Determination of the concentration of higher alcohols with enzyme coupled flow-injection analysis in model systems

Burfeind, Jens; Weigel, B.; Kretzmer, G.; Schügerl, K.; Huwig, Alexander; Giffhorn, Friedrich

doi:10.1016/0003-2670(95)00610-9

Cited by 8 publications

(6 citation statements)

References 19 publications

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“…Flow-injection systems for methanol determination have been reported in the literature [ 18 – 23 ]. However, the use of flow-injection for pectin analysis remains unexplored.…”

Section: Introductionmentioning

confidence: 99%

Rapid Enzymatic Method for Pectin Methyl Esters Determination

Łękawska-Andrinopoulou

Vasiliou

Georgakopoulos

et al. 2013

Journal of Analytical Methods in Chemistry

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Pectin is a natural polysaccharide used in food and pharma industries. Pectin degree of methylation is an important parameter having significant influence on pectin applications. A rapid, fully automated, kinetic flow method for determination of pectin methyl esters has been developed. The method is based on a lab-made analyzer using the reverse flow-injection/stopped flow principle. Methanol is released from pectin by pectin methylesterase in the first mixing coil. Enzyme working solution is injected further downstream and it is mixed with pectin/pectin methylesterase stream in the second mixing coil. Methanol is oxidized by alcohol oxidase releasing formaldehyde and hydrogen peroxide. This reaction is coupled to horse radish peroxidase catalyzed reaction, which gives the colored product 4-N-(p-benzoquinoneimine)-antipyrine. Reaction rate is proportional to methanol concentration and it is followed using Ocean Optics USB 2000+ spectrophotometer. The analyzer is fully regulated by a lab written LabVIEW program. The detection limit was 1.47 mM with an analysis rate of 7 samples h−1. A paired t-test with results from manual method showed that the automated method results are equivalent to the manual method at the 95% confidence interval. The developed method is rapid and sustainable and it is the first application of flow analysis in pectin analysis.

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“…Flow-injection systems for methanol determination have been reported in the literature [ 18 – 23 ]. However, the use of flow-injection for pectin analysis remains unexplored.…”

Section: Introductionmentioning

confidence: 99%

Rapid Enzymatic Method for Pectin Methyl Esters Determination

Łękawska-Andrinopoulou

Vasiliou

Georgakopoulos

et al. 2013

Journal of Analytical Methods in Chemistry

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“…Flow-injection systems for methanol determination have been reported in the literature (Almuzara et al, 2002;Burfeind et al, 1996;de María et al, 1 et al, 1993;Sekine et al, 1993;Yue and Zhang, 2007) injection for pectin analysis remains unexplored. The application of flow methods for pectin analysis offers novel, green technologies, which are in a with current sustainability trends.…”

Section: Methyl Esters Determinationmentioning

confidence: 99%

“…Pectin methyl esters can be hydrolyzed either by alkaline hydrolysis or enzymatically by pectin methylesterase. Released methanol can be quantified by means of (Bartolome and Hoff, 1972;Savary and Nuñez, 2003) (Klavons and Bennett, 1986) be related to formaldehyde (Mangos and Haas, 1996) of methylation include determin uronic content (Voragen et al, 1986) injection systems for methanol determination have been reported in the (Almuzara et al, 2002;Burfeind et al, 1996;de María et al, 1 et al, 1993;Sekine et al, 1993;Yue and Zhang, 2007) injection for pectin analysis remains unexplored. The application of flow methods for pectin analysis offers novel, green technologies, which are in a with current sustainability trends.…”

Section: Methyl Esters Determinationmentioning

confidence: 99%

“…When alcohol oxidase is used, the methanol content can (Anthon and Barrett, 2004) (Mangos and Haas, 1996). Other examples of methods for ation from the ratio of the methyl esters to the total en et al, 1986) or using FT injection systems for methanol determination have been reported in the (Almuzara et al, 2002;Burfeind et al, 1996;de María et al, 1 et al, 1993;Sekine et al, 1993;Yue and Zhang, 2007) injection for pectin analysis remains unexplored. The application of flow methods for pectin analysis offers novel, green technologies, which are in a…”

Section: Methyl Esters Determinationmentioning

confidence: 99%

“…Released methanol can be quantified by means of (Bartolome and Hoff, 1972;Savary and Nuñez, 2003) . When alcohol oxidase is used, the methanol content can (Anthon and Barrett, 2004) Other examples of methods for from the ratio of the methyl esters to the total or using FT-IR spectroscopy injection systems for methanol determination have been reported in the (Almuzara et al, 2002;Burfeind et al, 1996;de María et al, 1 et al, 1993;Sekine et al, 1993;Yue and Zhang, 2007). However, the use of flow injection for pectin analysis remains unexplored.…”

Section: Methyl Esters Determinationmentioning

confidence: 99%

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Green methods for methyl esters determination in foodstuffs

Lekawska-Andrinopoulou¹

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In this thesis development of novel, green methods for methyl esters in foodstuffs is described. Methods are based on enzymatic reactions and fluidics. Study focuses on two methyl esters: pectin methyl ester and aspartame, a methyl ester of aspartic acid/phenylalanine dipeptide. Pectin and aspartame are enzymatically hydrolysed by pectin methylesterase or _- chymotrypsin, respectively. Methanol is released and quantified. Several methanol determination methods have been tested, with the method with 4-AAP and phenol showing the best prospects for automation. Method was optimized and its robustness was investigated. Ascorbic acid interference removal with 4-hydroxy TEMPO was tested. Development of two automated methods for methyl esters determination is described: spectrophotometric pectin methyl esters determination and chemiluminescent aspartame determination.The method for pectin methyl esters is the first work on pectin analysis through flow injection. Detection limit down to 1.47 mM was achieved at the analysis rate of 7 samples h-1. The method provides identical results with manual off-line method. The development of the aspartame analyzer was preceded by the development of a spectrophotometric method, which showed good results in samples containing higher aspartame concentrations than expected in beverages. In order to improve method for possible application in beverages chemiluminescent detection was selected for the automated method. The chemistry from kinetic study was modified to accommodate luminol chemiluminescent detection and optimization of the system was performed. Several manifolds were constructed and the effect of following parameters was tested: flow rate, mixing coils length, location and number, preincubation time, alcohol oxidase concentration, use of separate solutions of AOX and HRP. 0.8 ml/min/line flow rate in combination with one 100 cm mixing coil, 60 s preincubation and use of separated solutions of AOX an HRP resulted in sufficient sensitivity that allowed for construction of a calibration curve within the range of aspartame concentration found in diet drinks. Additionally, following side projects related to the main topic of the study are described: development of PME activity assay and galacturonic acid determination.

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Flow injection system for on-line monitoring of fumaric acid in biological processes

Rhee¹,

Sohn²

2003

Analytica Chimica Acta

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Determination of the concentration of higher alcohols with enzyme coupled flow-injection analysis in model systems

Cited by 8 publications

References 19 publications

Rapid Enzymatic Method for Pectin Methyl Esters Determination

Rapid Enzymatic Method for Pectin Methyl Esters Determination

Green methods for methyl esters determination in foodstuffs

Flow injection system for on-line monitoring of fumaric acid in biological processes

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