Background: Vitamin B12 is a necessary in the methylation response, which results in the formation of myelin. A lack of vitamin B12 may result in demyelination of nerve cells in the cochlear nerve, resulting in hearing loss and tinnitus. Aim of this work: To analyze the effect of Vitamin B12 deficiency on auditory brain stem response and otoacoustic emission in patients suffering from tinnitus. Patients and Methods: This was a case-control study with 6 male (20.0%) and 24 female (80.0%) with tinnitus as cases (30 participants) and 7 male (23.3%) and 23 female (76.6%) as controls (30 participants). Pure tone audiometry, Auditory Brain Stem Response, Transient evoked otoacoustic emissions ,Detection of the serum levels of Vitamin B12 and Conventional MRI brain were performed . Results: There was a statistically significant delay at wave III latency in cases than the controls in the right side while in the left side, wave I and III latencies were statistically significant prolonged in cases than the controls..There was no statistically significant correlation between vitamin B12 level and the latency or amplitude of auditory brainstem responses. Conclusion:Our findings imply that auditory brain stem response results in patients with tinnitus linked with vitamin B12 deficiency can be impacted.