Enzymatic reverse FIA method for total phenols determination in urine samples

Area of Science:

• Analytical Chemistry
• Biochemistry
• Environmental Science

Background:

• Total phenols are important biomarkers in biological and environmental samples.
• Accurate quantification of total phenols in urine is crucial for health assessments.
• Existing methods may lack efficiency or require complex sample preparation.

Purpose of the Study:

• To develop and validate a novel reverse flow injection spectrophotometric enzymatic method for quantifying total phenols in urine.
• To utilize polyphenol oxidase from a readily available source (sweet potato) for enzymatic catalysis.
• To assess the method's performance in terms of detection limit, throughput, and precision.

Main Methods:

• A reverse flow injection analysis system was employed.
• Polyphenol oxidase (PPO) from sweet potato root (Ipomoea batatas) was used as the enzyme.
• Spectrophotometry was utilized for detection and quantification of the enzymatic reaction product.
• Method validation included recovery studies on real urine samples.

Main Results:

• The method achieved a detection limit of 7.7 mg/L for total phenols.
• A high sample throughput of 49 samples per hour was demonstrated.
• Excellent precision was observed with a relative standard deviation of 0.9%.
• Successful application and validation in real urine samples were confirmed through recovery studies.

Conclusions:

• The proposed reverse flow injection spectrophotometric enzymatic method is sensitive, rapid, and precise for total phenol determination in urine.
• The use of crude polyphenol oxidase from sweet potato offers a cost-effective enzymatic solution.
• This method provides a reliable tool for routine analysis of urinary total phenols.