High Performance Liquid Chromatography with Peroxyoxalate Chemiluminescence Detection of Synthetic Peptide, Ebiratide

Ebiratide [L-methionyl-L-glutamyl-L-histidyl-L-phenyl alanyl-D-lysyl- N-(8-aminooctyl)- L- phenylalanin -amide-S, S-dioxide] is one of the synthetic analogues of ACTH4-9 for Alzheimer disease. High performance liquid chromatography with peroxyoxalate chemiluminescence detection (HPLC-PO-CL) has been...

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Bibliographic Details
Published inJournal of liquid chromatography & related technologies Vol. 20; no. 15; pp. 2377 - 2387
Main Authors Hamachi, Y., Nakashima, K., Akiyama, S.
Format Journal Article
LanguageEnglish
Published Colchester Taylor & Francis Group 01.09.1997
Taylor & Francis
Subjects
Analysis
Biological and medical sciences
General pharmacology
Medical sciences
Pharmacology. Drug treatments
Trace analysis
Solid phase extraction
Biological fluid
Fluorescence detector
Rat
Alzheimer disease
Rodentia
Peptide hormone
HPLC chromatography
Heptapeptide
Derivatization
Blood
Blood plasma
Direct method
Vertebrata
Mammalia
Sulfur peptide
Animal
Ebiratide
Quantitative analysis
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Summary:Ebiratide [L-methionyl-L-glutamyl-L-histidyl-L-phenyl alanyl-D-lysyl- N-(8-aminooctyl)- L- phenylalanin -amide-S, S-dioxide] is one of the synthetic analogues of ACTH4-9 for Alzheimer disease. High performance liquid chromatography with peroxyoxalate chemiluminescence detection (HPLC-PO-CL) has been examined for the analysis of this compound. The fluorogenic reaction product of ebiratide with DBD-F [4-(N,N-dimethylaminosulphonyl)-7-fluoro-2,1,3-benzoxadiazole] was detected by the chemiluminescence generated with bis(2,4,6-trichlorophenyl)oxalate (TCPO) or bis[4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl]oxalate (TDPO) in the presence of hydrogen peroxide. TDPO gave about five times higher CL intensity than TCPO using 100mM hydrogen peroxide. The detection limit of DBD derivative of ebiratide was 25 fmol on column with TDPO. The HPLC-PO-CL was applied to the determination of ebiratide extracted from rat plasma by the solid phase extraction with Bond Elute C18. The method gave a good linearity over the range from 0.25 to 10 pmol on column with the recovery of 85% for the spiked rat plasma and was able to estimate the time course of ebiratide at a sub-pmol/mL level.
ISSN:1082-6076
1520-572X
DOI:10.1080/10826079708002709