Spectrum subtraction as a complementary method for six resolution techniques resolving overlapping spectra; application to multicomponent veterinary formulation with greenness and whiteness assessment

Table 1 Assay parameters and method validation obtained by applying the proposed spectrophotometric methods

Drug Name	TCB							LVM
Method	D⁰ (304 nm)	D⁰ (220 nm)	D¹ (310 nm)	D¹ (P_{228–216 nm})	DD¹ (P_{230–220 nm})	CNV	CV	D⁰ (214 nm)	D¹ (P_221 nm)	DD¹ (P_219 nm)
Range (µg/mL)	2.0–20.0	1.0 – 10.0	2.0–20.0	1.0–10.0	2.0–20.0	…	2.0–20.0	2.0–14.0
Slope	0.0533	0.1322	0.0049	0.0088	0.0306	…	1.0043	0.0834	0.0037	0.0099
Intercept	0.0011	− 0.0198	0.0001	− 0.0002	− 0.0031	…	0.0177	− 0.003	− 0.0002	− 0.0004
Accuracy (R% ± SD)	100.78 ± 0.61	101.05 ± 0.88	100.84 ± 0.46	100.002 ± 0.80	98.76 ± 0.59	101.01 ± 0.44	100.41 ± 0.37	99.35 ± 0.76	98.79 ± 0.94	100.54 ± 0.37
Correlation coefficient (r)	0.9998	1.0000	0.9999	0.9997	0.9999	…	0.9998	0.9997	0.9998	0.9998
Repeatability (RSD)^a	0.176	0.522	0.334	0.410	0.585	0.100	0.184	0.537	0.365	0.203
Intermediate precision (RSD)^b	0.554	1.159	0.386	0.531	0.821	0.216	0.259	0.565	0.383	0.769
LOD	0.28	0.08	0.26	0.14	0.52	…	0.29	0.27	0.19	0.23
LOQ	0.86	0.23	0.78	0.41	1.58	…	0.89	0.80	0.58	0.70

RSD^a &RSD^b: relative standard deviation for the intra-day & inter-day precision, respectively (n = 3) using concentrations 4, 6, 9 µg/ mL for TCB and 6, 9, 13 µg/ mL for LVM

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