Professional Documents
Culture Documents
ACIDITY
OBJECT
APPARATUS
REAGENTS
THEORY
The acidity of water may be used by presence in uncombined carbon-dioxide, mineral acids and salts of strong acid
and weak bases. It is defined as the capacity of a solution to neutralize a standard alkali.
It can be noted that for carbonic acid the starchiometric end point is not reached unti1 the pH has been raised to
about 8.5 which indicates that all waters, having a pH lower that 8.5 contains acidity. Usually, the phenolphthalein
end point at pH 8.2 to 8.4 is taken as the reference point. Inspection of curve further indicates that at pH 7.0
considerable carbon-dioxide remains is to be neutralized but alone CO2 will not depress the pH below a value of
about 4.5.
Considering the nature of the curve which is for a strong acid, it may be concluded that neutralization of acid is
essentially complete at pH 4.5. It is incidentally, the methyl orange end point. Thus, it becomes obvious that the
acidity of neural water is caused by carbon dioxide or by strong mineral acids. Titration to methyl orange end point
(pH 4.5) is defined as the acidity which gives a measures of relatively strong acids such as mineral acids and
titration to the phenolphthalein end point (pH 8.3) is defined as ‘totally acidity’ and it includes also the weak acids,
acids salts and some acidity due to hydrolysis.
1 2 3 4 5 6 7 8 9 10 11
PROCEDURE
Its determination should be made on spot on a fresh sample collected in a bottle and stoppered immediately to
prevent escape of carbon dioxide. Take 50 ml to 100 ml of the sample in an Erlenmeyer flask, add 3 drops of
phenolphthalein indicator and titrate over a white surface with 0.02 N NaOH until faint colour appears.
Take 50 mL or l00 ml of the sample in an Erlenmeyer flask, add 2 drops of Methyl orange indicator, and titrate over
a white surface with 0.02 N NaOH until colour changes to faint orange. Brome-Cresol green indicator can also be
used in place of methyl orange; it gives a sharp end point.
OBSERVATIONS
CALCULATION
RESULT
The total acidity of sample B with initial pH is found to be …………mg/lt. and mineral acidity is ……….mg /lt.
The acidity of sample A (Tap water) is ………..mg/lt. The total acidity is due to mineral acidity & weak aids. For
pH range more than 8.5 the acidity due to OH- ions
SIGNIFICANCE
Acidity is of little concern from a statutory or public health view point carbon dioxide is present in malt and
carbonate beverages in concentration greatly in excess of any concentration known in natural water and no
deleterious effects due to the carbon dioxide have been recognized. Water that contains mineral acidity is usually so
unpalatable that problems related to human consumption arc non-existent.
Acid water is of concern to Sanitary engineer because of their corrosive characteristics and the expenses involved in
removing or controlling the corrosion producing substances. The corrosive factor in most water is carbon dioxide,
but in many industrial wastes it is mineral acidity. Carbon dioxide must be reckoned with water softening problems
where the lime or lime soda ash method is employed.
EXPERIMENT NO. 3
ALKALINITY
REAGENTS: 0.02 NH2SO4 solution, phenolphthalein indicator and Methyl orange indicator.
THEORY: Alkalinity is measure of the basic constituents of water and is defined as the capacity of a solution to
neutralize a standard acid. In natural water it is usually present as the carbonate and bicarbonate salts of calcium,
magnesium, sodium and potassium.
Bicarbonates represent the major form of alkalinity since they are formed in considerable amounts from the action of
carbon dioxide upon basic materials in the soil. Under certain conditions natural water may contain appreciable
amount of carbon and hydroxide alkalinity. Chemically treated water may contain appreciable amounts of carbonate
and hydroxide alkalinity. Chemically treated water, particularly those produced in lime or lime soda ash softening of
water, contain carbonates and excess hydroxide.
Thus it is obvious that alkalinity is caused by three major classes of materials may he ranked in order of their effect
on pH as hydroxides, carbonates, bicarbonates and other salt of weak acids.
Alkalinity is determined by titration with a standard solution of a strong acid to certain end point as given by
indicator solutions. Phenolphthalein is satisfactory indicator for the first end point (pH approx 8.3) contributed by
hydroxide and carbonate. Methyl orange is used for the second end point (pH approx 4.5) contributed by
bicarbonates. The phenolphthalein end point of titration is defined as ‘P’ alkalinity and the end point observed by
continuing the titration with same solution using methyl orange indicator is known as total or T-alkalinity.
