Pharmaceutical Analysis

Second Year - Pharmacy

Water Analysis
Dissolved gases in water

1. Oxygen
2. Chloride
3. Hydrogen sulfide
4. Carbon dioxide
 (1) Dissolved Oxygen

• It measures the amount of dissolved oxygen present in

water due to its aeration.
• A good water should contain about 11 ppm of
dissolved oxygen.
• Bacteria is readily oxidized and destroyed by
dissolved oxygen, therefore it is a good sign for water.
 Determination
a) Winkler method:

• Water sample is treated with MnSO4, NaOH and KI

and set aside in a dark place for about 15 minutes.
MnSO4+ NaOH  Mn(OH)2 + 2Na2SO4
• In the presence of dissolved oxygen, the white
precipitate of Mn(OH)2 is oxidized to brown
Mn(OH)3.

2Mn(OH)2 + O + H2O  2Mn(OH)3

• Upon acidification, manganese hydroxide dissolves
and reacts with KI to liberate free iodine (I )
 Notes:

• The experiment is carried out in a closed bottle of

known volume and care should be taken to exclude
air bubbles.
• Blank experiment should be carried out.
 Interferences:

• Reducing agent, such as ferrous, sulfide and sulfite

reduce iodine to iodide leading in low results.
Solution: Dilute KMnO4 solution  faint pink color.
Excess KMnO4 is removed by sodium oxalate solution.
• Oxidising agent such as Ferric, nitrite react with iodide
and liberate free iodine, giving high results. The
difficulty caused by the presence of nitrite in the sample
can be avoided by addition of sodium azide.
NaN3 + H+  NH3 + Na+

NH3 + NO2- + H+  N2O + H2O + N2

 2- Rideal-Stewart modification :

• It is used to remove the undesirable effect of reducing

substances .
• It depends on removal of organic compounds by
addition of KMnO4 , then the excess KMnO4 is
removed by adding K-oxalate,
• After the removal of organic compounds, the
procedure is completed as in Winkler method .
 (2) Chlorine

• Chlorine is added to public water supplies, swimming pool

and sewage treatment plant effluents to destroy bacteria.
• 1 mg/L of free chlorine generally is sufficient to control
bacteria without causing noxious odor or taste to water.
• Chlorine can be present in water either as free chlorine or
combined available chlorine.
• Free chlorine is present as hypochlorous acid and or
hypochlorite ion.
• Combined chlorine exists as monochloramine, dichloramine,
nitrogen tetrachloride and other chloro derivatives.
• Both forms can exist in water and can be determined together
 Determination:

a) Iodometric method:
• Water sample is acidified and KI solution is added
then the liberated iodine is titrated with sodium
thiosulfate using starch as indicator.
• Many oxidizing substance can interfere with the
determination.
• To overcome this problem, the method is carried out in
neutral medium. In this case chlorine liberate iodine by
displacement not by oxidation
 b) O-toludine method:

• O-toludine solution in dilute hydrochloric acid is added to the

water sample in Nessler tube.
• The mixture is kept in dark place for about 5 minutes at pH
below 4.
• The color produced is varied from pale yellow to orange color
according to the amount of chlorine present in the sample.
• Since standard chlorine solution is difficult to be prepared the
comparison is carried out using “artificial standard color”
prepared by using CuSO4 and K2Cr2O7 solutions in different
proportion.
• Colored glass discs may be also used.
Ortho-toludine reaction with chlorine

O -toludine +
H

Pale yellow to orange color

 (3) Hydrogen sulfide

• Sulfide is poisonous by-product of the anaerobic

decomposition of organic matter. It is commonly found in
sewage and industrial waste waters.
• The toxicity of H2S is equivalent to that of HCN, but its
odor is detectable before toxic level is reached.
• Effects on human:
1. Hydrogen sulfide inhibits aerobic respiration,
2. inhibits muscle contractions, including breathing, and
3. promotes excess breakdown of glucose.
 Detection and determination:

• The test is based on the ability of hydrogen sulfide to

convert p-amino-dimethyl aniline into methylene blue
under suitable condition.
• The intensity of the formed blue color is directly
proportional to the amount of H2S present in the water
sample.
• Interferences:
Strong reducing agents such as sulfites, thiosulfate and
hydrosulfite interfere by reducing the intensity of the blue
color or prevent its development.
Reaction of p-aminodimethyl aniline
with H2S

p-aminodimethyl aniline HCl /FeCl3

Methyline blue
 (4) Carbon dioxide

• Carbon dioxide present in all surface water in amount

generally less than 10 ppm.
• Many ground water have high level of CO2.

