A Rapid Method for Determination of ATP and Its Related Compounds in Dried Fish and Shellfish Products Using HPLC

HPLC에 의한 시판수산건제품의 ATP분해생성물의 신속정량법

This paper deals with a rapid method for determination of ATP and its related compounds in fish and shellfish products using high performance liquid chromatography(HPLC). The HPLC used is a HPLC/ALC-224 equiped with UV-spectrophotometer (254 nm) as detector and integrator (Yanagimoto system-1000). The column used is a stainless steel tubing ($30.0\;cm{\times}3.9\;mm\;i.d.$) packed with ${\mu}-Bon-dapak\;C_{18}$. A mixture of $1\%$ triethylamine-phosphoric acid(pH6.5) was used as an eluent and the flow rate of the eluent was controlled at 2 ml/min. For the separation of ATP and its related compounds, a standard mixture of ATP, ADP, AMP, IMP, inosine and hypoxanthine was subjected to HPLC under the above mentioned conditions. Six peaks were obtained with retention times within 20 min, and elution order were hypoxanthine, IMP, inosine, AMP, ADP and ATP. But 5'-IMP and 5'-GMP fractions were not separated by this method. In generally, IMP content in boiled-dried fish and shellfish products purchased from the market was comparatively higher than that of other nucleotides. Especially, boiled-dried big eye herring marked higher value in IMP content than other boiled-dried ones. Hypoxanthine and inosine were major components of ATP-related compounds in dried products and seasoned-dried ones. And IMP content in seasoned-dried products was higher than that of dried ones. This fact is suggested that a part of IMP in seasoned-dried ones was derived from flavoring matter (MSG, 5'-IMP and 5'-GMP) which is added during the seasoning treatment.

보다 간편하게 핵산관련물질을 정량할 목적으로 고속액체크로마토그래피를 이용한 정량법을 검토하였으며, 이 방법을 이용하여 시판 수산건제품의 ATP분해생성물의 함량을 정량하였다. 장치는 HPLC/ALC-244(Waters Associates Inc.)를, 칼럼은 ${\mu}-Bondapak\;C_{18}$ stainless steel column($3.9mm\;i.d.{\times}30.0cm$)을, 검출기는 UV검출기(254nm)를, integrator는 Yanagimoto system-1000을 각각 사용하였으며, 이동상(移動相)으로서는 $1\%$ triethylamine${\codt}$phosphoric acid 혼액 (pH 6.5, $1\%$ triethylamine 용액을 phosphoric acid로써 pH조절)을 사용하고 2ml/min로 용리시키는 것이 가장 분리능이 좋았다. 용리되는 순서는 hypoxanthine, IMP, inosine, AMP, ADP 및 ATP의 순이었으며, retention time 20분이내에 분석이 완료되었다. 그러나 5'-IMP와 5'-GMP는 분리되지 않았다. 자건품(煮乾品)은 이노신산(IMP) 함량이 전반적으로 많았는데, 그 중에서도 벤뎅이가 가장 함량이 많았다. 소건품(素乾品)에는 inosine, hypoxanthine 함량이 많았으며, 조미건제품(調味乾製品)도 소건품과 비슷한 양상이었으나 IMP 함량이 비교적 높은 것은 조미할 때 핵산계조미료가 첨가되었기 때문이라고 볼 수 있었다.