Abstract

The development of biosensors using biological components has an important role in detecting the disease early because it has good selectivity and accuracy. In this study, a biosensor which made is a urea biosensor, based on immobilization urease in chitosan using adsorption techniques, to measure urea levels by colorimetric analysis with bromothymol blue (BTB) as an indicator. The purpose of this study was to find out how to measure urea levels using biosensors based on urease immobilization in chitosan and find out the biosensor performance including optimum enzymatic reaction time, linearity, the limit of detection, repetition, and determination of disrupting compounds. The study began with the making of an immobilization supporting matrix using chitosan which was made in the form of cryogel through an ionic gelation process which adsorbs the urease enzyme. Cryogel urease catalyzes the hydrolysis of urea into NH4+ and CO2-. The reaction product was added with the BTB indicator, and the color change formed was measured using a spectrophotometer. The results showed that the performance of urea biosensors was good enough for urea level detection systems by producing enzymatic reaction times at 15 minutes, linearity at 0.9951, detection limit at 0.018 mM, not affected by the addition of 0.05 mM ascorbic acid and 0.4 mM uric acid. This urea biosensor can be used up to 5 repetitions.

Highlights

  • IntroductionUrea in blood or urine is an important marker for the diagnosis of kidney disease

  • Urea is the final product of the nitrogen cycle in the liver

  • The chitosan cryogel was prepared by dissolving chitosan into the acetic acid solution

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Summary

Introduction

Urea in blood or urine is an important marker for the diagnosis of kidney disease. The normal urea levels in serum are about 2.5 mM to 6.7 mM (Saeedfar, Heng, Ling, & Rezayi, 2013), In patients with kidney failure, the urea levels in serum could increase to 50 – 150 mM (Velychko et al, 2016). It is important to develop or improve the urea detection methods as part of the preventive strategy of early detection of kidney failure. Urea concentration in blood could be determined in various methods such as chromatographic (Wang et al, 2016), spectrophotometric (Fuertes, Pérez, & Alonso, 2004), colorimetric (Fatoni, Anggraeni, & Zulhidayah, 2019) and electrochemical (Tiwari, Aryal, Pilla, & Gong, 2009). The recent study of urea determination was the use of biological sensing element, such as enzyme to detect the urea concentration

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