IGNOU BCHEL -150

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University of Calcutta
B.Sc. Chemistry (Hons.) Semester-3 (CBCS) CC-7P

B. Quantitative Estimations

1. Estimation of glycine
Determination of Theby Sörensen formol
Concentration methodSolution by Formylation Method
of Glycine
Principle:

In aqueous solution of amino acid, H2NCH(R)COOH, the following equilibrium exists:

The dipolar ion (zwitterions) structure accounts for the absence of any acidic or basic property of
aqueous solution of amino acid. For this reason amino acids cannot be titrated directly with an
alkali. But in presence of neutralized formalin solution, amino acids behaves as strong monoprotic
acids and can be quantitatively titrated with standard alkali. Formalin stabilises the amino (-NH2)
group by forming the Schiff base linkage (-N=CH-), as a result, zwitterions cannot be formed and
the carboxylate group of the amino acids can be titrated with a strong base (NaOH) using
phenolphthalein indicator.

But the reaction is more complex. As for example, formaldehyde reacts with-NH2 group of
glycine to form dimethylol glycine, which behaves as a strong monoprotic acid,

In either way,

1000 ml of (N) NaOH ≡ 1 g. equivalent of glycine ≡ 75 g. of glycine ≡ 1 F.W. of amino acid.

Working Formula:
Equivalent weight of oxalic acid (C2H2O4.2H2O) =126/2 g. = 63 g.
Thus,
1000 ml of (N) oxalic acid soln. ≡ 63.0 g of oxalic acid
 250 ml of (N/20) oxalic acid soln. ≡ 0.7875 g oxalic acid
𝑤 𝑁
 Strength of oxalic acid soln. = 0.7875 (20) = S1 (N) (w = weight taken)
So, Strength of NaOH solution = S (N)

Say, 25 ml of amino acid (say glycine) solution ≡ V ml of NaOH solution

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Now, 1000 ml of (N) NaOH ≡ 75 g. of glycine

V ml of S (N) NaOH solution ≡ 0.075 × V × S g of glycine / 25 ml soln.

 Strength of glycine solution = (0.075 × V × S × 40) g/lit.

= (3 × V × S) g/lit.

Chemicals required:

(i) Standard (N/20) oxalic acid solution: To be prepared by accurate weighing.

(ii) (~N/20) NaOH solution: ~ 0.5 to 0.6 g of (A. R.) NaOH per 250 ml solution.

(iii) Formalin solution: 40% aqueous solution of formaldehyde: To be neutralized with (~N/20)
NaOH using phenolphthalein indicator prior to estimation of glycine.

(iv) Phenolphthalein indicator: (0.4 ¬ 0.5)% solution in 1:1 alcohol.

(v) (~N/20) Glycine solution (unknown): 3.75 g.lit-1. Or, dissolve 0.9 ¬ 1.0 g of glycine in distilled
water and dilute to 250 ml in a volumetric flask.

Procedure:

1. Standardisation of (~N/20) NaOH solution against standard (N/20) oxalic acid solution
using phenolphthalein indicator.

2. Estimation of glycine with standard (~N/20) NaOH solution:

Pipette out 25 ml of glycine solution into a 250 ml conical flask, Add 25 ml of the neutralized
formalin solution and 2-3 drops of phenolphthalein indicator. Titrate the mixture with standard
(~N/20) NaOH solution till endpoint (colourless to pink). (Titrate slowly near the end point adding
the titrant NaOH dropwise with constant shaking).

Experimental Results:

(A) Preparation of 250 ml of standard (N/20) oxalic acid solution

Table 1
Initial wt. of Final wt. of Wt. of oxalic Wt. of oxalic Strength of oxalic acid
oxalic acid oxalic acid acid taken acid to be taken soln.
gm gm gm gm
W (N) = S1 (N)
W 0.7875 0.7875 20

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(B) Standardisation of NaOH soln. against standard (N/20) oxalic acid solution
Table 2
No. Vol. of Strength Burette reading Vol. of Mean vol. Strength of
of std. oxalic of oxalic NaOH soln. of NaOH NaOH
obs. acid soln. acid Initial Final reqd. soln soln.
ml soln. ml ml ml ml
1 25 S1 (N) S (N)
2 25 x

(C) Estimation of glycine with standard (N/20) NaOH solution

Table 3
No. Vol. of Vol. of Burette reading Vol. of NaOH Mean vol. of
of glycine soln. neutral soln. reqd. NaOH soln.
obs. Formalin Initial Final
soln.
ml ml ml ml ml ml
1 25 25 V
2 25 25

Calculation:

Strength of glycine solution = (0.075 × V × S × 40) g/lit.

