Simultaneous quantitation of five triazole anti-fungal agents by paper spray-mass spectrometry

Abstract

Introduction: Invasive fungal disease is a life-threatening condition that can be challenging to treat due to pathogen resistance, drug toxicity, and therapeutic failure secondary to suboptimal drug concentrations. Frequent therapeutic drug monitoring (TDM) is required for some anti-fungal agents to overcome these issues. Unfortunately, TDM at the institutional level is difficult, and samples are often sent to a commercial reference laboratory for analysis. To address this gap, the first paper spray-mass spectrometry assay for the simultaneous quantitation of five triazoles was developed. Methods: Calibration curves for fluconazole, posaconazole, itraconazole, hydroxyitraconazole, and voriconazole were created utilizing plasma-based calibrants and four stable isotopic internal standards. No sample preparation was needed. Plasma samples were spotted on a paper substrate in pre-manufactured plastic cartridges, and the dried plasma spots were analyzed directly utilizing paper spray-mass spectrometry (paper spray MS/MS). All experiments were performed on a Thermo Scientific TSQ Vantage triple quadrupole mass spectrometer. Results: The calibration curves for the five anti-fungal agents showed good linearity (R2 = 0.98 – 1.00). The measured assay ranges (LLOQ – ULOQ) for fluconazole, posaconazole, itraconazole, hydroxyitraconazole, and voriconazole were 0.5 – 50 μg/mL, 0.1 – 10 μg/mL, 0.1 – 10 μg/mL, 0.1 – 10 μg/mL, and 0.1 – 10 μg/mL, respectively. The inter- and intra-day accuracy and precision were less than 25% over the respective ranges. Conclusion: We developed the first rapid paper spray MS/MS assay for simultaneous quantitation of five triazole anti-fungal agents in plasma. The method may be a powerful tool for near point-of-care TDM aimed at improving patient care by reducing turnaround time and for use in clinical research.