Flow injection determination of ethanol in whole blood using immobilized enzymes

Enzymatic methods for the determination of ethanol in whole blood are proposed. They use different types of detection and flow injection analysis (FIA) modes: fluorometric detection (use of normal FIA and stopped-flow/FIA); amperometric detection by monitoring of NADH (use of normal amperometric and pulse mode) and with the aid of a coupled enzymatic reaction (2,6-dichlorophenolindophenol/diaphorase). Determination ranges between 0.1 and 30.0 μg/ml are obtained (which in all cases comprise the legal range of ethanol in blood), with good precision and sampling frequency. The sensitivity of the methods can be manipulated by changing the injected sample volume or the pH.