人血清蛋白键合固定相的合成及DL(±)氨基酸的手性HPLC分离  

Synthesis of human serum albumin immobilized stationary phase and chiral separation of DL(±) aminoacids by HPLC

作　　者：傅强[1] 贺浪冲[1] 常春[1] 

机构地区：[1]西安医科大学药学院,西安710061

出　　处：《西北药学杂志》1997年第3期99-101,共3页Northwest Pharmaceutical Journal

摘　　要：报道一种人血清蛋白（Humanserumalbumin，HSA）键合固定相的合成方法及DL（±）色氨酸等在HSA柱上的手性分离结果。硅胶与3-aminopropyltrimethoxysilane反应生成氨丙基硅胶，分散于乙腈中，用N，N’-disuccinimidylcarbonate活化。HSA与活化硅胶在20mmol／L磷酸钠缓冲液（pH6．6）中，于30℃反应20h。用RP-HPLC法监控固定相的合成过程。当硅胶与HSA重量比为10：1时，HSA的表面覆盖率为2.75μmol／g。用1％异丙醇的20mmol／LK2HPO4-KH2PO4。缓冲液（pH7．50）作流动相，DL（±）色氨酸可以得到较好的分离，D（＋）异构体先流出色谱柱，k为675，a为1．44，Rs为1．52。DL（±）苯丙氨酸、酪氨酸和组氨酸在上述条件下未得到分离。结果表明：HSA柱对结构相似的氨基酸显示出不同的分离能力。A method of synthesizing HSA (human serum albumin) bonded stationary phase and chiral HPLC separation of some DL (±) aminoacids were reprored. Silica gel reacted with 3-aminopropyltrimethoxylsilane to form aminopropyl silica gel, which was dispersed in acetonitrile and activated by N,N'-disuccinimidyl carbanate. HSA and the activated gel reacted at 30℃ for 20 h in 20 mmol/L phosphate buffer (pH 6.6). The synthetic procedure was monitored by a RP-HPLC programme. When the weight ratio of HSA to silica gel was 1: 10, HSA surface coverage was 2. 75μmol/g. The chiral separation of DL (±) tryptophan was obtained with k1 6. 75, α 1. 44 and Rs 1. 52 as the 20 mmol/L phosphate buffer (pH7.50) containing 1% iso-propanol was used as mobile phase. But chiral separation was not good for DL (±) phenylalanine, tyrosine and histidine in the same conditions. The results revealed that the immobilized HSA column showed different chiral separation ability to aminoacids with similar structures.

关 键 词：DL(±)色氨酸 HSA键合固定相 手性HPLC 

分 类 号：R977.6[医药卫生—药品]