Abstract The measurement of the relative concentrations of hydrogen, deuterium, tritium and helium is an important task in the nuclear fusion research area. Control of the deuterium-tritium (D-T) isotropic ratio and limiting the helium ash content in a fusion plasma are the key to optimizing the fuel burn in a fusion reactor such as ITER. A diagnostic technique has been developed to measure the D-T isotopic ratio in the divertor of a tokamak with a Penning vacuum gauge. The Penning discharge provides a source of electrons to excite the neutral deuterium and tritium in the pumping duct. Subsequently, the visible light from the hydrogen isotopes is collected in an optical fibre bundle, transferred away from the tokamak into a low radiation background area and detected in a high resolution Czerny-Turner spectrometer, equipped with a fast CCD (charge-coupled device) camera for optical detection. The intensity of the observed line emission (D α, 6561.03 Å; T α, 6560.44 Å) is directly proportional to the partial pressure of each gas found in the divertor. The line intensity of each isotope is calibrated as a function of pressure. The ratio of the line intensities thus provides a direct measurement of the D-T isotopic ratio. The lower limit for the determination of the D-T isotopic ratio is about 0.5%. This system is applicable for the pressure range from 10 −5 mbar to a few times 10 −2 mbar.