Accelerating isotope dilution LC-MS-based desmosine quantification for estimating elastin turnover.

IF 1.8 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS Bioanalysis Pub Date : 2025-01-01 Epub Date: 2025-02-01 DOI:10.1080/17576180.2025.2452723

Elena Kuzmanova, Angela McIntyre, Maaz Syed, Zaid Iskandar, David E Newby, Matt J Bown, Anna-Maria Choy, Jeffrey Tj Huang

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Abstract

Aims: Circulating total desmosine, representing endogenous systemic elastin degradation activity, is an emerging biomarker for mortality risk in several diseases and aging. However, the existing analytical method takes more than 23 hours to complete, limiting its potential applications. The objective of this study was to shorten the turnover time of a stable isotope dilution liquid chromatogram mass spectrometry-based desmosine assay.

Materials & methods: Plasma samples were analyzed using acid hydrolysis followed by solid-phase extraction and LC-MS. Two approaches to reduce assay time were tested: microwave-assisted acid hydrolysis and direct injection following solid-phase extraction.

Results: The combination of acid hydrolysis at 180°C for 8 minutes and a low-volume elution design for solid-phase extraction reduced the overall assay time to ~ 30 minutes. The assay was validated with intra-day precision and accuracy ranging from 4% to 14%, and -7% to 9%, respectively, while inter-day precision and accuracy were 0% to 9% and 1% to 3%, respectively. The assay was tested in a cohort of patients with acute aortic dissection and control subjects, where desmosine concentrations were approximately three-fold higher in patients.

Conclusions: These results demonstrated that rapid desmosine analysis can be achieved with the use of both microwave-assisted hydrolysis and streamlined solid-phase extraction.

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加速同位素稀释lc - ms定量测定弹性蛋白周转。

目的：循环总桥苷，代表内源性全身弹性蛋白降解活性，是几种疾病和衰老中死亡风险的新兴生物标志物。然而，现有的分析方法需要超过23小时才能完成，限制了其潜在的应用。本研究的目的是缩短稳定同位素稀释液相色谱-质谱法测定氨基葡萄糖的周转时间。材料与方法：血浆样品采用酸水解，固相萃取和LC-MS分析。试验了两种缩短测定时间的方法：微波辅助酸水解和固相萃取后直接进样。结果：180°C酸水解8分钟和固相萃取小体积洗脱设计相结合，将总检测时间缩短至~ 30分钟。日内精密度和准确度分别为4% ~ 14%和-7% ~ 9%，日内精密度和准确度分别为0% ~ 9%和1% ~ 3%。该试验在急性主动脉夹层患者和对照组的队列中进行了测试，其中患者的氨基葡萄糖浓度大约高出三倍。结论：这些结果表明，微波辅助水解和流线型固相萃取都可以实现快速分析。

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来源期刊

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.