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METHOD FOR THE ESTIMATION OF TOTAL PHENOLIC CONTENT USING THE FOLIN-
C ASSAY
Page
4
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10.0
Results of validation:
Following test were conducted to verify the performance of the
method as per the requirement of
AOAC SMPR 2015.009.
10.1.
Analytical Range:
Analytical range was demonstrated on reference standard.
Different concentrations ranging from 5-100ppm were prepared and measure the
absorbance.
Sr.no.
Concentration (ppm)
Absorbance
1
5
0.101
2
25
0.325
3
50
0.651
4
100
1.273
Conclusion:
The Co-relation coefficient (R
2
) of 0.999 confirms that the concentrations
ranges are linear and follows the Beers lambert’s laws.
10.2.
Limit of Quantification (LOQ):
Based on Linearity study a concentration of 5
ppm is found to be the LOQ. Six different preparation of 5ppm were analyzed and the
relative standard deviation (RSD) is within 10%. Confirming LOQ to be 5ppm
10.3.
Recovery:
Recovery study was conducting by adding known amount (80 to
120% of working range) of standard in both the raw material & extract. The samples
were analyzed using the methodology mentioned above (7.4). The recovery is within 80
to 110%.
y = 0.012x + 0.028
R² = 0.999
0
0.2
0.4
0.6
0.8
1
1.2
1.4
0
50
100
150
Absorbance
Concentartion in ppm
Calibration Curve of Gallic acid
Abosrabance