Volumetric Analysis

VOLUMETRIC ANALYSIS
CORE PRACTICAL I
COMPILED BY
Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI
DEPARTMENT OF BIOCHEMISTRY
ISLAMIAH COLLEGE (AUTONOMOUS)
VANIYAMBADI - 635751
FOR THE STUDENT OF IST B. Sc BIOCHEMISTRY
COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI
Page 1
ESTIMATION OF AMINO ACID (GLYCINE) BY SORENSONS FORMAL TITRIMETRIC METHOD
CALCULATION
Weight of oxalic acid = 1.576 g
Titration I
Standard oxalic acid Vs Sodium hydroxide
Indicator: phenolphthalein
S.NO
Volume of standard oxalic
Burette readings
acid (ml)
Initial (ml)
20 ml
0 ml
20 ml
0 ml
Volume of Sodium
Final (ml)
Volume of Standard oxalic acid solution
(V 1)
20 ml
Normality of Standard oxalic acid solution
(N1 )
0.1 N
Volume of Sodium hydroxide solution
(V2)
-------- ml
Normality of Sodium hydroxide solution
(N2)
--------------------------- N
We know that,
hydroxide (ml)
V1N1 = V2N2
N2
= V1N1
V2
N2 =
20 x 0.1/---------
N2 =
-----------------
(N2)
Page 2
Ex. No :
Date
ESTIMATION OF AMINO ACID (GLYCINE) BY SORENSONS FORMAL TITRIMETRIC METHOD
Aim
To estimate the amount of amino acid (Glycine) present in the whole of the given unknown
solution
Principle
Amino acid reacts with excess of formaldehyde to give free hydrogen ion and act as acidic
solution. This acidic solution is titrated against standard alkali (Sodium hydroxide) using phenolphthalein
as indicator.
Reagents required
i.
Standard oxalic acid solution (0.1 N)
ii.
Sodium hydroxide solution
iii.
Phenolphthalein as indicator.
iv.
Formaldehyde
v.
Amino acid (Glycine)
Page 3
Titration II (Test value)

Unknown amino acid (Glycine) Vs Standardized Sodium hydroxide solution
Indicator: phenolphthalein
S.NO
Contents taken in the

conical flask (ml)
Burette readings
Initial (ml)
Volume of Sodium
hydroxide (ml)
Final (ml)
20 ml amino acid + 5 ml
0 ml
1
formaldehyde
20 ml amino acid + 5 ml
0 ml
2
formaldehyde
Volume of Sodium hydroxide consumed by amino acid (Glycine)

and formaldehyde
(Test value)
------------------ ml
Titration III (Blank value)

Blank Vs Standardized Sodium hydroxide solution
S.NO
Contents taken in the

conical flask (ml)
Burette readings
Initial (ml)
Volume of Sodium
Final (ml)
hydroxide (ml)
20 ml distilled water + 5 ml
1
0 ml
formaldehyde
20 ml distilled water + 5 ml
0 ml
formaldehyde
Volume of Sodium hydroxide consumed by formaldehyde (Blank value) =
------------------ ml
Volume of Sodium hydroxide consumed by amino acid alone
Test value Blank value
= ------------------------ ml
Page 4
Procedure
Titration I
Standard oxalic acid Vs Sodium hydroxide solution
Weighed accurately 1.576 g of crystalline oxalic acid and transfer into a 250 ml of standard flask
then the volume is made up to 250 ml using distilled water. Pipette out exactly 20 ml of this solution
into a clean conical flask and two drops of phenolphthalein as indicator is added. This is titrated against
the Sodium hydroxide solution taken in the burette. The end point is the appearance of pale permanent
pink colour. The titrations are repeated for concordant values. From the titre value the normality of
Sodium hydroxide solution is calculated.
Titration II (Test value)

