Minimum segment width (MSW) plays a fundamental role in the shaping of optimized apertures and creation of segments of varying sizes and shapes in complex radiotherapy treatment plans. The purpose of this work was to study the effect of MSW on dose distribution in patients planned with VMAT for various treatment sites using dose volume histogram (DVH) analysis. Materials and methods: For the validation of optimum MSW, 125 clinical treatment plans were evaluated. Five groups were identified (brain, head and neck, thorax, pelvis, and extremity), and five cases were chosen from each group. For each case, five plans were created with different MSW (0.5, 0.8, 1.0, 1.25, and 1.5 cm). The quality of treatment plans created using different MSW were compared using dosimetric indicators such as target coverage (D 98 -dose to 98% of the planning target volume (PTV), maximum dose (D 2 -maximum dose to 2% of the PTV), monitor units (MU), and DVH parameters related to organs at risk (OAR). The effect of the MSW on delivery accuracy was quantitatively analyzed using the measured fluence utilizing ionization chamber-based transmission detector and model-based dose verification system. Traditional global gamma analysis (2%, 2 mm) and dose volume information was gathered for the PTV and organs at risk and compared for different MSWs. Results: A total of 125 plans were created and compared across five groups. In terms of treatment plan quality, the plans using MSW of 0.5 cm was found to be superior in all groups. PTV coverage (D 98 ) decreased significantly (p < 0.05) as the MSW increased. Similarly, the maximum dose (D 2 ) was found to be increased significantly (p < 0.05) as the MSW increased from 0.5 cm, with MSW of 1.5 cm being the least in terms of plan quality for both PTVs and OARs.In terms of plan deliverability using DVH analysis,treatment planning system (TPS) compared to measured fluence, VMAT plans produced with MSW of 0.5 cm showed a better dosimetric index and a smaller deviation for both PTVs and OARs. The deliverability of the plans deteriorated as the MSW increased. Conclusion: Dose volume histogram (DVH) analysis demonstrated that treatment plans with minimal MSW showed better plan quality and deliverability and provided clinical relevance as compared to gamma index analysis.