Internal stress in structural steel members is an important parameter for steel structures in their design, construction, and service stages. However, it is hard to measure via traditional approaches. Among the existing non-destructive testing (NDT) methods, the ultrasonic method has received the most research attention. Longitudinal critically refracted (Lcr) waves, which propagate parallel to the surface of the material within an effective depth, have shown great potential as an effective stress measurement approach. This paper presents a systematic non-destructive evaluation method to determine the internal stress in in-service structural steel members using Lcr waves. Based on theory of acoustoelasticity, a stress evaluation formula is derived. Factor of stress to acoustic time difference is used to describe the relationship between stress and measurable acoustic results. A testing facility is developed and used to demonstrate the performance of the proposed method. Two steel members are measured by using the proposed method and the traditional strain gauge method for verification. Parametric studies are performed on three steel members and the aluminum plate to investigate the factors that influence the testing results. The results show that the proposed method is effective and accurate for determining stress in in-service structural steel members.