AUTOMATED ENANTIOSEPARATION OF AMINO-ACIDS BY DERIVATIZATION WITH ORTHO-PHTHALDIALDEHYDE AND N-ACYLATED CYSTEINES
AUTOMATED ENANTIOSEPARATION OF AMINO-ACIDS BY DERIVATIZATION WITH ORTHO-PHTHALDIALDEHYDE AND N-ACYLATED CYSTEINES
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DOI：
10.1016/s0021-9673(01)93857-9
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发表时间：
1989-08-04
期刊：
影响因子：
--
通讯作者：
GODEL, H
中科院分区：
文献类型：
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作者：
BRUCKNER, H;WITTNER, R;GODEL, H
采用反相高效液相色谱法，以邻苯二甲醛（OPA）和一系列N-酰基-L-半胱氨酸（Acyl-Cys）为衍生化试剂，对蛋白质-氨基酸的非对映异构体异吲哚衍生物进行分离。使用在338 nm下操作的光电二极管阵列检测器进行检测。为了系统地评价巯基手性试剂中酰基结构的影响，合成了一系列新的N-酰基-L-半胱氨酸[酰基=正丁酰基、异丁酰基（i-But）、新戊酰基、苯甲酰基]，并与已知试剂N-乙酰基-L-半胱氨酸和N-叔丁基-L-半胱氨酸的色谱行为进行了比较。丁氧羰基-l-半胱氨酸。DL-氨基酸的所有酰基-Cys衍生物均被解析。特别地，与其他酰基-Cys相比，i-But-Cys对于大多数氨基酸对映体给出了最高的分辨率。酸奶使用OPA-乙酰-半胱氨酸的调查证明了该方法的适用性，以复杂的食品基质和发生的d-Asp，d-Glu和d-Ala在这种乳制品。
The enantioseparation of standard mixtures composed of proteindl-amino acids was performed by reversed-phase high-performance liquid chromatography of the corresponding diastereomeric isoindolyl derivatives, formed by automated precolumn derivatization witho-phthaldialdehyde (OPA) and a series of N-acyl-l-cysteines (Acyl-Cys). A photodiode-array detector, operating at 338 nm, was used for detection. In order to evaluate systematically the influence of the structures of the acyl group in the chiral thiol reagents, a series of novel N-acyl-l-cysteines was synthesized [acyl =n-butyryl, isobutyryl (i-But), pivaloyl, benzoyl) and the chromatographic behaviour of the diastereomers formed was compared with those of already known reagents, N-acetyl-l-cysteine and N-tert.-butyloxycarbonyl-l-cysteine. All Acyl-Cys derivatives ofdl-amino acids were resolved. In particular,i-But-Cys gave the highest resolutions for most of the amino acid enantiomers in comparison with the other Acyl-Cys. Investigation of yoghurt using OPA-acetyl-Cys demonstrated the applicability of the method to a complex food matrix and the occurrence ofd-Asp,d-Glu andd-Ala in this dairy product.