Purpose Midwives with high work stress are prone to suboptimal health status (SHS). However, there is a lack of unified measurement standard for SHS. Sub-Health Measurement Scale V1.0 (SHMS V1.0) is a multidimensional inventory that has been proved to have adequate psychometric properties in other populations. Therefore, this study aimed to evaluate validity and reliability of SHMS V1.0 in midwives.Methods In this cross-sectional study, 842 midwives from 46 midwifery institutions completed the SHMS V1.0. Internal consistency and test-retest reliability of SHMS V1.0 were assessed with Cronbach's alpha and Pearson correlation coefficient. Project analysis and confirmatory factor analysis (CFA) were performed to assess construct validity. Concurrent validity of SHMS V1.0 was evaluated by Pearson correlation analysis with the 14-item Fatigue Scale (FS-14).Results The total Cronbach's alpha and test-retest reliability coefficient of SHMS V1.0 was 0.908 and 0.804, respectively. The correlation coefficients between each item and its corresponding dimension, and those between each dimension and its corresponding subscale were all greater than 0.4. The CFA supported the structure of SHMS V1.0 with strong factor loadings and excellent fit indexes in the first-order and modified second-order factor model. The concurrent validity for SHMS V1.0 showed large correlation with FS-14, but social subscale showed medium correlations with total fatigue, physical fatigue and mental fatigue. Conclusions The SHMS V1.0 is a valid and reliable instrument for measuring SHS for midwives. Given the severity and harmfulness of SHS in midwives, SHMS V1.0 is recommended as an important tool for clinical practice and research.