Sensitive and eco-friendly extraction of antifungal drugs from artificial urine and saliva samples using MSPE coupled with HPLC-DAD

Abstract

The analysis of drug molecules in biological samples is a challenge for method development studies. The existence of matrix components and trace concentrations of target molecules in the samples make this analysis more difficult. Sample pre-treatment procedures, such as solid-phase micro extraction (SPME), facilitate and make this analysis possible using conventional analytical techniques such as HPLC. A newly synthesized trimesic acid (TMA)-modified magnetic adsorbent, Fe₃O₄@SiO₂–TMA, effectively retained drug molecules, enabling the successful determination of bifonazole (BFZ) and itraconazole (ITZ) residues in model solutions, including artificial saliva and urine samples. The characterization of the new adsorbent was performed by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) and Raman spectroscopy. The linear ranges for both target molecules were 15.00–1000.00 ng mL⁻¹ (R² > 0.9954), with a limit of detection lower than 5.00 ng mL⁻¹. The accuracy and repeatability of the proposed method were determined as recovery (R%) and relative standard deviation (RSD%) using spiked samples. This study presented the synthesis and characterization of Fe₃O₄@SiO₂–TMA, a new magnetic adsorbent, for the accurate and precise analysis of BFZ and ITZ, known as antifungal molecules, in biological samples. A straightforward and efficient determination of the target compounds was achieved using a magnetic solid-phase extraction (MSPE) procedure coupled with a high-performance liquid chromatography system equipped with a diode array detector (HPLC-DAD). The greenness and practical applicability of the method were assessed using BAGI and MoGAPI tools, with scores of 65 and 76, respectively. The developed method was successfully applied to real biological samples, yielding recovery values between 94.7% and 107.5%. These results demonstrate that the proposed approach is sensitive, reliable, and environmentally sustainable for routine antifungal drug analysis.