We herein developed a selective method for the determination of nucleotides by fluorous-assisted metal chelate affinity extraction followed by hydrophilic interaction liquid chromatography (HILIC) combined with tandem mass spectrometric (MS/MS) analysis. In this study, the nucleotides were selectively chelated by Fe(III)-immobilized perfluoroalkyliminodiacetic acid, and the resulting chelates were subsequently extracted into a fluorous solvent. The nucleotides present in the fluorous solvent were then back-extracted into a non-fluorous solution, such as a solution of ammonia in aqueous acetonitrile. The resulting non-fluorous solution containing the nucleotides was then directly injected into an amide-type HILIC column using a mixture of acetonitrile and aqueous ammonium bicarbonate as the mobile phase for gradient elution, and the nucleotides were detected using the negative electrospray ionization MS/MS mode. In this method, the extraction recoveries of the nucleotides ranged from 43.2 to 94.7% within a relative standard deviation of 17%. This method enabled the determination of intracellular concentrations of nucleotides.

我们在本文中开发了一种选择性方法，该方法通过氟辅助金属螯合亲和力提取，然后通过亲水相互作用液相色谱（HILIC）与串联质谱（MS / MS）分析相结合，确定核苷酸。在这项研究中，核苷酸被Fe（III）固定的全氟烷基亚氨基二乙酸选择性地螯合，然后将所得的螯合物提取到氟溶剂中。然后将含氟溶剂中存在的核苷酸反萃取到无氟溶液中，例如氨在乙腈水溶液中的溶液。然后使用乙腈和碳酸氢铵水溶液的混合物作为流动相，将所得的含有核苷酸的无氟溶液直接注入酰胺型HILIC色谱柱中，进行梯度洗脱，并使用负电喷雾电离MS / MS模式检测核苷酸。在这种方法中，核苷酸的提取回收率在43.2％至94.7％之间，相对标准偏差为17％。该方法能够确定细胞内核苷酸的浓度。