A Simple Stabilization Method of Reduced Albumin in Blood and Plasma for the Reduced/Oxidized Albumin Ratio Measurement
A Simple Stabilization Method of Reduced Albumin in Blood and Plasma for the Reduced/Oxidized Albumin Ratio Measurement
复制标题
用于测定还原型/氧化白蛋白比率的血液和血浆中还原型白蛋白的简单稳定方法
DOI：
--
复制
发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Ei
中科院分区：
文献类型：
--
作者：
K. Kubota;Akira Nakayama;K. Takehana;Asami Kawakami;N. Yamada;Ei
白蛋白 (Alb) 是还原形式和氧化形式的混合物。测定Alb(red)%，即还原后的Alb在Alb总和中所占的比例，具有重要的生理意义。然而，血液和血浆样本中的还原白蛋白很容易转化为氧化白蛋白。因此，样品中 Alb 的稳定对于确定精确的 Alb(red)% 值是必要的。通过 pH 控制和缓冲液稀释实现血液或血浆中白蛋白的稳定。人血浆至少需要用 50 mmol/l 磷酸盐缓冲液 (pH 6.0) 稀释 50 倍。对于人血，需要用 0.5 mol/l 柠檬酸钠缓冲液（pH 4.3）稀释 10 倍。为了测量 Alb(red)%，将处理过的样品应用于 HPLC 或 LC-ESI-TOFMS。我们还开发了“预孵育法”，通过37°C加热加速血浆中的氧化反应。 Alb(red)%值在稳定人血浆后48小时和稳定人血液后72小时维持在初始值附近。加速血浆中的氧化反应会导致正常样本和模型疾病样本之间的 Alb(red)% 值存在较大差异。在稳定控制下常规获得精确的Alb（红色）％值。此外，测量前对血浆进行预孵育有助于增强正常样本和疾病样本之间的差异。
Albumin (Alb) is mixture of reduced and oxidized forms. It is physiologically significant to determine Alb(red)%, which is the proportion of reduced Alb in the sum of Alb. However, reduced Alb in both blood and plasma samples is easily converted to oxidized Alb. Accordingly, the stabilization of Alb in samples is necessary to determine precise Alb(red)% values. Alb stabilization in blood or plasma was achieved by pH control and buffer dilution. At least a 50-fold dilution with 50 mmol/l phosphate buffer (pH 6.0) was required for human plasma. For human blood, a 10-fold dilution with 0.5 mol/l sodium citrate buffer (pH 4.3) was required. To measure Alb(red)%, treated samples were applied to HPLC or LC-ESI-TOFMS. We also developed a “pre-incubation method”, to accelerate the oxidative reaction in plasma by heating at 37°C. Alb(red)% values were maintained around the initial value for 48 h after stabilizing human plasma and 72 h after stabilizing human blood. Accelerating the oxidative reaction in plasma produced large differences in the Alb(red)% values between normal and model disease samples. Precise Alb(red)% values were routinely obtained under the stabilization control. Additionally, pre-incubation of the plasma before measurement is useful to enhance the difference between normal and disease samples.
DOI：
10.1073/pnas.89.16.7674
发表时间：
1992-08-15
影响因子：
11.1
作者：
STAMLER, JS;JARAKI, O;LOSCALZO, J
通讯作者：
LOSCALZO, J