Autonomic nervous system (ANS) specificity of emotion remains controversial in contemporary emotion research, and has received mixed support over decades of investigation. This study was designed to replicate and extend psychophysiological research, which has used multivariate pattern classification analysis (PCA) in support of ANS specificity. Forty-nine undergraduates (27 women) listened to emotion-inducing music and viewed affective films while a montage of ANS variables, including heart rate variability indices, peripheral vascular activity, systolic time intervals, and electrodermal activity, were recorded. Evidence for ANS discrimination of emotion was found via PCA with 44.6% of overall observations correctly classified into the predicted emotion conditions, using ANS variables (z=16.05, p<.001). Cluster analysis of these data indicated a lack of distinct clusters, which suggests that ANS responses to the stimuli were nomothetic and stimulus-specific rather than idiosyncratic and individual-specific. Collectively these results further confirm and extend support for the notion that basic emotions have distinct ANS signatures.