A validated method for measurement of serum total, serum free, and salivary cortisol, using high-performance liquid chromatography coupled with high-resolutionESI-TOF mass spectrometry

Gergely Montskó, Zita Tarjányi, Emese Mezosi, Gábor L. Kovács

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Blood cortisol level is routinely analysed in laboratory medicine, but the immunoassays in widespread use have the disadvantage of cross-reactivity with some commonly used steroid drugs. Mass spectrometry has become a method of increasing importance for cortisol estimation. However, current methods do not offer the option of accurate mass identification. Our objective was to develop a mass spectrometry method to analyse salivary, serum total, and serum free cortisol via accurate mass identification. The analysis was performed on a Bruker micrOTOF high-resolution mass spectrometer. Sample preparation involved protein precipitation, serum ultrafiltration, and solid-phase extraction. Limit of quantification was 12.5 nmol L-1 for total cortisol, 440 pmol L-1 for serum ultrafiltrate, and 600 pmol L-1 for saliva. Average intra-assay variation was 4.7 %, and inter-assay variation was 6.6 %. Mass accuracy was -1, and serum free cortisol levels were in the range 0.5-12.4 nmol L-1. Salivary cortisol levels were in the range 0.7-10.4 nmol L-1. Mass accuracy was equal to or below 2.5 ppm, resulting in a mass error less than 1 mDa and thus providing high specificity. We did not observe any interference with routinely used steroidal drugs. The method is capable of specific cortisol quantification in different matrices on the basis of accurate mass identification.

Original languageEnglish
Pages (from-to)2333-2341
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Volume406
Issue number9-10
DOIs
Publication statusPublished - 2014

Fingerprint

High performance liquid chromatography
Mass spectrometry
Hydrocortisone
Mass Spectrometry
High Pressure Liquid Chromatography
Serum
Assays
Solid Phase Extraction
Ultrafiltration
Mass spectrometers
Saliva
Immunoassay
Pharmaceutical Preparations
Medicine
Blood Proteins
Blood
Steroids

Keywords

  • High-resolution mass spectrometry
  • Liquid chromatography
  • Salivary cortisol
  • Serum cortisol

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Medicine(all)

Cite this

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title = "A validated method for measurement of serum total, serum free, and salivary cortisol, using high-performance liquid chromatography coupled with high-resolutionESI-TOF mass spectrometry",
abstract = "Blood cortisol level is routinely analysed in laboratory medicine, but the immunoassays in widespread use have the disadvantage of cross-reactivity with some commonly used steroid drugs. Mass spectrometry has become a method of increasing importance for cortisol estimation. However, current methods do not offer the option of accurate mass identification. Our objective was to develop a mass spectrometry method to analyse salivary, serum total, and serum free cortisol via accurate mass identification. The analysis was performed on a Bruker micrOTOF high-resolution mass spectrometer. Sample preparation involved protein precipitation, serum ultrafiltration, and solid-phase extraction. Limit of quantification was 12.5 nmol L-1 for total cortisol, 440 pmol L-1 for serum ultrafiltrate, and 600 pmol L-1 for saliva. Average intra-assay variation was 4.7 {\%}, and inter-assay variation was 6.6 {\%}. Mass accuracy was -1, and serum free cortisol levels were in the range 0.5-12.4 nmol L-1. Salivary cortisol levels were in the range 0.7-10.4 nmol L-1. Mass accuracy was equal to or below 2.5 ppm, resulting in a mass error less than 1 mDa and thus providing high specificity. We did not observe any interference with routinely used steroidal drugs. The method is capable of specific cortisol quantification in different matrices on the basis of accurate mass identification.",
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AU - Mezosi, Emese

AU - Kovács, Gábor L.

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