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In osteoporosis, total bone mass decreases and the thickness of the cortical layer diminishes in the shafts of the long bones. In this study, a simple ultrasonic in vivo method for determining the thickness of the cortical bone layer was applied, and the suitability of two different signal analysis techniques, i.e., envelope and cepstral methods, for measuring cortical thickness was compared. The values of cortical thickness, as determined with both techniques, showed high linear correlations (r ges 0.95) with the thickness values obtained from in vitro measurements with a caliper or in vivo measurements by peripheral quantitative CT (pQCT). No systematic errors that could be related to the cortical thickness were found. The in vivo accuracy of the measurements was 6.6% and 7.0% for the envelope and cepstral methods, respectively. Further, the in vivo precision for the envelope and cepstral methods was 0.26 mm and 0.28 mm, respectively. Although the results are similar for both of the techniques, the simplicity of the envelope method makes it more attractive for clinical applications. In conclusion, a simple ultrasound measurement provides an accurate estimate of the cortical bone thickness. The techniques investigated may have clinical potential for osteoporosis screening and therefore warrant more extensive clinical investigations with healthy and osteoporotic individuals.