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Home / Equine Research Bank / Anal Bioanal Chem / Determination of hair cortisol in horses: comparison of immu...
Analytical and bioanalytical chemistry2022; 414(28); 8093-8105; doi: 10.1007/s00216-022-04343-6

Determination of hair cortisol in horses: comparison of immunoassay vs LC-HRMS/MS.

Abstract: The measure of hair cortisol concentration (HCC) is becoming an emerging approach to monitor mid-/long-term stress in animals, so it is more and more important to develop accurate and reliable methods. In the light of this, the aim of the present study was to compare mane HCCs of 47 horses with different managements, by means of an immunoassay (ELISA) and liquid chromatography coupled to hybrid high-resolution mass spectrometry (LC-HRMS/MS). After the washing step, the ground hair was extracted with methanol. The extract was evaporated and redissolved in two different aqueous solutions, depending on the detection technique. The methods were validated according to EMA guideline for bioanalytical method validation, in the range 2-50 pg mg (ELISA) and 1-100 pg mg (LC-HRMS/MS). Satisfactory quantitative performances were obtained for both of the approaches, but this latter demonstrated better precision. The detected concentrations in real samples were encompassing the range 1.3-8.8 pg mg and 2.0-17.9 pg mg by means of LC-HRMS/MS and ELISA, respectively. Overall, HCCs measured with ELISA technique were 1.6 times higher. The overestimation of immunoassay results might be caused by cross-reactivity phenomena of laboratory reagents and other structurally similar hormones present in the mane.
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Publication Date: 2022-09-22 PubMed ID: 36136115PubMed Central: PMC9613578DOI: 10.1007/s00216-022-04343-6Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research aimed to assess the accuracy and reliability of two methods used to measure hair cortisol concentration (HCC) in horses. The two methods compared were an immunoassay (ELISA) and liquid chromatography coupled to hybrid high-resolution mass spectrometry (LC-HRMS/MS). The study indicated that while both methods offered satisfactory quantitative performance, the LC-HRMS/MS method demonstrated better precision.

Research Methodology

  • The researchers carried out the study on 47 horses with differing managements.
  • The extracted cortisol from the mane HCCs of the horses was tested by using immunoassay (ELISA) and LC-HRMS/MS.
  • Different aqueous solutions were used to redissolve the hair extracts, depending on the detection technique utilized.
  • The research methods were validated in accordance with the EMA guideline for bioanalytical method validation, with a range of 2-50 pg mg for ELISA and 1-100 pg mg for LC-HRMS/MS.

Results and Findings

  • The results revealed that both techniques had satisfactory quantitative performace, but LC-HRMS/MS showed better precision.
  • The concentrations detected in real samples ranged between 1.3-8.8 pg mg for LC-HRMS/MS and 2.0-17.9 pg mg for ELISA.
  • The measurements obtained with the ELISA technique were approximately 1.6 times higher than those achieved with LC-HRMS/MS. The overestimation possibly caused by immunoassay could be due to cross-reactivity phenomena of laboratory reagents and other hormones similar in structure present in the mane.

Conclusion and Implications

  • The research provides insights into the most effective methods of measuring HCC in horses.
  • The results suggest that while ELISA and LC-HRMS/MS both offer satisfactory results, the latter demonstrates better precision.
  • Moreover, this differentiation in the results may impact decision-making concerning which method is best suited for determining and monitoring mid-/long-term stress in animals.
  • As a consequence, more reliable results might be obtained with LC-HRMS/MS, although ELISA may still be useful in contexts where the higher concentration results are not a critical factor.

Cite This Article

APA
(2022). Determination of hair cortisol in horses: comparison of immunoassay vs LC-HRMS/MS. Anal Bioanal Chem, 414(28), 8093-8105. https://doi.org/10.1007/s00216-022-04343-6

Publication

Analytical and bioanalytical chemistry
ISSN: 1618-2650
NlmUniqueID: 101134327
Country: Germany
Language: English
Volume: 414
Issue: 28
Pages: 8093-8105

Researcher Affiliations

MeSH Terms

  • Horses
  • Animals
  • Hydrocortisone / analysis
  • Chromatography, Liquid / methods
  • Hair / chemistry
  • Mass Spectrometry
  • Enzyme-Linked Immunosorbent Assay
  • Immunoassay

Grant Funding

  • IZS AM PS/19 RC / Ministero della Salute

Conflict of Interest Statement

The authors declare no competing interests.

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Citations

This article has been cited 5 times.
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