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Related Experiment Video

Updated: Jun 23, 2026

Extraction and Analysis of Cortisol from Human and Monkey Hair

Extraction and Analysis of Cortisol from Human and Monkey Hair

Published on: January 24, 2014

Human saliva cortisone and cortisol simultaneous analysis using reverse phase HPLC technique.

Elio F De Palo¹, Giorgia Antonelli, Arianna Benetazzo

¹Section of Clinical Biochemistry, Department of Medical Diagnostic Sciences and Special Therapies, University of Padua, 35100 Padua, Italy. eliof.depalo@unipd.it

Clinica Chimica Acta; International Journal of Clinical Chemistry

|April 28, 2009

Summary

This study presents a validated method for simultaneously analyzing cortisol (F) and cortisone (E) in saliva using solid-phase extraction and RP-HPLC. The method is suitable for clinical diagnostics, showing significant diurnal variations in hormone levels.

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Last Updated: Jun 23, 2026

Extraction and Analysis of Cortisol from Human and Monkey Hair

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Published on: January 24, 2014

Cortisol Extraction from Sturgeon Fin and Jawbone Matrices

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Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Published on: December 18, 2013

Area of Science:

Endocrinology
Clinical Chemistry
Analytical Chemistry

Background:

Accurate cortisol (F) analysis is crucial for diagnosing hyper- and hypo-tension disorders like Cushing's syndrome.
Simultaneous measurement of cortisol and its inactive metabolite cortisone (E) in saliva aids in endocrine investigations.

Purpose of the Study:

To develop and validate a method for the simultaneous analysis of cortisol and cortisone in human saliva.
To assess the suitability of this method for routine clinical biochemistry laboratory use.

Main Methods:

Saliva samples were extracted using C18 solid-phase extraction (SPE) and analyzed by isocratic reversed-phase high-performance liquid chromatography (RP-HPLC).
Cortisol and cortisone were detected using UV detection.
Saliva specimens were collected from healthy volunteers in the morning and evening.

Main Results:

The method demonstrated reliable calibration curve ranges, low limits of detection (LOD), and acceptable intra- and inter-assay coefficients of variation (CVs).
Recovery rates for spiked saliva samples were 99% for cortisone and 88% for cortisol.
Significantly higher levels of both cortisone and cortisol were observed in morning saliva samples compared to evening samples (p<0.001).

Conclusions:

The developed RP-HPLC method is suitable for the simultaneous analysis of cortisone and cortisol in saliva specimens.
This validated method can be utilized for periodic analyses in clinical biochemistry laboratories for endocrinology investigations.