How we performed the estimation of total flavonoid content ?

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This time I am sharing a method for the estimation of total flavonoid content in our experiments. We
Flowchart. the process of estimating total flavonoids content.
followed a previously done standard protocol (Quettier et al. 2000 and Stankovic 2011) with slight modification. 

Firstly, we mixed each plant extract (2 ml; 0.5 mg/ml) separately  with aluminium chloride
solution (1 ml; 2 %). Then, we allowed the reaction mixture to stand for an hour at room temperature. Again, we measured the absorbance of the mixture at 415 nm using the UV-spectrophotometer. We obtained calibration curve (see Fig 1.) using quercetin solution series (25, 50 75, and 100 µg/ml) as a standard. Here, we used the reagent mixture with the same volume of ethanol instead of plant extract solution as a blank. Thus, the obtained curve was calibrated to estimate the concentration of total flavonoid in each plant extract. The total flavonoids content was expressed in terms of the milligram of quercetin equivalent per gram of the dry mass (mg QE/ g). The test was done in triplicates.


 References: 

  1. Quettier D C., Gressier B., Vasseur J., Dine T., Brunet C., Luyckx M C., Cazin M., Cazin J C., Bailleul F., Trotin F. 2000. Phenolic compounds and antioxidant activities of buckwheat (Fagopyrum esculentum Moench) hulls and flour. Journal of Ethnopharmacology 72: 35-42.
  2. Stanković M. S. 2011. Total phenolic content, flavonoid concentration and antioxidant activity of Marrubium peregrinum L. extracts. Kragujevac Journal of Science 33(2011): 63-72.

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