Chemical Tagging Assisted Mass Spectrometry Analysis Enables Sensitive Determination of Phosphorylated Compounds in a Single Cell
| Autor: | Wei-Hua Huang, Xiaoming Yin, Tiantian Ye, Xiao-Ke Yang, Bi-Feng Yuan, Fei-Long Liu, Jiang-Hui Ding, Yu-Qi Feng |
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| Rok vydání: | 2021 |
| Předmět: |
Indole test
Chromatography 010401 analytical chemistry 010402 general chemistry Mass spectrometry 01 natural sciences Gas Chromatography-Mass Spectrometry Mass Spectrometry 0104 chemical sciences Analytical Chemistry Isotopomers chemistry.chemical_compound Metabolomics chemistry Limit of Detection Nucleic acid Chemical stability Diazo Gas chromatography–mass spectrometry Chromatography Liquid |
| Zdroj: | Analytical chemistry. 93(17) |
| ISSN: | 1520-6882 |
| Popis: | Polar phosphorylated metabolites are involved in a variety of biological processes and play vital roles in energetic metabolism, cofactor regeneration, and nucleic acid synthesis. However, it is often challenging to interrogate polar phosphorylated metabolites and compounds from biological samples. Liquid chromatography-mass spectrometry (LC/MS) now plays a central role in metabolomic studies. However, LC/MS-based approaches have been hampered by the issues of the low ionization efficiencies, low in vivo concentrations, and less chemical stability of polar phosphorylated metabolites. In this work, we synthesized paired reagents of light and heavy isotopomers, 2-(diazomethyl)phenyl)(9-methyl-1,3,4,9-tetrahydro-2H-pyrido[3,4-b]indol-2-yl)methanone (DMPI) and d3-(2-(diazomethyl)phenyl)(9-methyl-1,3,4,9-tetrahydro-2H-pyrido[3,4-b]indol-2-yl)methanone (d3-DMPI). The paired reagents of DMPI and d3-DMPI carry diazo groups that can efficiently and selectively react with the phosphate group on polar phosphorylated metabolites under mild conditions. As a proof of concept, we found that the transfer of the indole heterocycle group from DMPI/d3-DMPI to ribonucleotides led to the significant increase of ionization efficiencies of ribonucleotides during LC/MS analysis. The detection sensitivities of these ribonucleotides increased by 25-1137-fold upon DMPI tagging with the limits of detection (LODs) being between 7 and 150 amol. With the developed method, we achieved the determination of all the 12 ribonucleotides from a single mammalian cell and from a single stamen of Arabidopsis thaliana. The method provides a valuable tool to investigate the dynamic changes of polar phosphorylated metabolites in a single cell under particular conditions. |
| Databáze: | OpenAIRE |
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