Background and Purpose: The revised European Working Group on Sarcopenia in Older People (EWGSOP2, version 2019) definition of sarcopenia differs with respect to the EWGSOP (version 2010) definition in applied criteria and their cut-off values. We aimed to investigate the impact of the new definition on sarcopenia prevalence in various populations of older adults. Methods: Eight cohorts, including community-dwelling older adults, geriatric outpatients and patients admitted to acute and subacute inpatient wards were assessed on sarcopenia prevalence. Results: A total of 2256 participants (56.4 % female) were included with a median age of the cohorts of 71.7–83.3 years. In males, sarcopenia prevalence was 31.9 % according to EWGSOP compared to 12.0 % according to EWGSOP2. In females, sarcopenia prevalence was 4.9 % and 6.1 % according to EWGSOP and EWGSOP2 respectively. Lower cut-off points for handgrip strength (27 kg versus 30 kg (males) and 16 kg versus 20 kg (females) for EWGSOP and EWGSOP2 respectively) resulted in the lower sarcopenia prevalence in males. Conclusions: According to the EWGSOP2 definition, the prevalence of sarcopenia in males is significantly lower compared to the EWGSOP definition, whereas the prevalence among women is slightly higher. The lower cut-off points for handgrip strength result in fewer adults being diagnosed with sarcopenia.