A horizontal diamond attenuated total reflection (ATR) element has been incorporated in a flow-through cell with low dead volume and used for on-line mid-IR detection in high-performance liquid chromatography. The chemical inertness of the ATR element permitted the use of a strongly acidic mobile phase in the isocratic separation. The hyphenation was used for the analysis of organic acids, sugars and alcohols in red wine. In the case of co-eluting analytes multivariate curve resolution-alternating least squares (MCR-ALS) was successfully employed for quantitative analysis.