Determination of human urinary cortisol, cortisone and relative phase II metabolites by dilute-and-shoot LC-MS/MS analysis: an application to adrenal disease

IF 3.2 3区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Clinica Chimica Acta Pub Date : 2025-04-20 DOI:10.1016/j.cca.2025.120324

Laura Leoni , Mirko Parasiliti Caprino , Giulia Montesano , Chiara Lopez , Martina Bollati , Fabio Settanni , Ezio Ghigo , Giulio Mengozzi , Federico Ponzetto

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引用次数: 0

Abstract

Background

Cortisol and its primary metabolite, cortisone, play key roles in managing stress responses and maintaining balance via the hypothalamic–pituitary–adrenal axis. Traditional immunoassays, often hindered by cross-reactivity, can yield inaccurate results. This study introduces a novel dilute-and-shoot using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for the rapid and accurate analysis of urinary free cortisol, cortisone, and their intact phase II metabolites.

Methods

A new LC-MS/MS method was developed to quantify five analytes, including intact glucuro- and sulfo-conjugated cortisol and cortisone metabolites. The method underwent validation assessments in accordance with ISO 17025 standards for quantitative methods. Its performance was compared with a commercial LC-MS/MS assay used in clinical laboratories, analyzing free cortisol and cortisone in 79 24-hour urine samples. Additionally, the novel method was applied to authentic urine samples from 168 adrenal disease patients.

Results

The validation protocol assessed method’s selectivity, matrix effects, quantitative performance and carry-over. The comparison showed excellent correlation with the commercial LC-MS/MS method for urinary free cortisol and cortisone measurement. Pathological sample analysis indicated that monitoring intact phase II metabolites, particularly sulfoconjugates, could aid in differentiating adrenal diseases.

Conclusion

The developed and validated dilute-and-shoot LC-MS/MS method demonstrated satisfactory performance, offering a faster, cost-effective solution for adrenal hormone analysis in clinical settings. This research also provided insights into adrenal function by examining the urinary excretion of target analytes in a pathological cohort, enhancing diagnostic precision for related disorders.

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稀释射击LC-MS/MS分析测定人尿皮质醇、可的松及相关II期代谢物：在肾上腺疾病中的应用

皮质醇及其主要代谢物可的松在管理应激反应和通过下丘脑-垂体-肾上腺轴维持平衡中发挥关键作用。传统的免疫测定常常受到交叉反应性的阻碍，可能产生不准确的结果。本研究采用液相色谱-串联质谱联用技术（LC-MS/MS）对尿液中游离皮质醇、可的松及其完整的II期代谢物进行快速、准确的分析。方法建立了一种新的LC-MS/MS方法，用于定量5种分析物，包括完整的葡萄糖和亚砜共轭皮质醇和可的松代谢物。该方法按照ISO 17025定量方法标准进行验证评估。将其性能与临床实验室使用的商用LC-MS/MS分析方法进行比较，分析79份24小时尿液样本中的游离皮质醇和可的松。此外，该方法还应用于168例肾上腺疾病患者的真实尿液样本。结果验证方案评价了方法的选择性、基质效应、定量性能和结转性。结果表明，该方法与商用LC-MS/MS法测定尿游离皮质醇和可的松具有良好的相关性。病理样本分析表明，监测完整的II期代谢物，特别是硫代偶联物，可以帮助鉴别肾上腺疾病。结论建立并验证的稀释-发射LC-MS/MS方法具有良好的性能，为临床肾上腺激素分析提供了一种快速、经济的解决方案。本研究还通过在病理队列中检测目标分析物的尿排泄，提供了对肾上腺功能的深入了解，提高了相关疾病的诊断精度。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinica Chimica Acta 医学-医学实验技术

CiteScore

10.10

自引率

2.00%

发文量

1268

审稿时长

23 days

期刊介绍： The Official Journal of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) Clinica Chimica Acta is a high-quality journal which publishes original Research Communications in the field of clinical chemistry and laboratory medicine, defined as the diagnostic application of chemistry, biochemistry, immunochemistry, biochemical aspects of hematology, toxicology, and molecular biology to the study of human disease in body fluids and cells. The objective of the journal is to publish novel information leading to a better understanding of biological mechanisms of human diseases, their prevention, diagnosis, and patient management. Reports of an applied clinical character are also welcome. Papers concerned with normal metabolic processes or with constituents of normal cells or body fluids, such as reports of experimental or clinical studies in animals, are only considered when they are clearly and directly relevant to human disease. Evaluation of commercial products have a low priority for publication, unless they are novel or represent a technological breakthrough. Studies dealing with effects of drugs and natural products and studies dealing with the redox status in various diseases are not within the journal''s scope. Development and evaluation of novel analytical methodologies where applicable to diagnostic clinical chemistry and laboratory medicine, including point-of-care testing, and topics on laboratory management and informatics will also be considered. Studies focused on emerging diagnostic technologies and (big) data analysis procedures including digitalization, mobile Health, and artificial Intelligence applied to Laboratory Medicine are also of interest.