not boiling, for an hour. By this time, dry a filter paper at 105 oC and take its weight using
analytical balance. Filter the solution through this filter paper (Whatman 40). Dry the BaSO4
precipitate with filter for about 40 minutes and weigh again accurately. Repeat the process of drying
and weighing until two nearly constant weights. Calculate the amount of sulfate in g/L unit

10. Experiment Name: Determination of Moisture and Ash content of leather sample
Moisture Content: About 1 g of finely powdered air-dried leather sample is weighed in a
crucible. The crucible is placed inside an electric hot air-oven, maintained at 105 to 110 0C for
one hour. The crucible is allowed to remain in oven for 1 hour and then taken out, cooled in
desiccators and weighed. Loss in weight is reported as moisture.

Ash: The residual leather sample taken in a crucible and then heated without lid in a muffle
furnace at 700 + 50 oC for ½ hour. The crucible is then taken out, cooled first in air, then in
desiccators and weighed. Hearing, cooling and weighing are repeated, till a constant weight is
obtained. The residue is reported as ash on percentage-basis.

Complexometric Titration
The formation of stable metal-complex ions by the titration of a metal ion solution with a suitable
standard solution of a complexing agent, or ligand, has gained increasing importance as a method
of volumetric analysis. The vast majority of complexation titrations are carried out using a
multidentate ligand, ethylenediaminetetraacetic acid, H4EDTA, as the complexone. A water-soluble
salt, Na2EDTA.2H2O (Na2H2Y) is preferred as the reagent for practical purposes. Na2EDTA.2H2O
ionizes in water to give 2Na+ and a strong chelating ion, H2Y2- (Y = EDTA4-) having six potential
groups – four carboxyl groups and two amine groups for bonding a metal ion. In fact, H2Y2- ion
forms very stable, one to one, six coordinated complexes with a wide range of metal ions.

Most metal-EDTA titrations are performed in neutral or alkaline

solutions. The prominent EDTA species in neutral or alkaline
media are the HY3- and Y4- ions and thus the net reactions
become,
(n-4)+
n+ 3- (n-4)+ +
M + HY ⇌ MY + H
Mn+ + Y4- ⇌ MY(n-4)+

depending on the exact solution pH.

The formation of hydrogen ions as a reaction product would cause the pH of the solution to decrease
and the desired titration reaction to cease or even reverse itself, unless the solution is buffered
against the pH change. Therefore, a buffered system is employed to keep the pH at the desired value
for the particular application. This buffer also serves as the auxiliary complexing agent in order to
prevent the undesired precipitation of metal hydroxides.
A very large number of metal ion indicators are employed for complexometric titrations. Two most
widely used metal ion indicators are,

Na+ -O
O- Na+

O-Na+

Eriochrome Black T Murexide Fast Sulphon Black F

Such indicators form complexes with specific metal ions at the desired pH; these differ in colour
from the free indicator and, in consequence, a sudden colour change occurs at the equivalence point.
The use of the metal ion indicator in an EDTA titration may be written as
From the above discussion, it reveals that complexation titration with EDTA fulfill the same
requirements as all other volumetric methods of analysis. It is rapid, proceeds according to well-
defined stoichiometry, and possesses the desired characteristics for the application of the various
end point detection systems.

11. Experiment No. : Determination of hardness of the supplied water sample.

Water samples that do not readily produce lather with soap are called hard water. Principle hardness
causing cations are Ca2+ and Mg2+. The portion of the total hardness that can be largely removed
by boiling is known as temporary hardness. Boiling converts the bicarbonates present into insoluble
carbonates and hydroxides, which can be removed by filtration.
Δ
Ca(HCO3)2 → CaCO3(s)↓ + H2O + CO2
Δ
Mg(HCO3)2 → Mg(OH)2(s)↓ + CO2
Permanent hardness is due to the presence of soluble calcium or magnesium sulfates in the water.
In contrast to temporary hardness, permanent hardness cannot be removed by boiling. The sum of
the permanent and temporary hardness is the total hardness of water.
Hardness of water can be determined complexometrically at pH 8-10, where EDTA (H2Y2-) is used
as complexing agent. It forms complexes with Ca2+ and Mg2+ represented by the reactions:
M2+ + HY3- → MY2- + H+
An ammonia-ammonium ion buffer is used to maintain the pH of the solution at approximately 8-
10. When a small amount of the indicator solution (Eriochrome Black T) is added to a hard water
sample whose pH has been controlled by the addition of the buffer solution, the indicator reacts
with Mg2+ to produce wine red colour.
Mg2+ + HIn3- → MgIn- + H+
(Blue) (Red)
As EDTA is added, free Ca2+ ions are first complexed to CaY2-, this being the most stable complex
]..]
Free Mg2+ ions then react to give MgY2- complex

