• Title/Summary/Keyword: nucleosidephosphorylase

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Enzymatic Method for Measuring ATP Related Compounds in Fish Sauces (효소법에 의한 액젓중의 ATP 관련물질 측정)

  • CHO Young Je;IM Yeong Sun;KIM Sang Moo;CHOI Young Joon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.385-390
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    • 1999
  • 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.

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