Objective: The difficult laparoscopic cholecystectomy (LC) is defined as the presence of one of the followingconditions including prolonged operative time, conversion to open cholecystectomy or significant blood loss. Atpresent, there is no evidence of predictive factors related to longer operative time in single-incision laparoscopiccholecystectomy (SILC). The aim of this study is to determine predictive factors associated with longer operativetime in SILC procedure.Materials and Methods: A retrospective study was conducted of patients with benign gallbladder disease whounderwent SILC in Thammasat University Hospital between October 2014 and December 2020. Patients’ recordswere reviewed. Primary outcomes were preoperative predictive factors associated with DSLC. Secondary outcomeswere perioperative and 3-month postoperative adverse outcomes.Results: 592 SILC procedures were categorized as 80 DSLC and 512 non-difficult SILC (NDSLC). The median(interquartile range) of operative time in all SILC procedure is 48 (38, 62) minutes. The threshold of operative timeof difficult SILC was 72 minutes. The multivariate analysis indicated 5 significant predictive factors. Obesity (bodymass index > 25 kg/m2)) and abdominal pain reflected the difficulty of SILC procedures (p = 0.041 and p = 0.009).Calcified gallbladder showed the highest RR of 14.08 (p = 0.011). Contracted gallbladder and chronic cholecystitiswere also predictive factors with RR of 13.79 and 3.64, respectively (p < 0.001 and p = 0.007).Conclusion: Obesity, abdominal pain, chronic cholecystitis, contracted gallbladder and calcified gallbladder werepreoperative predictive factors. Surgeons should perform the SILC procedure carefully when predictive factors areidentified.