Statistical Evaluation for Linearity

This report is applicable for statistical evaluation of the linearity studies performed in dissolution analytical
method validation of Acetate Abiraterone tablets USP 250 & 500 mg (PO/AMVR/ABIR/23/023/00). in
accordance with the requirements of RDC 166/2017. The statistical evaluation was conducted using the
software Action Stat Version 3.7.

1 EXPERIMENTAL DATA

Linearity Level (%) Concentration (ppm) Response (Area)

10% 0.0303 306445
10% 0.0302 304200
10% 0.0302 308670
20% 0.0605 627386
20% 0.0604 624887
20% 0.0605 624799
50% 0.1513 1549836
50% 0.1511 1543816
50% 0.1512 1548222
80% 0.242 2477506
80% 0.2418 2505857
80% 0.2418 2483734
100% 0.3025 3124953
100% 0.3022 3123228
100% 0.3023 3113161
120% 0.363 3762449
120% 0.3626 3758804
120% 0.3628 3745990
Table 1: Linearity Results

2 STATISTICAL LINEAR REGRESSION: ORDINARY LEAST SQUARE

METHOD

2.1 SIGNIFICANCE OF THE SLOPE (ANOVA: F-TEST)

Null hypothesis: slope is zero;

Alternative hypothesis: slope is different than zero.
Source DF Sum of Square Mean Square F-value P-value
Concentration (mg/mL) 1 28604170216879.625 28604170216879.625 294836.6835 0
Residuals 16 1552271983.3095 97016998.9568 NA NA
Table 2: ANOVA
As the P-value (0) obtained by the ANOVA F-Test is lower than 0.05. we reject the null hypothesis and it is concluded
that at a significance level of 5%. the slope is different than zero.
Regression analysis:
Coefficient Std Dev T-value P-value
Intercept -8215.8916 4326.4207 -1.899 0.0757
Slope 10352461.4991 19065.7048 542.9887 0
Table 3: Coefficient

Lower Limit Upper Limit

Intercept -17387.4938 955.7106
Slope 10312044 10392878.9879
Table 4: Confidence Interval for the parameters

2.2 SIGNIFICANCE OF THE INTERCEPT (STUDENT’S T-TEST)

Null hypothesis: intercept (linear coefficient) is zero;

Alternative hypothesis: intercept is different than zero.
As the P-value (0.0757) obtained by the Student’s t-Test is higher than 0.05. we do not reject the null hypothesis and
it is concluded that at a significance level of 5% the intercept (linear coefficient) is zero.

2.3 PEARSON LINEAR CORRELATION

Pearson’s correlation coefficient measures the degree of proportionality between the variables (concentration and
area). Data is having significant linear correlation if the Pearson’s correlation coefficient (r) between “concentration”
and “area” is above 0.99.

Residues Std Dev DF r2 r

9849.7208 16 0.9999 0.9999
Table 5: Correlation Coefficient

The coefficient of correlation (r) and determination (r2) obtained corresponds to 0.9999 and 0.9999. respectively.
Since the coefficient of correlation (0.9999) is higher than 0.99 (as proposed). it is concluded that the data is having
significant linear correlation.

3 GRAPHICAL ANALYSIS

3.1 SCATTER PLOT: VISUAL ANALYSIS OF THE CURVE

The scatter plot is a graph that allows the visualization of a possible association between quantitative variables.
 Figure 1: Scatter Plot

Observation: From the visual evaluation. it is observed that there is a linear relationship between the concentration
(mg/mL) and response (Area).

3.2 RESIDUAL PLOTS

Figure 2: Residual Plots

Statistic P-value
Shapiro-Wilk 0.9524 0.4643
Table 6: Normality Test
Null hypothesis: residuals are normally distributed;
Alternative hypothesis: residuals are not normally distributed.
As the P-value (0.4643) obtained through Shapiro-Wilk test is > 0.05. we do not reject the null hypothesis and it is
concluded that at a significance level of 5%. the residuals are normally distributed.

4 HOMOCEDASTICITY EVALUATION: BREUSCH PAGAN TEST

Null hypothesis: all variances are equal (data is homocedastic);

Alternative hypothesis: at least one variance is different (data is heterocedastic).

Statistic DF P-value
1.5178 1 0.2179
Table 7: Homocedasticity Test – Breusch Pagan

As the P-value (0.2179) obtained through Breusch Pagan test is not less than 0.05. we do not reject the null hypothesis
and it is concluded that at a significance level of 5%. all variances are equal and the data is homocedastic.

5 EXTREME VALUES IN THE RESPONSE

5.1 RESIDUALS
Observations with standardized and/or studentized residuals greater than 3 will be considered as an extreme value in
the response.
Concentration
S No. Residuals Standardized Residuals
(mg/mL)
1 0.0303 981.3082 0.1082
2 0.0302 -228.4457 -0.0252
3 0.0302 4241.5543 0.4679
4 0.0605 9277.9709 1.004
5 0.0604 7814.2171 0.8457
6 0.0605 6690.9709 0.7241
7 0.1513 -8275.5332 -0.8673
8 0.1511 -12225.0409 -1.2813
9 0.1512 -8854.2871 -0.928
10 0.242 -19573.7912 -2.0553
11 0.2418 10847.7011 1.139
12 0.2418 -11275.2989 -1.1839
13 0.3025 1549.2881 0.166
14 0.3022 2930.0266 0.3138
15 0.3023 -8172.2196 -0.8753
16 0.363 12721.3674 1.4141
17 0.3626 13217.352 1.4687
18 0.3628 -1667.1403 -0.1853
Table 8: Summary of Residual Analysis
 Figure 3: Residual Plot (Standardized)

Figure 4: Residual Plot

As can be seen in the graphs of “Standardized Residuals vs Fitted Values” and “Studentized Residuals vs Fitted
Values”. extreme values were not detected.

6 INDEPENDENCE TEST

Null hypothesis: Observations are independent;

Alternative hypothesis: Observations are not independent.

Statistic P-value
1.7902 0.4658
Table 9: Independence Test – Durbin-Watson
 Figure 5: Residual vs Observation Order

As the P-value (0.4658) obtained through Durbin-Watson test is > 0.05. we do not reject the null hypothesis and it is
concluded that at a significance level of 5%. observations are independent.

7 SUMMARY OF LINEARITY STUDIES

Test Conclusion
ANOVA: F-Test Significance of the linear model was not rejected
Intercept Test Significant Intercept equal to zero
Pearson correlation coefficient r = 1 is higher than the proposed (0.99)
Normal Distribution of Residues Residuals are normally distributed
Homoscedasticity Test The homoscedasticity of variance was not rejected
Extreme Value in Response Extreme values in response not detected
Independence Test Dependence of observations not detected

Based on the statistical evaluation. the linearity data found satisfactory and linear from 10 to 120% of the target concentration.