Enzymatic Microreactors for the Determination of Ethanol by an Automatic Sequential Injection Analysis System

Alhadeff, Eliana M.; Salgado, Andréa Medeiros; Cós, Oriol; Pereira, Nei; Valdman, Belkis; Valero, Francisco

doi:10.1007/978-1-60327-181-3_3

Cited by 4 publications

(10 citation statements)

References 10 publications

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“…The experiments were performed with FIA-IV-I and FIA-IV-II at controlled temperature of 25°C. For the integrated biosensor SIA-I system, the best profile of the response signal was obtained with 7.4 ml/min [12] with two microreactors working in series. The detection limit values have been calculated as three times the standard deviation of the background noise [12].…”

Section: Resultsmentioning

confidence: 99%

“…Each system had been tested, after calibration, on both response signal repeatability and stability of the immobilized enzymes, when reused in successive analysis. The results were compared to gas chromatography and high-performance liquid chromatography, having shown good agreement [3,4,12].…”

Section: The Sequential Injections Analysis System Of the Integrated mentioning

confidence: 99%

“…The colorimetric biochemical reaction results in a coloured stable product, which could be detected in accordance with Lambert-Beer law. Other advantages of working with FIA and SIA methods should be mentioned, such as the economic aspects inherent to the reduction of reagent and sample volumes required for each analysis [12].…”

Section: Introductionmentioning

confidence: 99%

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Integrated Biosensor Systems for Ethanol Analysis

Alhadeff

Salgado

Cós

et al. 2008

Appl Biochem Biotechnol

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Different integrated systems with a bi-enzymatic biosensor, working with two different methods for ethanol detection--flow injection analysis (FIA) or sequential injection analysis (SIA)--were developed and applied for ethanol extracted from gasohol mixtures, as well as for samples of alcoholic beverages and fermentation medium. A detection range of 0.05-1.5 g ethanol/l, with a correlation coefficient of 0.9909, has been reached when using FIA system, working with only one microreactor packed with immobilized alcohol oxidase and injecting free horseradish peroxidase. When using both enzymes, immobilized separately in two microreactors, the detection ranges obtained varied from 0.001 to 0.066 g ethanol/l, without on-line dilution to 0.010-0.047 g ethanol/l when a 1:7,000 dilution ratio was employed, reaching correlation coefficients of 0.9897 and 0.9992, respectively. For the integrated biosensor SIA system with the stop-flow technique, the linear range was 0.005-0.04 g/l, with a correlation coefficient of 0.9922.

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Section: Resultsmentioning

confidence: 99%

Section: The Sequential Injections Analysis System Of the Integrated mentioning

confidence: 99%

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Integrated Biosensor Systems for Ethanol Analysis

Alhadeff

Salgado

Cós

et al. 2008

Appl Biochem Biotechnol

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“…Flow analysis techniques have also been applied in method development for ethanol analysis. Alhadeff et al reported some enzymatic methods using flow-based techniques for determination of ethanol contents in gasohol fuels [8,9] and in fermentation bioprocess [10]. This paper reports the development of a new method for quantitative analysis of ethanol in gasohol.…”

Section: Introductionmentioning

confidence: 99%

“…In principle, the method in this paper should be more robust than use of the enzymes [8][9][10], and the method has a good potential in further development for making a portable device for quality control of gasohol.…”

Section: Introductionmentioning

confidence: 99%

Quality control of gasohol using a micro-unit for membraneless gas diffusion

Muncharoen

Sitanurak²,

Tiyapongpattana³

et al. 2008

Microchim Acta

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This work describes development of a new spectrophotometric flow analysis technique suitable for monitoring of ethanol content in gasohol fuel. In this technique, the concept of membraneless gas-diffusion (MBL-GD) was applied with one-step aqueous extraction of gasohol (1:2 gasohol:water). Segments of aqueous extract and color developing reagent were allowed to flow into two separate channels in the MBL-GD device. Inside the device, ethanol vapor can diffuse across a small headspace between the two channels (donor and acceptor). Introduction of an air-segment behind the zone of acceptor reagent to stop dispersion of the colored zone has greatly improved the rapidity of analysis using this MBL-GD technique. Two methods were developed for quality control of gasohol by measuring ethanol content. Method I is suitable for direct calibration of E5 and E10. Method II is recommended for E20. These methods have high accuracy with good precision (% RSD: 1 to 4.9, n=45) and have a sample throughput of 26 samples h -1 . E10 samples were compared with analysis using a standard GC method.

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High-throughput screening and characterization of xylose-utilizing, ethanol-tolerant thermophilic bacteria for bioethanol production

Zhang

et al. 2011

Journal of Applied Microbiology

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Aims: To develop a high‐throughput assay for screening xylose‐utilizing and ethanol‐tolerant thermophilic bacteria owing to their abilities to be the promising ethanologens. Methods and Results: Based on alcohol oxidase and peroxidase‐coupled enzymatic reaction, an assay was developed by the formation of the coloured quinonimine to monitor the oxidation of ethanol in the reaction and calculate the concentration of ethanol. This assay was performed in 96‐well microtitre plate in a high‐throughput and had a well‐linear detection range of ethanol from 0 up to 2·5 g l−1 with high accuracy. The assay was then verified by screening soil samples from hot spring for xylose‐utilizing and ethanol production at 60°C. Three isolates LM14‐1, LM14‐5 and LM18‐4 with 3–5% (v/v) ethanol tolerance and around 0·29–0·38 g g−1 ethanol yield from xylose were obtained. Phylogenetic and phenotypic analysis showed that the isolates clustered with members of the genus Bacillus or Geobacillus subgroup. Conclusions: The developed double enzyme‐coupled, high‐throughput screening system is effective to screen and isolate xylose‐utilizing, ethanol‐producing thermophilic bacteria for bioethanol production at the elevated temperature. Significance and Impact of the Study: Our research presented a novel high‐throughput method to screen thermophilic bacteria for producing ethanol from xylose. This screening method is also very useful to screen all kinds of ethanologens either from natural habitats or from mutant libraries, to improve bioethanol production from lignocellulosic feedstocks.

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Enzymatic Microreactors for the Determination of Ethanol by an Automatic Sequential Injection Analysis System

Cited by 4 publications

References 10 publications

Integrated Biosensor Systems for Ethanol Analysis

Integrated Biosensor Systems for Ethanol Analysis

Quality control of gasohol using a micro-unit for membraneless gas diffusion

High-throughput screening and characterization of xylose-utilizing, ethanol-tolerant thermophilic bacteria for bioethanol production

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