Implementation of different fluorogenic condensation reactions for the spectrofluorimetric detection of the banned growth promoter, bacitracin, in water samples: assessment of the greenness of methods

Bacitracin is among a group of antibiotic growth promoters banned in the European Union since the 1999 Council. In this study, two novel spectrofluorimetric methods were designed, developed, and validated for determining bacitracin by turning on its fluorescence activity. The turning on strategies are based on the condensation of its 1ry amine moiety either with acetylacetone and formaldehyde (Ac-Fr method), or with ninhydrin and phenylacetaldehyde (Nin-Ph method). The produced fluorescence intensity was measured at an emission of 482.5 nm (excitation = 421.5 nm) for Ac-Fr method and measured at an emission of 479.5 nm (excitation = 383.5 nm) for Nin-Ph method. A thorough investigation and optimization of the reaction conditions for both strategies was carried out. The selectivity of the established methodologies towards bacitracin was studied. The established linear ranges were 0.1–1.0 and 1.0–5.0 µg/mL for Ac-Fr method and Nin-Ph method, respectively. The LOD values were 31.2 and 163 ng/mL for Ac-Fr method and Nin-Ph method, respectively. Furthermore, the established methods were applied to determine bacitracin in water samples successfully. The recommended process for comprehensive greenness evaluations was implemented using Modified NEMI, AGREEprep, ComplexGAPI, and the newest SPMS tools. Moreover, the most recently introduced RGB12 and BAGI algorithms were employed in the concepts of “whiteness” and “blueness” inspection. This emphasizes the significance of doing a holistic sustainability assessment that takes into account greenness, analytical excellence, and practicality while creating new techniques. The suggested technique provides sensitive, cost-effective, and sustainable substitutes for traditional analysis methods.