Visible and near infrared reflectance spectroscopy (Vis-NIR) has been widely used by the industry research-base for large-scale meat quality evaluation to predict the chemical composition of meat quickly and accurately. Meat tenderness is measured by means of slow and destructive methods (e.g. Warner-Bratzler shear force). Similarly, sensory analysis, using trained panellists, requires large meat samples and is a complex, expensive and time-consuming technique. Nevertheless, these characteristics are important criteria that affect consumers’ evaluation of beef quality. Vis-NIR technique provides information about the molecular bonds (chemical constituents) and tissue ultra-structure in a scanned sample and thus can indirectly predict physical or sensory parameters of meat samples. Applications of Vis-NIR spectroscopy in an abattoir for prediction of physical and sensory characteristics have been less developed than in other fields. Therefore, the aim of this study was to test the on-line Vis-NIR spectroscopy for the prediction of beef quality characteristics such as colour, instrumental texture, water holding capacity (WHC) and sensory traits, by direct application of a fibre-optic probe to the M. longissimus thoracis with no prior sample treatment.