Unknown amino acid (Glycine) Vs Standardized Sodium hydroxide solution
The given unknown amino acid (Glycine) solution is made up to 100 ml standard flask using
distilled water. Pipette out exactly 20 ml of this solution
into a clean conical flask to this 5 ml of
formaldehyde and two drops of phenolphthalein as indicator is added and shaken well for 5 minutes.
This is titrated against the standardized Sodium hydroxide solution taken in the burette. The end point is
the appearance of pale permanent pink colour. The titrations are repeated for concordant values.
Titration III (Blank value)
Blank Vs Standardized Sodium hydroxide solution
Pipette out exactly 20 ml of distilled water into a clean conical flask to this 5 ml of formaldehyde
and followed by two drops of phenolphthalein as indicator is added. The contents are mixed well for 5
minutes. This is titrated against the standardized Sodium hydroxide solution taken in the burette. The
end point is the appearance of pale permanent pink colour. The titrations are repeated for concordant
values.
Page 5
Calculation
Volume of Sodium hydroxide solution
(V 1)
-------------- ml
(N1)
-------------- N
Volume of amino acid (Glycine) solution
(V2)
20 ml
Normality of amino acid (Glycine) solution
(N2)
We know that,
V1N1 = V2N2
N2
= V1N1
V2
N2 =
----------------
Normality of amino acid (Glycine)
(N2)
= ------------------ N
Equivalent weight of Glycine
75
The amount of amino acid (Glycine) present in the whole of the given unknown solution
Normality of amino acid (Glycine) x Equivalent weight of Glycine x 100

1000
----------------------------- grams
Page 6
Test value Blank value will give the actual amount of Sodium hydroxide consumed by the
amino acid solution. From this value the strength of amino acid is calculated, and from this strength the
amino acid present in the whole of the given unknown solution is calculated
Equivalent weight of Glycine
75
Result
The amount amino acid (Glycine) present in the whole of the given unknown solution is
=
----------------------- grams
Page 7
Ex. No :
Date
ESTIMATION OF SUGAR (GLUCOSE) BY BENEDICTS TITRIMETRIC METHOD
:
CALCULATION
Standard Glucose
250 mg of Glucose/250 ml of distilled water
Titration I
Standard Glucose Vs Benedicts Reagent
S.NO
Volume of Benedicts
Burette readings
Reagent (ml)
Initial (ml)
5 ml
0 ml
5 ml
0 ml
Final (ml)
Volume of Standard
Glucose (ml)
5 ml of Benedicts reagent consumes --------------------------- ml of Standard Glucose solution
Titration II
Unknown Glucose Vs Benedicts Reagent
S.NO
Volume of Benedicts
Burette readings
Volume of Unknown
Glucose (ml)
Reagent (ml)
Initial (ml)
5 ml
0 ml
5 ml
0 ml
Final (ml)
5 ml of Benedicts reagent consumes --------------------------- ml of unknown Glucose solution
Page 8
ESTIMATION OF SUGAR (GLUCOSE) BY BENEDICTS TITRIMETRIC METHOD

Aim
To estimate the amount of Sugar (Gluose) present in the whole of the given unknown solution
Principle
When glucose is heated with an alkaline solution of copper (Cu2+) ions, the copper (Cu2+) ions is
reduced to cupric (Cu+) ion, which is precipitated as copper oxide CuO2. This is the basic for the
estimation of reducing sugar.
Reagent required
Benedicts quantitative Reagent
Crystalline copper sulphate 18 grams, anhydrous sodium carbonate 200 grams, potassium
thio cyanate 125 grams, sodium nitrite 200 grams are added to 250 ml of distilled water and
dissolved with the aid of heat. The contents are finally made up to 800 ml using distilled water.
Standard glucose solution
Weighed accurately 250 mg of Glucose and transfer into a 250 ml of standard flask then the volume is
made up to 250 ml using distilled water. (concentration 1 mg/1 ml)
Procedure
Titration I
Standard Glucose solution Vs Benedicts quantitative Reagent
Pipette out exactly 5 ml of Benedicts quantitative reagent into a clean conical flask. To this add 1 gram
of sodium carbonate. The contents are mixed well and heated in boiling water bath till the first bubble
appearance. This is titrated against the Standard Glucose solution taken in the burette. The end point is
the disappearance of blue colour. The titrations are repeated for concordant values
Page 9
Volume of standard Glucose
Volume of unknown Glucose
ie. --------------------- ml of standard Glucose