• Although CO2 is not harmful for human, high

concentration of dissolved CO2 in water have a
corrosive effect and have been known to kill fish.
 Determination:
• The analysis for CO2 is similar to that for acidity
determination.
• Water sample is titrated with standard sodium
hydroxide solution using ph. ph as indicator.
• This will give both mineral acidity and acidity due to
dissolved CO2.
• Mineral acidity can be determined alone by using
M.O. as indicator.
• Care must be taken during the analysis to minimize
the loss of CO2 from water sample as a result of
aeration, during sample collection and swirling.
METALS IN WATER.

• Iron
• Copper
• Zinc
• Lead
• Manganese
 (1) Iron 0.3 mg/l (0.3 ppm)
• Natural water contain variable but minor amounts of
iron despite its universal distribution and abundance.
• Iron in natural water may exist in ferrous or ferric form.
• Ferrogenous sand is the source of iron in natural water.
• High concentrations can indicate runoff from mining
operations or industrial effluent.
• Concentrations of iron above 1 mg/L will impart a foul
taste to the water.
• The level of iron in drinking water must not exceed 0.3
mg/L (0.3 ppm).
• Water containing higher level should be rejected.
 a) Phenanthroline method

• Iron is brought into solution by boiling with acid.

• Any ferric ions present is reduced to ferrous by addition of
hydroxylamine.
4 Fe3+ + 2H2N-OH  4 Fe2+ + H2O + N2O
• Finally the solution is treated with 1,10-phenanthroline
solution.
• A red color is produced due to the formation of complex
between three molecules of phenantholine and one ion of
ferrous.
 Phenanthroline reaction

N
N
3 + Fe2- Fe
N N

1,10-phenanthroline Red colored complex

 b) Thioglycolic acid method:

• Thioglycolic acid (mercapto acetic acid) reacts with

ferric ions to form a red purple color which can be
measured spectrophotometrically at 535 nm.
• This reaction of a great importance because it is
relatively free from interferences.
• Precipitation of Al3+ and Cr3+ ions are prevented by the
addition of ammonium citrate.
Fe3+ + 2HS-CH2-COOH + 3OH- 
Fe(OH)(SCH2COO)2 + 2H2O
 c) Other colorimetric methods:

• Iron in the ferric state can be determined colorimetric

methods as a red soluble complex by using thiocyanate
in acid medium or as a blue precipitate by using
ferrocyanide.
• For determination of total iron, ferrous should be first
oxidized to ferric state by the addition of very dilute
KMnO4 solution which is added to the acidified sample
till faint pink color.
• Excess permanganate is removed by repeated filtration
of the treated sample through a filter paper which act as
reducing agent.
 (2) Zinc 1 mg/l (1 ppm)

• The average acceptable level of zinc in potable water is 1

ppm.
• The average of zinc concentration in most water supplies is
about 1 ppm, but in some area it may be as high as 50 ppm.
• The deterioration of galvanic iron and leaching of brasses can
add substantial amount of zinc to water.
• Zinc may also be found in spring and stream waters in
mining districts.
• Industrial effluents may also contribute large amount of zinc.
 Effect of Zinc:

• Zinc is essential to human metabolism.

• High concentration of zinc have irritant effect on
stomach but this effect is temporary.
• Concentration above 5 ppm show no harmful
physiological effect but can cause a bitter taste and/ or
an opalescence in alkaline drinking water.
 Determination

• Zinc ions react with ferrocyanide in acid medium to produce

a white precipitate of zinc ferrocyanide
• This precipitate can be measured turbidimetrically or by
comparison with the precipitate formed by using standard
zinc solution.
• Iron, lead and copper ions interfere with the test.
 (3) Copper 0.03 mg/l (0.03 ppm)
The average acceptable concentration of zinc in potable water is
0.03 ppm.
Sources:
Copper may be present in water due to:
4. Solvent effect of acid water in copper pipe
5. Copper sulfate added to control algae and bacteria in water.
Effects:
• Traces of copper are necessary for normal body metabolism
and its absence causes nutritional anemia in children.
• Large oral doses of copper can cause emesis and may
eventually result in liver damage.
Determination of copper

a) Diethyldithiocarbamate method:
b) ferrocyanide method:
c) Hydrogen sulfide method:
d) Dithizone method:
 a) Diethyldithiocarbamate method:
• Copper form a yellowish brown colloidal chelate
compound with sodium diethyldithiocarbamate.
• In the presence of traces amount of copper a golden yellow
color is obtained. The reagent is not specific for copper.
• Interference caused by the presence of iron can be
eliminated by:
1. extracting the formed complex with chloroform (iron
complex is insoluble in chloroform),
2. precipitating iron as hydroxide.
3. improving the selectivity of the reagent by using
masking agent particularly EDTA.
Continued

C2H5 S
N C + Cu+2
C2H5 SNa
diethyldithiocarbamate

S C2H5
C2H5 S
C N
N C Cu
C2H5 S C2H5
S

Yellowish brown colloidal chelate

 b) ferrocyanide method:

• Copper react with ferrocynide in dilute acid solution

with the formation of brown precipitate.
• In the presence of traces of copper the precipitate
appears as yellowish brown color.
• Interference can arise from the presence of iron and
zinc.
 c) Hydrogen sulfide method:

• Copper ions form a black precipitate with hydrogen sulfide.

