Rapid separation of tritiated thyrotropin-releasing hormone and its catabolic products from mouse and human central nervous system tissues by high-performance liquid chromatography with radioactive flow detection.
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.
Turner
J G
JG
Neurology and Research Services, William S. Middleton Memorial VA Hospital, Madison, WI 53705.
Schwartz
T M
TM
Brooks
B R
BR
D000818:Animals; D002490:Central Nervous System / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003500:Cyclization; D004151:Dipeptides / Q000378:metabolism; D006801:Humans; D051379:Mice; D010456:Peptides, Cyclic / Q000378:metabolism; D010879:Piperazines / Q000378:metabolism; D013045:Species Specificity; D013973:Thyrotropin-Releasing Hormone / Q000302:isolation & purification*
D004151:Dipeptides; D010456:Peptides, Cyclic; D010879:Piperazines; C035699:histidylproline; D013973:Thyrotropin-Releasing Hormone; C015924:histidyl-proline diketopiperazine