--------------------- ml of unknown Glucose
100 ml of unknown solution contain
X
X 100
Y
= -------------------- mg
Page 10
Titration II
Unknown Glucose Vs Benedicts Reagent
The given unknown Glucose solution is made up to 100 ml standard flask using distilled water.
The burette is rinsed with unknown Glucose solution and filled with the same unknown Glucose
solution. Pipette out exactly 5 ml of Benedicts quantitative Reagent into a clean conical flask to this
1gram of sodium carbonate added. The contents are mixed well and heated till the first bubble
appearance. This is titrated against the unknown Glucose solution taken in the burette. The end point is
the disappearance of blue colour. The titrations are repeated for concordant values.
Precaution
i.
Excess amount Sodium carbonate will cause frothing
ii.
Benedicts quantitative Reagent is kept at boiling throughout in the titration
iii.
Porcelain bits are added to prevent bumping
Result
The amount of Sugar (Glucose) present in the whole of the given unknown
Solution ------------------------ grams
Page 11
ESTIMATION OF ASCORBIC ACID USING 2, 6 DICHLORO PHENOL INDOPHENOL

Titration I
Standard Ascorbic acid Vs Dye
S.NO
Volume of Standard
Burette readings
Ascorbic acid (ml)
Initial (ml)
10 ml
0 ml
10 ml
0 ml
Volume of Dye
(ml)
Final (ml)
Volume of the Dye
--------------------------- ml
10 ml of standard ascorbic acid consumes
------------------------ ml of the dye
Titration II
Unknown Ascorbic acid Vs Dye
S.NO
Volume of Unknown
Burette readings
Ascorbic acid (ml)
Initial (ml)
10 ml
0 ml
10 ml
0 ml
Volume of Dye
Final (ml)
Volume of the Dye
--------------------------- ml
X - mg of ascorbic acid consumes
------------------------ ml of the dye
(ml)
Page 12
Ex. No :
Date
ESTIMATION OF ASCORBIC ACID USING 2, 6 DICHLORO PHENOL INDOPHENOL
:
Aim
To estimate the amount of ascorbic acid present in the whole of the given unknown solution
Principle
Ascorbic acid is oxidized by the colourD dye 2, 6 Dichloro phenol indophenol to dehydro
ascorbic acid. At the same time the dye is reduced to colourless compound, so that, the end point of the
reaction can be easily determined.
Page 13
Calculation
Working standard ascorbic acid
10 mg ascorbic acid/100 ml of 0.6 % oxalic acid

[Concentration 0.1 mg/1 ml]
Ie. 1 ml of standard contains 0.1 mg of ascorbic acid

10 ml standard contains [0.1 mg x 10] 1 mg of ascorbic acid
10 ml standard ascorbic acid contains -------------------------- ml of Dye
Ie. 1 mg of ascorbic acid consumes ---------------------------- ml of Dye
But X of ascorbic acid consumes ------------------------ ml of Dye
The amount of ascorbic acid present in the 100 ml of unknown solution

Volume of dye consumed by unknown solution
=
Volume of dye consumed by standard solution
=
-------------------- mg
The amount of ascorbic acid present in the whole of the given unknown solution
------------------ mg x 10
------------------- mg
Page 14
Reagent required
2, 6 Dichloro phenol indophenols
Dissolve 42 grams of Sodium bi carbonate and 52 grams of Dichloro phenol indophenols in 50 ml
of distilled water and finally dilute to 250 ml using distilled water.
Stock standard ascorbic acid solution
100 mg of ascorbic acid is weighed exactly and carefully transfer in to 100 ml standard flask and
made up to 100 ml using 0.6 % oxalic acid.
Working standard ascorbic acid solution
10 ml of Stock standard ascorbic acid solution is pipette out in to a 100 ml standard flask and
made up to 100 ml using 0.6 % oxalic acid.
Procedure
Titration I
Standard Ascorbic acid Vs Dye
Pipette out exactly 10 ml of working standard ascorbic acid solution into a clean conical flask
and it is titrated against the dye taken in the burette. The end point is the appearance of pale
permanent pink colour. The titrations are repeated for concordant values.
Titration II
Unknown Ascorbic acid Vs Dye
The given unknown ascorbic acid solution is made up to 100 ml standard flask using 0.6 % oxalic
acid. Pipette out exactly 10 ml of this unknown ascorbic acid solution into a clean conical flask and it is
titrated against the dye taken in the burette. The end point is the appearance of pale permanent pink
colour. The titrations are repeated for concordant values.
Page 15
Result
The amount of ascorbic acid present in the whole of the given unknown
Solution ------------------------ mg
Page 16
DETERMINATION OF ACID NO OF FAT

