Ann Lab Med.  2021 Jan;41(1):108-113. 10.3343/alm.2021.41.1.108.

A Sensitive and Specific Liquid ChromatographyTandem Mass Spectrometry Assay for Simultaneous Quantification of Salivary Melatonin and Cortisol: Development and Comparison With Immunoassays

• Shin S
• Oh H
• Park HR
• Joo EY
• Lee SY

• KMID: 2512740
• DOI: http://doi.org/10.3343/alm.2021.41.1.108

Abstract

Melatonin and cortisol are clinically important for diagnosing sleep and mood disorders. We developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/ MS) assay for simultaneous measurement of salivary melatonin and cortisol concentrations according to the Clinical and Laboratory Standards Institute guidelines. Additionally, we compared the LC-MS/MS assay with immunoassays, ELISA (Direct Salivary Melatonin Elisa EK-DSM, Bühlmann Laboratories AG, Schönenbuch, Switzerland) and electrochemiluminescence immunoassay (Cortisol II, Roche, Mannheim, Germany), using 121 saliva samples. The LC-MS/MS assay exhibited good performance in terms of linearity, precision, accuracy, limit of detection, lower limit of quantification, extraction recovery, carry-over, and matrix effect. The LC-MS/MS assay and immunoassays showed strong correlation (Pearson’s r = 0.910 for melatonin, r = 0.955 for cortisol), but demonstrated a significant mean bias of 23.2% (range 54.0–143.7%) for melatonin and 48.9% (range 59.7–184.7%) for cortisol. Our LC-MS/MS assay provided more sensitive and reliable salivary melatonin and cortisol quantification results compared with immunoassays.

Keyword

Melatonin; Cortisol; Liquid chromatography-tandem mass spectrometry (LCMS/MS); Immunoassay; Saliva; Performance

Figure

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  Fig. 1 Comparison plots between LC-MS/MS and immunoassays. Top: (A) Passing-Bablok regression and (B) Bland-Altman plot to compare salivary melatonin concentrations measured by LC-MS/MS assay and ELISA. Bottom: (C) Passing-Bablok regression and (D) Bland-Altman plot to compare salivary cortisol concentrations measured by LC-MS/MS assay and ECLIA. Abbreviations: LC-MS/MS, liquid chromatography-tandem mass spectrometry; ECLIA, electrochemiluminescence immunoassay.

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  Fig. 2 Melatonin and cortisol profiles of two participants measured by LC-MS/MS assay and immunoassays. Abbreviations: ECLIA, electrochemiluminescence immunoassay; LC-MS/MS, liquid chromatography-tandem mass spectrometry.

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