Rapid UHPLC-MS/MS measurement of pregnanediol 3-glucuronide in spot urine samples for detecting ovulation.

Autor: Leoni L; Clinical Biochemistry Laboratory, Department of Medical Sciences, University of Turin, Turin, Italy., Rosmini F; Endocrinology, Diabetes and Metabolism, Department of Medicine, University of Verona, Verona, Italy., Ledda F; Endocrinology, Diabetes and Metabolism, Department of Medical Sciences, University of Turin, Turin, Italy., Parasiliti-Caprino M; Endocrinology, Diabetes and Metabolism, Department of Medical Sciences, University of Turin, Turin, Italy., Settanni F; Clinical Biochemistry Laboratory, City of Health and Science University Hospital, Turin, Italy., Nonnato A; Clinical Biochemistry Laboratory, City of Health and Science University Hospital, Turin, Italy., Ghigo E; Endocrinology, Diabetes and Metabolism, Department of Medical Sciences, University of Turin, Turin, Italy., Moghetti P; Endocrinology, Diabetes and Metabolism, Department of Medicine, University of Verona, Verona, Italy., Mengozzi G; Clinical Biochemistry Laboratory, Department of Medical Sciences, University of Turin, Turin, Italy.; Clinical Biochemistry Laboratory, City of Health and Science University Hospital, Turin, Italy., Ponzetto F; Endocrinology, Diabetes and Metabolism, Department of Medical Sciences, University of Turin, Turin, Italy.
Jazyk: angličtina
Zdroj: Biomedical chromatography : BMC [Biomed Chromatogr] 2024 Oct; Vol. 38 (10), pp. e5982. Date of Electronic Publication: 2024 Aug 16.
DOI: 10.1002/bmc.5982
Abstrakt: Biochemical confirmation of ovulation typically involves measuring serum progesterone levels during the mid-luteal phase. Alternatively, this information could be obtained by monitoring urinary excretion of conjugated metabolites of ovarian steroids such as pregnanediol 3-glucuronide (PDG) using immunoassay techniques that have methodological limitations. The aim of the present study was to develop a mass spectrometry (MS)-based method for the rapid and accurate measurement of urinary PDG levels in spot urine samples. A "dilute and shoot" ultra-high-performance liquid cromatography tandem mass spectrometry (UHPLC-MS/MS) method was developed for measuring PDG urinary concentration with a 6-min analysis time. The method underwent validation in accordance with ISO 17025 documentation for quantitative methods, proving an efficient separation of PDG from other structurally similar glucuro-conjugated steroid metabolites and ensuring a sufficient sensitivity for detecting the target analyte at concentrations as low as 0.01 μg/mL. The validation protocol yielded satisfactory results in terms of accuracy, repeatability, intermediate precision, and combined uncertainty. Additionally, the stability of both the samples and calibration curves was also conducted. The application to real urine samples confirmed the method's capability to measure PDG levels throughout an entire menstrual cycle and detecting ovulation. The rapidity of the analytical platform would therefore enable high throughput analysis, which is advantageous for large cohort clinical studies.
(© 2024 The Author(s). Biomedical Chromatography published by John Wiley & Sons Ltd.)
Databáze: MEDLINE
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