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KMID : 1059519950390040257
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Journal of the Korean Chemical Society
1995 Volume.39 No. 4 p.257 ~ p.265
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Micro-Determination of D-Amino Acids in Milk by using Column Switching System
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Lee Sun-Haing
Kim Kyoung-Hee
Lee Young-Cheol
Kim Sang-Tae
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Abstract
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Free amino acids were isolated from milk and their absolute amounts were determined by reversed phase high performance liquid chromatography after derivatization with dansyl chloride. The determination of D- and L-amino acids was based on achiral separation on a C18 column. It was found that milk contained totally 41.00 mg DL-amino acids in 100 mL milk. The level of D-amino acids to L-amino acids was determined by a column-switching system combining an achiral reversed phase separation and chiral chelate additive. The chiral separation was carried out with addition of the chiral Cu(N-benzyl-L-proline)2 chelate to the mobile phase in reversed phase liquid chromatography. It was found that the determination of 16 different amino acids is feasible in the milk sample with a C18 column separation and 12 D-amino acids out of the 16 amino acids can be determined via the column-switching system with chiral separation. 2.05% of D-glutamic acid and 2.93% of D-alanine were found in milk.
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