= (3× V × S) g/lit.

2. Estimation of glucose by titration using Fehling’s solution

Principle:

Glucose is a reducing sugar. Fehling’s solution is composed of (i) copper(II) sulfate

(Fehling’s solution A) and (ii) sodium potassium tartarate rendered strongly alkaline with NaOH
(Fehling’s solution B). A mixture of equal volumes of these two solutions is the actual regent.

Under boiling condition, glucose is quantitatively oxidized by Fehling’s solution to

gluconate with precipitation of red cuprous oxide:

CHO COO
H OH O CHCOO Na H OH HO CHCOO Na
OH
HO H + 2Cu HO H +
O CHCOO K H2O
+ Cu2O
H OH H OH HO CHCOO K
H OH H OH
CH2OH CH2OH
D-glucose

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 3rd Semester, Organic Chemistry

2. Determination of Saponification Value of an Oil or Fat

Estimation of Saponification value of Oil/ Fat/Ester

[I] Principle

Fats and oils are esters of long chain fatty acids and glycerol called triglycerides occurs in
natural products (like seeds , nuts etc). The basic difference between fats and oils is that fats
are solid at room temperature while oils are liquid at room temperature.

The alkaline hydrolysis of esters/ fats/oils is called saponification. Saponification numbers is

defined as the numbers of milligrams of caustic potash (KOH) required to saponify 1.0 gm of
esters/ fats/ oils.

The saponification reaction is shown below

Thus, the saponification value can be determined by completely hydrolysis a known weight
of esters/ fats/oils with the measured excess of standard alcoholic KOH solution. The
unreacted KOH is back titrated with standard oxalic acid solution, KOH consumed for
hydrolysis can be calculated and thus saponification value is obtained.

[II] Reagents Required

(i) Supplied sample =Mustered oil(supposed)

(ii) 250 mL (N/2) Oxalic acid solution
(iii) 250 mL (N/2) alcoholic KOH solution
(iv) 250 mL (N/2) HCl solution

[III] Preparation of Reagents

(i) 250 mL (N/2) Oxalic acid solution

7.879 g of oxalic acid(accurately weighted) is transferred from weighing bottle to 250 mL volumetric
flask which is then dissolved and diluted to 250mL

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(ii) 250 mL (N/2) alcoholic KOH solution

Approximately, 7 gm of KOH is transferred to a 250 mL volumetric flask and dissolved in 50%
aqueous alcohol and diluted to 250mL adding 50% aqueous alcohol.
(iii) 250 mL (N/2) HCl solution
10.5 mL of conc. HCl is added to 200 mL of distilled water taken in a volumetric flask then diluted
250 mL by adding water.
[IV] Experimental Section

[A] Standardization of (N/2) KOH soln. against standard Oxalic acid soln

10 mL of (N/2) KOH(alcoholic) solution is transferred into a 250 mL conical flask and 50 ml of

distilled water is added followed by 2 drops of phenolphthalein indicator is added to it.
Then it is titrated against (N/2) oxalic solution from the burette and the end point is
indicated by the colour changes from pink to colourless.

[B] Standardization of (N/2) HCl against standardized (N/2) KOH solution

10 mL of (N/2) KOH(alcoholic) solution is transferred into a 250 mL conical flask and 50 ml of

distilled water is added followed 2 drops of phenolphthalein indicator is added to it. Then it
is titrated against (N/2) HCl solution from the burette and the end point is indicated by the
colour changes from pink to colourless.

[C] Blank Titration

Then 25 mL (by using pipette) of alcoholic solution of (N/2) KOH is taken in a 250 mL conical
flask and 50 ml of distilled water is added to it followed by the solution is titrated against
standardized (N/2) HCl solution using phenolphthalein indicator ( at end point colour
changes from pink to colourless). The volume of HCl is recorded as (VHCl/B) V3.

