解决通过LC/MS检测和量化D-2-基酸盐和L-2-基酸盐的挑战
bioRxiv : the preprint server for biology
|June 21, 2024
概括
精确量化D-2-Hydroxyglutarate (D-2HG) 和L-2-Hydroxyglutarate (L-2HG) 对了解疾病至关重要. 一种新的高通量LC/MS方法能够精确,同时测量这些反体和其他代谢物.
科学领域:
- 生物化学 生物化学
- 代谢学 代谢学 代谢学
- 分析化学 分析化学
背景情况:
- D-2-Hydroxyglutarate (D-2HG) 和L-2-Hydroxyglutarate (L-2HG) 是代谢物,它们的积累与癌症和酸性等疾病有关.
- 这些氧氨酸酶体干扰了参与细胞代谢和表观遗传调节的关键酶.
- 准确量化D-2HG和L-2HG对于理解它们在疾病发病过程中的作用至关重要.
研究的目的:
- 开发和验证一种可靠的高通量方法,用于同时量化D-2HG和L-2HG反体.
- 为了能够在疾病相关的样本中准确测量这些生物标志物.
- 为了促进对失调的代谢途径的调查.
主要方法:
- 检查现有的D-2HG/L-2HG衍生和液态染色学-质谱学 (LC/MS) 量化方法.
- 开发一种高分辨率的四极飞行时间LC/MS (HR-QTOF-LC/MS) 方法.
- 使用二甲基-L-酸盐无水化物 (DATAN) 进行基因分离的基因衍生.
主要成果:
- 为了同时量化D-2HG和L-2HG,建立了一个强大的,高通量HR-QTOF-LC/MS方法.
- 该方法表现出极好的线性 (r2 ≥ 0.995),精度 (日内CV% ≤ 8.0,日间CV% ≤ 6.3) 和准确性 (相对误差% ≤ 2.7).
- 达到了D-2HG和L-2HG反体的成功分辨率 (Rs = 1.6),允许对其他途径中间体进行量化.
结论:
- 开发的HR-QTOF-LC/MS方法提供了对D-2HG和L-2HG的准确和精确的反选择性量化.
- 这种方法适用于分析与疾病相关的样本,并且可以扩展到大型临床队列.
- 能够量化和其他代谢物的能力为与疾病相关的代谢失调提供了宝贵的见解.
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