Chemomechanical root canal preparation (CMRCP) is an important step in root canal treatment. However, one of its negative consequences is apical extrusion of debris of the root canal system contributing to treatment failure and flare-ups. Glide path preparation (GPP) is the initial phase of CMRCP and is crucial for assessing root canal anatomy and establishing unobstructed access to the apical part of the canal. Materials and methods: Forty human mandibular permanent central and lateral incisors were selected; the debris collection apparatus was prepared and the teeth were then divided into four groups: Group 1: Rotary glide path preparation with ProGlider followed by instrumentation with Wave One files. Group 2: Rotary glide path preparation with ProGlider followed by instrumentation with One Shape files. Group 3: Hand glide path preparation with K-file followed by instrumentation with Wave One files. Group 4: Hand glide path preparation with K-file followed by instrumentation with One Shape files. The collected debris was weighed in an analytical digital balance and the collected data were statistically analyzed. Results: No significant difference was present between groups with the same method of glide path preparation or between Wave One and One Shape files. Rotary glide path preparation produced less debris than hand preparation (p≤0.05). Conclusions: Extrusion of debris was observed in all test groups. Rotary glide path preparation could be preferred in clinical practice as it is associated with less debris extrusion than the manual method.