Background:Most of the academic teachers use four or five options per item of multiple choice question (MCQ) test as formative and summative assessment. Optimal number of options in MCQ item is a matter of considerable debate among academic teachers of various educational fields. There is a scarcity of the published literature regarding the optimum number of option in each item of MCQ in the field of medical education.Objectives:To compare three options, four options, and five options MCQs test for the quality parameters – reliability, validity, item analysis, distracter analysis, and time analysis.Materials and Methods:Participants were 3rd semester M.B.B.S. students. Students were divided randomly into three groups. Each group was given one set of MCQ test out of three options, four options, and five option randomly. Following the marking of the multiple choice tests, the participants’ option selections were analyzed and comparisons were conducted of the mean marks, mean time, validity, reliability and facility value, discrimination index, point biserial value, distracter analysis of three different option formats.Results:Students score more (P = 0.000) and took less time (P = 0.009) for the completion of three options as compared to four options and five options groups. Facility value was more (P = 0.004) in three options group as compared to four and five options groups. There was no significant difference between three groups for the validity, reliability, and item discrimination. Nonfunctioning distracters were more in the four and five options group as compared to three option group.Conclusion:Assessment based on three option MCQs is can be preferred over four option and five option MCQs.