摘要
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 by combination of trichloroacetic acid（TCA） precipitation and curcumin colorimetry for the determination of residual boron content in Hemoglobin-Based Oxygen carrier（HBOC） was developed.Free boride was separated from HBOC by precipitating the polymerized hemoglobin（PolyHb） by TCA with the ratio of HBOC： TCA=9： 1（V/V） and centrifuging at 5000 r/min for 10 min.The loss of volume caused by PolyHb precipitation was compensated by normal saline.Free boron compound was transformed to boron acid after sequential addting of the mixed solution of sulfuric acid and ice acetic acid and acetic acid solution of curcumin,and then chelated with curcumin for 2.0 h.The chelation was stabled for 20 min by adding the phosphoric acid.At last,the color of the solution was shifted into the orange for 20 min after adding the buffer of ammonium acetate and acetic acid,and quantitative determination could be performed at the maximal absorption wavelength（λ538）.The linear range of this method was from 0.05～5.0 μg/mL（R2=0.9993） with relative standard deviation of 2.5%（n=6）,the standard addition recoveries were（97.6±2.3）%.
出处
《分析试验室》
CAS
CSCD
北大核心
2011年第3期28-31,共4页
Chinese Journal of Analysis Laboratory
基金
国家"863计划"(2006AA02A143)
陕西省教育厅专项科研计划项目(09JK784)资助
关键词
三氯乙酸
姜黄素
血红蛋白氧载体
硼
Trichloroacetic acid
Curcumin
Hemoglobin-based oxygen carrier
Boron
