Micro-Determination of D-Amino Acids in Food by Using Achirai/Chiral Coupled Column Method

Achiral/Chiral Coupled Column법에 의한 식품 중의 D-아미노산의 정량분석

Detectable levels of several free D-amino acids were found in some food. This was accomplished by using a column switching method. The determination of total amount of D- and L-amino acids was based on an achiral separation with a $C_{18}$ column. The level of D-amino acids to L-amino acids was determined by the column switching system including the postcolumn reaction detection of the amino acids derivatized with O-phthalaldehyde. The chiral separation of the postcolumn detection system was carried out with chiral crown ether column. This system was applied for the micro-determination of D-amino acids in food such as soy sauce, fermented soy bean and beans. It turned out that the sampling process is critical for the trace analysis of D-amino acids under this achiral / chiral coupled-column system. It was found that commercial soy sauce contained 42ppm, conventional soy sauce 102ppm, fermented soy bean 8.34mg per 1g and bean 2.87mg per 1g sample for phenylalanine. D-phenylalanine was found 0.67% in commercial soy sauce, 0.34% in conventional soy sauce, less than 1.81% in fermented soy bean, and Jess than 2.82% in bean.

식품 중의 몇몇 유리 D-아미노산이 column switching method에 의해 검출되었다. D-아미노산과 L-아미노산의 총량의 정량은 $C_{18}$ 컬럼을 사용한 비키랄 분리에 근거한다. L-아미노산에 대한 D-아미노산의 수준은 o-phthalaldehyde로 유도체화된 아미노산의 postcolumn reaction detection을 포함하는 column switching system에 의해 정량되었다. Postcolumn detection system의 키랄 분리는 chiral crown ether 컬럼에 의해 실행되었다. 이 system은 간장과 발효된 콩, 그리고 콩 같은 식품 중에 있는 D-아미노산의 정량에 적용된다. 이 achiral/chiral coupled-column system하에서는 시료 전처리과정이 중요함을 알게 되었다. Phenylalanine은 시판용 간장 속에 242ppm, 재래식 간장 속에는 102ppm, 발효된 콩에는 1g당 8.34mg, 콩에는 1g당 2.87mg이 함유된 것으로 밝혀졌다. D-phenylalanine은 시판용 간장 속에는 0.67%, 재래식 간장에는 0.34%, 발효된 콩에는 1.81% 미만, 콩에는 2.82% 미만이 검출되었다.