This study concerns the establishment of a simple testing method for breath concentration of hydrogen and methane in horses. Twenty-eight healthy thoroughbreds and 24 Arabians were used. Breath samples were collected using one-minute closed circulatory respiration through an aluminum bag filled with 10 liters of pure oxygen, which was mounted on the subjects by means of a face mask. Breath samples obtained, were analyzed by gas chromatography. A significant correlation in both hydrogen and methane levels was observed for samples collected at separate times. These findings confirmed the usefulness of our approach for testing breath concentrations of hydrogen and methane in horses.