Accelerator Mass Spectrometry-Based Signal Generation for Thin Layer Chromatography

A highly sensitive accelerator mass spectrometry (AMS)based assay for determination of the 14 C content in the scraped silica gel after thin layer chromatography (TLC) has been established. In the present study, 14 C-labeled inositol or its cellular derivatives was selected as a model analyte to evaluate the designed assay, since a series of phosphatidyl inositols (PIs) are known to be derived from inositol and to play pivotal roles as precursors or second messengers for several cell signaling processes. After incubating HeLa cells with 14 C-labeled inositol, allowing the dosed 14 C-labeled inositol to be converted to a series of 14 C-labeled PIs and phosphoinositides through biochemical processes, the phospholipids were extracted and separated by the conventional TLC. The resulting TLC plates were scraped and graphitized for AMS measurement. The obtained data demonstrates the intracellular distribution of the phosphatidyl inositols, implying that the developed assay is better and more conv