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Table 4 Assay and method validation parameters for the determination of flutamide and its metabolites in pure samples by the proposed methods

From: Determination of flutamide and two major metabolites using HPLC–DAD and HPTLC methods

Parameters

HPTLC method

HPLC method

Pure samples

Pure samples

Flutamide

Flu-1

Flu-3

Flutamide

Flu-1

Flu-3

Rangea

0.1–3

0.3–2.5

0.3–3.5

2–50

1–50

5–200

Slope

− 645.07b

4902.40c

− 2070.03c

13121d

− 101.45c

825.60d

43.6020

34.9780

6.4161

Intercept

464.92

2938.80

1275.60

− 17.9250

− 5.1212

5.4658

Correlation (r)

0.9998

0.9998

0.9999

0.9998

0.9999

0.9998

Accuracy Precision (SD)d

99.98

100.99

100.67

99.86

99.45

99.57

 Repeatabilitye, f

1.12

0.35

2.47

0.863

1.224

1.016

 Intermediateprecisione, g

1.59

1.37

2.86

1.144

1.334

1.027

LODh

0.03

0.09

0.09

0.45

0.31

1.65

LOQi

0.09

0.28

0.27

1.35

0.93

4.95

  1. aRange: for HPTLC, it is measured by µg/band and for HPLC method in µg/mL
  2. The linearity was achieved using the polynomial regression equation: A = aX2 + bX + C
  3. bCoefficient 1, c Coefficient 2 X = concentration µg/band C = intercept
  4. dAccuracy: a Mean of 9 concentrations of each component
  5. eAverage of three experiments
  6. fStandard deviation of 3 concentrations of each component (0.5, 1.5 and 2 µg/band) for HPTLC method and 10, 20 and 30 µg/band (for flutamide and Flu-1), 50, 100 and 150 µg/mL (for Flu-3) for HPLC method on the same day
  7. gStandard deviation of 3 concentrations of each component (0.5, 1.5 and 2 µg/band) for HPTLC method and 10, 20 and 30 µg/band (for flutamide and Flu-1), 50, 100 and 150 µg/mL (for Flu-3) for HPLC method on three successive days
  8. hLOD = (3.3 X SD)/slope (SD of the intercept using the lower part of the calibration graph, the slope of the calibration curve)
  9. iLOQ = (10X SD)/slope (SD of the intercept using the lower part of the calibration graph, the slope of the calibration curve)