Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1243502 | Talanta | 2010 | 5 Pages |
Abstract
A new kinetic method is proposed for the simultaneous determination of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) based on the different rate of the 3-hydroxybutyrate dehydrogenase-catalysed reactions of these compounds with coenzyme NAD+. A flow injection system with two reactors of immobilised 3-hydroxybutyrate dehydrogenase and dual detection is used. The concentrations of NADH produced after two different reaction times are measured by fluorometry or spectrophotometry and multivariate linear calibration is applied for quantification. Concentrations of 3HB and 3HV between 1Â ÃÂ 10â6 and 1Â ÃÂ 10â4Â M can be determined at an average sampling frequency of 20Â hâ1. In contrast to usual methods, the proposed here makes possible the discrimination of 3HB and 3HV without previous separation so that usual extraction with chlorinated solvents and/or chromatographic separation is not required. The method is of interest in a wide variety of fields concerning PHAs, as it can provide information on the degradation rate and mechanism, composition and structure of these polymers. Its applicability has been proved through the determination of 3HB and 3HV in the digests of some chemically degraded commercial PHAs.
Keywords
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
C. GarcÃa de MarÃa, K.B. Hueso DomÃnguez,