Advanced search
1 file | 2.17 MB Add to list

Optimization and validation of metabolomics methods for feline urine and serum towards application in veterinary medicine

Ellen Vanden Broecke (UGent) , Laurens Van Mulders (UGent) , Ellen De Paepe (UGent) , Sylvie Daminet (UGent) and Lynn Vanhaecke (UGent)
Author
Organization
Project
Abstract
Background Metabolomics is an emerging and powerful technology that offers a comprehensive view of an organism's physiological status. Although widely applied in human medicine, it is only recently making its introduction in veterinary medicine. As a result, validated metabolomics protocols in feline medicine are lacking at the moment. Since biological interpretation of metabolomics data can be misled by the extraction method used, species and matrix-specific optimized and validated metabolomic protocols are sorely needed. Results Systematic optimization was performed using fractional factorial experiments for both serum (n = 57) and urine (n = 24), evaluating dilution for both matrices, and aliquot and solvent volume, protein precipitation time and temperature for serum. For the targeted (n = 76) and untargeted (n = 1949) validation of serum respectively, excellent instrumental, intra-assay and inter-day precision were observed (CV ≤ 15% or 30%, respectively). Linearity deemed sufficient both targeted and untargeted (R2 ≥ 0.99 or 0.90, respectively). An appropriate targeted recovery between 70 and 130% was achieved. For the targeted (n = 69) and untargeted (n = 2348) validation of the urinary protocol, excellent instrumental and intra-assay precision were obtained (CV ≤ 15% or 30%, respectively). Subsequently, the discriminative ability of our metabolomics methods was confirmed for feline chronic kidney disease (CKD) by univariate statistics (n = 41 significant metabolites for serum, and n = 55 for urine, p-value<0.05) and validated OPLS-DA models (R2(Y) > 0.95, Q2(Y) > 0.65, p-value<0.001 for both matrices). Significance This study is the first to present an optimized and validated wholistic metabolomics methods for feline serum and urine using ultra-high performance liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry. This robust methodology opens avenues for biomarker panel selection and a deeper understanding of feline CKD pathophysiology and other feline applications.
Keywords
Biofluids, Metabotyping, UHPLC-Orbitrap-HRMS, Feline nephrology, CHRONIC KIDNEY-DISEASE, MASS-SPECTROMETRY, PLASMA, DIAGNOSIS, COVERAGE, GRAVITY, FECES

Downloads

  • (...).pdf
    • full text (Published version)
    • |
    • UGent only
    • |
    • PDF
    • |
    • 2.17 MB

Citation

Please use this url to cite or link to this publication:

MLA
Vanden Broecke, Ellen, et al. “Optimization and Validation of Metabolomics Methods for Feline Urine and Serum towards Application in Veterinary Medicine.” ANALYTICA CHIMICA ACTA, vol. 1310, 2024, doi:10.1016/j.aca.2024.342694.
APA
Vanden Broecke, E., Van Mulders, L., De Paepe, E., Daminet, S., & Vanhaecke, L. (2024). Optimization and validation of metabolomics methods for feline urine and serum towards application in veterinary medicine. ANALYTICA CHIMICA ACTA, 1310. https://doi.org/10.1016/j.aca.2024.342694
Chicago author-date
Vanden Broecke, Ellen, Laurens Van Mulders, Ellen De Paepe, Sylvie Daminet, and Lynn Vanhaecke. 2024. “Optimization and Validation of Metabolomics Methods for Feline Urine and Serum towards Application in Veterinary Medicine.” ANALYTICA CHIMICA ACTA 1310. https://doi.org/10.1016/j.aca.2024.342694.
Chicago author-date (all authors)
Vanden Broecke, Ellen, Laurens Van Mulders, Ellen De Paepe, Sylvie Daminet, and Lynn Vanhaecke. 2024. “Optimization and Validation of Metabolomics Methods for Feline Urine and Serum towards Application in Veterinary Medicine.” ANALYTICA CHIMICA ACTA 1310. doi:10.1016/j.aca.2024.342694.
Vancouver
1.
Vanden Broecke E, Van Mulders L, De Paepe E, Daminet S, Vanhaecke L. Optimization and validation of metabolomics methods for feline urine and serum towards application in veterinary medicine. ANALYTICA CHIMICA ACTA. 2024;1310.
IEEE
[1]
E. Vanden Broecke, L. Van Mulders, E. De Paepe, S. Daminet, and L. Vanhaecke, “Optimization and validation of metabolomics methods for feline urine and serum towards application in veterinary medicine,” ANALYTICA CHIMICA ACTA, vol. 1310, 2024.
@article{01HY01095GVCKJ1A59GTWYAE48,
  abstract     = {{Background
Metabolomics is an emerging and powerful technology that offers a comprehensive view of an organism's physiological status. Although widely applied in human medicine, it is only recently making its introduction in veterinary medicine. As a result, validated metabolomics protocols in feline medicine are lacking at the moment. Since biological interpretation of metabolomics data can be misled by the extraction method used, species and matrix-specific optimized and validated metabolomic protocols are sorely needed.

Results
Systematic optimization was performed using fractional factorial experiments for both serum (n = 57) and urine (n = 24), evaluating dilution for both matrices, and aliquot and solvent volume, protein precipitation time and temperature for serum. For the targeted (n = 76) and untargeted (n = 1949) validation of serum respectively, excellent instrumental, intra-assay and inter-day precision were observed (CV ≤ 15% or 30%, respectively). Linearity deemed sufficient both targeted and untargeted (R2 ≥ 0.99 or 0.90, respectively). An appropriate targeted recovery between 70 and 130% was achieved. For the targeted (n = 69) and untargeted (n = 2348) validation of the urinary protocol, excellent instrumental and intra-assay precision were obtained (CV ≤ 15% or 30%, respectively). Subsequently, the discriminative ability of our metabolomics methods was confirmed for feline chronic kidney disease (CKD) by univariate statistics (n = 41 significant metabolites for serum, and n = 55 for urine, p-value<0.05) and validated OPLS-DA models (R2(Y) > 0.95, Q2(Y) > 0.65, p-value<0.001 for both matrices).

Significance
This study is the first to present an optimized and validated wholistic metabolomics methods for feline serum and urine using ultra-high performance liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry. This robust methodology opens avenues for biomarker panel selection and a deeper understanding of feline CKD pathophysiology and other feline applications.}},
  articleno    = {{342694}},
  author       = {{Vanden Broecke, Ellen and Van Mulders, Laurens and De Paepe, Ellen and Daminet, Sylvie and Vanhaecke, Lynn}},
  issn         = {{0003-2670}},
  journal      = {{ANALYTICA CHIMICA ACTA}},
  keywords     = {{Biofluids,Metabotyping,UHPLC-Orbitrap-HRMS,Feline nephrology,CHRONIC KIDNEY-DISEASE,MASS-SPECTROMETRY,PLASMA,DIAGNOSIS,COVERAGE,GRAVITY,FECES}},
  language     = {{eng}},
  pages        = {{9}},
  title        = {{Optimization and validation of metabolomics methods for feline urine and serum towards application in veterinary medicine}},
  url          = {{http://doi.org/10.1016/j.aca.2024.342694}},
  volume       = {{1310}},
  year         = {{2024}},
}

Altmetric
View in Altmetric
Web of Science
Times cited: