Global LC-MS/MS targeted metabolomics using a combination of HILIC and RP LC separation modes on an organic...

The paper presents an LC-MS/MS-based approach to targeted screening of both polar and non-polar metabolites using a synthesized monolithic column which is a copolymer of styrene, divinylbenzene, and 1-vinyl-1,2,4-triazole. It was shown that this column in combination with eluents 20 mM (NH₄)₂CO₃ + NH₃ (pH = 9.8, eluent A) and ACN (eluent B) allows for separation of metabolites of different nature in two modes, HILIC and RP LC, and these methods are mutually complementary. A combination of analyses based on these two modes was proposed, allowing detection of about 400 metabolites in a total time of less than 30 min. Comparison of the developed method with those utilizing commercially available columns with sorbents of various types showed that it could provide a broader metabolite coverage. Using the developed approach, metabolomic screening of dried blood spots samples of mice exposed with X-ray was performed, and metabolites that could be considered as possible markers of irradiation exposure and organ tissue damage were detected. Analysis of marker metabolites revealed metabolic pathways that were altered by radiation exposure. Comparison of the results with literature data showed the effectiveness of the developed metabolomic screening approach.