Impact of glucuronide interferences on therapeutic drug monitoring of posaconazole by tandem mass spectrometry.
BACKGROUND
Posaconazole is a novel antifungal drug for oral application intended especially for therapy of invasive mycoses. Due to variable gastrointestinal absorption, adverse side effects, and suspected drug-drug interactions, therapeutic drug monitoring (TDM) of posaconazole is recommended.


METHODS
A fast ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for quantification of posaconazole with a run-time <3 min was developed and compared to a LC-MS/MS method and HPLC method with fluorescence detection.


RESULTS
During evaluation of UPLC-MS/MS, two earlier eluting peaks were observed in the MRM trace of posaconazole. This was only seen in patient samples, but not in spiked calibrator samples. Comparison with LC-MS/MS disclosed a significant bias with higher concentrations measured by LC-MS/MS, while UPLC-MS/MS showed excellent agreement with the commercially available HPLC method. In the LC-MS/MS procedure, comparably wide and left side shifted peaks were noticed. This could be ascribed to in-source fragmentation of conjugate metabolites during electrospray ionisation. Precursor and product ion scans confirmed the assumption that the additional compounds are posaconazole glucuronides. Reducing the cone voltage led to disappearance of the glucuronide peaks. Slight modification of the LC-MS/MS method enabled separation of the main interference, leading to significantly reduced deviation.


CONCLUSIONS
These results highlight the necessity to reliably eliminate interference from labile drug metabolites for correct TDM results, either by sufficient separation or selective MS conditions. The presented UPLC-MS/MS method provides a reliable and fast assay for TDM of posaconazole.
Krüger
Ralf
R
Institute of Clinical Chemistry and Laboratory Medicine, Medical Center of the Johannes Gutenberg University, Mainz, Germany. krueger@zentrallabor.klinik.uni-mainz.de
Vogeser
Michael
M
Burghardt
Stephan
S
Vogelsberger
Rita
R
Lackner
Karl J
KJ
D000935:Antifungal Agents; D016903:Drug Monitoring / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D020719:Glucuronides / Q000737:chemistry*; D006801:Humans; D015588:Observer Variation; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards; D014230:Triazoles / Q000032:analysis* / Q000627:therapeutic use
D000935:Antifungal Agents; D020719:Glucuronides; D014230:Triazoles; C101425:posaconazole