Abstract:Potentiometric creatinine microsensors were fabricated by use of a composite film consisting of polyion complex (PlC), which contains creatinine iminohydrolase (CIH), and the electrochemically inactive polypyrrole (iPPy). The PPy/PIC-CIH composite film electrode displayed potential response to creatinine concentration owing to a pH change occurring during the enzymatic reaction. The sensitivity was varied by using different polyions of the PlC. With poly-Llysine and sebacic acid, the iPPy/PIC-CIH electrode dem… Show more
“…After preparing PIC membranes containing enzymes, polypyrrole can be synthesized in the membranes electrochemically. [48][49][50] Even though other polymers can be synthesized in the PIC, the PIC membranes are not destroyed, but can be used for biosensors.…”
The immobilization of biomolecules is an important technique for bio-analysis, and can be applied to biosensors with both high selectivity and high sensitivity. Many researchers have developed immobilization techniques to optimize these characteristics. In the last two decades, an immobilization technique that meets the desired requirements was developed by using polyelectrolytes to form complexes, based on the electrostatic binding between polycations and polyanions. This review summarizes the techniques used for the immobilization of biomolecules by polyelectrolyte complexes; it also discusses related subjects.
“…After preparing PIC membranes containing enzymes, polypyrrole can be synthesized in the membranes electrochemically. [48][49][50] Even though other polymers can be synthesized in the PIC, the PIC membranes are not destroyed, but can be used for biosensors.…”
The immobilization of biomolecules is an important technique for bio-analysis, and can be applied to biosensors with both high selectivity and high sensitivity. Many researchers have developed immobilization techniques to optimize these characteristics. In the last two decades, an immobilization technique that meets the desired requirements was developed by using polyelectrolytes to form complexes, based on the electrostatic binding between polycations and polyanions. This review summarizes the techniques used for the immobilization of biomolecules by polyelectrolyte complexes; it also discusses related subjects.
“…16 Entrapment of CIH has been achieved within a composite film consisting of a polyion complex and pH-sensitive, inactive polypyrrole prepared by electropolymerization. 17 The microsensor measures pH changes that accompany the enzyme-catalyzed reaction. The sensitivity of the modified electrode was relatively high and could be controlled with different polyions in the polyion complex.…”
Section: Enzyme Immobilization For Potentiometric Biosensorsmentioning
To measure creatinine, electrochemical techniques have been coupled with a range of biological recognition elements in a variety of sensor configurations. (To listen to a podcast about this feature, please go to the Analytical Chemistry website at pubs.acs.org/ac.The measurement of creatinine levels in human blood or urine is clinically essential because the levels partially reflect the state of renal and muscle function. Creatinine is naturally produced by the body and is filtered from the bloodstream by the kidneys in relatively constant amounts every day. The normal physiological concentration is 40-150 µM, but it can exceed 1000 µM in certain pathological conditions. Blood levels >150 µM indicate the need to perform tests such as creatinine clearance. Values >500 µM indicate severe renal impairment, ultimately leading to dialysis or transplantation; 1 levels <40 µM indicate decreased muscle mass.The methods most often used for the clinical determination of creatinine are based on colorimetry. 2 However, the methods are affected by numerous metabolites and drugs found in biological samples, such as glucose, fructose, ketone bodies, ascorbic acid, and cephalosporins. 3,4 Introducing enzymes has increased specificity, but the methods also became complicated and less reliable. Large and expensive benchtop analyzers incorporating a number of electrochemical electrodes have been used in central clinical laboratories. Portable and handheld devices incorporating a single-use creatinine biosensor cartridge also have been used.The goal of biosensor engineering in the clinical laboratory setting is to reduce cost, time, and complexity of routine analysis of biological fluids; to enable near-patient testing of blood, urine, and saliva in medical centers; and ultimately to enable home testing by individuals. 5 This article looks at the developments in electrochemical creatinine biosensor research in terms of sensor design and analytical performance on the basis of the recognition element and the nature of transducer. Parameters and specific performance characteristics to consider include cost (<$10/ sensing strip), response time (<1.5 minutes [min]), detection limit (e10 µM), linear range (10-1000 µM), and lifetime (>1 year).
“…7,8 To enhance the specificity, enzymatic methods are often used. [9][10][11][12] The most popular among them is the three-enzyme method, in which creatinine amidohydrolase (CA), creatine amidinohydrolase (CI), and sarcosine oxidase (SO) can be employed to catalyze the hydrolysis of creatinine to hydrogen peroxide (H 2 O 2 ), which can then be detected amperometrically. 9 Besides, creatinine iminohydrolase (CIH) was utilized to catalyze the hydrolysis of creatinine to ammonia (NH 3 ); the NH 3 was then detected by potentiometry.…”
Section: Introductionmentioning
confidence: 99%
“…9 Besides, creatinine iminohydrolase (CIH) was utilized to catalyze the hydrolysis of creatinine to ammonia (NH 3 ); the NH 3 was then detected by potentiometry. 10 Although enzymatic methods are much more specific, they are usually expensive and suffer from instability.…”
A method based on disposable pipette extraction (DPX) was successfully applied to creatinine determination in urine samples analysis using liquid chromatography with ultraviolet spectrophotometric detection (DPX/LC-UV). DPX variables, number of draw/eject cycles, sample pH, and type of the desorption solvent, were employed in a factorial experimental design to optimize the sorption equilibrium and time analysis. Among the evaluated DPX variables, the highest extraction efficiency was obtained with 500 µL of urine sample mixed with 1 mL of borate solution (pH 9) with one draw/eject cycle followed by liquid desorption of 1 mL of methanol in seven draw/eject cycles. The developed DPX/LC-UV method showed a linear response from the limit of quantification of 0.317 to 3.390 g L -1 with r 2 = 0.996 and inter-day precision with a coefficient of variation below 8.8%. Based on these results, the proposed method can be a useful tool for determining the creatinine levels in urine samples.
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