Quenching rate is an important parameter to include in fluorescence measurements that are aimed at quantifying the thermochemical field of a reacting flow. Traditionally, the quenching measurements were obtained at low pressures using the direct measurements of quenching times followed by a linear scaling to the desired pressure. This approach, however, cannot account for the possible deviation from the linear pressure scaling at elevated pressures due to three and multi-body collisions. Furthermore, the best accuracy on the quenching rate is obtained with ultra-short pulse lasers that are typically not readily available. This study offsets these limitations by demonstrating a new approach for making direct quenching measurements at atmospheric conditions and using nanosecond lasers. The quenching measurements are demonstrated in a krypton-perturber system, and the 5p[32]2←←4p 6 1 S 0 two-photon electronic transition is accessed. A theoretical construct is presented that relates the absorption spectral parameters and the integrated fluorescence signal to the quenching rate, referenced to a given species and conditions. Using this formulation, the relative quenching rates for different perturber species, namely, air, C H 4, C 2 H 4, and C O 2, are reported as measured at 1 atm and 300 K. As such, the present technique is limited to the measurement of the relative quenching rate, unlike the previous studies where absolute quenching rates are measured. Nonetheless, when the reference quenching rate is independently measured, the relative quenching rates can be converted to absolute values.