Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS
A simple and efficient validated assay for quantifying 21-deoxycortisol (21-DOC), 17-hydroxyprogesterone (17-OHP), cortisol, and cortisone in human plasma has been developed using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Analysis of plasma sam...
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| Format: | Article |
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Wiley
2025-01-01
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| Series: | Advances in Pharmacological and Pharmaceutical Sciences |
| Online Access: | http://dx.doi.org/10.1155/adpp/3859670 |
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| author | Syed N. Alvi Saleh Al dgither Ali Al-Odaib |
| author_facet | Syed N. Alvi Saleh Al dgither Ali Al-Odaib |
| author_sort | Syed N. Alvi |
| collection | DOAJ |
| description | A simple and efficient validated assay for quantifying 21-deoxycortisol (21-DOC), 17-hydroxyprogesterone (17-OHP), cortisol, and cortisone in human plasma has been developed using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Analysis of plasma samples were performed on Atlantis dC18 (3 μm) column using a mobile phase of 20.0 mM ammonium acetate and acetonitrile (50:50, v:v) that was delivered at isocratic flow rate 0.3 mL/minute. After addition of d4-cortisol as an internal standard (IS), plasma samples containing 21-DOC, 17-OHP, cortisol, and cortisone were extracted with mixture of dichloromethane and tert-butylmethyl ether 1:2 (v/v). Analytes were detected and quantified in the positive ion mode of electrospray ionization using multiple reaction monitoring transition set at mass to charge (m/z): 347.17 ⟶ 311.12, 331.17 ⟶ 96.97, 363.11 ⟶ 121.00, 361.18 ⟶ 163.11, and 367.19 ⟶ 121.24 for 21-DOC and 17-OHP, cortisol, cortisone, and cortisol-d4 (IS), respectively. The relationship between concentration and peak area response (analyte/IS) were linear over the range of 0.25–50, 0.5–100, 1–200, and 2–400 ng/mL for 21-DOC, 17-OHP, cortisone, and cortisol, respectively. The mean extraction recovery of the analytes was in the range of 83%–96%. The coefficient of variation within and between days was less than 13.6%, and the bias ranged from −9.2% to 12%. The measured level of cortisol, cortisone, and 17-OHP was in the range of 21.9–110, 4.33–12.71, and 0.37–1.4 ng/mL, respectively. Furthermore, the measured value of cortisone–cortisol ratio was in the range of 0.08–0.21. |
| format | Article |
| id | doaj-art-583e6897e8744538984c91da7643c7a3 |
| institution | DOAJ |
| issn | 2633-4690 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advances in Pharmacological and Pharmaceutical Sciences |
| spelling | doaj-art-583e6897e8744538984c91da7643c7a32025-08-20T03:12:35ZengWileyAdvances in Pharmacological and Pharmaceutical Sciences2633-46902025-01-01202510.1155/adpp/3859670Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MSSyed N. Alvi0Saleh Al dgither1Ali Al-Odaib2Environmental Health ProgramEnvironmental Health ProgramResearch Administrative OperationsA simple and efficient validated assay for quantifying 21-deoxycortisol (21-DOC), 17-hydroxyprogesterone (17-OHP), cortisol, and cortisone in human plasma has been developed using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Analysis of plasma samples were performed on Atlantis dC18 (3 μm) column using a mobile phase of 20.0 mM ammonium acetate and acetonitrile (50:50, v:v) that was delivered at isocratic flow rate 0.3 mL/minute. After addition of d4-cortisol as an internal standard (IS), plasma samples containing 21-DOC, 17-OHP, cortisol, and cortisone were extracted with mixture of dichloromethane and tert-butylmethyl ether 1:2 (v/v). Analytes were detected and quantified in the positive ion mode of electrospray ionization using multiple reaction monitoring transition set at mass to charge (m/z): 347.17 ⟶ 311.12, 331.17 ⟶ 96.97, 363.11 ⟶ 121.00, 361.18 ⟶ 163.11, and 367.19 ⟶ 121.24 for 21-DOC and 17-OHP, cortisol, cortisone, and cortisol-d4 (IS), respectively. The relationship between concentration and peak area response (analyte/IS) were linear over the range of 0.25–50, 0.5–100, 1–200, and 2–400 ng/mL for 21-DOC, 17-OHP, cortisone, and cortisol, respectively. The mean extraction recovery of the analytes was in the range of 83%–96%. The coefficient of variation within and between days was less than 13.6%, and the bias ranged from −9.2% to 12%. The measured level of cortisol, cortisone, and 17-OHP was in the range of 21.9–110, 4.33–12.71, and 0.37–1.4 ng/mL, respectively. Furthermore, the measured value of cortisone–cortisol ratio was in the range of 0.08–0.21.http://dx.doi.org/10.1155/adpp/3859670 |
| spellingShingle | Syed N. Alvi Saleh Al dgither Ali Al-Odaib Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS Advances in Pharmacological and Pharmaceutical Sciences |
| title | Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS |
| title_full | Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS |
| title_fullStr | Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS |
| title_full_unstemmed | Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS |
| title_short | Method Development and Clinical Utility for Simultaneous Measurement of 21-Deoxycortisol, 17-Hydroxyprogesterone, Cortisol, and Cortisone Levels in Human Plasma Using UHPLC-MS/MS |
| title_sort | method development and clinical utility for simultaneous measurement of 21 deoxycortisol 17 hydroxyprogesterone cortisol and cortisone levels in human plasma using uhplc ms ms |
| url | http://dx.doi.org/10.1155/adpp/3859670 |
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