开发一种准确的双同位素标准LC-MS/MS方法,用于生物矩阵中的氨酸量化
Simone Manzi1, Alessandra Altomare1, Giacomo Mosconi1
1Department of Pharmaceutical Sciences (DISFARM), Università degli Studi di Milano, Via Mangiagalli 25, Milan 20133, Italy.
Analytical chemistry
|February 11, 2026
概括
通过新的LC-MS/MS方法,在生物样本中量化氨酸 (HA) 现在更容易. 这种方法使用酶解水解和标记标准,用于在复杂矩阵中准确测量.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 生物材料科学 生物材料科学
背景情况:
- 氨酸 (HA) 对于组织水分,修复和细胞信号传递至关重要.
- 在生物样本中精确量化HA是很困难的,因为其内源性质和低可检测性质谱学.
- 现有的方法经常与矩阵效应和可变性作斗争.
研究的目的:
- 开发和验证一种可靠的LC-MS/MS方法,用于在复杂的生物矩阵中量化氨酸.
- 克服与HA内源性存在和质谱检测能力差相关的挑战.
- 为临床和制药应用提供可靠的工具.
主要方法:
- 使用复合氨基酶对HA的酶化水解产生可量化的Δ4-mer寡合体.
- 使用双C标记的氨酸内部标准 (100%和50%),以纠正酶消化和MS检测器变异性.
- 整合了标准添加方法,以有效地减轻生物样本中的矩阵效应,如牛玻璃体和人类突液.
主要成果:
- 开发的方法显示出出色的线性 (r2>0.99) 和高的恢复率 (>90%).
- 达到低检测极限 (LOD) 和量化极限 (LOQ),适合生理度.
- 观察到显著的矩阵效应,验证了标准加法技术的必要性.
结论:
- 经过验证的LC-MS/MS策略提供了一种可靠和准确的方法,用于在具有挑战性的生物样本中定量氨酸.
- 这种方法解决了以前方法的关键局限性,使得更精确的测量.
- 这些发现支持该方法在临床诊断和涉及氨酸的药物研究中的应用.
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