Kuzmanova Elena, McIntyre Angela, Syed Maaz, Iskandar Zaid, Newby David E, Bown Matt J, Choy Anna-Maria, Huang Jeffrey Tj
Systems Medicine, School of Medicine, University of Dundee, Dundee, UK.
Queen's Medical Research Institute, MRC/University of Edinburgh Centre for Inflammation Research, Edinburgh, UK.
Bioanalysis. 2025 Jan;17(2):99-104. doi: 10.1080/17576180.2025.2452723. Epub 2025 Feb 1.
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.
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.
These results demonstrated that rapid desmosine analysis can be achieved with the use of both microwave-assisted hydrolysis and streamlined solid-phase extraction.
循环总去甲二氢愈创木酸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，限制了其潜在应用。本研究的目的是缩短基于稳定同位素稀释液相色谱-质谱的去甲二氢愈创木酸检测的周转时间。
血浆样品采用酸水解，然后进行固相萃取和液相色谱-质谱分析。测试了两种减少检测时间的方法：微波辅助酸水解和固相萃取后直接进样。
180℃酸水解8分钟与固相萃取的小体积洗脱设计相结合，将总检测时间缩短至约30分钟。该检测方法的日内精密度和准确度分别为4%至14%和-7%至9%，日间精密度和准确度分别为0%至9%和1%至3%，得到了验证。该检测方法在一组急性主动脉夹层患者和对照受试者中进行了测试，患者的去甲二氢愈创木酸浓度约为对照受试者的三倍。
这些结果表明，使用微波辅助水解和简化的固相萃取可以实现快速的去甲二氢愈创木酸分析。
