DETERMINATION OF D-AMINO ACIDS .2. USE OF A BIFUNCTIONAL REAGENT, 1,5-DIFLUORO-2,4-DINITROBENZENE
DETERMINATION OF D-AMINO ACIDS .2. USE OF A BIFUNCTIONAL REAGENT, 1,5-DIFLUORO-2,4-DINITROBENZENE
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DOI：
10.1007/bf02908688
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
MARFEY, P
中科院分区：
文献类型：
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作者：
MARFEY, P
由1,5 - 二氟 - 2,4 - 二硝基苯和L - 丙氨酸 - 氨合成了1 - 氟 - 2,4 - 二硝基苯基 - 5 - L - 丙氨酸酰胺，产率较高（76%）。该化合物含有一个活性氟原子，可用于与L - 和D - 氨基酸的混合物发生反应。所得到的非对映异构体产率为定量，可通过高效液相色谱（HPLC）进行分离和测定。对于所研究的5种氨基酸（丙氨酸、天冬氨酸、谷氨酸、蛋氨酸和苯丙氨酸），L - 非对映异构体在反相柱上比D - 非对映异构体先洗脱出来。这种现象可以用后者（D - 非对映异构体）中更强的分子内氢键来解释。当用该试剂对含有已知比例L - 和D - 异构体的5种氨基酸的人工混合物进行衍生化，并通过HPLC分析反应产物时，可以测定每种异构体在纳摩尔范围内的相对含量。
1-Fluoro-2,4-dinitrophenyl-5-L-alanine amide was synthesized in high yield (76%) from 1,5-difluoro-2,4-dinitrobenzene and L-Ala-NH2. This compound contains a reactive fluorine atom which can be used for the reaction with a mixture of L- and D-amino acids. The resulting diastereomers which are obtained in quantitative yield can be separated and estimated by HPLC [high performance liquid chromatography]. With the 5 amino acids studied (Ala, Asp, Glu, Met and Phe), L-diastereomers were eluted from the reverse-phase column before D-diastereomers. This behavior can be explained by a stronger intramolecular hydrogen bonding in the latter diastereomer. When artificial mixtures of the 5 amino acids containing known proportions of L- and D-isomers were derivatized with the reagent and the reaction products analyzed by HPLC, it was possible to determine the relative content of each isomer in a nanomole range.