The present study investigated the effect of tonal changes on voice onset time (VOT) between normal laryngeal (NL) and superior esophageal (SE) speakers of Mandarin Chinese. VOT values were measured from the syllables /pha/, /tha/, and /kha/ produced at four tone levels by eight NL and seven SE speakers who were native speakers of Mandarin. Results indicated that Mandarin tones were associated with significantly different VOT values for NL speakers, in which high-falling tone was associated with significantly shorter VOT values than mid-rising tone and falling-rising tone. Regarding speaker group, SE speakers showed significantly shorter VOT values than NL speakers across all tone levels. This may be related to their use of pharyngoesophageal (PE) segment as another sound source. SE speakers appear to take a shorter time to start PE segment vibration compared to NL speakers using the vocal folds for vibration.