Hyphenated affinity capillary electrophoresis with a high-sensitivity cell for the simultaneous binding study of retinol and retinoic acid in nanomolars with serum albumins
Hyphenated affinity capillary electrophoresis with a high-sensitivity cell for the simultaneous binding study of retinol and retinoic acid in nanomolars with serum albumins
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DOI：
10.1016/j.jchromb.2012.11.007
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发表时间：
2012-12-12
影响因子：
3
通讯作者：
Albishri, H. M.
中科院分区：
文献类型：
--
作者：
Abd El-Hady, D.;Albishri, H. M.
视黄醇和视黄酸是维生素A的成分，对许多生物过程至关重要。它们都能与血清白蛋白强络合，常数为10(5)Lmol（-1）或更高。鉴于这一事实，亲和毛细管电泳（ACE）并不适用于其常用的形式。因此，我们首次建立了带高灵敏度细胞的连字ACE，并应用于生理条件下研究纳米分子中视黄醇和视黄酸与人血清白蛋白（HSA）和牛血清白蛋白（BSA）的结合。ACE/高灵敏度偶联细胞有助于加快分析物在纳摩尔尺度上的缔合和解离速度，确保在较短的电泳时间内建立动态平衡。此外，这种连字符导致血清白蛋白作为添加剂在背景电解质中的浓度降低，除了适当的冲洗方案外，它们吸附的可能性可以忽略不计。基于非线性回归分析的迁移率可以精确地推导出分析物与血清白蛋白的结合常数。视黄醇与HSA和BSA的结合常数（K, L mol(-1)）分别为1.28 × 10(5)和5.25 × 10(6)，视黄酸与HSA和BSA的结合常数分别为3.29 × 10(5)和2.27 × 10(6)。研究了分析物的位移和相互竞争结合，表明维甲酸能够取代HSA中的视黄醇，反之亦然。(C) 2012 Elsevier B.V.版权所有
Retinol and retinoic acid are Vitamin A components that are critical for many biological processes. Both of them are strongly complexing with serum albumins giving constants of the order of 10(5) Lmol(-1) or higher. With respect to this fact, affinity capillary electrophoresis (ACE) is not applicable in its commonly used form. Therefore, for the first time, the hyphenated ACE with a high-sensitivity cell was developed and employed to investigate the binding of retinol and retinoic acid in nanomolars with human serum albumin (HSA) and bovine serum albumin (BSA) under physiological conditions. ACE/high-sensitivity coupled cell had contributed to fast the association and dissociation rates of the complexes in nanomolar scale of analytes ensuring the establishment of a dynamic equilibrium within a short electrophoresis time. In addition, this hyphenation led to reduce the concentrations of serum albumins as additives in background electrolyte making a sense beside the proper rinsing protocol for the negligible possibility of their adsorption. The mobility ratio based on nonlinear regression analysis was used to deduce precise binding constants of analytes with serum albumins. The binding constants (K, L mol(-1)) of retinol were 1.28 x 10(5) and 5.25 x 10(6) and retinoic acid were 3.29 x 10(5) and 2.27 x 10(6) with HSA and BSA, respectively. The displacement and reciprocal competitive binding of analytes were investigated and indicated that retinoic acid was able to replace retinol from HSA and vice versa in the case of BSA. (C) 2012 Elsevier B.V. All rights reserved.