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Updated: Feb 17, 2026

Time-Resolved Fluorescence Anisotropy from Single Molecules for Characterizing Local Flexibility in Biomolecules
Published on: April 25, 2025
Simplex optimization of the variables influencing the determination of pefloxacin by time-resolved chemiluminescence
José A Murillo Pulgarín1, Aurelia Alañón Molina1, Elisa Jiménez García1
1Department of Analytical Chemistry and Food Technology, University of Castilla-La Mancha, 13071 Ciudad Real, Spain.
Abstract:
A new chemiluminescence (CL) detection system combined with flow injection analysis (FIA) for the determination of Pefloxacin is proposed. The determination is based on an energy transfer from Pefloxacin to terbium (III). The metal ion enhances the weak CL signal produced by the KMnO4/H2SO3/Pefloxacin system. A modified simplex method was used to optimize chemical and instrumental variables. The influence of the interaction of the permanganate, Tb (III), sodium sulphite and sulphuric acid concentrations, flow rate and injected sample volume was thoroughly investigated by using a modified simplex optimization procedure. The results revealed a strong direct relationship between flow rate and CL intensity throughout the studied range that was confirmed by a gamma test. The response factor for the CL emission intensity was used to assess performance in order to identify the optimum conditions for maximization of the response. Under such conditions, the CL response was proportional to the Pefloxacin concentration over a wide range. The detection limit as calculated according to Clayton's criterion 13.7μgL-1. The analyte was successfully determined in milk samples with an average recovery of 100.6±9.8%.

