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An improved method for measuring catalase activity in biological samples.

Mahmoud Hussein Hadwan1, Marwah Jaber Hussein1, Rawa M Mohammed2

  • 1Department of Chemistry, College of Science, University of Babylon, Hilla 51002, Iraq.

Biology Methods & Protocols
|March 25, 2024
PubMed
Summary
This summary is machine-generated.

A new, safer spectrophotometric method accurately measures catalase (CAT) enzyme activity in urine. This simple assay can detect pathological urine, aiding in clinical diagnostics.

Keywords:
Bland–Altman plotferrous ammonium sulfatemicroplate protocolspectrophotometrysulfosalicylic acid

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Area of Science:

  • Biochemistry
  • Enzymology
  • Analytical Chemistry

Background:

  • Catalase (CAT) is a crucial enzyme protecting against oxidative damage by decomposing hydrogen peroxide (H₂O₂).
  • While present in most organisms, CAT is typically absent in normal human urine but indicates pathological conditions.
  • Accurate CAT detection in urine is vital for diagnosing certain medical conditions.

Purpose of the Study:

  • To develop and validate a novel, simple, sensitive, and rapid spectrophotometric method for quantifying catalase activity.
  • To establish a safer alternative to existing methods that avoids concentrated acids.
  • To assess the reliability and applicability of the new method for clinical and research use.

Main Methods:

  • A spectrophotometric assay was developed involving sample incubation with H₂O₂, followed by termination with ferrous ammonium sulfate (FAS) and sulfosalicylic acid (SSA).
  • The formation of a maroon-colored ferrisulfosalicylate complex, measured at 490 nm, quantifies residual H₂O₂ and thus CAT activity.
  • The new method was compared against the standard ferrithiocyanate method using correlation analysis.

Main Results:

  • The novel method demonstrated simplicity, sensitivity, specificity, and rapidity in determining CAT activity.
  • The assay produced a stable, measurable complex with a maximum absorbance at 490 nm.
  • A high correlation coefficient (0.99) was observed when compared to the established ferrithiocyanate method, indicating comparable accuracy and reliability.

Conclusions:

  • A safe, interference-free, and reliable spectrophotometric protocol for assessing urine catalase activity has been successfully developed.
  • This method, adaptable for cuvette or microplate formats, offers considerable accuracy and precision for research and clinical analysis.
  • The new assay provides a valuable tool for detecting pathological urine based on catalase presence.