Objectives: The main objective of the current study is to examine the items using the item analysis to assess their competency of knowledge in human anatomy. Methods: This is a quantitative descriptive cross-sectional study conducted at the Najran University faculty of Medicine, in the department of anatomy. A 31-second year MBBS students of level three took the multiple-choice question exam comprising 100 questions in one session. Each item was analyzed by item analysis. The planned period of data collection is the first semester of the academic year 2019-2020. The data were analyzed using SPSS version 25, and Excel Software. All analyses were performed using the descriptive frequency, explore and correlation probabilities a p-value, p<0.01 (2-sided) was considered to indicate statistical significance. Results: The assessment of test items use the item analysis that is very difficult question: 25; 25%, difficult question: 8; 8%, average question: 46; 46%, easy questions; 9; 9% and very easy: 9; 9%. The discrimination indices, which are poor discrimination; 34; 34%, acceptable discrimination; 11; 11%,good discrimination; 6; 6%, very good discrimination; 4; 4% and excellent discrimination; 45; 45%. The performance of students in this test showing that a large number of items have a role in distinguishing students knowledge in the anatomy test, but in Simple parts of the scale that the items did not succeed in assessing students. Items of anatomy exams displayed a strong relationship of difficulty (P) and discrimination (D) indices. Conclusion: The MCQs items assess their competency of knowledge in human anatomy. The competence of students in the knowledge of anatomy measured at a high rate of A-type context-rich MCQs can be stem is case scenario, lead-in; question and options; distractors, and key. These items can differentiate good and poor accomplishment students..