Liquid chromatography-mass spectrometry method development for monitoring stress-related corticosteroids levels in pig saliva

J Chromatogr B Analyt Technol Biomed Life Sci. 2015 May 15:990:158-63. doi: 10.1016/j.jchromb.2015.03.021. Epub 2015 Apr 2.

Abstract

Biochemical response stressors results in an increase of adrenocortical activity. Before knowing the corticosteroid levels in saliva in a stressful situation, baselines salivary levels should be established. A method for simultaneous determination of five corticosteroids was developed, validated and applied to pig saliva at farms. The method employs solid-phase extraction (SPE) coupled with clean-up extraction step using silica cartridge in the same step followed by liquid chromatography/tandem mass spectrometry (LC-MS/MS), using electrospray ionization (ESI) in positive mode. The overall method quantification limits range from 0.050 to 0.30μg/L for the enrichment of 1.0mL saliva samples and analyte recoveries are between 60 and 90% (RSD<11%). Some factors studied were: pig sex, breeds, and time at farm. The analytical method clearly shows that CRL and CRS levels of, respectively, 3.0 and 4.0μg/L in saliva can be indicative of maxima non-stress levels in different pig breeds at farm.

Keywords: Corticosteroids; Liquid chromatography/tandem mass spectrometry; Pig breeds; Saliva; Solid-phase extraction; Stress hormones.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenal Cortex Hormones / analysis*
  • Animals
  • Chromatography, Liquid / methods*
  • Female
  • Linear Models
  • Male
  • Reproducibility of Results
  • Saliva / chemistry*
  • Sensitivity and Specificity
  • Sus scrofa
  • Swine
  • Tandem Mass Spectrometry / methods*

Substances

  • Adrenal Cortex Hormones