血小板胞内海藻糖测定方法的建立和评价

本研究的目的是建立一种适用、方便和快速的血小板胞内海藻糖浓度的测定方法，用于海藻糖负载技术的优化和血小板冻干保存。采用三氯乙酸沉淀细胞蛋白，硫酸-蒽酮法测定血小板胞内海藻糖的浓度，同时作高效液相色谱分析。结果表明，负载液中海藻糖浓度为17％，硫酸蒽酮法测定血小板胞内海藻糖浓度为0．22％，高效液相色谱测定为0.20％。硫酸-蒽酮法测定海藻糖的回收率平均为100．7％。稳定性和重复性较好。结论：该方法测定血小板胞内海藻糖较准确，可靠，适用于细胞内海藻糖浓度的测定。

Establishment and Evaluation on the Method for Platelet Intracellular Trehalose Determination

This study was aimed to establish an applicable, convenient and rapid method for platelet intracellular trehalose determination, so that the technique of trehalose loading could be optimized and used in research on freeze drying platelets. Protein from loaded trehalose platelets was deposited by using trichloroacetic acid, trechalose concentration in platelets was determinated by sulfuric anthrone reaction and was analyzed by high performance liquid chromatography (HPLC). The results showed that when platelets were loaded with 1.7% of trehalose in loading solution, intracellular concentration determinated by sulfuric anthrone reaction was 0.22% and derterminated by HPLC was 0.2%. The recovery rate of trehalose determinated by sulfuric anthrone reaction was 100.7% and stability and repeatability of results were good. In conclusion, the method is convenient, rapid, accurate and highly sensitive for determination of platelet intracellar trehalose.