Side effects of amiodarone, an effective antiarrhythmic drug, include anemia, which may be caused by decreased formation or accelerated death of erythrocytes. Suicidal erythrocyte death (eryptosis) is characterized by cell shrinkage and cell membrane scrambling leading to phosphatidylserine exposure at the cell surface. Stimulators of erythrocyte membrane scrambling include increase of cytosolic Ca2+ concentration ([Ca2+]i) following activation of Ca2+-permeable cation channels. Moreover, eryptosis is triggered by ceramide. The present study has been performed to test for an effect of amiodarone on eryptosis. Erythrocytes from healthy volunteers were exposed to amiodarone and phosphatidylserine exposure (annexin V binding), cell volume (forward scatter), [Ca2+]i (Fluo3-dependent fluorescence), and ceramide formation (anti-ceramide-FITC antibody and radioactive labelling) determined by flow cytometry. Exposure of erythrocytes to amiodarone (1 microM) increased [Ca2+]i and triggered annexin V binding, but did not significantly decrease forward scatter and did not significantly influence ceramide formation. Amiodarone augmented the increase of annexin binding following hypertonic shock (addition of 550 mM sucrose) but did not significantly alter the enhanced annexin binding following Cl- removal (replacement with gluconate). Amiodarone did not significantly modify the decrease of forward scatter following hypertonic shock or Cl- removal. The present observations disclose a novel action of amiodarone which may contribute to the side effects of the drug.