RT info:eu-repo/semantics/article
T1 HPLC‐QTOF method for quantifying 11‐ketoetiocholanolone, a cortisol metabolite, in ruminants' feces: Optimization and validation
A1 Molina García, Lucía
A1 Pérez, Jesús M.
A1 Sarasa, Mathieu
A1 Ureña Gutiérrez, Benjamín
A1 Espinosa Cerrato, José
A1 Azorit, Concepción
A2 Sanidad Animal
K1 Sanidad animal
K1 Veterinaria
K1 11-ketoetiocholanolone
K1 Capra pyrenaica
K1 Fecal cortisol metabolites
K1 HPLC-QTOF
K1 Noninvasive monitoring
K1 Ruminant stress
K1 Solid phase extraction
K1 3109 Ciencias Veterinarias
AB [EN]Studies of animal ecology can benefit from a quantified understanding of eco-physiologicalprocesses and, in particular, of the physiological responses in free-ranginganimals to potential stressors. The determination of fecal cortisol metabolitesas a noninvasive method for monitoring stress has proved to be a powerful tool.High-performanceliquid chromatography coupled with tandem mass spectrometry(HPLC-MS/MS) has emerged as the most accurate method for avoiding problemsrelated to the nonspecificity of immunoassays. In this study, we optimize and validatea reliable method using HPLC-MS/MS for quantifying 11-ketoetiocholanolone(11-k),a representative fecal cortisol metabolite in ruminants. An appropriate extractionand purification procedure was developed taking into account the complex nature offeces. The final extract obtained was then analyzed with HPLC-MS/MS using aquadrupole-time-of-fly(QTOF) tandem mass spectrometer with an electrospray ionizationinterface operating in positive mode, which allowed an unequivocal determinationof the metabolite due to its accurate mass capabilities. After rigorousoptimization of both sample extraction and the HPLC-QTOFparameters, making useof feces from free-rangingIberian ibex, ideal conditions were established. Matrix-matchedstandards were used to calibrate the method. The limit of detection andquantification was 13-and40-ng/g, respectively. The validation of the method wasperformed with recoveries in the range of 85–110%, a figure much higher than the60% obtained with the previous extraction methods used in our laboratory, and withrelative standard deviations (RSDs) no higher than 15% for the complete analyticalprocedure, including extraction and analysis. The time required for the fecal 11-kanalysis was greatly reduced in comparison with the previous work carried out in ourlaboratory. This is the first time that QTOF mass detection coupled with HPLC hasbeen validated for 11-kquantification in feces from free-rangingruminants such asIberian ibex. Given the high selectivity and sensitivity attained, our method couldbecome a useful tool for noninvasive stress quantification in ruminants.
PB Wiley Open Access
SN 2045-7758
LK https://hdl.handle.net/10612/19504
UL https://hdl.handle.net/10612/19504
NO Molina-García, L., Pérez, J. M., Sarasa, M., Ureña-Gutiérrez, B., Espinosa, J., & Azorit, C. (2018). HPLC-QTOF method for quantifying 11-ketoetiocholanolone, a cortisol metabolite, in ruminants” feces: Optimization and validation. Ecology and Evolution, 8(18), 9218-9228. https://doi.org/10.1002/ECE3.4285
NO This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. © 2018 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 01-jun-2024