Enantiomer separation of alpha-hydroxy acids in high-performance immunoaffinity chromatography.
Enantiomer separation of alpha-hydroxy acids in high-performance immunoaffinity chromatography.
复制标题
高效免疫亲和色谱法中 α-羟基酸的对映体分离。
DOI：
10.1016/j.jpba.2007.10.004
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发表时间：
2008
影响因子：
3.4
通讯作者：
Hofstetter,Oliver
中科院分区：
文献类型：
--
作者：
Franco,ElliottJ;Hofstetter,Heike;Hofstetter,Oliver
将抗d-羟基酸单克隆抗体固定在合成的高流通色谱支持材料上，制备了一种适用于游离α-羟基酸对映体分离的手性固定相。在温和的等度缓冲液条件下，使用磷酸盐缓冲盐水（pH 7.4）作为移动的相，在HPLC中实现了此类化合物的几种脂肪族和芳香族成员的手性分离。由于固定化抗体所表现出的高度立体选择性，在所有情况下，l-对映异构体以空隙体积洗脱，而d-对映异构体被保留并第二次洗脱。考察了移动的相参数流速、温度、pH和离子强度对模型分析物扁桃酸对映体分离的影响。虽然它被发现，在流速的变化并没有改变保留因子k2，显着的影响固定化抗体和d-扁桃酸之间的相互作用时，观察到任何其他移动的相参数进行调制。
In this study, a monoclonal anti-d-hydroxy acid antibody was immobilized onto a synthetic high-flow-through chromatographic support material to produce a chiral stationary phase suitable for enantiomer separation of free α-hydroxy acids. Chiral separation of several aliphatic and aromatic members of this class of compounds was achieved in HPLC under mild isocratic buffer conditions using phosphate buffered saline, pH 7.4, as mobile phase. Due to the high degree of stereoselectivity exhibited by the immobilized antibody, in all cases the l-enantiomer eluted with the void volume, while the d-enantiomer was retained and eluted second. The effect of the mobile phase parameters flow rate, temperature, pH, and ionic strength on the enantiomer separation of the model analyte mandelic acid was investigated. While it was found that variations in the flow rate did not change the retention factor k2, dramatic effects on the interaction between the immobilized antibody and d-mandelic acid were observed when any of the other mobile phase parameters were modulated.