Analytical Chemistry p. 6848 - 6856 (2021)
Update date:2022-08-11
Topics:
Ding, Jiang-Hui
Feng, Yu-Qi
Huang, Wei-Hua
Liu, Fei-Long
Yang, Xiao-Ke
Ye, Tian-Tian
Yin, Xiao-Ming
Yuan, Bi-Feng
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.
View MoreContact:+852-8198 2399
Address:9E, Leapont Industrial Building, 18-28 Wo Liu Hang Road, Shatin, New Territories, Hong Kong
Contact:86-898-65311214
Address:Room 102, BLDG. 68 Jiangnan City, No. 66 Heping Road,Haikou, Hainan, China
Contact:+86 21 34123252
Address:14, 4580 Dushi, Shanghai, China
website:http://www.uvchemkleys.com
Contact:+86 21 3868 0172
Address:RM-1203, Ziguang Building, #341 Shangcheng Road, Pudong Dist, Shanghai
Huaian Double Win Chemicals Co.,Ltd.
Contact:+86-13511538872
Address:Blk 43,Greenland Century Town,Huaian District, Huaian City,Jiangsu Province,China
Doi:10.1021/jf60163a024
(1969)Doi:10.3184/174751913X13730099043760
(2013)Doi:10.1016/j.bioorg.2019.03.052
(2019)Doi:10.1002/chir.22751
(2017)Doi:10.1039/c39740000384
(1974)Doi:10.1002/jhet.5570450308
(2008)