Following table can be used for working out OH, CO3 and HCO3 alkalinity individually after completing titration.
Table
PROCEDURE:
Phenolphthalein alkalinity
The 50 or 100 ml of sample in an Erlenmeyer flask, add two drops phenolphthalein indicator and titrate over a white
tile with 0.02 N l-{2S04 until the pink colour just disappears.
Total or methyl orange alkalinity
Add two drops methyl orange indicator to the same sample in which phenolphthalein alkalinity has been determined
previously and titrated with 0.02 N H2S04 until the colour changes from yellow to faint orange.
OBSERVATIONS:
S. No. Volume of sample Initial burette Final burette reading Volume of H2S04
reading
1
2
3
S. No. Volume of sample Initial burette Final burette reading Volume of H2S04
reading
1
2
3
CALCULATIONS:
RESULT:
Total alkalinity due to bicarbonate is got by using methyl orange indicator, it comes …………………mg/It.
CONCLUSION:
Since alkalinity of tap water is …………..mg/It, which is very large/moderate/low. Thus it can be used/not used as
drinking water because according to IS-10500:1991 range of alkalinity for drinking water is 200-600 mg/It. OH- ion
is mainly responsible for alkalinity. Due to only OH- ion alkalinity has range of pH 8.3 to 14 and practical range of
alkalinity comes pink to colorless solution of alkalinity above 600 mg/lt, is not good for human point of view.
SIGNIFICANCE: Within regional limit alkalinity has sanitary significance, but it is very important in connection
with coagulation, softening and corrosion preservation, Alum used in coagulation is an acid salt which when added
in small quantity to natural water, reacts with alkalinity present to form flocs. If insufficient alkalinity is present to
react with all the alum, coagulation will be incomplete and soluble alum will be left in the water. It may thereafter be
necessary to add alkalinity in the form of soda ash or lime to complete the coagulation or to maintain sufficient
alkalinity to prevent the coagulated water for being corrosive. Ordinarily the total alkalinity determined with methyl
orange indicator; gives sufficient information for the control of coagulation and corrosion prevention when pH is
also determined.
Many regulatory agencies prohibit the discharge of waste containing caustic alkalinity to receiving water. Municipal
authorities usually prohibit discharge of waste containing caustic alkalinity to sewers. Alkalinity as well as pH is an
important factor in determining the amenability of waste water to biological treatment.
Lastly from public health point of view, alkaline water is usually unpalatable and consumer tends to seek other
supplies. Chemically treated water some time has rather high pH values, which have met with some objections on
the part of consumers. For these reasons, standards are sometimes established on chemically treated water. Where
biological processes of treatment are used the pH must ordinarily be maintained whim the range of 6 to 9.5. This
criterion often requires adjustment of pH to favorable levels and calculations of the amount of chemical needed is
based upon acidity values in most cases.
EXPERIMENT 5
HARDNESS TEST
OBJECT: To determine the total hardness and calcium hardness of a given sample of water.
REAGENTS: Standard EDTA solution (N/50), Ammonia buffer solution and NaOH solution, Eriochrome black T
indicator and Murex indicator (dry power), inhibitor.
THEORY
Water that consumes considerable quantity of soap to produce lather and or produces scale in hot water pipes,
heater, boilers and utensils used for cooking is called hard water.
Hardness is caused by divalent metallic anions that are capable of reacting with soap to form precipitates with
cations present in water to form scale. Principal actions causing hardness and the major anions associated with them
are as listed below:
CATIONS ANIONS
Ca++ HCO3-
Mg++ S04--
Sr++ Cl-
Fe++ N03-
Mn++ Si03--
Calcium and magnesium are primarily the constituents of chalk and limestone. When rain falls it takes up carbon
dioxide from the atmosphere and forms a weak acid and this percolates underground, it then dissolves calcium and
magnesium forming hard water. In general hard water originates in the areas where the topsoil is thick and limestone
formation are present. Soft water originates in areas where the topsoil is thin and limestone is either sparse or absent.
