Development and validation of a stability-indicating reverse-phase high-performance liquid chromatography method for simultaneous determination of antibiotic residues in hospital wastewater from dehradun, uttarakhand with SIX sigma and blue applicability grade index evaluation
Abstract
Objectives: The present study aimed to develop and validate a green and stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method for simultaneous determination of metronidazole, ornidazole, ofloxacin, chloramphenicol, and moxifloxacin hydrochloride in hospital wastewater samples collected from Dehradun, Uttarakhand. Methods: Chromatographic separation was achieved using an Accellis C18 column (150 × 4.6 mm, 5 μm) with gradient elution employing buffer (pH 2.5±0.05) and acetonitrile as mobile phase at a flow rate of 1.0 mL/min with ultraviolet-photodiode array detector detection at 280 nm. Wastewater samples were subjected to membrane filtration followed by solid-phase extraction before analysis. The developed RP-HPLC method was validated according to International Council for Harmonization Q2(R1) guidelines. Environmental sustainability was evaluated using analytical GREEnness metric approach, green analytical procedure index, blue applicability grade index, analytical method greenness score, and high-performance liquid chromatography environmental assessment tool assessment tools. Results: The developed RP-HPLC method demonstrated satisfactory specificity, precision, and linearity for all investigated antibiotics. System suitability results showed percentage relative standard deviation values below 1.1%, tailing factors between 1.012 and 1.125, and theoretical plate counts above 3400. Excellent linearity was obtained with regression…
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