Abstract
Intracellular water and electrolyte balance is regulated in large part by Na/K ATPase. Inhibition of this enzyme leads to cellular depolarization, a rise in extracellular K and intracellular Na, cellular swelling, and has been associated with shock, and failure to restore blood volume after hemorrhage. Bufadienolides are a strong inhibitor of Na/K ATPase and have recently been identified in plasma and urine of swine and man. To identify which organs synthesize bufadienolides, various organs from rat were homogenized. Bufadienolides were extracted using 1x volume ethyl acetate/ 4x volume acetonitrile, dried, and extracted by affinity chromatography (DigiFabTM, Fougera Pharmaceuticals INC). Extracts were run on Reverse Phase High Pressure Liquid Chromatography (Beckman Coulter). Chromatographic peaks were identified as Telocinobufagin, marinobufogenin, bufalin, and resibufogenin from testis, adrenal and brain by 1) co‐elution of standards, 2) UV spectrum and 3) mass spectroscopy of collected fractions. These data suggest that bufadienolides, potent inhibitors of Na/K ATPase, are synthesized in testis, adrenal and brain of rat. Supported by the ARMY MRMC and DMRDP.
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