Analysis of bioactive compounds

Total fenolic content

Total phenolic
 Phenolic compounds are important plant
constituents with redox properties responsible
for antioxidant activity
 The hydroxyl groups in plant extracts are
responsible for facilitating free Plants radical
scavenging.
 As a basis, phenolic content was measured
using the Folin–Ciocalteu reagent in each
extract
Materials
 Apparatus and Glass ware
 Spectrofotometer (single beam or double beam)
 Cuvete
 Analytical balance
 Test tube
 Vortex
 Spatula
 Chemicals
 Sodium carbonate
 Folin–Ciocalteu reagent
 Metanol pro analysis
 aquadeionized
Procedure
 Make a mother solution with concentration of extract
approximately 100–500 µg/mL
 One mL of extract (100–500 µg/mL) solution was mixed with
2.5 mL of 10% (w/v) Folin–Ciocalteu reagent.
 After 5 min, 2.0 mL of Na2CO3 (75%) was subsequently
added to the mixture and incubated at 50 °C for 10 min with
intermittent agitation. Afterwards, the sample was cooled
 the absorbance was measured utilizing a UV
Spectrophotometer at 765 nm
 against a blank without extract.
 The outcome date were then calculated at standard curve of
gallic acid that also measured with Folin–Ciocalteu method.
 The outcome data were expressed as mg/g of gallic acid
equivalents in milligrams per gram (mg GAE/g) of dry extract.
Calculation
 The calculation is based on the utilization of linear regression
equation.
 First, develop a graph that consist of gallic acid concentration
(X) and gallic acid absorbance (Y)
 Then find out the regression linear equation.
 Y = aX+b
 The linearity of the measurement should be in the range of
0.98 to 1. if less than 0.98 you should repeat the
measurement.
 Then put the absorbance of your sample into the obtained
equation as Y.
 You will get the X and express as the data of phenolic content
(the data is expressed as mg/g of gallic acid equivalents in
milligrams per gram (mg GAE/g) of dry extract)