FOL-02

METHOD FOR THE ESTIMATION OF TOTAL PHENOLIC CONTENT USING THE FOLIN- C ASSAY

bath at 80 0 C for 5min. Decent the supernant in to 100mL volumetric Flask containing previous solution. Allow to cool the solution and make up to the mark with methanol (Solution-A). Pipette out 2.0mL of Solution-A and transfer it into 25mL volumetric flask Make up to the mark with methanol (Solution-B) Further pipette out 1mL from Solution-B in to a 15mL test tubes and add 5 ml of Folin-Ciocalteu’s phenol reagent followed by 4 mL of 7.5% Sodium Carbonate solution. Close & Keep the test tubes in dark condition up to 1hour for colour development. Take the absorbance at 765nm against Blank using UV-Visible spectrophotometer. • Extract containing 15% Polyphenols: Weigh accurately 150-200mg of Sample extract in 50mL volumetric flask and add 25mL of methanol , Sonicate for 10min and make up to the mark with Methanol(Solution-A) Pipette out 2.0mL of Solution-A and transfer it into 25mL volumetric flask. Make up to the mark with methanol (Solution-B). Further pipette out 1mL from Solution-B in to a 15mL test tubes and add 5 ml of Folin-Ciocalteu’s phenol reagent followed by 4 mL of 7.5% Sodium Carbonate solution. Keep the test tubes in dark condition for 1hour for colour development. Take the absorbance at 765nm against Blank using UV- visible spectrophotometer.

8.0 Calculation: Total Phenolic content in Salacia Species is calculated by below mentioned formula,

A Sample

D standard

% Purity of Standard

%Total phenolic content = ---------- x ----------- x A Standard D sample

Where: A Sample A Standard D sample D standard

= Absorbance of sample at 765nm = Absorbance of standard at 765nm = Dilution of sample in mg per mL = Dilution of standard in mg per mL

9.0 Controls (Precaution):

• Prepared Folin-Ciocalteu’s phenol reagent can be used for 15 days if it stored under dark condition.

• Sodium carbonate solution should be prepared freshly and used.

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