Abstract Species concentration (e.g. CO) and temperature measurements in the combustion field require fast-response technique without interfering species. In the last decade, tunable diode lasers have been established as strong technique to measure species such as CO, CO2, and H2O as well as temperature with high sensitivity. The drawback is the degree of interference that might hamper the robustness of the technique. In this work simultaneous measurements of temperature and CO concentration were carried out using an interference-free mid-infrared laser-based absorption technique behind reflected shock waves. Two transition lines of CO (P(v″ = 0, J″ = 21) and P(v″ = 1, J″ = 21)) in the fundamental vibrational band near 4.87 and 4.93 μm, respectively, were selected. Absorbance interferences from CO2 and H2O at room and high temperatures were evaluated. Spectroscopic parameters for the development of the system were measured: line strengths and collisional broadening coefficients (in Ar) of both lines were obtained at 1020–1950 K by using the scanned-wavelength direct-absorption method. The technique was demonstrated for non-reactive and reactive mixtures. For the non-reactive case, temperature and CO concentration were measured at 1030–1910 K and 1.0–3.7 bar. For the reactive case, oxidation of i-C8H18/O2/Ar and i-C8H18/C2H5OH/O2/Ar mixtures were investigated at three equivalence ratios of 2.0, 1.0, and 0.5. The two newly adopted lines exhibited good performance in the detection of CO concentration and are immune to interferences from CO2 and H2O. In addition, the simulated data from the state-of-the-art isooctane/ethanol mechanisms in literature were compared with the measured data, showing overall good agreement.