Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Enzymatic Method for Measuring ATP Related Compounds in Fish Sauces

효소법에 의한 액젓중의 ATP 관련물질 측정

  • CHO Young Je (Faculty of Food Science and Biotechnology, Food Science and Technology major, Pukyong National University) ;
  • IM Yeong Sun (Faculty of Food Science and Biotechnology, Food Science and Technology major, Pukyong National University) ;
  • KIM Sang Moo (Faculty of Marine Bioscience and Technology, Marine Food Science and Technology major, Kangnung National University) ;
  • CHOI Young Joon (Division of Marine Bioscience, Marine Food Manufacturing major, Gyeongsang National University)
  • 조영제 (부경대학교 식품생명공학부) ;
  • 임영선 (부경대학교 식품생명공학부) ;
  • 김상무 (강릉대학교 해양생명공학부) ;
  • 최영준 (경상대학교 해양생물이용학부)
  • Published : 1999.07.01
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Abstract

HPLC method usually has been used for the determination of ATP and its related compounds in fish muscle and fish sauce. But, total amount of ATP related compounds in fish sauce is determined less than that of fish muscle. In order to establish the extract analysis method for ATP related compounds in fish sauce, a new enzymatic method was developed and compared with existing HPLC method. Fish sauce was extracted with chilled perchloric acid and neutralized to Ph 7.0 with potassium hydroxide solution, the extract was used as sample analyzed by HPLC as usual. On the other hand, for sample analyzed by enzymatic method, 1 ml extract solution was pipetted into test tube. To the tube, 0.5ml of mixed suspension adenosinedeaminase (4U), nucleosidephosphorylase (0.02U) and xanthineoxidase (0.03U) suspended in 2.0ml of 1/15 M sodium phosphate buffer solution pH 7.6 and 1.5ml deionized water wereadded for the decomposition of IMP, HxR and Hx to uric acid at $37^{\circ}C$ for 40 minutes. Total uric acid was determined by measuring optical density at 290nm. In HPLC method, salt decreased the total amount of ATP related compounds by $13.6\~16.2\%$ at $2.5\%$ concentration, but no effect in enzymatic method. IMP, HxR and Hx were detected at 254nm, while uric acid at only 290nm. The ratio of the total amount of ATP related compounds by HPLC method was about $45\%$ of that by enzymatic method in fish sauce. Form these results, enzymatic method is more accurate and simple than HPLC method for analysis of ATP related compounds in fish sauce.

액젓중의 ATP 관련물질의 정확한 분석방법을 확립하기 위하여 현재 사용되고 있는 HPLC법과 새로운 분석방법인 효소법을 비교$\cdot$검토한 결과는 다음과 같다. 1. HPLC 분석법에 의한 멸치 및 까나리육중의 ATP 관련물질총량은 각각 9.332$\mu$mo1/g 및 9.468$\mu$mo1/g이었고, 멸치 및 까나리 액젓중의 ATP 관련물질총량은 각각 3.989$\mu$mol/g 및 4.145$\mu$mol/g 으로 원료육의 $42.7\%$ 및 $43.8\%$였다. 그리고, Hx량은 ATP 관련물질총량의 $85\%$ 정도를 차지하였다. HPLC 법으로 ATP 관련물질을 분석시에 식염의 영향은, 식염농도가 $2.5\%$일 때, IMP, HxR 및 Hx 표준물질의 검출율은 $83.8\~86.4\%$였다. 그리고, IMP, HxR 및 Hx는 254nm에서, 그리고 uric acid는 290nm의 파장에서만 검출되었다. 2. 효소법으로 ATP관련물질 분석시에는 IMP, HxR 및 Hx 표준물질의 검출율이 거의 100\%$였으며, 식염의 영향은 없었다. 멸치 및 까나리 액젓중의 ATP 관련물질총량은 각각 8.942$\mu$mol/g 및 9.424$\mu$mol/g으로, 원료 멸치육 및 까나리육보다 약간 적었다. 효소법으로 분석한 멸치 및 까나리 액젓중의 요산량은 각각 4.357$\mu$mol/g 및 4.632$\mu$mol/으로, 총ATP관련물질의 $48.7\%$ 및 $49.2\%$를 차지하였다. 3. 이상의 결과로부터, 액젓중의 ATP 관련물질을 분석시에는 HPLC 법이 적당하지 못하며, ATP 관련물질을 이들 분해효소를 사용하여 인위적으로 요산으로 전부 분해시켜 측정하는 효소법으로 분석해야할 것이다.

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