Total phenolics:

Following Julkenen-Titto (1985), fresh leaf 0.5g was ground in 10ml of 80% of methanol. Centrifugation was done at 10,000rpm for 10 minutes.  Then 0.25ml of plant extract, along with 1.25ml of Folin-Phenol Ciocalteus reagent was mixed. The addition of 1.25ml of sodium carbonate was the last step. The total phenolics in leaf samples were anticipated at an absorbance range of 750nm with the help of a spectrophotometer.

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