Abstract
Reversed-phase high-performance liquid chromatography with radioactive flow detection was utilized to investigate the catabolism of thyrotropin-releasing hormone (TRH) in central nervous system (CNS) tissues. Two different column/gradient solvent systems were tested: (1) octadecylsilane (ODS) with an acetic acid-acetonitrile gradient and (2) poly (styrenedivinylbenzene) (PRP-1) with a trifluoroacetic acid-acetonitrile gradient. Both systems used 1-hexanesulfonic acid as the second ion-pairing reagent and yielded excellent separation of TRH and its catabolic products, TRH acid, cyclo (histidyl-proline), histidyl-proline, proline, and prolinamide, produced in CNS tissue homogenates. The PRP-1 column with a trifluoroacetic acid-acetonitrile solvent system produced a better and more reproducible separation of TRH catabolic products than the ODS column with the acetic acid-acetonitrile solvent system. This PRP-1 technique was utilized to demonstrate different rates and products of TRH catabolism in mouse and human spinal cord compared with cerebral cortex.
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