I have done TFC. The procedure is as follow:
# I dissolved 1 mg extract in 1 ml MeOH.
# 100 ul of extract + 100 ul of 2% AlCl3 was mixed in 96 well and incubate for 15 min.
# Absorbance was taken at 435 nm.
# Quercetin was used as standard.
Now the average absorbance was 1.1795 after deducting the abs of blank. The calibration curve equation is y= 0.0106 x + 0.0052 and R square = 0.9995. When I calculate the TFC in equation C = cV/M, the value was 110800 mg/g. This looks weird and I may made some mistake. Can anyone help me to calculate it properly?
First, to help you in the calculation, you should give more details about the calibration curve equation (the concentration against the absorbency or vice versa), the units used for the axis, and finally have you followed exactly the same protocol for the standard solutions of different concentrations ? I mean if you have taken 100µl of each quercetin concentration and followed exactly the other steps?
yes I followed the same protocol for standard. the concentration for standard was 10 mg/ml, 5 mg/ml, 2.5 mg/ml, 1.25 mg/ml, 0.625 mg/ml. X axis was concentration and Y was abs.
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