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[Purpose] The characteristics of neck and shoulder pain (NSP) in different age populations have not been sufficiently examined. Therefore, the purpose of this study was to compare and verify the cervicothoracic spinal alignment and neck flexor muscle endurance of young and older adult females with and without NSP. [Participants and Methods] We assessed 72 female participants (39 young participants, 33 elderly participants, 43 NSP, 29 non-NSP) aged 18-82 years who were recruited for this study. Cervicothoracic spinal alignment measurements were obtained with forward head alignment (FHA) along with the upper thoracic angle. The neck flexor endurance test was performed. [Results] There were no significant age-by-group interactions for any of the assessment variables. However, the upper thoracic angle and neck flexor muscle endurance showed significant effects in the groups. Age also had significant effects on FHA and upper thoracic angle. [Conclusion] These results suggested that the neck flexor muscle endurance was more appropriate as an evaluation tool for older adult females with NSP. It was also suggested that the cervical flexor muscle endurance and upper thoracic angle were more appropriate as evaluation tools for young adult females with NSP.