Development and Validation of UPLC-MS/MS Method for Simultaneous Determination of Multi-Class Antibiotics, Antivirals and Antifungals in Water

Abstract

According to a WHO report, antimicrobial resistance is one of the top 10 health threats. Environmental sources are extremely contaminated with high levels of antibiotics, antivirals and antifungals due to their excessive and unsafe usage. Hence, there is a need to detect and quantitate the antimicrobial agents in water. Consequently, a simultaneous analytical method is developed and validated, employing a solid-phase extraction methodology with an HLB cartridge. This method was utilized to quantify a minimum of twenty different chemicals derived from antibiotics, antivirals and antifungal drugs. Antibiotics include quinolones, isonicotinic acid, macrolides, oxazolidinones, penicillin, sulphonamides, antivirals that contain nucleotide class compounds and antifungals that comprising of triazoles and imidazoles. Ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC- S/MS) was used to quantify the analytes. Detection was achieved using multiple reaction monitoring with positive electron spray ionization. All the compounds were well separated with the column, Xterra MS C18 4.6 × 50 mm 2.5 μm, using water and acetonitrile with 0.1% formic acid as a mobile phase. The calibration curve range was set at 50 to 2000 ng mL-1. Validation parameters like precision and accuracy, matrix effect, sensitivity, autosampler stability, the limit of detection and recovery were performed.