Method for the estimation of Total Polyphenols content

Method for the estimation of Total Polyphenols content 

Reagent Required:

Distilled water

Folin –Ciocalteau reagent

Sodium carbonate

Gallic acid Standard

Glass wares, Apparatus & Instruments required:

Volumetric Flasks          100mL, 50mL, 10mL

Pipettes –                     5mL, 2mL,1ml

U-V Spectrophotometer

STANDARD PREPARATION:

Weigh accurately about 50mg of Gallic acid standard into a 50mL volumetric flask. Add about 50mL of distilled water.Ultrasonicate till completely dissolved and make up the volume with water.(1000µg/ml)(Stock solution)

Transfer 1 ml of Stock solution into a 100 ml volumetric flask and dilute Upto the mark with distilled water.  

from the above solution,transfer 1,2,3,4,& 5 ml to 10 ml of volumetric flask, add 1ml of 1N Folin-Ciocalteau reagent and 2ml sodium carbonate solution and dilute upto the mark with distilled water.

SAMPLE PREPARATION:

Weigh accurately about 500mg of sampled hydrolysed with 10 ml dil. HCL for 1 hour.

Transfer to 100 ml of volumetric flask and dilute upto mark with distilled water and filter.

Transfer 1 ml of the above solution to 10 ml of volumetric flask, add 1ml of 1N Folin-Ciocalteau reagent and 2ml sodium carbonate solution and dilute upto the mark with distilled water.

Mix well and measure the absorbance  after 30 mins at 750nm.

                                          Absorbance of sample X Concentration of std X   purity of std

% of Polyphenols =----------------------------------------------------------------------------------------------

                                             Absorbance of standard X Concentration of sample