An automatic flow procedure for the determination of 3-hydroxybutyrate in animal serum and plasma

• Type: Article in International Scientific Journal
• Date: 2003

Authors

Pires, CK (Author) Other; Martelli, PB (Author) Other; Reis, BF (Author) Other; Lima, JLFC (Author) FFUP; Saraiva, MLMFS (Author) FFUP

Journal

Title: Journal of Agricultural and Food Chemistry

Vol. 51

Pages: 2457-2460

ISSN: 0021-8561

Publisher: American Chemical Society

Indexing

ISI Web of Knowledge - 13 Citations

ISI Web of Science

Scopus - 14 Citations

Scientific classification

FOS: Agrarian Sciences > Other Agrarian Sciences

CORDIS: Health sciences

Other information

Authenticus ID: P-000-H19

DOI: 10.1021/jf021024n

Resumo (PT): An automatic flow procedure based on the multicommutation concept, comprising three-way solenoid valves, for the spectrophotometric determination of 3-hydroxybutyrate in animal serum and plasma is proposed. The 3-hydroxybutyrate was enzymatically converted to acetoacetate with the reduction of NAD+ to NADH monitored at 340 nm. It was possible to carry out up to 600 determinations without a significant decrease in the analytical signal, with 5 mg of 3-hydroxybutyrate dehydrogenase immobilized on porous silica beads and packed in a column. The system enabled 60 determinations/h of 3-hydroxybutyrate in the range of 10−150 mg L-1, with a consumption of 0.9 mg of NAD+ and 200 μL of sample per determination. A detection limit of 2 mg L-1 for both animal serum and plasma and coefficients of variation of 1.4% and 1.2% (n = 17), respectively, were determined. Animal serum and plasma samples were analyzed without previous treatment, the results of which agreed with those obtained using the conventional method (UV kit, Sigma). <br> <br> Keywords: Flow analysis; multicommutation; 3-hydroxybutyrate; spectrophotometry; enzyme reactor; plasma and serum <br> <a target="_blank" href="http://pubs.acs.org/doi/abs/10.1021/jf021024n "> Texto integral </a> <br> <br>

Abstract (EN): An automatic flow procedure based on the multicommutation concept, comprising three-way solenoid valves, for the spectrophotometric determination of 3-hydroxybutyrate in animal serum and plasma is proposed. The 3-hydroxybutyrate was enzymatically converted to acetoacetate with the reduction of NAD(+) to NADH monitored at 340 nm. It was possible to carry out up to 600 determinations without a significant decrease in the analytical signal, with 5 mg of 3-hydroxybutyrate dehydrogenase immobilized on porous silica beads and packed in a column. The system enabled 60 determinations/h of 3-hydroxybutyrate in the range of 10-150 mg L-1, with a consumption of 0.9 mg of NAD(+) and 200 muL of sample per determination. A detection limit of 2 mg L-1 for both animal serum and plasma and coefficients of variation of 1.4% and 1.2% (n = 17), respectively, were determined. Animal serum and plasma samples were analyzed without previous treatment, the results of which agreed with those obtained using the conventional method (UV kit, Sigma).

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 4