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METHOD FOR THE ESTIMATION OF TOTAL PHENOLIC CONTENT USING THE FOLIN-
C ASSAY
Page
2
of
5
•
Water Bath (Make: PCI analytics)
•
Weighing Balance (Make: Sartorius BSA224S-CW)
•
100 mL Volumetric Flask (Make: Borosil Class-A)
•
50 mL Volumetric Flask (Make: Borosil Class-A)
•
25 mL Volumetric Flask (Make: Borosil Class-A)
•
15 mL Test tubes (Make: Borosil Class-A)
•
10mL,5 mL, 2mL, 1mL Pipettes (Make: Borosil Class-A)
6.0
Calibration:
All Equipments and glassware’s used for performing the analysis should be
calibrated.
7.0
Procedure:
7.1.
Preparation of reagents:
•
10% Folin-Ciocalteu’s Phenol reagent:
Pipette out 10mL of Folin-Ciocalteu’s
phenol reagent in 100 mL Volumetric Flask and Make up to volume with water
and Sonicate & stored under dark. This solution can be stored for 15 days.
•
7.5% Sodium carbonate Solution:
Weigh accurately 7.5gm of sodium
carbonate in 100mL volumetric flask , Add 50mL of Water , Shake and Sonicate
till it dissolve , then make up the volume up to the mark with water.
7.2.
Preparation of Blank:
•
Take 1mL of Water in to 15mL test tube and add 5 ml of 10%Folin-Ciocalteu’s
phenol reagent and add 4 mL of 7.5% Sodium Carbonate solution. Keep the test
tubes in dark condition for 1hour.This blank solution should be used for auto
zeroing the UV-Visible spectrophotometer at 765nm.
7.3.
Preparation of Standard Solution:
•
Weigh accurately 50mg of Gallic acid in to a 50mL volumetric Flask, add 25mL
of Methanol and Sonicate for 5 min & make up the volume up to the mark with
methanol
. (Stock –A Concentartion1000ppm).
Pipette out solution from stock-A and prepare a Four different concentration
ranges from 5-100ppm and plot the linearity graph. The R
2
value (Co-relation
coefficient) should be greater than 0.99.
7.4.
Preparation of Sample:
•
Raw Material Solution :
Weigh accurately 2.00gm of Powdered sample passed
through 40mesh (425µm) in Round bottom flask and add about 50mL of
Methanol and Sonicate for 10min, followed by Reflux in water bath at 80
0
C for
5min using Air condenser. Decent the upper layer in to 100mL Volumetric
Flask. Again add 40mL of methanol in round bottom flask & Reflux in water