Mg2+ + HY3- → MgY2- + H+

which is less stable than CaY2- complex but more stable than MgIn- complex. Therefore, if an extra
drop of EDTA is added after all the free Ca2+ and Mg2+ ions have been complexed, EDTA takes up
Mg2+ from the weak MgIn- complex to form stable MgY2- complex simultaneously liberating the
indicator in the free form:
MgIn- + HY3- → MgY2- + HIn3-
(Red) (Blue)
Completion of the above reaction makes the end point of the titration.
Parts per million (ppm) is the most commonly used unit of hardness and represent the number of
parts of calcium carbonate, equivalent to hardness causing ions present, per millions of water.

(a) Determination of total hardness

Procedure: Take 50.0 mL of the supplied water sample into a conical flask. Acidify it with dilute
HCl and heat for a couple of minutes to drive off carbon dioxide. Cool and neutralize the solution
with NaOH solution. Add 1 mL ammonia-ammonium chloride buffer and 3-4 drops of Eriochrome
Black T indicator. Then titrate the solution with a standard 0.01 M EDTA solution until the colour
changes from wine red to pure blue. Calculate the total hardness of the water sample in parts of
CaCO3 per million of water.

(a) Determination of permanent hardness

Procedure: Take 250 mL of sample water in a 500 mL beaker. Heat the water to boiling for about
20-30 minutes. Cool and filter the water and collect the filtrate into a 250 mL volumetric flask. Do
not wash the filter paper. Dilute the filtrate to a constant volume with distilled water and mix well.
Then titrate 50.0 mL of the filtrate the same way as was done in the case of total hardness. Calculate
the permanent hardness of the supplied water as parts of CaCO3 per million of water.

Note: One mole of EDTA is equivalent to one mole of calcium carbonate. The molar mass of CaCO3
is 100 g mol-1.

(c) Determination of temporary hardness

Procedure: Calculate the temporary hardness by subtracting the permanent hardness from the total
hardness.

12.Experiment No.: Determination of sulphate.

The sulfate is precipitated as barium sulphate from the acid solution. The precipitate is filtered off
and dissolved in a measured excess of standard EDTA solution in the presence of aqueous ammonia.
The excess of EDTA is then back titrated with standard magnesium chloride solution using
Eriochrome Black T indicator. Aqueous ammonia also maintains the pH 10 of the solution. At this
pH, the predominant EDTA species is HY3-. The reactions are:
SO42-(aq) + BaCl2(aq) → BaSO4(s)↓ + 2Cl-(aq)
NH3(aq)
BaSO4(s)↓ + HY (excess) → BaY2- + HY3-(residual) + SO42-
3-

HY3-(residual) + Mg2+ → MgY2- + H+

Mg2+ + HIn3- ⇌ MgIn- + H+

Wine-red

Procedure: Pipette 20.0 mL of the supplied sulphate solution in a beaker and dilute it with distilled
water to about 50 mL. Adjust the pH of the solution to ~1 with 2 M HCl. Heat the solution to
boiling. Add rapidly 15 mL hot BaCl2 solution (~0.05 M) with vigorous stirring. Heat the solution
on a steam bath for at least one hour. Cool the solution and filter the precipitate through a filter
paper (Whatman No. 42). Wash the precipitate with cold distilled water and then transfer the
precipitate quantitatively to a beaker. Add 40.0 mL standard EDTA solution and 5 mL concentrated
ammonia solution to dissolute the precipitate, and boil gently if needed. Cool the clear resultant
solution. Make the solution to a constant volume. Then titrate a known volume of the solution by
adjusting the pH to 10 using 10 mL buffer solution. Add a few drops of Eriochrome Black T
indicator and titrate the excess of EDTA with a standard magnesium chloride solution (~0.05 M) to
a clear red colour. Calculate the amount of sulphate (g/L) in the supplied solution.