Development of an HPLC method for the determination of amines in a leukemia mouse model

Abstract

This study developed a high-performance liquid chromatography coupled with fluorescence detection (HPLC-FLD) method for the simultaneous determination of 19 amino acids in the cortex, hippocampus, and bone marrow samples of BALB/C-nu/nu and NOD/SCID mice. We measured the changes in the levels of 19 amino substances in the cortex, hippocampus, and bone marrow samples of mice in the blank model group, acute leukemia model group, chronic leukemia model group, and treatment group. The chromatographic column used was a COSMOSIL 5 C18-MS-II analytical column (150 mm × 4.6 mm, 5 μm), with the column temperature set at 35 °C. The mobile phase was a methanol-ammonium acetate buffer solution (30 mmol L⁻¹ ammonium acetate solution, adjusted to pH 6.40 with acetic acid), at a flow rate of 0.9 mL min⁻¹. The single analysis time was 46 minutes. By optimizing the analysis conditions, the newly established HPLC-FLD method exhibits high sensitivity, good accuracy, and good reproducibility. This method can be successfully applied to determine 19 amino substances in the cortex, hippocampus, and bone marrow of BALB/C-nu/nu and NOD/SCID mice. The differences in amino acid content between different groups can be used to further study the development process of leukemia, disease diagnostic markers, and personalized treatment plans.