UHPLC-QTOF-HRMS法测定干血斑中磷脂酰乙醇(PEth):方法验证及微创进样新酒精滥用生物标志物的实际应用

IF 1.8 4区 医学 Q4 BIOCHEMICAL RESEARCH METHODS
Christina Ververi, Marta Massano, Eugenio Alladio, Alberto Salomone, Marco Vincenti
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引用次数: 0

摘要

我们的研究目的是开发和验证一种简单、快速、灵敏的方法来检测干血斑(DBS)中的磷脂酰乙醇(PEth)。取30 μ l血样于DBS卡上,室温干燥。然后,将斑点切割并转移到干净的管中,加入内标(PEth-D5)和1 ml己烷,搅拌,超声,室温离心。用30 μ l乙腈复配干燥后的上清液,采用UHPLC-HRMS-QTOF分析。分别在20、50、100、200、300、500 ng/mL下建立校准曲线;在5 ng/mL (S/N > 3)下计算出检出限,并成功评估了准确度、精密度、回收率和基质效应,以及分析物在不同时间间隔和温度下的稳定性。该研究表明,使用UHPLC-QTOF或UHPLC-QqQ仪器定量DBS卡上的PEth 16:0/18:1是可行的,而QTOF方法被验证并证明用于PEth检测来评估过量饮酒和戒酒是可靠的,符合当前的指南。对真实样本的初步数据证实了该方法在简便、可持续性和速度方面的性能,支持其在慢性酒精滥用的常规毒理学诊断方面的巨大潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Determination by UHPLC-QTOF-HRMS of Phosphatidylethanol (PEth) in Dried Blood Spots: Method Validation and Practical Application of a Rising Alcohol Abuse Biomarker With Minimally Invasive Sampling

Determination by UHPLC-QTOF-HRMS of Phosphatidylethanol (PEth) in Dried Blood Spots: Method Validation and Practical Application of a Rising Alcohol Abuse Biomarker With Minimally Invasive Sampling

The goal of our study was to develop and validate a simple, quick, and sensitive method to detect phosphatidylethanol (PEth) in dried blood spots (DBS). A 30-μL aliquot of blood was collected on a DBS card and allowed to dry at room temperature. Then, the spot was cut and transferred into a clean tube where the internal standard (PEth-D5) and 1-mL hexane were added followed by stirring, sonication, and centrifugation at room temperature. The dried supernatant was reconstituted with 30-μL acetonitrile and analyzed by UHPLC-HRMS-QTOF. Calibration curve was created at 20, 50, 100, 200, 300, and 500 ng/mL; the limit of detection was calculated at 5 ng/mL (S/N > 3) while accuracy, precision, recovery, and matrix effect were successfully evaluated, along with the analyte stability at different time intervals and temperatures. The study demonstrates that quantifying PEth 16:0/18:1 from DBS cards is feasible using UHPLC-QTOF or UHPLC-QqQ instrumentation while the QTOF method was validated and proved reliable for PEth detection to assess both excessive alcohol consumption and alcohol abstinence, matching current guidelines. Preliminary data on authentic samples confirmed the method's performance in terms of ease, sustainability, and speed, supporting its great potential for routine toxicological diagnosis of chronic alcohol abuse.

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来源期刊
Biomedical Chromatography
Biomedical Chromatography 生物-分析化学
CiteScore
3.60
自引率
5.60%
发文量
268
审稿时长
2.3 months
期刊介绍: Biomedical Chromatography is devoted to the publication of original papers on the applications of chromatography and allied techniques in the biological and medical sciences. Research papers and review articles cover the methods and techniques relevant to the separation, identification and determination of substances in biochemistry, biotechnology, molecular biology, cell biology, clinical chemistry, pharmacology and related disciplines. These include the analysis of body fluids, cells and tissues, purification of biologically important compounds, pharmaco-kinetics and sequencing methods using HPLC, GC, HPLC-MS, TLC, paper chromatography, affinity chromatography, gel filtration, electrophoresis and related techniques.
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