CALCULATION
Standard 0.1 N oxalic acid = 1.576 g of oxalic acid/250 ml of distilled water
Titration I
Standard oxalic acid Vs Potassium hydroxide
S.NO
Volume of standard oxalic
Burette readings
acid (ml)
Initial (ml)
20 ml
0 ml
20 ml
0 ml
Final (ml)
Volume of Standard oxalic acid solution
(V 1)
20 ml
Normality of Standard oxalic acid solution
(N1 )
0.1 N
Volume of Potassium hydroxide solution
(V2)
-------- ml
Normality of Potassium hydroxide solution
(N2)
--------------------------- N
Volume of Potassium
hydroxide (ml)
We know that,
V1N1 = V2N2
N2 = V1N1
V2
N2 =
20 x 0.1/---------
N2 =
-----------------
Normality of Potassium hydroxide solution
(N2)
Page 17
Ex. No :
Date
DETERMINATION OF ACID NO OF FAT
Aim
To estimate the amount of Acid no of the given Fat
Principle
During storage of fat become rancid. As a result the peroxide formation of the double bond by
atmospheric oxygen and or hydrolyzed by micro organism with liberation of free fatty acids. The amount
of acid present gives the indication of age and quality of the fat.
Acid value is the number of milligrams of KOH required to neutralize the free fatty acids in one gram of a
fat or oil. It is a measure of free fatty acid contents in a fat or oil.
Reagents required
i.
Fat
ii.
Fat solvent
iii.
Standard 0.1 N oxalic acid
iv.
Potassium hydroxide solution
v.
Phenolphthalein as indicator.
vi.
Methanol / Ethanol
Procedure
Titration I
Standard oxalic acid Vs Potassium hydroxide solution
Weigh accurately 1.575 g of oxalic acid and transfer into a 250 ml of standard flask then the
volume is made up to 250 ml using distilled water. Pipette out exactly 20 ml of this solution into a clean
conical flask and two drops of phenolphthalein as indicator is added. This is titrated against the
Potassium hydroxide solution taken in the burette. The end point is the appearance of pale permanent
pink colour. The titrations are repeated for concordant values. From the titre value the normality of
Potassium hydroxide solution is calculated.
Page 18
Weight of the oil

Weight of the weighing bottle + Oil
Weight of the weighing bottle
Weight of the Oil transferred
(-)
Titration II
Test value
S.NO
Contents taken in the conical

flask
Burette readings
Initial (ml)
Volume of Potassium
hydroxide (ml)
Final (ml)
-------------- gram of Oil + 50 ml

1
of alcohol + 3 drops of
0 ml
Phenolphthalein
Titration III
Blank Value
Contents taken in the conical
Burette readings
Volume of Potassium
S.NO
flask
Initial (ml)
50 ml of alcohol + 3 drops of
0 ml
Final (ml)
hydroxide (ml)
Phenolphthalein
Volume of Potassium hydroxide consumed
By Oil alone
------------------ ml
Page 19
Titration II
Test Value
Weigh about one gram of edible oil and carefully transfer in to a clean dry conical flask. Then 50
ml of alcohol is added followed by two drops of phenolphthalein as indicator is added. The contents are
mixed well for 20 minutes. This is titrated against the standardized Potassium hydroxide solution taken
in the burette. The end point is the appearance of pale permanent pink colour and persisting up to
20 30 seconds.
Titration III
Blank Value
50 ml of alcohol is taken in a conical flask and three drops of phenolphthalein as indicator is
added. The contents are mixed well. This is titrated against the standardized Potassium hydroxide
solution taken in the burette. The end point is the appearance of pale permanent pink colour and
persisting up to 20 30 seconds.
Page 20
100 ml of 1 N Potassium hydroxide contain 56 grams of Potassium hydroxide
56 x Strength of Potassium hydroxide x Test value Blank value

1000
Acid no of Fat =
--------------------- grams
---------------------- grams x 1000

Weight of Oil
----------------------------- Acid no of Fat (Oil)
Result
Acid No of the given Fat
------------------------------
Page 21
DETERMINATION OF IODINE NO OF FAT

