Simple assay for quantifying xanthine oxidase activity

Azeez, A.; Hadwan, Mahmoud Hussein

doi:10.1016/j.ab.2023.115192

Cited by 2 publications

(1 citation statement)

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“…The overproduction of uric acid, attributed to hyperuricemia, leads to conditions such as gout [4]. To counter this, XO inhibitors such as allopurinol block the final step of uric acid metabolism and are commonly employed for gout treatment [5]. Hyperuricemia, a condition arising from an imbalance in uric acid excretion, can be induced by excessive consumption of certain dietary components, such as meats, select seafood, and nucleic acid-rich foods.…”

Section: Introductionmentioning

confidence: 99%

Inhibitory effects of Moringa oleifera seed extracts on crude xanthine oxidase

Abdullahi Abdulkadir,

Nafisat Ozavize Abdullahi,

Sakariyau Adio Waheed

et al. 2023

GSC Biol. Pharm. Sci.

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The quest for novel xanthine oxidase (XO) inhibitors, characterized by reduced side effects and heightened therapeutic efficacy, holds significant importance in addressing hyperuricemia and gout. This study was undertaken to assess the inhibitory properties of Moringa oleifera seed extracts on crude XO. The isolation of crude XO from bovine (cow) milk was achieved through ammonium sulfate precipitation techniques, and its activity was subsequently quantified using spectrophotometric measurements at a wavelength of 293 nm. The impact on xanthine oxidase inhibition was investigated utilizing varying concentrations of Moringa oleifera seed extracts in methanol, ethyl acetate, and aqueous solutions. Among these extracts, the ethyl acetate variant displayed the most robust inhibitory effect, with a notable inhibition rate of 89.16% ± 0.42, followed by the methanol extract at 65.59% ± 0.79 and the aqueous extract at 48.77% ± 0.85. For comparison, the positive control (allopurinol) exhibited an inhibition rate of 94.32% ± 0.66. The kinetic parameters of the enzyme were determined, revealing Vmax and km values of 0.05 U/ml and 131.35 µM, respectively. When the enzyme was exposed to the inhibitory extracts, the resulting Vmax and km values were as follows: ethyl acetate extract; 0.03 U/ml and 77.12 µM, methanol extract; 0.04 U/ml and 131.35 µM, and aqueous extract; 0.02 U/ml and 74.23 µM. This research work establishes a foundation for the potential utilization of Moringa oleifera seed extracts in addressing hyperuricemia and, consequently, gout.

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Section: Introductionmentioning

confidence: 99%

Inhibitory effects of Moringa oleifera seed extracts on crude xanthine oxidase

Abdullahi Abdulkadir,

Nafisat Ozavize Abdullahi,

Sakariyau Adio Waheed

et al. 2023

GSC Biol. Pharm. Sci.

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An improved method for measuring catalase activity in biological samples

Hadwan,

Hussein,

Mohammed

et al. 2024

Biology Methods and Protocols

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Catalase is an important enzyme that protects biomolecules against oxidative damage by breaking down hydrogen peroxide into water and oxygen. Catalase is present in all aerobic microbes, animals, and plants. It is, however, absent from normal human urine but can be detected in pathological urine. Catalase testing can thus help to detect such urine. This study presents a novel spectrophotometric method for determining catalase activity characterized by its simplicity, sensitivity, specificity, and rapidity. The method involves incubating enzyme-containing samples with a carefully-chosen concentration of hydrogen peroxide for a specified incubation period. Subsequently, a solution containing ammonium ferrous sulfate (FAS) and sulfosalicylic acid (SSA) is added to terminate the enzyme activity. A distinctive maroon-colored ferrisulfosalicylate complex is formed. The formation of this complex is a direct result of the reaction between ferrous ammonium sulfate (FAS) and any residual peroxide present. This leads to the generation of ferric ions when coordinated with SSA. The complex has a maximum absorbance of 490 nm. This advanced method eliminates the need for concentrated acids to stop catalase activity, making it safer and easier to handle. A comparative analysis against the standard ferrithiocyanate method showed a correlation coefficient of 0.99, demonstrating the new method's comparable effectiveness and reliability. In conclusion, a simple and reliable protocol for assessing catalase activity, which utilizes a cuvette or microplate, has been demonstrated in this study. This interference-free protocol can easily be used in research and clinical analysis with considerable accuracy and precision.

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Simple assay for quantifying xanthine oxidase activity

Cited by 2 publications

References 50 publications

Inhibitory effects of Moringa oleifera seed extracts on crude xanthine oxidase

Inhibitory effects of Moringa oleifera seed extracts on crude xanthine oxidase

An improved method for measuring catalase activity in biological samples

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