TY - JOUR

T1 - Assay validation for vilanterol and its metabolites in human urine with application for doping control analysis

AU - Panchal, Tina

AU - Baldwin, Sandra

AU - Østergaard, Martin

AU - Hansen, Erik Sören Halvard

AU - Backer, Vibeke

AU - Hostrup, Morten

AU - Daley-Yates, Peter

N1 - This article is protected by copyright. All rights reserved.

PY - 2023

Y1 - 2023

N2 - A bioanalytical method for detecting the ultra-long-acting beta2-agonist (U-LABA), inhaled vilanterol and its metabolites, GSK932009 and GW630200, in urine was developed to potentially monitor permitted therapeutic versus prohibited supratherapeutic use in sport. The World Anti-Doping Agency (WADA) has established urinary concentration thresholds for beta2-agonists salbutamol and formoterol. Therapeutic use of vilanterol (25 μg once-daily) was recently permitted by WADA, however, there is no established decision limit for adverse analytical findings due to insufficient urine concentration data. In this study, we validated an assay to detect vilanterol in urine collected from four healthy male and female athletes 0 - 72 h who received inhaled corticosteroid fluticasone furoate/U-LABA vilanterol (800/100 μg) combination, four times the normal therapeutic dose. After administration, subjects performed one hour of bike ergometer exercise. The experiment was conducted again after repeat dosing for one week. Our method utilised liquid chromatography with tandem mass spectrometry and was validated over urine concentrations of 5 - 5000 pg/mL (vilanterol), and 50 - 50,000 pg/mL (GSK932009 and GW630200). Plasma samples were analysed for vilanterol, using a previously validated assay. The C max for urine vilanterol, GSK932009 and GW630200 were 9.5, 10.4 and 0.17 ng/mL, for single dosing, and 18.6, 19.5, and 0.20 ng/mL, for repeat dosing. Urine samples from four volunteers using the final validated method are reported, demonstrating this assay has sensitivity to detect vilanterol or GSK932009 in urine for ≥ 72 hours post single or repeat dosing with 800/100 μg fluticasone furoate/vilanterol whereas GW630200 was quantifiable ≤ 4 hours post-dose.

AB - A bioanalytical method for detecting the ultra-long-acting beta2-agonist (U-LABA), inhaled vilanterol and its metabolites, GSK932009 and GW630200, in urine was developed to potentially monitor permitted therapeutic versus prohibited supratherapeutic use in sport. The World Anti-Doping Agency (WADA) has established urinary concentration thresholds for beta2-agonists salbutamol and formoterol. Therapeutic use of vilanterol (25 μg once-daily) was recently permitted by WADA, however, there is no established decision limit for adverse analytical findings due to insufficient urine concentration data. In this study, we validated an assay to detect vilanterol in urine collected from four healthy male and female athletes 0 - 72 h who received inhaled corticosteroid fluticasone furoate/U-LABA vilanterol (800/100 μg) combination, four times the normal therapeutic dose. After administration, subjects performed one hour of bike ergometer exercise. The experiment was conducted again after repeat dosing for one week. Our method utilised liquid chromatography with tandem mass spectrometry and was validated over urine concentrations of 5 - 5000 pg/mL (vilanterol), and 50 - 50,000 pg/mL (GSK932009 and GW630200). Plasma samples were analysed for vilanterol, using a previously validated assay. The C max for urine vilanterol, GSK932009 and GW630200 were 9.5, 10.4 and 0.17 ng/mL, for single dosing, and 18.6, 19.5, and 0.20 ng/mL, for repeat dosing. Urine samples from four volunteers using the final validated method are reported, demonstrating this assay has sensitivity to detect vilanterol or GSK932009 in urine for ≥ 72 hours post single or repeat dosing with 800/100 μg fluticasone furoate/vilanterol whereas GW630200 was quantifiable ≤ 4 hours post-dose.

KW - Faculty of Science

KW - Vilanterol

KW - Beta2-agonist

KW - WADA

KW - Doping

KW - Assay validation

U2 - 10.1002/dta.3433

DO - 10.1002/dta.3433

M3 - Journal article

C2 - 36581315

VL - 15

SP - 495

EP - 505

JO - Drug Testing and Analysis

JF - Drug Testing and Analysis

SN - 1942-7603

IS - 5

ER -