University of Cincinnati/Agilent Technologies Metallomics Center of the Americas, Department of Chemistry, University of Cincinnati, Cincinnati, OH 45221-0172, USA.
Analyst. 2010 Oct;135(10):2560-5. doi: 10.1039/c0an00399a. Epub 2010 Sep 7.
A method for the separation and detection of oligonucleotides utilizing hydrophilic interaction liquid chromatography (HILIC) with inductively coupled plasma mass spectrometry (ICPMS) is described. Polythymidylic acids of various lengths (10, 15, 20 and 30 nucleotides) were separated under gradient HILIC conditions. Selective detection of oligonucleotides was possible through monitoring m/z 47, corresponding to (31)P(16)O(+), using ICPMS. Oxygen was used as a reaction gas in the collision/reaction cell to produce PO(+) by reacting with phosphorus in the gas phase, thereby effectively eliminating the interferences for phosphorus normally seen at m/z 31. Limits of detections (LODs) were determined to be 1.69 pmol, 1.21 pmol, 1.0 pmol and 0.55 pmol loaded on column for the 10, 15, 20 and 30 mer, respectively.
本文描述了一种利用亲水作用色谱（HILIC）与电感耦合等离子体质谱（ICPMS）分离和检测寡核苷酸的方法。在梯度 HILIC 条件下，分离了不同长度的聚胸腺嘧啶核苷酸（10、15、20 和 30 个核苷酸）。通过使用 ICPMS 监测 m/z 47，对应于（31）P（16）O（+），可以实现对寡核苷酸的选择性检测。在碰撞/反应池中使用氧气作为反应气体，通过与气相中的磷反应生成 PO（+），从而有效地消除了通常在 m/z 31 处观察到的磷干扰。对于 10、15、20 和 30 mer，分别在柱上加载的检测限（LOD）为 1.69 pmol、1.21 pmol、1.0 pmol 和 0.55 pmol。
