Establishment of Platelet Membrane Chromatography and Column Temperature Study / 中国药学杂志

ABSTRACT

OBJECTIVE: To establish a platelet cell membrane chromatographic model and investigate the retention behaviors of anti-platelet aggregation drugs on chromatographic column at different temperatures, and simulate the interactions between drugs and platelets under normal and febrile pathological conditions. METHODS: The platelet cell membrane chromatographic stationary phase was constructed by physical adsorption method. The column was packed with wet method. The protein content was determined by BCA protein concentration assay kit. The biological activity was determined by Na+, K+-ATPase assay kit. The chromatographic model was used to investigate the specificity of the column and the retention characteristics of drugs in the temperature range of 35.0-42.0℃. RESULTS: The activity of platelet ATPase was 0.214, and the concentration of platelet membrane protein was 0.340 9 mg•mL-1 before bonding and 0.080 5 mg•mL-1 after bonding. The retention characteristics of clopidogrel, dipyridamole and cilostazole on platelet cell membrane chromatographic column and blank silica gel column were quite different. The retention time of the three drugs on platelet cell membrane chromatographic column was the maximum at 36.0℃, and then decreased with the increase of temperature. CONCLUSION: A platelet cell membrane chromatographic model is successfully constructed, and the retention characteristics of antiplatelet aggregates at different temperatures are studied for the first time. The chromatographic retention behaviors of antiplatelet aggregates at normal and febrile body temperatures are simulated.