Integrating Electrostatic Adsorption-Laser Desorption/Ionization-MS with a Derivative Full-Spectrum Internal Standard Approach for Acylcarnitine Quantification in Clinical Samples

IF 2.7 2区 化学 Q2 BIOCHEMICAL RESEARCH METHODS
Shuo Cao, Xinrong Jiang, Xiaoming Chen, Xuetong Qu and Jianmin Wu*, 
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引用次数: 0

Abstract

Acylcarnitines (ACs), essential intermediates in fatty acid β-oxidation, are increasingly recognized as biomarkers for inborn errors of metabolism (IEM), insulin resistance, heart failure, and neurodegenerative diseases. However, conventional liquid chromatography–mass spectrometry (LC-MS) and flow injection–mass spectrometry (FI-MS) methods are hindered by complex sample preparation, limited throughput, and matrix interferences. Meanwhile, matrix-assisted laser desorption/ionization–MS (MALDI-MS) suffers from low-mass background noise and poor reproducibility. Here, we present a dual-innovation mass spectrometry platform combining a substrate composed of sulfonic acid–gold nanoparticle-decorated silicon nanowires (Sulfo-Au-SiNWs) for electrostatic adsorption-laser desorption/ionization (EALDI) and a full-spectrum internal standard (FS IS) strategy based on d6-ethanol derivatization. The optimized Sulfo-Au-SiNWs offer uniform crystallinity, in situ electrostatic enrichment, efficient thermal desorption, and gold-mediated charge–hole separation, resulting in a ∼300% increase in AC signal intensity. This enabled the selective detection of over ten medium- and long-chain ACs that were previously undetectable using unmodified SiNWs. The FS IS method incorporates over 30 derivatized AC species, enabling simultaneous quantification with detection limits below 0.01 μmol/L, surpassing the sensitivity of traditional FI-MS. The platform delivers spike recovery rates of 96–112% and supports high-throughput analysis, processing each sample within seconds. Validation against FI-ESI-MS/MS in both urine and dried blood spot samples demonstrated excellent agreement (ρ > 0.95) and effectively identified IEM patient profiles via statistically significant AC elevations. With strong intra- and interbatch precision (median RSD < 12.5%) and substrate stability exceeding one month, this EALDI-MS/FS IS workflow provides a high-throughput, high-coverage, and quantitatively robust solution for clinical AC profiling and newborn screening.

Abstract Image

结合静电吸附-激光解吸/电离-质谱与导数全光谱内标法定量临床样品中的酰基肉碱。
酰基肉碱(ACs)是脂肪酸β氧化的重要中间体,越来越多地被认为是先天性代谢错误(IEM)、胰岛素抵抗、心力衰竭和神经退行性疾病的生物标志物。然而,传统的液相色谱-质谱(LC-MS)和流动注射-质谱(FI-MS)方法受到复杂的样品制备、有限的通量和基质干扰的阻碍。同时,基质辅助激光解吸/电离-质谱法(MALDI-MS)存在质量背景噪声低、重现性差的问题。在这里,我们提出了一个双创新的质谱平台,结合了由磺酸-金纳米颗粒-装饰硅纳米线组成的静电吸附-激光解吸/电离(EALDI)衬底和基于d6-乙醇衍生化的全光谱内标(FS IS)策略。优化后的Sulfo-Au-SiNWs具有均匀的结晶度、原位静电富集、高效的热解吸和金介导的电荷-空穴分离,导致交流信号强度增加~ 300%。这使得选择性检测超过十种中链和长链ac成为可能,而以前使用未修饰的sinw无法检测到这些ac。FS - IS方法包含30多种衍生AC,可同时定量,检出限低于0.01 μmol/L,灵敏度优于传统的FI-MS。该平台提供96-112%的峰值回收率,支持高通量分析,在几秒钟内处理每个样品。对尿液和干血斑样本的FI-ESI-MS/MS验证显示出极好的一致性(ρ > 0.95),并通过统计学上显着的AC升高有效地识别了IEM患者的特征。EALDI-MS/FS IS工作流程具有强大的批内和批间精度(中位RSD < 12.5%)和超过一个月的底物稳定性,为临床AC分析和新生儿筛查提供了高通量、高覆盖率和定量可靠的解决方案。
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来源期刊
CiteScore
5.50
自引率
9.40%
发文量
257
审稿时长
1 months
期刊介绍: The Journal of the American Society for Mass Spectrometry presents research papers covering all aspects of mass spectrometry, incorporating coverage of fields of scientific inquiry in which mass spectrometry can play a role. Comprehensive in scope, the journal publishes papers on both fundamentals and applications of mass spectrometry. Fundamental subjects include instrumentation principles, design, and demonstration, structures and chemical properties of gas-phase ions, studies of thermodynamic properties, ion spectroscopy, chemical kinetics, mechanisms of ionization, theories of ion fragmentation, cluster ions, and potential energy surfaces. In addition to full papers, the journal offers Communications, Application Notes, and Accounts and Perspectives
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