Objectives: The aim of the present study is to define breath-holding in spontaneous respiration and to observe the difference of respiratory variables, EEG and HRV. Methods: 46 healthy young volunteers (M:F=31:15) were recruited in the study. By measuring and analysing respiration, EEG and HRV in the spontaneous respiration. We segment subjects by 100% of coefficient of variation in the breath-holding. Results & Conclusions: 1. There is a period of breath-holding after expiration and before endeavored inspiration, in the course of respiration. The greater coefficient of variation in the breath-holding, longer respiration period mean, inspiration period mean and breath-holding mean in respiratory variable. 2. There was no significant difference between HRV parameters. 3. The greater coefficient of variation in the breath-holding, the higher ${eta}$ frequency and ${gamma}$ frequency in the left prefrontal lobe.