Despite longstanding recognition that the adrenal medulla is a major source of catecholamines and opioid peptides, the dynamic co-release of these signaling molecules—and the full complexity of their molecular forms—remains poorly understood. This study addresses this gap by integrating real-time electrochemical detection with tandem mass spectrometry to map and quantify the neurochemical complexity of opioid peptide signaling from adrenal chromaffin cells. Using FSCV, we provide evidence for stimulus-evoked co-release of met-enkephalin and catecholamines from single chromaffin cells (and adrenal slices?). With MS, we show that proenkephalin-derived peptides dominate the adrenal peptidome and that extended enkephalin peptides—such as BAM-18 and BAM-22—are not only present in cell content but are actively released.