[D] Saponification of Oil

Approximately, 2 gm (accurately weighted) of mustered oil sample is taken in a 250 mL

standard joint round bottomed flask fitted with air/ water condenser. Then 50 mL of
alcoholic solution of KOH is added (by using pipette) to the oil and the condenser is fitted.
The mixture is then heated on a boiling water bath until the oil is completely saponified
(indicated by the disappearance of droplets of oil over the solution). The saponified solution
is then cooled at room temperature and titration of excess KOH is performed with the
standardized (N/2) HCl solution taken in a burette using phenolphthalein as indicator until
the pink colour is just discharged (colourless). The volume of HCl consumed is recorded as
(VHCl/S) V4.

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[V] Results
Table 1 Weight of Mustard oil used in the experiment

Initial Weight( W1) Final Weight (W2) g Weight taken (W1-W2)= W g

8.531 6.530 2.01g

Table 2 Preparation of 250 mL (N/2) standard Oxalic acid solution

Initial Weight( W1) Final Weight (W2) g Weight taken (W1-W2)= W g

28.082 20.156 7.926

Table 3 Standardization of (N/2) alcoholic KOH soln. against standard Oxalic acid solution

No. of Titration Volume of Burette Reading KOH Volume of Mean

Oxalic acid solution (mL) KOH Volume
taken (mL) required of KOH
Initial Final (mL) (mL)

1 10 0 12.5 12.5 V1 =12.5

2 10 12.5 25 12.5

3 10

Table 4 Standardization of (N/2) HCl against standardized (N/2) alcoholic KOH solution

No. of Volume of Burette Reading KOH Volume of Mean

Titration HCl taken solution (mL) KOH required Volume of
(mL) (mL) alcoholic
Initial Final KOH
(mL)
1 10 0 11.8 11.8 V2=11.8
2 10 11.8 23.8 11.8

Table 5 Blank titration of (N/2) alcoholic KOH soln. against standardized (N/2) HCl solution

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No. of Titration Volume of Burette Reading(HCl) mL Volume Mean

alcoholic KOH of HCl Volume
solution taken Initial Final required of HCl
(mL) (mL)
(mL)

1 25 0 22.5 22.5 (V3) =22.45

2 50 22.5 44.9 22.4

3 50

Table 6 titration of saponified soln. by standardized (N/2) HCl solution

No. of Titration Volume of Burette Reading(HCl) mL Volume Mean

Saponified of HCl Volume
solution(Oil+Alc. Initial Final required of HCl
(mL) (mL)
KOH) taken (mL)

1 25 0 7.9 7.9 (V4) =7.9

2 25 7.9 15.8 7.9

[VI] Calculation

[A] Weight of Oil taken = W g=2.01g

[B] Strength of standard Oxalic Acid=S1 (N) = Wox / 7.879 (N/2)=0.503N
[C] Strength of alcoholic KOH solution= S2 (N)= S1 x 10/V1 (N)
=(0.503x10)/12.5=0.4N
[D] Strength of standard HCl solution= S (N)= S2 x V2 / 10 (N)
=(0.4x11.8)/10=0.47(N)
[E] Blank titration HCl for 25 ml KOH=22.45 ml
[F] titre value for saponified = 7.9ml
Amount of KOH consumed by 2.01 g standard oil=(22.45-7.9)ml 0.47(N) of HCl
=(22.45-7.9)x 0.47 ml (N) of HCl
=(22.45-7.9)x 0.47 ml (N) of KOH solution
Since , 1000 ml (N) KOH solution=56.1 g of KOH
(22.45-7.9) x 0.47 ml (N) of KOH solution≡0.0561x(22.45-7.9) x 0.47g of KOH

჻ 2.01g oil≡0.0561 x (22.45-7.9) x 0.47 g of KOH≡ [56.1 x (22.45-7.9) x 0.47] mg

of KOH
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 3rd Semester, Organic Chemistry

჻ 1g of oil ≡ [56.1 x (22.45-7.9) x 0.47]/2 mg of KOH

=191.8

Saponified value of the mustered oil=191.8

Saponification values of some common Fats/ Oils

Fats/ Oils Saponification values Fats/ Oils Saponification values
Butter 220-233 Mustered Oil 188-193
Corn Oil 188-195 Olive Oil 185-195
Coconut Oil 269-271 Palm Oil 196-205
Cotton Seed Oil 246-260 Peanut Oil 190-196
Linseed Oil 192-195 Rapeseed Oil 170-179

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