NMR Analysis of Thiodiglycol Oxidation by Mammalian Alcohol Dehydrogenases
Abstract
This unit describes a proton nuclear magnetic resonance (1H NMR)-based methodology for investigation of thiodiglycol (TDG; Fig. 4.20.1A) transformation by human and equine alcohol dehydrogenases (ADH). An ultraviolet/visible (UV/vis) spectrophotometric method is also described in support of the NMR protocol. Its purpose is to initially indicate if TDG is actually a substrate for the various ADH isozymes before the more complicated NMR protocol is undertaken. Once verified spectrophotometrically that TDG is a substrate for a particular ADH isozyme, 1H NMR is used to obtain structural and kinetic data on the products as they form. Although interest was established for the study of TDG metabolism with regard to how the hydrolysis of sulfur mustard (Fig. 4.20.lA) may contribute to its toxicity, the protocols in this unit can be applied as general methods for the metabolic study of different water-soluble xenobiotics using ADMs. Access to NMR depends upon the facility in which an individual researcher works. Most large institutions have a centralized NMR facility where samples are submitted to an appointed scientist or technician. Because the personnel of the NMR facility have control of the instrumentation, they are responsible for acquiring data. This takes the operational aspects of running the NMR out of the hands of the researcher. Smaller research institutions usually have NMR as standard equipment, but they rely on the individual principal investigators and/or students to personally carry out their experiments as operators of the instrument. As such, this unit is being written in context of aiding an investigator who has to personally acquire and interpret the data. There are also multiple manufacturers of NMR spectrometers, each having a unique operational protocol. Therefore, this unit is also written from the standpoint that the specific operational steps in acquiring a spectrum are omitted.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2006
- Accession Number
- ADA468601
Entities
People
- Alan A. Brimfield
- Mark J. Novak
Organizations
- United States Army Medical Research Institute of Chemical Defense