0 50 ppm Soft
In alkaline condition EDTA (Ethylene-diamine tetra acetic acid) or its sodium salt forms a soluble chelated complex,
which is stable with Ca and Mg. Also Ca and Mg form a weak complex with the indicator Eriochrome black T,
which has wine red color. During titration when all free hardness ions are complexed by Eriochrom black T
indicator end point. The pH has to maintain at 10±0.1.
At higher pH i.e. about 12.0 mg ion precipitates and only Ca++ ions remain in solution. At this pH murex indicator
from a pink colour with Ca++, gets complexed resulting in a change from pink to purple, which indicates and point
of the reaction.
INTERFERENCE
PROCEDURE
A. TOTAL HARDNESS
4. Add a pinch of Eriochrome black T and titrate with standard EDTA solution till wine colour changes to blue.
Note down the volume of EDTA required.
B. CALCIUM HARDNESS
2. Add 3 drops of NaOH (N/10) to raise pH to 12 and a pinch of indicator. Note initial burette readings.
3. Titrate with EDTA till pink colour changes to purple Note the final burette readings.
4. Repeat the procedure for other sample s till concurrent readings are obtained.
C. MAGNESIUM HARDNESS
1. Take l00 ml of sample ,add 1.5 ml of the buffer solution and 2.3 ml of a saturated solution of ammonia oxalate.
2. Mix the solution and allow it to stand for two hours or overnight if possible.
4. Pipette cut 25 ml from the filtered solution and add Eriochrome black T indicator (1-2 drops) and titrate with
EDFA solution till the colour changes from wine red to blue.
5. Take two concurrent readings.
S. no. Volume of Initial reading Final reading Net volume of Total hardness
sample EDTA (ml) mg/lt as CaCO3
1
2
3
4
SAMPLE CALCULATIONS
ml. of sample
ml. of sample
RESULT
For the given tap water sample the hardness is found to be mg/lt., Calcium hardness is mg/lt. and Magnesium
hardness is mg/lt.
CONCLUSION
As the total hardness and calcium hardness arc below/above the maximum tolerable value i.e. 600 and 200
respectively. The water can be used/not used for domestic purposes.
SIGNFICANCE
The determination of hardness is helpful in deciding the suitability of water for domestic and industrial purpose. The
design of softening process depends upon the relative amounts of carbonate and non-carbonate hardness present in
water. The amount of calcium and magnesium hardness decides the suitability of water for boiler use.
1.
calcium hardness
mg/It.
EXPERIMENT 6
DISSOLVED OXYGEN
OBJECT
APPARATUS
500 ml ground glass, stopper bottles, nesslers tubes, pipettes, DO bottles, etc.
REAGENTS
MnSO4 solution concentrated H2S04, Alkali iodide azide solution, N/40, Na2S2O3 solution.
THEORY
Adequate dissolved oxygen is necessary for the use of fish and aquatic life .The dissolved oxygen concentration may
also he associated with corrositivity of water and its septicity. The D.O. test is used in B.O.D. determinations.
Determination of dissolved oxygen has no importance from the point of view of potability, but the dissolved oxygen
will indicate the power of self purification of water .Amount of dissolved oxygen in water varies with temperature
and also with dissolved salts present in water. Greater the temperature, lesser the dissolved oxygen and greater the
percentage of dissolved salts, lesser the dissolved oxygen.
There are two methods of determination of dissolved oxygen in a given sample of water.
1. winkler method
2. poplarograplic method
In determining the dissolved oxygen, the various ions and compounds present in water cause interference in desired
reactions and hence a specific modification is desired to be adopted in every case depending upon the type of ions
present in water sample. The most common interference is caused by nitrites ferrous and ferric ions in the surface
water and incubated waste water sample for B.O.D. hence the modification known as Winkler’s Azide modification
for nitrite is used.
In all the modification the principle is dependent on the reaction which releases free iodine from potassium iodine
and the amount of iodine released is directly proportional to the amount of oxygen originally presented. In this
method Fe++ and NO2 will oxide iodide to free iodine. Mn++ (manganese) salts are oxidized to Mn+++ salts in alkaline
solution. Mn+++ is capable of oxidized from iodide to iodine in acidic conditions. The reducing agent used in this
case is sodium thiosulphate, with starch as indicator. As stated above to avoid interferences of NO2 and Fe+++, S, SO3
etc. The alkali azide reagent is added, the titration is then proceeded with.
