The affinity of saxitoxin binding to cardiac sarcolemmal and cytosolic fractions was examined across species. In amphibia (frog) the plasma membrane site demonstrated a high affinity (Kd approx. 5 X 10(-9) M) but the majority of the total sites in the homogenate appeared to be high affinity soluble sites (Kd approx. 2 X 10(-9) M). Chicken and turtle cardiac plasma membrane fractions bound [3H]saxitoxin with 500-fold less affinity (Kd values of approx. 2 X 10(-6) M). No binding was seen in the cytosol. The affinity of cardiac sarcolemmal binding in amphibians correlates quantitatively with the K0.5 for the inhibition of sodium currents. Physiological correlation of the low affinity saxitoxin sites in chicken and turtle with toxin concentrations necessary to inhibit the sodium current remains unclear. The hypothesis that frog cytosolic saxitoxin binding sites originated from sarcolemma during homogenization is examined. The presence of three types of saxitoxin binding sites in cardiac preparations supports the existence of sodium channel subtypes.