Integrating green analytical chemistry and analytical quality by design: an innovative approach for RP-UPLC method development of ensifentrine in bulk and inhalation formulations

Table 2 ANOVA test results for Retention Time (a), Number of Theoretical Plates (NTP, b), and Tailing Factor (Symmetry) (TF, c) using CCD for various responses

(a) Source	Sum of squares	df	Mean square	F-value	p-value	Result
Model	0.1615	9	0.0179	407.02	< 0.0001	Significant
A-FR	0.1404	1	0.1404	3186.06	< 0.0001	Significant
B-MP	0.0023	1	0.0023	51.27	< 0.0001	Significant
C-TF	0.0171	1	0.0171	388.09	< 0.0001	Significant
AB	5.000E−07	1	5.000E−07	0.0113	0.9173	Significant
AC	0.0003	1	0.0003	6.53	0.0286	Significant
BC	4.500E−06	1	4.500E−06	0.1021	0.7559	Significant
A²	0.0012	1	0.0012	26.87	0.0004	Significant
B²	0.0003	1	0.0003	6.53	0.0286	Significant
C²	8.283E−06	1	8.283E−06	0.1879	0.6739	Significant
Residual	0.0004	10	0.0000			Significant
Lack of fit	0.0003	5	0.0001	1.41	0.3574	Not Significant
Pure error	0.0002	5	0.0000
Cor Total	0.1619	19

(b) Source	Sum of squares	df	Mean square	F-value	p-value	Results
Model	6.294E + 06	9	6.994E + 05	113.82	< 0.0001	Significant
A-FR	3.880E + 06	1	3.880E + 06	631.48	< 0.0001	Significant
B-MP	1719.08	1	1719.08	0.2798	0.6084	Significant
C-TF	1.131E + 06	1	1.131E + 06	184.13	< 0.0001	Significant
AB	0.4512	1	0.4512	0.0001	0.9933	Significant
AC	6.424E + 05	1	6.424E + 05	104.54	< 0.0001	Significant
BC	1267.56	1	1267.56	0.2063	0.6594	Significant
A²	5.857E + 05	1	5.857E + 05	95.31	< 0.0001	Significant
B²	4069.73	1	4069.73	0.6623	0.4347	Significant
C²	89,888.72	1	89,888.72	14.63	0.0033	Significant
Residual	61,446.60	10	6144.66			Significant
Lack of fit	49,555.27	5	9911.05	4.17	0.0717	Not significant
Pure error	11,891.33	5	2378.27
Cor total	6.356E + 06	19

(c) Source	Sum of Squares	df	Mean square	F-value	p-value	Result
Model	0.0184	9	0.0020	75.89	< 0.0001	Significant
A-FR	0.0058	1	0.0058	214.64	< 0.0001	Significant
B-MP	0.0072	1	0.0072	269.12	< 0.0001	Significant
C-TF	0.0000	1	0.0000	1.09	0.3215	Significant
AB	0.0032	1	0.0032	118.87	< 0.0001	Significant
AC	0.0002	1	0.0002	7.43	0.0214	Significant
BC	0.0002	1	0.0002	7.43	0.0214	Significant
A²	0.0001	1	0.0001	2.64	0.1351	Significant
B²	0.0005	1	0.0005	17.74	0.0018	Significant
C²	0.0010	1	0.0010	37.64	0.0001	Significant
Residual	0.0003	10	0.0000			Significant
Lack of fit	0.0002	5	0.0000	2.23	0.1996	Not significant
Pure error	0.0001	5	0.0000
Cor total	0.0187	19

A, Band C coded levels of independent variables; AB, AC, and BC are interaction terms; A², B², and C² quadratic terms

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