Pedagogical content knowledge (PCK) is an important target of science teacher knowledge assessment. Most studies that have assessed the PCK across a large sample of science teachers used a text‐based approach to elicit and assess the more declarative and static form of teachers' PCK. Recently, small‐scale qualitative studies have adopted a novel video‐based approach to characterize the more situated and dynamic form of PCK underpinning a teacher's in‐the‐moment pedagogical reasoning. In this relatively large‐scale mixed‐methods study, these two approaches to assessing PCK were compared. Specifically, a counterbalanced design was adopted to compare the test scores of 147 Grade 7 biology teachers in a video‐based PCK test with those of the same teachers in a text‐based PCK test with the same content. A subgroup of teachers (N = 30) with different levels of PCK was interviewed to further understand their reactions to the test and how these might have affected their test performance. The results of paired‐samples t tests showed that the teachers' PCK scores in the two types of test were significantly correlated, but their scores were significantly lower in the video‐based test. In terms of teacher reactions, although the teachers considered both types of test to be fair, they regarded the video‐based test as more authentic and more closely related to their job demands. The teachers reported putting more effort into the video‐based test and feeling more immersed and engaged, but also experiencing greater cognitive demand and a higher level of anxiety. A multiple regression analysis revealed that the cognitive demand perceived by the teachers significantly predicted their PCK scores in the video‐based test. The findings have implications for the valid design of video‐based PCK tests.