Determination of [D-Ala2,D-Leu5]enkephalin and the metabolites containing C-terminal D-leucine by high-performance liquid chromatography.
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.
Hasegawa
H
H
School of Pharmacy, Tokyo University of Pharmacy and Life Science, Japan.
Nakamura
A
A
Shinohara
Y
Y
Baba
S
S
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016308:Enkephalin, Leucine-2-Alanine / Q000008:administration & dosage / Q000097:blood*; D007930:Leucine / Q000097:blood; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D016308:Enkephalin, Leucine-2-Alanine; D007930:Leucine