CHEMICAL REACTIONS
Winkler’s method
2. Mn(OH)2 + O2 = 2MnO(OH)2
(D.O.) (brown ppt)
PROCEDURE
1. Sample is collected in D.O. bottle from source and air bubbles removed.
2. D.O. is fixed by adding 2 ml of each aikali-azide and manganese sulphates solution to the sample filled in bottle
water cause interference in desired.
5. Iodine is liberated from KI. The amount of I2 liberated is proportional to amount of D.O.
6. 100 ml of solution thus obtained is taken and two drops of starch is added as an indicator.
OBSERVATION
CALCULATION
The ml of N/40 Na2S2O3 (sodium thiosulphate ) used for 200 ml of sample original is equivalent to D.O. in mg/l as
oxygen.
1)D.O. = …………..mg/l
N1V1 = N2V2
N/40X x = N2 X 200
N2 = x/40 X 200
= x X 8 X 1000/40 X 200
RESULTS
COMMENT
In the experiment when 2 ml of MnSO4 followed by 2 ml of NaOH + KI + NaN3 is added to the sample 4 ml of
original sample is lost. Thus 203 ml taken for titration corresponding to 200 ml of original sample
SIGNIFICANCE
Living organisms require oxygen to maintain their metabolic process. Dissolved oxygen is very important in
precipitating and dissolution of organic substances in water depends upon its temperature. Analysis of dissolved
oxygen is the main key test in sanitary engineering.
Surface water contains dissolved oxygen from atmosphere. This oxygen is used by the aquatic animals for their life.
Also this dissolved oxygen gives freshness and sparkling to water. Thus the determination of D.O. is necessary. It
helps in determination of B.O.D. greater D.O. may use corrosively to pipe materials. D.O. decreases with rise in
temperature. Water used for domestic purposes DO. should be 5 - 6 ppm.
PRECAUTIONS
1. The initial pH should be taken.
OBJECT
APPRATUS
Evaporating dishes, water bath, hot air oven a weighing balance, filter
paper No.42
THEORY
The impurities in water or wash water, whether solids, liquid or gas are
disperse in three progressively finer states, names suspended, colloidal
and dissolved. T he state of sub division is of utmost importance since
it determines the method required for removal.
1. Fixed
2. Volatile
Water is never found nature in its pure form. Impurities pf organic and
inorganic nature are associated with it and they contribute colour,
odour, turbidity and taste etc. to it and make it unfit for drinking. To
make it fit for drinking, the water requires to be treated and bringing to
the standard levels. According to U.S.P.H.S. the total solid contents
should not be more than 500mg/lit for portable water, while both
W.H.O. & G.O.I. have laid down the limit as 500mg/lit. (excessive)
PROCEDURE
Total solids
Ignite the residue remaining in the dish at a low red heat over burner,
or in the muffel furnace at 500 ̊ to constant weight, cool in desiccators
and weigh. Record the loss in weight on ignition on total volatile solids.
Suspended solids
Dissolved solids
CALCULATION
= ______ mg/lit
Settable solids
(a)By volume: Fill in Imhoff cone to the liter mark with a thoroughly
mixed sample. Allow to settle for 45 minutes, gently stir the sides of
the cone with a rod or by spinning, and settle for a further period of 15
minutes. Record the volume in ml of the settable matter in the cone.
(b)By weight: This technique defines settable solids as that matter in
sewage which will not stay in suspension during the settling period
either by virtue of settling to the bottom or floating to the top.
Form a well mixed sample into a glass vessel not less than 9cm in
diameter, using a quantity of samples not less than 1 liter and
sufficient to ensure a depth of 20cm. A glass vessel of greater
diameter and larger volume may be used.
Allow the sample top stand quiescent for 1 hour and siphon 250ml
from the center of container at a point half way between the surface of
the settled sludge and the liquid surface with out disturbing the settled
material or that which may be floating. Determined the suspended
matter in mg/lit in all or in a adequate proportion of this supernatant
liquor. This value mg/lit is equivalent to the non settling solids.
OBSERVATION TABLE
Sample = artificial
CALCULATION
Total solids in mg/lit =(__________) x 1000 / ml of
sample
= _____________ gm x 1000 / 10
= _____________ mg/lit
= _____________
= _____________ mg/lit
RESULT
CONCLUSION
SIGNIFICANCE
EXPERIMENT 10
OBJECT
1. To determine the available chlorine in bleaching powder.
2. To determine the residual chlorine in given sample of water.
APPARATUS
Burette, pipette, conical flask, stirrer.
