A novel fully-automated method to measure steroids in serum by liquid chromatography-tandem mass spectrometry

IF 3.1 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY Journal of Mass Spectrometry and Advances in the Clinical Lab Pub Date : 2023-01-01 DOI:10.1016/j.jmsacl.2022.12.004

François Fraissinet , Tony Pereira , Alizée Violin , Guillaume Feugray , Kalyane Bach-Ngohou , Valéry Brunel

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引用次数: 3

Abstract

Background

Steroids play a key role in numerous physiological processes. Steroid determination is a useful tool to explore various endocrine diseases. Because of its specificity, mass spectrometry is considered to be a reference method for the determination of steroids in serum compared to radioimmunoassay. This technology could progress towards more automation for the optimal organization of clinical laboratories and ultimately for the benefit of patients.

Methods

A fully automated ultra-high-performance liquid chromatography-tandem mass spectrometry method was developed and fully validated to determine five steroids in serum. Sample preparation was based on protein precipitation with filtration followed by online solid phase extraction. Chromatographic separation was performed using a biphenyl stationary phase.

Results

The method was successfully validated according to European Medicine Agency guidelines. Coefficients of variation did not exceed, respectively, 8.4% and 8.1% for intra- and inter-assay precision. Method comparison with radioimmunoassay showed a proportional bias for all compounds, except for testosterone in men. Comparison with another LC-MS/MS method demonstrated acceptable concordance for all steroids, although a small bias was observed for androstenedione.

Conclusion

The novelty of this method is that it has been fully automated. Automation provides benefits in traceability and allows significant savings in cost and time.

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一种新型全自动液相色谱-串联质谱法测定血清中类固醇的方法

背景类固醇在许多生理过程中起着关键作用。类固醇测定是探索各种内分泌疾病的有用工具。由于其特异性，与放射免疫分析法相比，质谱法被认为是测定血清中类固醇的参考方法。这项技术可以朝着更加自动化的方向发展，以优化临床实验室的组织，并最终造福于患者。方法建立了全自动超高效液相色谱-串联质谱法测定血清中5种甾体激素的方法，并进行了验证。样品制备基于蛋白质沉淀和过滤，然后在线固相萃取。使用联苯固定相进行色谱分离。结果该方法根据欧洲药品管理局的指导方针得到了成功验证。批内和批间精密度的变异系数分别不超过8.4%和8.1%。方法与放射免疫分析法的比较显示，除男性睾酮外，所有化合物都存在比例偏差。与另一种LC-MS/MS方法的比较表明，所有类固醇的一致性都是可以接受的，尽管雄烯二酮的偏差很小。结论该方法的新颖性在于它已经实现了全自动化。自动化提供了可追溯性方面的优势，并显著节省了成本和时间。

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