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简化维生素D代谢物指纹分析使用同位素编码多重复合MS,具有成本效益的单双衍生

Pascal Schorr¹, Caroline S Stokes², Dietrich A Volmer¹

¹Department of Chemistry, Humboldt Universität zu Berlin, Brook-Taylor-Str. 2, Berlin 12489, Germany.

|January 1, 2025

概括

这项研究引入了对维生素D代谢物的常规同位素编码多重衍生,使两个样本的同时分析成为可能. 该方法改善了用于临床应用的标准C-18列上的关键维生素D体的分离.

科学领域:

临床化学临床化学
分析化学分析化学
质谱测量质量谱测量

背景情况:

对维生素D及其代谢物进行准确的分析对于临床诊断至关重要.
目前用于维生素D代谢物分析的方法在复杂化和分离关键表层时面临挑战.
传统的分离技术往往需要专门的列 (例如,PFP) 具有有限的长期稳定性.

研究的目的:

为维生素D及其代谢物开发第一个常规的同位素编码多重衍生方法.
为了使两个样品在一次液体染色学-并联质谱 (LC-MS/MS) 运行中同时进行分析.
为了改善25-基维生素D3在常规C-18列上的染色分离.

主要方法:

使用商业试剂扩展了一个单双导化反应.
采用迪尔斯-阿尔德反应与4--1,2,4-triazoline-3,5-dione (PTAD),然后用乙酸 anhydride (或其d3-同位素) 进行乙化.
使用C-18柱与甲醇/水梯度化,以增强维生素D3代谢物的分离.

主要成果:

建立了一种常规的同位素编码多重衍生,使两个样品的同时测量成为可能.
在C-18列上实现了关键维生素D3代谢物 (24,25(OH) 2D3,3α-25(OH) D3,3β-25(OH) D3和维生素D3) 的基线分离.
通过对来自慢性肝病 (CLD) 患者的40个血清样本进行测试,证明了方法的适用性.

结论:

开发的双重衍生方法为临床维生素D分析提供了强大且常规的方法.
这种方法克服了专业列的局限性,并增强了具有挑战性的epimers的分离.
同位素编码复合策略促进了临床实验室更高的吞吐量和效率.

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