Background: The association of sarcopenia with disability with ambulatory status is uncertain because most studies have targeted people who could walk independently. This study explored the prevalence of sarcopenia regardless of ambulatory status and the impact of ambulatory status on sarcopenia. Materials and Methods: In total, 778 consecutive patients, aged ≥65 years and admitted to a hospital, were enrolled. Ambulatory status was divided into 4 grades according to mobility as described in the Barthel index. Sarcopenia was defined as a loss of appendicular muscle mass index (AMI) with bioelectrical impedance and decreased muscle strength with handgrip strength (HGS); cutoff values were adopted from the Asian Working Group for Sarcopenia. Results: The mean patient age was 83.2 ± 8.3 years; 37.8% were male patients. Mobility limitation was associated with higher age, underweight body mass index, malnourishment, and comorbidities (all P < .001). AMI and HGS gradually decreased with declining ambulatory status (P < .001). The prevalence of sarcopenia in the independent walk, walk with help, wheelchair, and immobile groups was 57.9%, 76.1%, 89.4%, and 91.7%, respectively. AMI prevalence declined and sarcopenia drastically increased in patients who were unable to walk independently compared with those who could walk independently (P < .001). Multivariate regression analyses showed that mobility limitation was an independent indicator of decreasing AMI and sarcopenia after adjustment for confounders. Conclusion: Patients with dependent ambulatory status experienced a higher prevalence of sarcopenia compared with those with ambulation; in addition, decline in ambulatory status was an independent indicator for the presence of sarcopenia after adjustment for potential confounders.