Nitric oxide (NO) in exhaled breath has gained increasing interest in recent years mainly driven by the clinical need to monitor inflammatory status in respiratory disorders, such as asthma and other pulmonary conditions. Mid-infrared cavity ring-down spectroscopy (CRDS) using an external cavity, widely tunable continuous-wave quantum cascade laser operating at 5.3 µm was employed for NO detection. The detection pressure was reduced in steps to improve the sensitivity, and the optimal pressure was determined to be 15 kPa based on the fitting residual analysis of measured absorption spectra. A detection limit ($$1\sigma$$ 1σ , or one time of standard deviation) of 0.41 ppb was experimentally achieved for NO detection in human breath under the optimized condition in a total of 60 s acquisition time (2 s per data point). Diurnal measurement session was conducted for exhaled NO. The experimental results indicated that mid-infrared CRDS technique has great potential for various applications in health diagnosis.