• If traces of copper is present in the solution a brown or
yellow colloidal precipitate will appear.
• Interference from iron can be eliminated by the addition of
acetic acid.
• Lead ions will produce similar precipitate with hydrogen
sulfide. Lead sulfide is insoluble in potassium cyanide,
while, copper sulfide is soluble forming cupper cyanide
complex.
• Therefore the determination is carried out in absence and
presence of KCN, any difference in the color will be
 d) Dithizone method:

• Copper gives an orange complex with dithizone in

acid medium.
• The complex is extractable in chloroform.

Phe Phe
NH NH Phe NH N N N
S + Cu2+ S C Cu C S
N N Phe N N N NH
Phe Phe

Dithizone orange complex

 (4) LEAD 15 µg/l (15 ppb)

• Lead is a poison whose effects are cumulative. Drinking

water should not exceed 15 ppb.
• Natural water contain very low level of lead due to its
tendency to be precipitated by a large number of substances.
• When groundwater contains a higher level, it may indicate
contamination from the discharges of smelting or mining
operations, or leachate from municipal sewage sludge
fertilizer.
 The source of lead in drinking water

• Materials that contain lead which have frequently been

used in the construction of water supply distribution
systems and plumbing systems, also
• Water storage tanks which have been painted with lead-
based paint.
• Lead in these materials can contaminate drinking water as a
result of the corrosion that takes place when water comes
into contact with those materials.
 Lead effects in human health

• Lead can cause a variety of adverse health effects in humans.

• At relatively low levels of exposure, these effects may
include

1. interference in red blood cell chemistry,

2. delays in normal physical and mental development in
babies and young children,
3. slight deficits in the attention span, hearing, and
learning abilities of children,
4. and slight increases in blood pressure of some adults.
 Steps to be taken to minimize your exposure
to lead

1. Flush your plumbing to counteract the effects of "contact time."

Flushing involves allowing the cold faucet to run until a change
in temperature occurs (minimum of one minute).
2. Do not consume hot tap water.
3. Steps should be taken to make water non corrosive. Water
treatment devices for individual households include calcite
filters and other devices to lessen acidity.
4. Insist on lead-free materials for use in repairs and newly
installed plumbing.
5. Lead can be removed from your tap water by installing point-of-
use treatment devices now commercially available, which
include: ion-exchange filters, reverse osmosis devices, and
 Determination
a) Hydrogen sulfide method:
• Lead + hydrogen sulfide  a black precipitate.
• Interference from copper can be removed by the addition of
potassium cyanide which produce a soluble complex with
copper.
d) Dithizone method:
• The reagent in ammoniacal medium gives a red complex with
lead. The selectivity of the method may be enhanced by the
addition of potassium cyanide.
• Sodium citrate may be added to prevent the precipitation of iron
as hydroxide in the ammoniacal medium.
• The structure of the complex is similar to that of copper
• (5) Manganese
Manganese 0.05
is present in mg/l
ground water(0.05
as the ppm)
divalent ion
(Mn2+) due to the lack of surface oxygen. Surface water
may contain combination of various oxidation states as
soluble complexes or as suspended particles.
• Manganese is a minor constituent of many rocks and soils.
It present in slight amount in water particularly acid water.
• The presence of manganese in public water is of economic
problem rather than a potential health hazards. Manganese
causes dark stain in laundry and imparts an objectionable
taste to beverages such as coffee and tea. Level of 0.1 mg/L
is sufficient to cause taste and staining problems.
• The maximum allowable level of manganese in public
water is 0.05 mg/L (0.05 ppm) with the total iron and
manganese content not to exceed 0.3 mg/L.
 Determination

• The determination is based on the oxidation of manganese

ion into permanganate using strong oxidizing agent such as
persulfate and periodate.
• Silver nitrate is used as a catalyst in the case of persulfate.
• Interference from chloride can be avoided by the addition
of mercuric ion which form poorly ionized mercuric
chloride (HgCl2).

Mn2+ + K2S2O8  MnO4- +

 Fluoride

• Fluoride is found in some ground water.

• A level of 1 mg/L is maintained normally in public
drinking water supplies for the prevention of dental caries.
• Increased amount of fluoride causes objectionable
discoloration of tooth enamel called fluorosis.
• Level of 8 mg/L is found to be physiologically harmful for
human.
 Determination

• Fluoride in water is determined by colorimetric method.

• It is based on the reaction of fluoride with red zirconium
alizarin.
• Fluoride forms a colorless complex anion (ZrF6) and
liberates free alizarin sulphonic acid which have yellow
color in acid medium.