CALCULATION
Standard 0.1 N potassium dichromate solution = 1.225 g of potassium dichromate solution /250
ml of distilled water.
Titration I
Standard potassium dichromate solution Vs Sodium thio cyanate solution
S.NO
Volume of potassium
Volume of Sodium thio
Burette readings
dichromate solution (ml)
Initial (ml)
20 ml
0 ml
20 ml
0 ml
Final (ml)
cyanate solution (ml)
Volume of Standard potassium dichromate solution
(V 1)
20 ml
Normality of Standard potassium dichromate solution
(N1)
0.1 N
Volume of Sodium thio cyanate solution
(V2)
-------- ml
Normality of Sodium thio cyanate solution
(N2)
--------------------------- N
We know that,
V1N1 = V2N2
N2 = V1N1
V2
N2 =
20 x 0.1/---------
N2 =
-----------------
Normality of Sodium thio cyanate solution
(N2)
Page 22
Ex. No :
DETERMINATION OF IODINE NO OF FAT
Date
Aim
To estimate the amount of Iodine no of the given Fat
Principle
Iodine no of fat is defined as the no of grams of iodine absorbed by 100 gram of fat or oil. It is
a measure of degree unsaturation of the fatty acids in a fat or oil. Unsaturated fatty acids, either free or
combined in lipids react with halogens like bromine and iodine which get decolorized. These halogens
add at the carbon carbon double bond.
Hanes method is used for the determination of Iodine number. About 1 gram of the fat is taken in a well
cleaned dry iodine flask. To this 20 ml of chloroform is added to dissolve the fat. The contents are
shaken well and kept for 30 minutes. Then 20 ml of potassium iodide is added to liberate the iodine and
it is titrated against standard sodium thio cyanate solution. From this titration iodine number of fat is
calculated.
Reagents required
i.
Hanes solution
ii.
Fat
iii.
Standard 0.1 N potassium dichromate solution
iv.
Sodium thio cyanate solution
v.
10 % potassium iodide
vi.
1 % Starch
vii.
Chloroform
Preparation of Hanes solution

3.3 grams of iodine is dissolved in 200 ml of acetic acid by constant shaking and heating. It is cooled to
room temperature, to this 50 ml of glacial acetic acid containing 0.75 grams of Bromine is added and
mixed well and stored in brown bottle.
Page 23
Titration I
Standard potassium dichromate solution Vs Sodium thio cyanate solution
Weighed accurately 1.225 g of potassium dichromate solution and transfer into a 250 ml of
standard flask then the volume is made up to 250 ml using distilled water. Pipette out exactly 20 ml of
this solution into a clean conical flask to this 5 ml of Conc. Hydrochloric acid is added, followed by 10 ml
of 10 % potassium iodide is added. This contents are mixed well and titrated against the Sodium thio
cyanate solution taken in the burette, the titration is continued until a pale brown colour is appears. At
the time 1 ml of 1 % Starch solution is added. And the titration is continued till to get the end point
appearance of emerald green colour, it is the end point. The titrations are repeated for concordant
values. From the titre value the normality of Sodium thio cyanate solution is calculated.
Titration II
Determination of iodine no of fat (Test value)
Weigh about one gram of edible oil and carefully transfer in to a clean dry iodine flask. Then
20 ml of Chloroform is added, the contents are mixed well to dissolve the oil. To this 20 ml of Hanes
solution is added, shaken well and kept in dark for 30 minutes with occasional shaking. Then the flask is
taken out to this 20 ml of 10 % potassium iodide is added to liberate iodine. Except the iodine that is
absorbed by the oil. To this mixture 100 ml of distilled water is added, so the liberated iodine is nicely
disturbed in the solvent then it is titrated against the Sodium thio cyanate solution taken in the burette,
the titration is continued until a pale brown colour is appears. At the time 1 ml of 1 % Starch solution is
added. And the titration is continued till to get the end point disappearance of blue colour it is the end
point.
Blank value
Blank value is also done without oil
Equivalent weight of iodine
127
Page 24
Titration II
Determination of iodine no of fat (Test value)
Burette readings
Titration
Volume of Sodium
Contents taken in the conical flask

