A new, sensitive and versatile assay for quantitative determination of α-fluoro-β-alanine (AFBA) in human urine by using the reversed-phase ultrahigh performance-tandem mass spectrometry (rp-UHPLC–MS/MS) system

•Small and polar molecules, such as secondary metabolites, need a study of the most appropriate mode of chromatography.•The quantification of AFBA was performed in the reversed phase of chromatography by using a UHPLC-MS/MS system.•The method was applied to the monitoring of health care workers invo...

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Bibliographic Details
Published in:	Toxicology letters Vol. 298; pp. 164 - 170
Main Authors:	Sottani, Cristina, Grignani, Elena, Zaratin, Laura, Santorelli, Donatella, Studioso, Emanuele, Lonati, Davide, Locatelli, Carlo A., Pastoris, Ornella, Negri, Sara, Cottica, Danilo
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01-12-2018
Subjects:	AFBA urinary excretion Occupational exposure rp-UHPLC–MS/MS Validation study α-Fluoro-β-alanine (AFBA) α-Fluoro-β-alanine (AFBA) rp-UHPLC–MS/MS AFBA urinary excretion Occupational exposure Validation study
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Summary:	•Small and polar molecules, such as secondary metabolites, need a study of the most appropriate mode of chromatography.•The quantification of AFBA was performed in the reversed phase of chromatography by using a UHPLC-MS/MS system.•The method was applied to the monitoring of health care workers involved in the handling of antineoplastic agents.•The sampling procedure was discussed in relation to the PK parameters of 5-FU. A method for the quantitation of α-fluoro-β-alanine (AFBA), the main metabolite of capecitabine (Cape) and 5-fluoruracil (5-FU), is described. Among antineoplastic drugs (ADs), 5-FU and Cape (the new oral prodrug) are the most commonly applied drugs in cancer therapy. The main objective of this study was to develop a reliable method that would be easy to run on a reversed-phase UHPLC system coupled to tandem mass spectrometry. AFBA was derivatized with Sanger's reagent to ensure complete yield of a stable 2,4 dinitrophenil-α-fluoro-β-alanine derivative. This method was based on the use of a mixed-mode anion exchange solid phase extraction enabling urinary extracts to be clear of endogenous interferences affecting quantitative results. The assay was validated in human urine according to FDA criteria with the use of a labeled internal standard (β-alanine-d4) to minimize experimental error. Good accuracy and precision were demonstrated by determining spiked urine QC samples in four consecutive days. The recovery of AFBA was between 70.0 and 82.6%, with a matrix effect that was 12.8%–18.5%. The lower limit of quantitation (LOQ) was 0.5 ng/mL with a coefficient of variation of 5.3%. This assay was successfully applied to determine the levels of this metabolite in a large number of urine samples taken from personnel who were occupationally exposed to ADs.
ISSN:	0378-4274 1879-3169
DOI:	10.1016/j.toxlet.2018.10.007