In this study, we have successfully established a high-performance thin-layer chromatography (HPTLC) method for the quality assessment of Actinidiae Fructus Vermicultus, known as Mokcheonryo(ja) in Korea. This is the dried vermiculate fruit of Actinidia polygama and A. kolomikta, as stipulated by the Korean Herbal Pharmacopoeia (KHP). However, the Korean herbal market often witnesses the inclusion and distribution of ‘Mihudo’, an alternative herbal product sourced from the dried fruits of A. arguta, belonging to the same botanical genus. This confluence has raised substantial apprehensions concerning the veracity of quality. In response to this concern, we have meticulously developed an HPTLC analytical methodology capable of differentiation between Mokcheonryo and Mihudo by exploiting their distinct chemical profiles. We identified umbelliferone as a key marker compound for Mokcheonryo and quantified the content of umbelliferone in each sample using a TLC scanner. Throughout this study, we confirmed distinct fingerprints for Mokcheonryo and Mihudo, providing a reliable means to differentiate between these two herbal medicines. Furthermore, the presence of umbelliferone in Mokcheonryo serves as an indicator compound for quality assessment. The proposed HPTLC method offers a practical and effective tool for ensuring the quality and authenticity of Mokcheonryo in the herbal market.