An enzymatic method for zinc determination in serum

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Diagnostics

Background:

• Serum zinc (Zn2+) is crucial for various physiological processes.
• Accurate measurement of Zn2+ is essential for diagnosing and managing certain health conditions.
• Existing methods for Zn2+ determination may have limitations in specific clinical settings.

Purpose of the Study:

• To develop and validate a new enzymatic method for determining serum Zn2+ content.
• To compare the efficacy of the new enzymatic method with atomic absorption spectroscopy.
• To assess the utility of the enzymatic method in healthy individuals and patients with specific diseases.

Main Methods:

• Purification of carbonic anhydrase from bovine erythrocytes using affinity chromatography.
• Preparation of apocarbonic anhydrase by removing Zn2+ via dialysis.
• Enzymatic activity assay using 4-nitrophenyl acetate as a substrate.
• Determination of serum Zn2+ in healthy subjects and patients (cirrhosis, diabetes, chronic renal failure) using the enzymatic method and atomic absorption.

Main Results:

• A novel enzymatic method based on apocarbonic anhydrase reactivation was established.
• The enzymatic method demonstrated a good correlation with atomic absorption for serum Zn2+ determination.
• High precision was observed with low intra-assay and inter-assay coefficients of variation for the enzymatic method.

Conclusions:

• The developed enzymatic method provides a reliable and accurate alternative for quantifying serum Zn2+.
• This method holds potential for routine clinical use in assessing Zn2+ status.
• The study validates the enzymatic assay's performance across diverse patient populations.