Background: Medical documentation is an essential component of health care practice that has a Potential to improve patient care outcomes. Various studies have shown that documentation is still a critical issue in both high- and low-income countries, especially in Sub-Saharan Africa like Ethiopia. However, the status of medical documentation practice and its challenges are not well known by research in the study area. Objective: The study aimed to assess medical documentation practice and associated factors among health professionals in Tibebe Ghion specialized hospitals, Northwest Ethiopia,2022. Methods: An institutional based explanatory sequential mixed study was conducted from November 20 to December 30, 2022/2023G.C. Stratified sampling technique was used to select 422 study participants. Bi variable and multivariable logistic regression analyses were performed using SPSS version 25. In-depth interviews were conducted until saturation point using semi-structured questionnaires. Purposive sampling technique was used to enroll participants for qualitative part. Qualitative Respondent’s response was analyzed using Open Code version 4.03 thematically. Results: The mean age of the respondents was 29.7 (SD =± 2.88) years with the majority of the age group were from 26 to 30 years which was 304(73.1%). The overall medical documentation practice score was 49.5% (95% CI:25.4-26.3). Age belongs to 21-25 (AOR=.103, CI: .012,.870) ,sex(male) of participants (AOR=1.497, CI:.967,2.317), working unit belongs to inpatient (AOR=1.534, CI:919,2.562), familiarity with medical documentation standards (AOR=3.050, CI:1.951,4.766), feedback on documentation (AOR=1.961, CI:1.070,3.594) and knowledge (AOR=2.518, CI:.1.630,3.889) were factors significantly associated with medical documentation practice. The in-depth interview revealed that shortage of standards, training, lack of supervision with auditing, shortage of staff(workload), lack of computerized system, non-conducive environment, and poor attention for documentation by HPs were identified as factors affecting medical documentation practice. Conclusions and Recommendations: The overall medical documentation practice score was poor. Age of participants, being male of participants, working setting (inpatient department), familiarity with standard sheet, feedback on documentation and being knowledgeable on documentation were significantly associated with medical documentation practice. So, Hospital managers better to put further effort toward improving training, accessing standards, give supportive supervision and feedback, improving clarity of registration, making the documentation computerized, and give attention to medical documentation. Keywords: Medical documentation, Documentation practice, factors, Health professionals, Tibebe Ghion Specialized Hospital, Ethiopia.