通过UHPLC-MS3测试同时确定甲和7-基甲,并加上多个阶段的碎片化来提高灵敏度
Huaqiang Li1, Xujian Duan2,3, Feifei Wu1
1College of Biological and Food Engineering, Guangdong University of Petrochemical Technology, Maoming, Guangdong, P.R. China.
Journal of separation science
|November 8, 2024
概括
一种新的UHPLC/MS3测定准确地测量了甲基 (MTX) 和其代谢物7-基甲基 (7-OH-MTX) 在人体血中. 这种方法为临床应用提供了增强的选择性和灵敏性.
科学领域:
- 分析化学 分析化学
- 药理动力学 药理动力学
- 临床化学 临床化学
背景情况:
- 甲基 (MTX) 是一种重要的化疗剂.
- 监测MTX及其代谢物7-基甲 (7-OH-MTX) 对于优化治疗和最大限度地降低毒性至关重要.
- 现有的分析方法可能缺乏必要的选择性和灵敏性,以在复杂的生物矩阵中准确量化.
研究的目的:
- 开发和验证一种选择性的超高性能液体染色学双重质谱立方 (UHPLC/MS3) 试验.
- 为了使MTX和7-OH-MTX在人体血中的同时确定.
- 提供一种增强的分析方法,具有更好的选择性和灵敏度.
主要方法:
- 使用甲醇对人体血样本的蛋白质沉.
- 使用Waters AcQuity UPLC-BEH柱与渐变化进行染色分离.
- 通过对MTX和7-OH-MTX的多重反应监测立方 (MRM3) 过渡进行检测.
主要成果:
- 在UHPLC/MS3测定显示高选择性和灵敏度.
- 确定了线性范围:MTX的0.5-300 ng/mL (R2 ≥0.99) 和7-OH-MTX的5-1500 ng/mL (R2 ≥0.99). 线性范围是:MTX的0.5-300 ng/mL (R2 ≥0.99) 和7-OH-MTX的5-1500 ng/mL (R2 ≥0.99).
- 该方法在真实的人类血样本中成功应用了量化MTX和7-OH-MTX的方法.
结论:
- 一种新的和验证的UHPLC/MS3试验允许同时确定MTX和7-OH-MTX.
- 与现有技术相比,开发的方法提供了更高的选择性和灵敏度.
- 这种测定适用于常规临床分析和甲索特雷克萨特治疗的药理动力学研究.
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