Quantification of Carbonic Anhydrase Inhibitors and Metabolites in Urine and Hair of Patients and Their Relatives

Alfredo Fabrizio Lo Faro; Anastasio Tini; Giulia Bambagiotti; Filippo Pirani; Andrea Faragalli; Flavia Carle; Elena Pacella; Artan Ceka; Marco Moretti; Massimo Gottardi; Nicola Vito Lassandro; Michele Nicolai; Marco Lupidi; Cesare Mariotti; Francesco Paolo Busardò; Jeremy Carlier

doi:10.3390/biology11101379

Biology (Sep 2022)

Quantification of Carbonic Anhydrase Inhibitors and Metabolites in Urine and Hair of Patients and Their Relatives

Alfredo Fabrizio Lo Faro,
Anastasio Tini,
Giulia Bambagiotti,
Filippo Pirani,
Andrea Faragalli,
Flavia Carle,
Elena Pacella,
Artan Ceka,
Marco Moretti,
Massimo Gottardi,
Nicola Vito Lassandro,
Michele Nicolai,
Marco Lupidi,
Cesare Mariotti,
Francesco Paolo Busardò,
Jeremy Carlier

Affiliations

Alfredo Fabrizio Lo Faro: Unit of Forensic Toxicology, Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Anastasio Tini: Unit of Forensic Toxicology, Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Giulia Bambagiotti: Unit of Forensic Toxicology, Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Filippo Pirani: Unit of Forensic Toxicology, Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Andrea Faragalli: Center of Epidemiology Biostatistics and Information Technology, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Flavia Carle: Center of Epidemiology Biostatistics and Information Technology, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Elena Pacella: Department of Sense Organs, Faculty of Medicine and Dentistry, Sapienza University of Rome, Viale del Policlinico 155, 00161 Rome, RM, Italy
Artan Ceka: Department of Laboratory Medicine, Reunited Hospitals Torrette of Ancona, Via Conca 71, 60126 Ancona, AN, Italy
Marco Moretti: Department of Laboratory Medicine, Reunited Hospitals Torrette of Ancona, Via Conca 71, 60126 Ancona, AN, Italy
Massimo Gottardi: Comedical srl, Via della Cooperazione 29, 38123 Trento, TN, Italy
Nicola Vito Lassandro: Eye Clinic, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Michele Nicolai: Eye Clinic, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Marco Lupidi: Eye Clinic, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Cesare Mariotti: Eye Clinic, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Francesco Paolo Busardò: Unit of Forensic Toxicology, Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy
Jeremy Carlier: Unit of Forensic Toxicology, Section of Legal Medicine, Department of Biomedical Sciences and Public Health, Marche Polytechnic University, Via Tronto 10/a, 60126 Ancona, AN, Italy

DOI: https://doi.org/10.3390/biology11101379
Journal volume & issue: Vol. 11, no. 10
p. 1379

Abstract

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Carbonic anhydrase inhibitors (CAIs) are prescription drugs also used in doping to dilute urine samples and tamper with urinalyses. Dorzolamide, brinzolamide, and acetazolamide are prohibited by the World Anti-Doping Agency. Detecting CAIs and their metabolites in biological samples is crucial to documenting misuse in doping. We quantified dorzolamide, brinzolamide, acetazolamide, and their metabolites in the urine and hair of 88 patients under treatment for ocular hypertension or glaucoma. Samples of the patients’ relatives were analyzed to assess potential for accidental exposure. After washing, 25 mg hair was incubated with an acidic buffer at 100 °C for 1 h. After cooling and centrifugation, the supernatant was analyzed by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Urine (100 μL) was diluted and centrifuged before UHPLC-MS/MS analysis. Run time was 8 min through a reverse-phase column with a mobile phase gradient. MS/MS analysis was performed in a multiple-reaction monitoring mode after positive electrospray ionization. Median urinary concentration was 245 ng/mL (IQR: 116.2–501 ng/mL) for dorzolamide, 81.1 ng/mL (IQR: 35.9–125.3 ng/mL) for N-deethyl-dorzolamide, 0.77 ng/mL (IQR: 0.64 ng/mL–0.84 ng/mL) for N-acetyl-dorzolamide, 38.9 ng/mL (IQR: 20.4–79.2 ng/mL) for brinzolamide, and 72.8 ng/mL (IQR: 20.7–437.3 ng/mL) for acetazolamide. Median hair concentration was 0.48 ng/mg (IQR: 0.1–0.98 ng/mg) for dorzolamide, 0.07 ng/mg (IQR: 0.06–0.08 ng/mg) for N-deethyl-dorzolamide, 0.40 ng/mL (IQR: 0.13–1.95 ng/mL) for brinzolamide. Acetazolamide was detected in only one hair sample. Dorzolamide and brinzolamide were detected in the urine of three and one relatives, respectively. Cutoff concentrations of urinary dorzolamide and brinzolamide are necessary to preclude false positives due to contamination or passive exposure. We reported the first concentrations of brinzolamide in hair.

Published in Biology

ISSN: 2079-7737 (Online)
Publisher: MDPI AG
Country of publisher: Switzerland
LCC subjects: Science: Biology (General)
Website: https://www.mdpi.com/journal/biology

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