Echo intensity determined by muscle ultrasound has been proposed as an efficient method for the assessment of muscle quality. The influence of changing ultrasound parameter settings on echo intensity values was assessed using a standardized approach. In this repeated measures cross-sectional study, sixteen repeated scans of rectus femoris, gracilis, and rectus abdominis were taken in 21 middle-aged persons with a portable Mindray M7 premium ultrasound machine equipped with a linear 5.0–10.0 MHz transducer. The settings of three parameters were fixed: gain, depth, and frequency. The settings of the following adjustable parameters were changed over their entire range: dynamic range, gray map, line density, persistence, and IClear. Repeated measures analyses were performed to evaluate the effect of changing the settings on echo intensity values. In all three muscles, dynamic range, gray map, and IClear correlated significantly (rrm-values ranging between −0.86 and 0.45) with echo intensity. In all three muscles, the echo intensity values differed significantly across the dynamic range (p < 0.013), gray map (p < 0.003), and IClear (p < 0.003). In middle-aged subjects, echo intensity values of lower limb and trunk muscles are significantly related to ultrasound parameters and significantly differ across their respective setting range. For the assessment of muscle quality through ultrasound, it is suggested to fix parameter settings within their midrange in order to minimize the effect of setting-dependent factors on EI values.