Linked Life Data

18501250

Source:http://linkedlifedata.com/resource/pubmed/id/18501250

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2008-5-26
pubmed:abstractText
In this paper, we compare and evaluate the applicability of three UV-VIS absorbance based assays for high-throughput quantification of ascorbic acid in horticultural products. All the methods involve the use of a common enzyme (ascorbate oxidase) in combination with a different indicator molecule. The three methods were retrieved from literature: a direct oxidase-method, an OPDA coupled oxidase-method and a PMS-method, which is commercially available. The analysis in high-throughput context involved the analysis in microplates in combination with the use of an automated liquid handling system. We checked (i) the performance factors of the selected methods on standard solutions, (ii) the applicability of the defined methods in high-throughput context, and, (iii) the accuracy of the methods on real samples using HPLC as a reference technique. The OPDA-method was found to be the most appropriate method for the quantification of ascorbic acid in high-throughput context with a linear range between 7.0 and 950 mgL(-1) and excellent correlation parameters (slopes close to 1, intercepts close to 0, R(2)>0.91) with the reference technique when real samples were analyzed. Finally, this method was optimized for assay cost and assay time. Hereto the enzymatic reaction was mathematically described using a model for enzyme kinetics, which was then used to calculate the optimal concentrations of ascorbate oxidase and OPDA. As a result of the modeling the amount of enzyme in the assay could be reduced with a factor 2.5 without affecting significantly the reaction time. In a last step the optimal concentrations were used for a successful validation with the HPLC-reference technique.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
1873-4324
pubmed:author
pubmed:issnType
Electronic
pubmed:day
16
pubmed:volume
618
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
94-101
pubmed:meshHeading
pubmed:year
2008
pubmed:articleTitle
Evaluation and optimization of high-throughput enzymatic assays for fast l-ascorbic acid quantification in fruit and vegetables.
pubmed:affiliation
BIOSYST-MeBioS, Katholieke Universiteit Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium. steven.vermeir@biw.kuleuven.be
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Evaluation Studies