Sustainable chromatographic quantitation of multi-antihypertensive medications: application on diverse combinations containing hydrochlorothiazide along with LC–MS/MS profiling of potential impurities: greenness and whiteness evaluation

Table 3 Validation and Regression parameters of the proposed HPLC–DAD and LC–MS/MS methods

Method Parameter	HPLC–DAD method								LC–MS/MS method
Method Parameter	HCT	CT	DSA	VAL	AML	ATN	AMI	CAN	CT	SAL
Linearity range^a	0.10–100.00	0.05–8.00		0.10–100.0		0.05–100.00		0.10–100.00	1.0–200.0	5.0–200.0
Regression equation parameters
Slope	0.0530	0.4789	0.4957	0.0449	0.0493	0.0492	0.4940	0.0502	92.062	1.4412
Intercept	0.0051	0.0343	0.0134	0.0889	0.0169	0.0084	0.0155	0.0020	29.337	0.8209
Correlation Coefficient (r)	1.0000	1.0000	1.0000	1.0000	1.0000	1.0000	1.0000	1.0000	0.9999	0.9999
Accuracy ^b (Mean ± SD)	99.51 ± 0.68	100.44 ± 0.59	100.52 ± 0.69	100.18 ± 0.86	100.57 ± 1.05	99.25 ± 0.77	99.97 ± 0.69	99.03 ± 0.75	99.92 ± 0.60	99.64 ± 0.77
Precision (± %RSD)
Repeatability ^c	0.85	0.38	0.69	0.75	0.48	0.37	0.25	0.55	0.88	0.61
Intermediate precision ^d	0.91	0.80	0.86	0.87	0.53	0.48	0.48	0.76	1.00	0.78
LOD ^e	0.01	0.01	0.01	0.02	0.01	0.01	0.01	0.03	0.19	0.14
LOQ ^e	0.05	0.02	0.02	0.06	0.02	0.03	0.03	0.08	0.59	0.41
Robustness ^f	0.96	0.79	0.88	0.79	0.88	1.15	0.85	0.98	0.65	0.52

^a For HPLC–DAD method: in μg/mL and for LC–MS/MS method: in ng/mL
^b Accuracy was checked using concentrations for HPLC–DAD method: 0.2, 40.0 and 80.0 µg/mL for HCT, VAL, AML, ATN, AMI and CAN and 0.3, 3.0 and 7.0 µg/mL for CT and SAL while for LC–MS/MS method: 40.0, 80.0 and 120.0 ng/mL for CT and SAL
^c and ^d are repeatability and intermediate precision, respectively (n = 9) percentage relative standard deviation of three different concentrations in triplicate for HPLC–DAD method: 0.5, 20.0 and 60.0 µg/mL for HCT, VAL, AML, ATN, AMI and CAN and 0.1, 2.0 and 5.0 µg/mL for CT and SAL while for LC–MS/MS method: 5.0, 50.0 and 150.0 ng/mLfor CT and 10.0, 50.0 and 150.0 ng/mL for SAL
^e LOD and LOQ were calculated as per ICH, 3.3 × SD of the intercept/slope and 10 × SD of the intercept/slope, respectively
^f For HPLC–DAD: average %RSD of the change in detection wavelength (± 1 nm) and flow rate (± 0.1 mL/min), while for LC–MS/MS: average %RSD of the change in the mobile phase composition (± 2%)

ISSN: 2661-801X