Determination of [d-Ala2, d-Leu5]enkephalin and the metabolites containing C-terminal d-leucine by high-performance liquid chromatography |
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| Affiliation: | 1. Geology Department, Salamanca University, Pza/Caídos, s/n, Salamanca, Spain;2. Geological Institute, ETH Zürich, Sonneggstr. 5, 8092, Zuerich, Switzerland;3. Institute of Geosciences, University of Kiel, Olshausenstr. 40, 24098, Kiel, Germany;4. Laboratory of Ion Beam Physics, ETHZ, Otto-Stern Weg 5, HPK, 8093, Zurich, Switzerland;1. College of Electrical Engineering, Zhejiang University, Hangzhou, China;2. Economy Research Institute of State Grid Zhejiang Electric Power Company, Hangzhou, China |
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| Abstract: | A high-performance liquid chromatographic procedure has been developed for the determination of [d-Ala2, d-Leu5]enkephalin (DADLE) and the fragments containing d-leucine in rat blood. The procedure was applied to the determination of blood levels of [3H-d-Leu5]DADLE and the C-terminal fragments after intravenous administration of [3H-d-Leu5]DADLE to a rat. Unlabelled DADLE and the C-terminal fragments were spiked as carriers to rat blood samples and the blood samples were extracted with 1% trifluoroacetic acid in methanol. The recoveries from rat blood were quantitative for all compounds. DADLE and the C-terminal four fragments were well separated on a reversed-phase column with gradient elution using a mobile phase composed of 0.14% HClO4 and acetonitrile. |
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