A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human Saliva
A simple ultraperformance liquid chromatography-tandem mass spectrometry assay for measurement of cortisol level in human saliva was developed and validated. Saliva samples containing cortisol were spiked with tolperisone as internal standard (IS) and extracted with a mixture of methyl tert-butyl et...
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| Format: | Article |
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Wiley
2019-01-01
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| Series: | International Journal of Analytical Chemistry |
| Online Access: | http://dx.doi.org/10.1155/2019/4909352 |
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| author | Syed N. Alvi Muhammad M. Hammami |
| author_facet | Syed N. Alvi Muhammad M. Hammami |
| author_sort | Syed N. Alvi |
| collection | DOAJ |
| description | A simple ultraperformance liquid chromatography-tandem mass spectrometry assay for measurement of cortisol level in human saliva was developed and validated. Saliva samples containing cortisol were spiked with tolperisone as internal standard (IS) and extracted with a mixture of methyl tert-butyl ether and hexane (8:2, v:v). After solvent evaporation, residue was reconstituted in 100 μl mobile phase. Analysis was performed on Atlantis dC18 column (2.1 × 100 mm, 3 μm particle size) with a mobile phase composed of acetonitrile and 2 mM ammonium acetate (50:50, v:v) and delivered at a flow rate of 0.3 ml/minute. Mass spectrometry acquisition was performed with multiple reaction monitoring in positive-ion mode for cortisol and IS (m/z: 363.1 → 121.0 and 246.0 → 97.9, respectively). Retention times of cortisol and IS were about 1.35 and 2.45 minutes, respectively. The relationship between cortisol level and peak area ratio of cortisol to IS was linear in the range of 0.5-100 ng/ml. Intra- and interday coefficient of variation and bias were ≤ 9.0% and ≤12.0%, respectively. Mean extraction recoveries of cortisol and IS from saliva samples were 92% and 94%, respectively. Using the method, cortisol was found to be ≥ 86% stable in processed (24 hours at room temperature or 48 hours at -20°C) and ≥ 91% stable in unprocessed (24 hours at room temperature or 20 weeks at -20°C) saliva samples. Further, the method was successfully applied to determine daily cortisol profile in saliva samples of a healthy volunteer. |
| format | Article |
| id | doaj-art-9a09a47112424748b1d10708b8bef97d |
| institution | OA Journals |
| issn | 1687-8760 1687-8779 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
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| series | International Journal of Analytical Chemistry |
| spelling | doaj-art-9a09a47112424748b1d10708b8bef97d2025-08-20T02:20:16ZengWileyInternational Journal of Analytical Chemistry1687-87601687-87792019-01-01201910.1155/2019/49093524909352A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human SalivaSyed N. Alvi0Muhammad M. Hammami1Clinical Studies and Empirical Ethics Department, King Faisal Specialist Hospital & Research Center, MBC-03, P.O. Box 3354, Riyadh 11211, Saudi ArabiaClinical Studies and Empirical Ethics Department, King Faisal Specialist Hospital & Research Center, MBC-03, P.O. Box 3354, Riyadh 11211, Saudi ArabiaA simple ultraperformance liquid chromatography-tandem mass spectrometry assay for measurement of cortisol level in human saliva was developed and validated. Saliva samples containing cortisol were spiked with tolperisone as internal standard (IS) and extracted with a mixture of methyl tert-butyl ether and hexane (8:2, v:v). After solvent evaporation, residue was reconstituted in 100 μl mobile phase. Analysis was performed on Atlantis dC18 column (2.1 × 100 mm, 3 μm particle size) with a mobile phase composed of acetonitrile and 2 mM ammonium acetate (50:50, v:v) and delivered at a flow rate of 0.3 ml/minute. Mass spectrometry acquisition was performed with multiple reaction monitoring in positive-ion mode for cortisol and IS (m/z: 363.1 → 121.0 and 246.0 → 97.9, respectively). Retention times of cortisol and IS were about 1.35 and 2.45 minutes, respectively. The relationship between cortisol level and peak area ratio of cortisol to IS was linear in the range of 0.5-100 ng/ml. Intra- and interday coefficient of variation and bias were ≤ 9.0% and ≤12.0%, respectively. Mean extraction recoveries of cortisol and IS from saliva samples were 92% and 94%, respectively. Using the method, cortisol was found to be ≥ 86% stable in processed (24 hours at room temperature or 48 hours at -20°C) and ≥ 91% stable in unprocessed (24 hours at room temperature or 20 weeks at -20°C) saliva samples. Further, the method was successfully applied to determine daily cortisol profile in saliva samples of a healthy volunteer.http://dx.doi.org/10.1155/2019/4909352 |
| spellingShingle | Syed N. Alvi Muhammad M. Hammami A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human Saliva International Journal of Analytical Chemistry |
| title | A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human Saliva |
| title_full | A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human Saliva |
| title_fullStr | A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human Saliva |
| title_full_unstemmed | A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human Saliva |
| title_short | A Simple Ultraperformance Liquid Chromatography-Tandem Mass Spectrometry Method for Measurement of Cortisol Level in Human Saliva |
| title_sort | simple ultraperformance liquid chromatography tandem mass spectrometry method for measurement of cortisol level in human saliva |
| url | http://dx.doi.org/10.1155/2019/4909352 |
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