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Estimation of Pectin As Calcium Pectate by Gravimetric Method

This document outlines a gravimetric method for estimating pectin content in fruits and vegetables as calcium pectate. Pectin is extracted from plant material using acid and alkali solutions, then precipitated as calcium pectate by adding calcium chloride. The calcium pectate precipitate is filtered, dried, and weighed to calculate the percentage of pectin and protopectin in the original sample.

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Samiksha Singh
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444 views2 pages

Estimation of Pectin As Calcium Pectate by Gravimetric Method

This document outlines a gravimetric method for estimating pectin content in fruits and vegetables as calcium pectate. Pectin is extracted from plant material using acid and alkali solutions, then precipitated as calcium pectate by adding calcium chloride. The calcium pectate precipitate is filtered, dried, and weighed to calculate the percentage of pectin and protopectin in the original sample.

Uploaded by

Samiksha Singh
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Exercise No.

: __________ Date: _____________

Estimation of Pectin as Calcium Pectate by Gravimetric method

Test Sample : ____________________.

Objective : To understand the process for estimation of pectin content in fruits and
vegetables.

Relevance : Pectic substances abundantly exist in the middle lamella of the plant cells.
There are three types of pectic substances – pectic acids, pectinic acids and
protopectin. Pectic acid is an unbranched molecule made up of about 100
units of D-galacturonic acid residues. The monomers are linked through ∞-
1, 4- glycosidic linkage. Pectinic acid is an extensively esterified pectic acid.
Several carboxyl groups exist as methyl esters. Pectic acid is water soluble
whereas pectinic acid forms a colloidal solution. Protopectin is a larger
molecule than pectic and pectinic acid. During ripening of fruits, conversion
of protopectin into pectic acid and pectinic acid takes place. The pectinic
acids in fruits vary in their methoxyl content and in jellying power.

Principle : Pectin is extracted from plant material and saponified with alkali. It is
precipitated as calcium pectate by the addition of calcium chloride to an acid
solution. The calcium pectate precipitated is washed thoroughly to eliminate
chloride ions, dried and then weighed.

Reagents : 1. 1 N Acetic acid: Dilute 30 ml of glacial acetic acid to 500 ml with


distilled water.
2. 1 N Calcium chloride solution: Dissolved 27.5 g anhydrous CaCl2 in
distilled water and dilute to 500 ml.
3. 1% Silver Nitrate: Dissolve 1 g AgNO3 in 100 ml distilled water.
4. 0.01 N HCl
5. 0.05 N HCl
6. 0.30 N HCl
7. 1 N NaOH
8. Whatman No. 1 filter paper

Procedure : (a) Weigh 50 g of blended sample into a 1 L beaker and add 300 ml 0.01 N
HCl. Boil for 30 min and filter. Wash the residue with hot water and
collect the filtrate.
(b) To the residue add 100 ml 0.05 N HCl; boil for 20 min, filter, wash and
collect the filtrate.
(c) To the residue now add 100 ml 0.3 N HCl; boil for 10 min, filter, wash
and collect the filtrate.
(d) Pool the filtrates. Cool and make up the volume to 500 ml.
(e) Pipette out 200 ml aliquot into 1 L beaker.
(f) Add 250 ml distilled water and neutralize the acid with 1 N NaOH using
phenolphthalein indicator. Add an excess of 10 ml 1 N NaOH with
constant stirring and allow it to stand overnight.
(g) Add 50 ml 1 N acetic acid and after 5 min, add 25 ml 1N calcium
chloride solution with stirring. Allow it to stand for 1 hour.
(h) Boil for 1-2 min.
(i) Filter through a pre-weighed Whatman No. 1 filter paper (Wet a
Whatman No. 1 filter paper in hot distilled water, keep it in a covered
Petri dish, dry in oven at 102oC for 2 h. Cool in a desiccator and weigh).
(j) Wash the precipitate with almost boiling water until the filtrate is free
from chloride.
(k) Take the filtrate in a test tube and test with silver nitrate for chloride.
(l) Once the filtrate is free of chloride, transfer the filter paper containing
the calcium pectate to the original Petri dish, dry overnight at 100o C,
cool in a desiccator and weigh.

Work Sheet : (a) Wt. of the Petri dish (g) =


(b) Wt. of the Petri dish + dried filter paper (g) =
(c) Wt. of the dried filter paper (g) = (b) - (a) =
(d) Wt. of the Petri dish + dried filter paper containing Ca-pectate (g) =
(e) Wt. of the dried filter paper containing Ca-pectate (g) = (d) - (a) =
(f) Wt. of the Ca-pectate = (e) - (c) =

Calculation : Wt. of Calcium Pectate (f) x Vol made up (500) x 100


% Calcium Pectate =
ml of filtrate taken (200) x Wt. of sample for estimation

% Calcium pectate
% Protopectin =
1.10

Results : % Calcium Pectate =

% Protopectin =

Conclusion :

Remarks :

Signature :

*The empirical formula for calcium pectate is C17H22O16Ca. The calcium pectate yield of
highly purified pectinic acid is usually about 110% of the pectinic acid.

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