OBJECTIVE To assess in a laboratory setting the amount of apically extruded debris associated with different single-file nickel-titanium instrumentation systems compared to one multiple-file rotary system. METHODS Eighty human mandibular central incisors were randomly assigned to four groups (n = 20 teeth per group). The root canals were instrumented according to the manufacturers' instructions using the reciprocating single-file system Reciproc, the single-file rotary systems F360 and OneShape and the multiple-file rotary Mtwo instruments. The apically extruded debris was collected and dried in pre-weighed glass vials. The amount of debris was assessed with a micro balance and statistically analysed using anova and post hoc Student-Newman-Keuls test. The time required to prepare the canals with the different instruments was also recorded. RESULTS Reciproc produced significantly more debris compared to all other systems (P < 0.05). No significant difference was noted between the two single-file rotary systems and the multiple-file rotary system (P > 0.05). Instrumentation with the three single-file systems was significantly faster than with Mtwo (P < 0.05). CONCLUSIONS Under the condition of this study, all systems caused apical debris extrusion. Rotary instrumentation was associated with less debris extrusion compared to reciprocal instrumentation.