REAGENTS
KI solution, Glacial acetic acid, Distilled water, Starch, Sodium thiosulphate solution.
THEORY
In small water works, chlorine required for disinfections is usually obtained from bleaching powder. For this
purpose iodometric method i.e. oxidation-reduction method is employed. The iodometric method is more precise
when the residual chlorine concentration is greater than one ppm. Chlorine will liberate free iodine from KI solution
when its pH is 8 or less. The liberated iodine is titrated against standard solution of sodium thiosulphate using starch
as indicator. When blue colour disappears then all the liberate I iodine will have reacted. This indicates the end
point.
PROCEDURE
1.5 gram bleaching powder is taken and dissolved in 1 liter of distilled water. The solution thus prepared is to be
tested for available chlorine. 20 ml. of 10% KI is taken in i clean, dry conical flask, 2 ml of glacial acetic acid is
added into the flask to reduce 1 3 to 4.12. 12 5 ml of bleaching powder solution is then pipetted out and is added into
the flask. The colour of the solution will be brown. Titrate this solution against N/40 sodium thiosulphate solution,
till pale or straw yellow colour is developed. At this stage, add 2 drops of freshly prepared starch solution, which
results in appearance of blue colour. The titration against sodium thiosulphate solution is continued till the blue
colour disappear. This indicates the end point. Initial reading and final reading of sodium thiosulphate solution in the
burette is noted, the difference is then found out. The whole experiment is repeated and the mean difference is taken.
OBSERVATIONS
CALCULATIONS
mg/lt n the water sample N/40 normality remove the blue colour
= mg/lt. as chlorine
CHLORINATION
Chlorine is widely used for disinfection of water for removing odour since it is a powerful oxidizing agent and is
cheaply available. It can be used in molecular from or in hypochlorite form. For effective disinfection, dose of
chlorine, optimum contact period and residual chlorine are required to be found out.
PRINCIPLE
Chlorine combines with water to form hypochlorous and hydrochloric acid. Hypochlorous acid dissociates to give
OCl- ion. Quantities of OCl- and HOCl- depend on pH of solution. Hypochlorides also gives the OCl- ions, HOCl-
rupture the cell membrane of microbes, the disease producing organisms. These also reacts with the impurities like
ammonia, oxidisable inorganic matter like ferrous ion, nitrites etc. to from chloramines and stable ions of the latter
respectively.
INTERFERENCE
REAGENTS
1. Bleaching powder
4. Standard sodium thiosulphate 0.1 N-- Dissolve 25g Na2S2O3.5 H20 and dilute to 1000 ml in freshly boiled and
cooled distilled water. Add about 5ml chloroform as preservative.
5. Starch indicator: Prepare slurry by adding small quantity of water to 1 g starch powder. Add it to 100 ml boiling
water and continue boiling for few minutes then cool and uses.
REACTION
Hypochlorous acid is unstable and may break into hydrogen ions and hypochlorite ions.
Moreover the chlorine will immediately react with ammonia present in water to form various chloramines
PROCEDURE
2. Add standardized chlorine solution in ascending order. If chlorine demand of treated water is being estimated,
doses from 0 to 300mg Cl2/1. I will be found useful. However, if the sample is polluted, doses from 0.1 mg to 3 mg
C12/l may be required as in case of treated effluents etc.
3. A low a contact period of 30 minutes of probable water and suitably higher for poliute:1 ‘ater, or secondary
effluents.
5. Plot residual chlorine versus chlorine added. In case of organically polluted samples, a distinct break point can be
obtained. But in case of treated water sample, it is possible at only a straight line is obtained in absence of any
ammonium. A residual 0.2 mg Cl2, it. after the break point is recommended.
CONCLUSION
We have found the result in the above test is ……….. mg/lt. chlorine that chlorine is satisfied/not satisfied as
compared to permissible limit. The permissible limit is 0.2 according to IS: 10500, hence given sample of water
does not fit for drinking purpose The amount of available chlorine in a sample indicates that bacteria are reduced up
to safer limits but when it increases above the permissible limit. It may lead to the water born disease. In this water
sample the chlorine is below / average / above/ as per permissible limit. So it is fit / not fit for drinking and can be
accepted/ rejected.