Initial (ml)
Final (ml)
thio cyanate solution

(ml)
-------------- gram of Oil + 20 ml of

chloroform + 20 ml of Hanes Solution
Test
0 ml
+ 100 ml of distilled water + 20 ml of
10 % Potassium iodide + 1 ml of 1 %
Starch
Blank
20 ml of chloroform + 20 ml of Hanes
Solution + 100 ml of distilled water +
0 ml
20 ml of 10 % Potassium iodide + 1 ml
of 1 % Starch
Weight of the oil

Weight of the weighing bottle + Oil
Weight of the weighing bottle
Weight of the Oil transferred
Blank value - Test value
(-)
-------------------- ml
Page 25
Determination of iodine no of fat
Equivalent weight of iodine x Blank value - Test value x Normality of Sodium thio cyanate solution x 100
1000 x Weight of the Oil
= ---------------------------------- Iodine no of fat (oil)
Result
Iodine No of the given Fat (Oil)
------------------------------
Page 26
Analytical contents of oils and fats
OILS
ACID VALUE
SAPONIFICATION VALUE
IODINE VALUE
Coconut oil
5 13
250 264
8.0 9.5
Sesame oil
1 10
188 193
103 117
Castor oil
0.3 4
178 188
80 88
Linseed oil
18
190 196
170 203
Ground nut oil
26
186 196
83 105
0.4 2
220 241
26 38
Ghee (cow)
225.5 236
31.5 45
Ghee (Buffalo)
228.5 236
26.5 - 44
Butter
Page 27

Volumetric Analysis - Glycine Acid NO Io PDF

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FOR THE STUDENT OF IST B. Sc BIOCHEMISTRY

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

ESTIMATION OF AMINO ACID (GLYCINE) BY SORENSONS FORMAL TITRIMETRIC METHOD

Volume of standard oxalic

Volume of Standard oxalic acid solution

Normality of Standard oxalic acid solution

Volume of Sodium hydroxide solution

Normality of Sodium hydroxide solution

Normality of Sodium hydroxide solution

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

ESTIMATION OF AMINO ACID (GLYCINE) BY SORENSONS FORMAL TITRIMETRIC METHOD

Standard oxalic acid solution (0.1 N)

Sodium hydroxide solution

Amino acid (Glycine)

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

Titration II (Test value)

Contents taken in the

Volume of Sodium hydroxide consumed by amino acid (Glycine)

Titration III (Blank value)

Contents taken in the

Volume of Sodium hydroxide consumed by formaldehyde (Blank value) =

Volume of Sodium hydroxide consumed by amino acid alone

Test value Blank value

Titration II (Test value)

into a clean conical flask to this 5 ml of

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

Normality of Sodium hydroxide solution

Volume of amino acid (Glycine) solution

Normality of amino acid (Glycine) solution

Normality of amino acid (Glycine)

Equivalent weight of Glycine

Normality of amino acid (Glycine) x Equivalent weight of Glycine x 100

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

Test value Blank value

Equivalent weight of Glycine

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

ESTIMATION OF SUGAR (GLUCOSE) BY BENEDICTS TITRIMETRIC METHOD

250 mg of Glucose/250 ml of distilled water

5 ml of Benedicts reagent consumes --------------------------- ml of Standard Glucose solution

5 ml of Benedicts reagent consumes --------------------------- ml of unknown Glucose solution

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

ESTIMATION OF SUGAR (GLUCOSE) BY BENEDICTS TITRIMETRIC METHOD

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

Volume of standard Glucose

Volume of unknown Glucose

ie. --------------------- ml of standard Glucose

100 ml of unknown solution contain

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

Excess amount Sodium carbonate will cause frothing

Benedicts quantitative Reagent is kept at boiling throughout in the titration

Porcelain bits are added to prevent bumping

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI

ESTIMATION OF ASCORBIC ACID USING 2, 6 DICHLORO PHENOL INDOPHENOL

Ascorbic acid (ml)

Volume of the Dye

10 ml of standard ascorbic acid consumes

------------------------ ml of the dye

Ascorbic acid (ml)

Volume of the Dye

X - mg of ascorbic acid consumes

------------------------ ml of the dye

COMPILED BY Dr. A.LIYAKATH ALI & Mr. V. MAGENDIRA MANI