Total phenolic content
Totalphenolic compounds (TPC)were determined according
the protocol usedbyMuanda et al. (2011). 40 μLof samples
was addedto test tubes containing 3.16 mL of distilled water
followed by an addition of 200 μLof Folin-Ciocalteu reagent
(1 N) and 600μL of sodium carbonate (Na2CO3,20%).
Samples and blank were thoroughly mixed and vortexed.
After 40 min of incubation at room temperature, the absor-
bance was measured at 725 nm. For the calibration curves,
dilutedsolutions of gallic acid (solutions at 1mg/mL, 1.5mg/
mL, 2mg/mL, 2.5mg/mL and3mg/mL) were used andtotal
phenolic content was expressed in terms of equivalent
amounts of gallic acid per 100 g of dry matter (GAE/100 g
DM). Absorbance measurements were carried out with UV–
visiblespectrophotometer Cary 50 Scan(Agilent,SantaClara,
California, USA).
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