RESULT
If the amount of available chlorine present in water is more than the permissible limit then the water should have
unpleasant taste characteristics. But if it is present within limit it forms hydrochlorous acid and kills the bacteria
present in the water as it has been described earlier.
EXPERIMENT 11
OBJECT: To determine the break point chloride demand of given sample of water.
APPARATUS: A number of bottles with stoppers, pipette, O.T. test comparator, stock solution of Cl2 of known
strength.
THEORY: The determination of break point chlorine demand of water is in effect the extension of experiment
already performed for determination of chlorine demand of water. As discussed already in the said experiment the
residual chlorine appears only after the demand has been met after adding a particular dose of chlorine, at definite
period of contact.
In the absence of ammonia or its derivatives in water, the residual chlorine is the free available chlorine in the form
of H0CL and/or OOL which are oxidants and react with orthotolidine to show residual chlorine. Thus once the
residual appears, it will go on increasing with increase in applied dose.
However, when ammonia is present, the hypochlorus acid i.e. HOCL reacts with it form chlorine’s first mono and
then dichlormine if excess of HOCL is available
HOCl+NH3 = NH2Cl+H20
and HOCl+NH3Cl = NHCl2+H20
Both dichloromine and monochloromine are oxidizing agents as if HOCl and react with orthotolidine to show
residual chlorine. Hence, in the presence of ammonia in water, the residual chlorine is the sum total of the action of
HOCl and chloromines is called the ‘Combined available chlorine” and lie total residual of chloride is both due to
free and combined available chlorine. An interesting stage comes in when all the ammonia present has been
converted into monochloromine with addition of Cl2. At this stage further addition of chlorine will not results in
increase of residual because of the following reaction:
NHCl2+HOCl =NCl3+H20
The conversion of NHCI2 to NCl3 rather results in drop of residual chlorine from the level already attained. This is
due to the fact that NCl3 is non-oxidizing and does not react with O.T. and thus the drop in NHCl2 results in the drop
of residual chlorine with added dose of chlorine. Thus, once again the chlorine demand goes on increasing because
of consumption of chlorine in producing NCl3, which does not show any residue. A stage reaches when all the
NCHCl2 present is converted into NCl3 and there is no combined available chloride at all, the chlorine demand is
maximum at this product. This is called the ‘break point’ in chlorination. Further addition of Cl2 will again show
residue but this will be in the form of free chlorine i.e. HOCl and OCl as all the NH3 has been converted into NCl3 or
oxidized to free nitrogen or other gases at the break point.
The form of chlorine available as residue after break point is entirely free available chlorine and under controlled
conditions of pH it may be entirely available as HOCL (and not as OCl because at lower pH values, the ionization of
HOCl will be minimum).Which is the most efficient as bactericide, the chloramines and OCl being poor in action as
disinfectants. The break point chlorination therefore achieves:
d) Removal of manganese
PROCEDURE
A number of sample bottles are taken and to each of them water sample (containing NH3 and its derivatives) in equal
quantity, say 100 ml is put. to those sample are added at 5 minutes interval one by one gradual increasing does of Cl
solution from a stock solution of chlorine of known strength. Residual chlorine is tested in each bottle after a
contact time of 10 minutes by means of O.T. reagent and comparators.
OBSERVATIONS
Sample No. ml of Cl2 Solution added Dose of Cl2 in ppm Residual chlorine
after 10 minutes
1 2 3 4 5
CALCULATIONS
SIGNIFICANCE
EXPERIMENT No. 18
BIOCHAMICAL OXYGEN DEMAND
OBJECT
APPARATUS
B.O.D. incubator, Burette, Pipette and flasks.
REAGENTS
THEORY
The kinetics of B.O.D, reaction indicate that they are first order
reaction, in which the rate of the reaction is proportion to the
amount if oxidisable organic matter at any time as modified by
population of active organisms. This can be express in the from of
an equation , Y=L(1-10-kt) , where Y=B.O.D. at any time and Y=(I-
10-kt).
PROCEDURE
Initial Final
reading
reading
1.
2.
3.
4.
Initial Final
reading
reading
1.
2.
3.
4.
CALCULATION
RESULTS
COMMENTS
REACATION
SIGNIFICANCE
LIST OF CHEMICAL
1. Ferric chloride FeCl3
5. Phosphate Buffer.