Trace Phosphate Improves ZIC-pHILIC Peak Shape, Sensitivity, and Coverage for Untargeted Metabolomics.
Trace Phosphate Improves ZIC-pHILIC Peak Shape, Sensitivity, and Coverage for Untargeted Metabolomics.
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DOI：
10.1021/acs.jproteome.8b00487
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发表时间：
2018-10-05
影响因子：
4.4
通讯作者：
Patti GJ
中科院分区：
文献类型：
--
作者：
Spalding JL;Naser FJ;Mahieu NG;Johnson SL;Patti GJ
现有的亲水相互作用液相色谱(HILIC)方法，单独考虑，每个方法都表现出对很大一部分极性代谢物的较差的层析性能。除了限制代谢组的覆盖范围外，这些缺陷还使自动化数据处理复杂化。在这里，我们展示了一些这样的分析挑战可以通过添加微量的磷酸盐来解决，这是一种常用于代谢组学的两性离子固定相。具体地说，微摩尔磷酸盐扩大了代谢组的覆盖范围，增加了数百个可信的特征，改善了峰的形状，并减少了信息处理过程中的峰检测错误。虽然添加高水平的磷酸盐(毫摩尔)作为HILIC流动相缓冲剂之前已经被探索过，但这样的浓度干扰了质谱学(MS)的检测。我们发现，在微摩尔浓度下使用磷酸盐作为痕量添加剂可以改善电喷雾质谱仪的分析，增加不同极性标准的信号。考虑到所需的少量磷酸盐，在重建过程中将磷酸盐直接添加到样品中也可以实现类似的色谱分析改进。我们的结果表明，ZIC亲水性的缺陷主要是由静电相互作用驱动的，而静电相互作用可以被磷酸盐调制。这些发现既是对非靶向代谢组学方法的改进，也是我们对限制HILIC复盖范围的机制的理解的进步。
Existing hydrophilic interaction liquid chromatography (HILIC) methods, considered individually, each exhibit poor chromatographic performance for a substantial fraction of polar metabolites. In addition to limiting metabolome coverage, such deficiencies also complicate automated data processing. Here we show that some of these analytical challenges can be addressed for the ZIC-pHILIC, a zwitterionic stationary phase commonly used in metabolomics, with the addition of trace levels of phosphate. Specifically, micromolar phosphate extended metabolome coverage by hundreds of credentialed features, improved peak shapes, and reduced peak-detection errors during informatic processing. Although the addition of high levels of phosphate (millimolar) as a HILIC mobile phase buffer has been explored previously, such concentrations interfere with mass spectrometric (MS) detection. We show that using phosphate as a trace additive at micromolar concentrations improves analysis by electrospray MS, increasing signal for a diverse set of polar standards. Given the small amount of phosphate needed, comparable chromatographic improvements were also achieved by direct addition of phosphate to the sample during reconstitution. Our results suggest that defects in ZIC-pHILIC performance are predominantly driven by electrostatic interactions, which can be modulated by phosphate. These findings constitute both a methodological improvement for untargeted metabolomics and an advance in our understanding of the mechanisms limiting HILIC coverage.
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DOI：
10.1016/s0021-9673(00)96972-3
发表时间：
1990-01-19
期刊：
JOURNAL OF CHROMATOGRAPHY
影响因子：
--
作者：
ALPERT, AJ
通讯作者：
ALPERT, AJ
影响因子：
16.6
作者：
Lu W;Su X;Klein MS;Lewis IA;Fiehn O;Rabinowitz JD
通讯作者：
Rabinowitz JD
影响因子：
3.1
作者：
Fredriksson, Mattias J.;Petersson, Patrik;Bylund, Dan
通讯作者：
Bylund, Dan
影响因子：
7.4
作者：
Alpert, Andrew J.
通讯作者：
Alpert, Andrew J.
DOI：
10.1016/j.jchromb.2005.08.008
发表时间：
2005-11-05
影响因子：
3
作者：
Bahrami, G;Mohammadi, B;Kiani, A
通讯作者：
Kiani, A