Ethylene glycol (EG) and isopropanol (ISO) are among the major toxic alcohols that pose a risk to human health. However, it is important to distinguish them, since EG is more prone to cause renal failure, and can thus be more dangerous when ingested than ISO. Analysis of alcohols such as isopropanol and ethylene glycol generally can be performed with a complex chromatographic method. Here, we present an optical method based on absorption spectroscopy, performed remotely on EG-ISO mixtures filling a microchannel. Mixtures of ethylene glycol in isopropanol at different volume concentrations were analyzed in a contactless manner in a rectangular-section glass micro-capillary provided with integrated reflectors. Fiber-coupled broadband light in the wavelength range 1.3–1.7 µm crossed the microchannel multiple times before being directed towards an optical spectrum analyzer. The induced zig-zag path increased the fluid–light interaction length and enhanced the effect of optical absorption. A sophisticated theoretical model was developed and the results of our simulations were in very good agreement with the results of the experimental spectral measurements. Moreover, from the acquired data, we retrieved a responsivity parameter, defined as power ratio at two wavelengths, that is linearly related to the EG concentration in the alcoholic mixtures.