title: Enzymatic detection of γ-hydroxybutyrate using aldo-keto reductase 7A2.
abstract: Gamma-hydroxybutyrate (GHB) is a prescribed medication as well as a drug of abuse. Its detection in various matrices for in-field forensic scientists remains a challenge. We have developed an assay that uses aldo-keto reductase 7A2 (AKR7A2) for the specific determination of GHB in various drinks. AKR7A2 was purified using Ni-affinity chromatography. The Michaelis-Menten constant for the GHB oxidation reaction was 10 mM, and the minimum detection limit was 4 mM. Ethanol was not a substrate for AKR7A2. In a coupled reaction with NADP(+), phenazine methosulfate (PMS), and 2,6-dichlorophenolindophenol, various beverages (orange juice, milk, soda, and numerous alcoholic drinks) containing GHB turned from blue to light yellow. In a second coupled reaction where diaphorase replaced PMS, the presence of GHB also caused the expected change of color in various beers.
authors.affiliation: Department of Chemistry, SUNY-Oswego, Oswego, NY 13126, USA. kestutis.bendinskas@oswego.edu
authors.forename: Kestutis
authors.initials: K
authors.lastname: Bendinskas
authors.forename: Patricia
authors.initials: P
authors.lastname: Sattelberg
authors.forename: Daniel
authors.initials: D
authors.lastname: Crossett
authors.forename: Andrew
authors.initials: A
authors.lastname: Banyikwa
authors.forename: Daniel
authors.initials: D
authors.lastname: Dempsey
authors.forename: James A
authors.initials: JA
authors.lastname: MacKenzie