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建立了一种三氯乙酸沉淀法结合姜黄素法测定血红蛋白氧载体中硼残余含量的方法。以血红蛋白氧载体:三氯乙酸=9∶1(V/V)的比例用三氯乙酸沉淀血红蛋白氧载体中聚合血红蛋白,离心5000 g×10 min将溶液中的沉淀分离,获得游离硼化物,聚合血红蛋白沉淀引起体积的缺失由生理盐水补齐,依次加入硫酸冰乙酸混合酸、姜黄素冰乙酸溶液将硼化物转化成硼酸并与姜黄素络合显色2.0 h,再加入磷酸稳定显色反应20 min,最后加入乙酸铵-冰乙酸缓冲溶液,将颜色转化为桔黄色20 min,在538 nm处进行定量测定,同时对测定条件进行了优化。硼的浓度在0.05～5.0μg/mL范围内相关系数为0.9993,重复测定6次的相对标准偏差为2.5%,回收率为(97.6±2.3)%。
A method for the determination of residual boron in hemoglobin oxygen carrier by trichloroacetic acid precipitation combined with curcumin was established. Hemoglobin in the hemoglobin oxygen carrier was precipitated with trichloroacetic acid at a ratio of hemoglobin oxygen carrier: trichloroacetic acid = 9: 1 (V / V), and the precipitate in the solution was centrifuged at 5000 g for 10 min to obtain the free boride. The polymerization Hemoglobin precipitation caused by the lack of volume filled with saline, followed by adding glacial acetic acid mixed acid, curcumin glacial acetic acid solution of boride into boric acid and curcumin coloration for 2.0 h, then add phosphoric acid stable color reaction 20 min. Finally, ammonium acetate-glacial acetic acid buffer solution was added to convert the color to orange for 20 min, and the determination was carried out at 538 nm. Meanwhile, the determination conditions were optimized. The correlation coefficient of boron concentration in the range of 0.05-5.0μg / mL was 0.9993. The relative standard deviation (RSD) was 2.5% and the recovery rate was (97.6 ± 2.3)%.