Comparative performance evaluation of triple quadrupole tandem mass spectrometry and orbitrap high-resolution mass spectrometry for analysis of antibiotics in creek water impacted by CETP discharge

Abstract

The widespread detection of antibiotics in aquatic environments, particularly in effluent-receiving surface waters, poses significant ecological and public health concerns due to their role in promoting antimicrobial resistance. Accurate trace-level antibiotic measurement is essential for environmental risk assessment and for improving wastewater treatment strategies. This study presents the development, optimization, and validation of two complementary liquid chromatography-mass spectrometry (LC-MS) workflows for the simultaneous quantification of nine antibiotics across five therapeutic classes in creek water impacted by a Common Effluent Treatment Plant (CETP). The performance of a triple quadrupole LC-MS/MS system (LC-QqQ-MS) was compared to that of a high-resolution Orbitrap mass spectrometer (LC-Orbitrap-HRMS). Both instruments demonstrated excellent linearity (R² > 0.99) and satisfactory recoveries (70–90%) across a wide concentration range. The method detection limits ranged from 0.11 to 0.23 ng L⁻¹ for LC-QqQ-MS and from 0.02 to 0.13 ng L⁻¹ for LC-Orbitrap-HRMS, confirming the superior sensitivity of the high-resolution system approach. Application to real-world creek water samples revealed the ubiquitous presence of multiple antibiotics, with azithromycin and enrofloxacin dominating the detected concentrations, particularly near the CETP discharge point and a nearby waste dumping site. A three-way ANOVA confirmed that antibiotic concentrations were significantly affected by instrument type, sampling site, and antibiotic class along with their interactions. Additionally, non-target screening performed using LC-Orbitrap-HRMS enabled the detection of additional antibiotics belonging to quinolones, sulfonamides and aminoglycosides, further demonstrating the broader analytical scope of high-resolution mass spectrometry. The study highlights the necessity of using advanced analytical tools for the accurate quantification of antibiotics in complex matrices and underscores the environmental risks posed by pharmaceutical pollution in industrial discharge-impacted water bodies.