Flow Injection Analysis Biosensor for Urea Analysis in Urine Using Enzyme Thermistor

There is a need for analytical methods capable of monitoring urea levels in urine for patients under clinical monitoring to appraise renal function. Herein, we present a practical method to quantify levels of urea in human urine samples using flow injection analysis-enzyme thermistor (FIA-ET) biosen...

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Bibliographic Details
Published in:Applied biochemistry and biotechnology 2014-10, Vol.174 (3), p.998-1009
Main Authors: Mishra, Geetesh K, Sharma, Atul, Deshpande, Kanchanmala, Bhand, Sunil
Format: Article
Language:English
Subjects:
Biochemistry
Biological and medical sciences
Biosensing Techniques
Biosensors
Biotechnology
Body Fluids
Canavalia ensiformis
Chemistry
Chemistry and Materials Science
Enzymes
Enzymes, Immobilized - chemistry
Flow Injection Analysis
Fundamental and applied biological sciences. Psychology
Humans
immobilized enzymes
Injection
Kidneys
Methods. Procedures. Technologies
monitoring
Nanotechnology
patients
Renal function
Silica
Silicon Dioxide - chemistry
temperature
Urea
Urea - isolation & purification
Urea - urine
urease
Urease - chemistry
urinalysis
Urine
Various methods and equipments
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Summary:There is a need for analytical methods capable of monitoring urea levels in urine for patients under clinical monitoring to appraise renal function. Herein, we present a practical method to quantify levels of urea in human urine samples using flow injection analysis-enzyme thermistor (FIA-ET) biosensor. The biosensor comprises a covalently immobilized enzyme urease (Jack bean) on aminated silica support, which selectively hydrolyzes the urea present in the sample. Under optimized conditions, the developed biosensor showed a linear response in the range of 10–1,000 mM, R ² = 0.99, and response time of 90 s in 100 mM phosphate buffer (PB) (flow rate of 0.5 mL/min, sample volume of 0.1 mL, and pH 7.2). The urea-spiked human urine samples showed minimal matrix interference in the range of 10–1,000 mM. Recoveries were obtained (92.26–99.80 %) in the spiked urine samples. The reliability and reproducibility of the developed biosensor were found satisfactory with percent relative standard deviation (% RSD) = 0.741. The developed biosensor showed excellent operational stability up to 30 weeks with 20 % loss in original response when used continuously at room temperature. These results indicate that the developed biosensor could be very effective to detect low and high levels of urea in urine samples.
ISSN:0273-2289
1559-0291
DOI:10.1007/s12010-014-0985-0