Biochemistry is an important and necessary subject in professional training in some undergraduate courses, but it is difficult to assimilate and can generate repulsion due to the lack of prior knowledge of Chemistry in elementary school. For this reason, a high level of dedication is required from educators for methodological and pedagogical practices that arouse curiosity and interest in students and improve the teaching process. In order to identify the limitations of this process, this study investigated the main subjects that generate difficulties in teaching and learning Biochemistry and collected videos published on YouTube on the different topics of Biochemistry. The greatest difficulty pointed out were “assimilation” (56.8%), “learning” (32.4%) and “density of the course program” (31.1%). Furthermore, “class notes” (71.6%) and “textbooks” (66.2%) are the materials that students most often use to study. Furthermore, the citric acid cycle (55.4%), electron transport chain (52.7%), bioenergetics (45.9%) and lipids (45.9%) are the topics that generate the most difficulty in understanding. On YouTube, explanatory videos in different languages were raised for all the content indicated in the survey. Thus, it was possible to identify limitations in the teaching and learning of Biochemistry, such as the deficiency acquired at the basic level in organic chemistry, the lack of laboratory structure that makes micro visualization difficult, making videos a facilitator in the classroom. However, the use of videos depends on the student's concentration on the digital medium, access to the Internet, equipment and teaching planning, and does not replace teacher clarifications.