Analysis The Products Of Glucose Metabolite In The Leukemia Cells

The reserach work is to establish a method to analyze the products of glucose metabolism:glucose-6-phosphate, fiructose-6-phosphate, ribose-5-phosphate, fructose-1,6-diphosphate, glucose, phosphoenolpyruvic acid, pyruvic acid, lactic acid, citric acid, and serine which may be generated in the cell during the glucose metabolism process. HILIC column ZIC-HILIC is better in retention and separation effect than Phenomenex luna NH2 column and Venusil XBP silica column. High concentration of buffer salt and high proportion of organic phase is conducive to retention on the ZIC-HILIC column, while high proportion of organic phase in the initiation gradient concentration is against the separation and detection of the 11 compounds. When the ratio of the organic phase to water phase is 40:60, the 11 compounds can be separated. Reducing the increase rate of water phase ratio contributes to the separation of the selected compounds. They all can be detected by the charged aerosol detector (CAD). Uv detector can only detect phosphoenolpyruvic acid and pyruvic acid at 220 nm, while evaporative light-scattering detector (ELSD) can detect 10 compounds except pyruvic acid.Used a high performance liquid chromatography-electrospray ionization mass spectrometric methods for analysis and identification of corresponding chromatographic peaks of the mixture of 11 standard samples, and determine the mass-to-charge ratio of the molecular ion peak of each standard compound. Using the method to detect the leukemia cell extracts to see if they contain the selected 11 kinds of metabolites. Detecting the extract of leukemia cells found it contains the selected compound fructose-6-phosphate, fructose-1,6-diphosphate, and lactic acid in which the lactic acid content is significantly higher than the other two compounds. Detecting large amounts of lactic acid in the extract of leukemia cells verifies the existence of "Warburg effect" in the leukemia cells.