|Hämmerle, M; Hilgert, K; Horn, MA; Moos, R: Analysis of volatile alcohols in apple juices by an electrochemical biosensor measuring in the headspace above the liquid, Sensors and Actuators B Chemical, 158, 313-318 (2011)|
An electrochemical biosensor was optimised for the analysis of volatile alcohols directly from the gas phase without prior absorption or pre-concentration. The sensor is based on the alcohol oxidase (Pichia pastoris) catalyzed conversion of ethanol and the amperometric detection of the generated hydrogen peroxide. Key part of the three-electrode set-up was a gas-diffusion working electrode (potential: +600 mV vs. Ag/AgCl) that consisted of a porous Teflon membrane coated with a thin platinum layer. Headspace samples were analysed for alcohols and used to derive alcohol concentrations in the liquid phase. The biosensor had a sensitivity of 3.43 µA/mM for ethanol, a response time of 69 s, a linear dynamic range of 0.10–30 mM, a theoretical detection limit (3 < S/N) of 9.9 µM, and a stability of 86% during continuous operation (18 h @ 1 mM ethanol). Using one sensor on three consecutive days, the mean coefficient of variation was 1.3% (three measurements each day @ 10 mM ethanol). Alcohol contents of three apple juices determined with the biosensor were in the range 0.30 g/l–0.67 g/l (equivalent to 6.51 mM–14.5 mM). However, ethanol contents determined by high pressure liquid chromatography coupled to refractive index detection (HPLC-RI) and by a commercial enzyme test kit based on alcohol dehydrogenase ranged from 0.12 g/l to 0.38 g/l (equivalent to 2.60 mM–8.25 mM). Both indicate that the biosensor detected alcohols other than ethanol in the apple juices. HPLC-RI coupled to the biosensor in a flow-through configuration demonstrated that the biosensor detected methanol concomitant to ethanol. Thus, the biosensor could perform a qualitative analysis of the total content of volatile alcohols in apple juices by analysing the gas phase above the sample. This offers the additional advantage that possible, non-volatile interfering substances in the liquid sample cannot impair the measurement.