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논문 기본 정보

자료유형
학술저널
저자정보
Ahlam Majid Azeez (University of Babylon) Mahmoud Hussain Hadwan (University of Babylon)
저널정보
한국분석과학회 분석과학 분석과학 제36권 제1호
발행연도
2023.2
수록면
44 - 52 (9page)

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초록· 키워드

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This protocol clarifies a simple and precise method for measuring the activity of xanthine oxidase (XO) enzyme inhibitor. XO enzyme, which accelerates oxidative stress-related disorders through its capacity to generate hydrogen peroxide and superoxide anion radicals (O₂·˙⁻), has been found to be inhibited by several plant extracts. Enzyme samples were incubated with a suitable buffer containing adequate amounts of xanthine as a substrate to determine XO activity. The method depends on direct measurements of uric acid and hydrogen peroxide production to test XO with and without interference. The CUPRAC reagent (Cu(Nc)₂<SUP>2+</SUP>) was used to inhibit enzyme reaction after incubation was complete. The generated urate and peroxide reduced the Cu(II)-neocuproine complex (Cu(Nc)₂<SUP>2+</SUP>) to a brightly colored Cu(I)-neocuproine complex (Cu(Nc)₂⁺), which was assessed with a spectrophotometer at 450 ㎚. XO activity was found to be directly related to the increased absorbance of the colored Cu(I)-neocuproine complex (Cu(Nc)₂⁺). To eliminate catalase enzyme interference, the proposed method used sodium azide and was validated against XO activity using the UV method in matched samples with t-test analysis. The proposed assay can determine XO activity with high precision, as indicated by the correlation coefficient (R² = 0.9935) from comparison with the reference protocol.

목차

Abstract
1. Introduction
2. Materials and Methods
3. Results and Discussion
4. Conclusions
References

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