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A rapid and accurate assay for monitoring ethanol production is required to control the progress of fermentation in various industrial-related research and processes. In this study, a spectrophotometric assay to measure ethanol concentration in fermentative sample was developed. Ethanol in unknown aqueous solution was extracted using tri-n-butyl phosphate (TBP) and subsequently oxidized by dichromate reagent. Oxidation product of ethanol with dichromate reagent could be visualized as blue green-color. The A595 value detected by spectrophotometer and ethanol concentration between 0.7%-8.0% was plotted in linear regression with high correlation coefficiency (R2). In addition, concentration of methanol, propanol and butanol were determined as did in ethanol suggesting the broad application of this assay. Our established method was applied to commercial wine and fermentative products from yeast culture broths and the results were compared with Gas Chromatography Mass Spectrophotometry (GCMS) method. In this study, this assay was demonstrated as a cheap, rapid, and high accuracy method for determination of ethanol concentration in unknown solutions.
Sriariyanun et al. (2019) studied this question.