EXPERIMENT NO. 3
EXTRACTION AND ISOLATION OF PECTIN FROM CITRUS PEELSIntroductionPectins are polysaccharides composed mainly of polymers of D-galacturonic acid whose monomers are joined by (1,4)-linkages. They also contain arabinose and galactose. The carboxyl group of galacturonic acid can be esterified to varying extents with methanol. The hydroxyl groups in the 2nd and 3rd positions menwhile can be acetylated to a lesser extent.
In strong acidic medium, the glycosidic linkages are hydrolyzed. Pectin is most stable at pH 3-4. In basic medium, the ester and glycosidic linkages are split to the same extent. The latter is eliminated because the hydrogen atom in theC-5 position is more acidic compared to the galacturonic acid with free carboxyl groups.
The degree of esterification is determined by the saponification of pectin followed by titration of the free carboxyl acid groups. The volumes obtained is used to calculate % methoxy content which is equal to the total mass of methoxy over the mass of the pectin sample multiplied by 100.
ProcedureA. Isolation
B. Determination of methoxy content
air-dried pectin weigh not more than 0.25 g + 2.5 ml EtOH + 0.5 g NaCl + 100 ml CO2-free distilled water + 6 drops phenol red |
| yellow solutiontitrate with 0.1 N NaOH |
pink solution(V1)+ 12.5 ml 0.25 N NaOH, shake stand for 30 minutes at room temperature in stoppered flask + 0.25 N HCl dropwise |
yellow solution titare with 0.1 N NaOH |
| pink solution (V2) |