5-二乙氨基萘-1-磺酰氯（DensCl）：一种新型的三链同位素标记试剂，用于通过液相色谱-质谱联用进行定量代谢组学分析。
5-Diethylamino-naphthalene-1-sulfonyl chloride (DensCl): a novel triplex isotope labeling reagent for quantitative metabolome analysis by liquid chromatography mass spectrometry.
机构信息
Department of Chemistry, University of Alberta , Edmonton, Alberta, Canada.
出版信息
Anal Chem. 2013 Dec 3;85(23):11532-9. doi: 10.1021/ac403000n. Epub 2013 Nov 19.
We describe a new set of isotope reagents, (12)C4-, (12)C2(13)C2-, and (13)C4-5-diethylamino-naphthalene-1-sulfonyl chloride (DensCl), in combination with liquid chromatography Fourier-transform ion cyclotron resonance mass spectrometry (LC-FTICR-MS), for improved analysis of the amine- and phenol-containing submetabolome. The synthesis of the reagents is reported, and an optimized derivatization protocol for labeling amines and phenols is described. To demonstrate the utility of the triplex reagents for metabolome profiling of biological samples, urine samples collected daily from a healthy volunteer over a period of 14 days were analyzed. The overall workflow is straightforward, including differential isotope labeling of individual samples and a pooled sample that serves a global internal standard, mixing of the isotope differentially labeled samples and LC-MS analysis for relative metabolome quantification. Comparing to the dansyl chloride (DnsCl) duplex isotope reagents, the new triplex DensCl reagents offer the advantages of improved metabolite detectability due to enhanced sensitivity (i.e., about 1000 peak pairs detected by DensCl labeling vs about 600 peak pairs detected by DnsCl labeling) and analysis speed (i.e., simultaneous analysis of two comparative samples by DensCl vs only one comparative sample analyzed by DnsCl).
我们描述了一组新的同位素试剂，（12）C4-、（12）C2（13）C2-和（13）C4-5-二乙氨基萘-1-磺酰氯（DensCl），结合液相色谱傅里叶变换离子回旋共振质谱（LC-FTICR-MS），用于改善含胺和酚的亚代谢组分析。报道了试剂的合成，并描述了用于标记胺和酚的优化衍生化方案。为了证明三联试剂在生物样品代谢组分析中的实用性，对一名健康志愿者在 14 天期间每天收集的尿液样本进行了分析。整个工作流程简单明了，包括对单个样本和混合样本进行差异同位素标记，然后混合同位素差异标记的样本进行 LC-MS 分析，以进行相对代谢组定量。与丹磺酰氯（DnsCl）双试剂相比，新的三联 DensCl 试剂具有提高的代谢物可检测性的优点，因为其灵敏度增强（即，通过 DensCl 标记检测到约 1000 个峰对，而通过 DnsCl 标记检测到约 600 个峰对）和分析速度（即，通过 DensCl 同时分析两个比较样本，而通过 DnsCl 仅分析一个比较样本）。
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