Background: The reproducibility of left ventricular (LV) mass measurement by two-dimensional (2-D) echocardiography is inadequate for individual assessments. Hypothesis: This study was undertaken to evaluate the potential of LV mass determination with a new three-dimensional (3-D) echocardiographic method compared with 2-D measurements. Methods: Pot-cine agarose-filled left ventricles (n = 15, true mass 61-511 g) of different shapes were measured by a multiplane 3-D method based on 90 images acquired by probe rotation axis (1) perpendicular and (2) parallel to the ventricular long axis [''parasternal'' (the left sternal border was not present as a reference point in this study) and apical views]. Mass was also obtained using (3) the biplane truncated ellipsoid and (4) area-length methods, as well as (5) the modified cube formula. Five hearts were not analyzed with the apical 3-D technique because of insufficient image quality. Results: Systematic deviation from true mass was small with all methods (<5.3%). Accuracy, expressed as 1 standard deviation of individual estimates around this systematic bias, was 7.7, 13.6, 8.2, 11.9, and 11.9% of true mass for the methods 1-5, respectively. Interobserver reproducibility, expressed as the coefficient of variation, was 4.7, 8.8, 8.1, 8.9, and 9.4% for the same methods. Conclusion: Limits fur individual accuracy and reproducibility of LV mass estimates are nearly doubled using apical compared with ''parasternal'' 3-D echocardiography in vitro. A main advantage of ''parasternal'' 3-D compared with 2-D LV mass estimates is better reproducibility, but at the expense of greater time consumption. Apical 3-D technique is not superior to simpler 2-D methods based on ''parasternal'' short axis imaging.