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Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney

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Flow injection analysis biosensor for urea analysis in urine using enzyme thermistor.

Geetesh K Mishra¹, Atul Sharma, Kanchanmala Deshpande

¹Biosensor Laboratory, Department of Chemistry, Birla Institute of Technology and Science, Pilani, KK Birla Goa Campus, Goa, 403726, India, geeteshmishra26@gmail.com.

Applied Biochemistry and Biotechnology

|June 8, 2014

Summary

A new flow injection analysis-enzyme thermistor (FIA-ET) biosensor effectively quantifies urea levels in urine. This practical method aids in monitoring renal function by accurately detecting urea in patient samples.

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Area of Science:

Analytical Chemistry
Biomedical Engineering
Biotechnology

Background:

Clinical monitoring of renal function requires accurate urea level assessment in urine.
Existing analytical methods may lack the practicality or sensitivity needed for routine clinical use.
Urea quantification is crucial for diagnosing and managing kidney-related conditions.

Purpose of the Study:

To develop and validate a practical flow injection analysis-enzyme thermistor (FIA-ET) biosensor for quantifying urea in human urine.
To assess the biosensor's performance characteristics, including linearity, response time, and interference.
To evaluate the reliability, reproducibility, and operational stability of the developed urea biosensor.

Main Methods:

A biosensor was constructed using covalently immobilized urease enzyme on an aminated silica support.
Flow injection analysis coupled with an enzyme thermistor (FIA-ET) was employed for urea detection.
Optimized conditions included a 100 mM phosphate buffer at pH 7.2, a flow rate of 0.5 mL/min, and a sample volume of 0.1 mL.

Main Results:

The FIA-ET biosensor demonstrated a linear response for urea detection in the range of 10-1,000 mM (R² = 0.99) with a response time of 90 seconds.
Urea-spiked human urine samples exhibited minimal matrix interference, with recovery rates between 92.26% and 99.80%.
The biosensor showed high reliability (% RSD = 0.741) and excellent operational stability, retaining 80% of its response after 30 weeks.

Conclusions:

The developed FIA-ET biosensor provides a practical and effective method for quantifying urea levels in human urine.
This biosensor is suitable for monitoring patients' renal function due to its accuracy, stability, and minimal interference.
The technology holds potential for widespread clinical application in kidney function assessment.