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使用GC-ToF-MS和UHPLC-MS对突液代谢概况的分析前变量的系统评估
Yumna Ladha1,2,3, Adam Burke3, Nigel Gotts3
1Oswestry Keele Orthopaedic Research Group (OsKOR), Robert Jones and Agnes Hunt Orthopaedic Hospital Foundation Trust, Oswestry, Shropshire, UK. c.hulme1@keele.ac.uk.
标准化突液 (SF) 样本处理对于可靠的代谢分析至关重要. 稀释显著减少了代谢物检测,而血液染色改变了个人资料,影响了疾病生物标志物的发现.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 临床诊断 临床诊断 临床诊断
背景情况:
- 突液 (SF) 对于通过代谢学来研究关节疾病至关重要.
- 它的粘弹性和缺乏标准化的预分析协议使分析复杂化,并引入了变性.
- 这种变异性可能会损害数据的可靠性,并阻碍疾病机制的解释.
研究的目的:
- 系统地评估常见样本处理变量对SF代谢概况的影响.
- 调查稀释,冷解,血液染色和粘度降低 (氨酸酶,珠子打) 的影响.
- 评估代谢物数量,类分布和相对丰度的变化.
主要方法:
- 利用飞行质谱法 (GC-ToF-MS) 的非定向气体染色学时间和超高性能液体染色学-MS (UHPLC-MS).
- 采用电喷离子阳性 (ESI+) 和阴性 (ESI-) 模式,用于全面的极性和非极性代谢物分析.
- 评估了多个样本处理变量,包括稀释,结解周期,血液污染和粘度降低技术.
主要成果:
- 样品稀释显著降低了可检测代谢物的数量.
- 血液染色引入了独特的代谢物和人工升高的腺素,素和4--DL-三甲水平.
- 珠子敲击显著增强了脂类物种的检测,特别是在UHPLC ESI+.
- 冷-解循环和氨基酶治疗对整体代谢物概况的影响很小.
结论:
- 交尾液样本的处理显著影响了代谢数据.
- 标准化预分析步骤,如稀释和处理血染样本,对于可重复性至关重要.
- 优化协议对于准确的生物标志物发现和理解关节疾病机制至关重要.
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