Determination of a cyclic heptapeptide, a novel fibrinogen receptor antagonist, in human plasma by high-performance liquid chromatography with automated pre-column derivatization, column switching and fluorescence detection.
A sensitive high-performance liquid chromatographic (HPLC) assay for the determination of the cyclic heptapeptide Ac-Cs-Asn-Dtc-Amf-Gly-Asp-Cys-OH (Dtc = beta,beta-dimethylthioproline, Amf = p-aminomethylphenylalanine) in human plasma has been developed. The key steps in the assay include: solid-phase extraction of the drug from plasma, chemical derivatization of the primary amino group with naphthalene-2,3-dicarboxyaldehyde in the presence of N-acetyl-D-penicillamine as a nucleophile to form a fluorescent benzo[f]isoindole derivative, and HPLC with column switching to provide the necessary chromatographic separation of the derivative from endogenous plasma components. The assay has been validated in the concentration range 1-10 ng/ml of plasma.
Kline
W F
WF
Merck Sharp & Dohme Research Laboratories, West Point, PA 19486.
Matuszewski
B K
BK
Hsieh
J Y
JY
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D009281:Naphthalenes; D010456:Peptides, Cyclic / Q000097:blood* / Q000302:isolation & purification / Q000494:pharmacology; D010975:Platelet Aggregation Inhibitors / Q000097:blood; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors*; D018000:Receptors, Peptide
D009281:Naphthalenes; D010456:Peptides, Cyclic; D010975:Platelet Aggregation Inhibitors; D011956:Receptors, Cell Surface; D018000:Receptors, Peptide; C048650:fibrin-fibrinogen degradation products receptor; C077476:cyclo(N-acetylcysteinyl-asparaginyl-dimethylthioprolyl-(4-aminomethyl)phenylalanyl-glycyl-aspartyl-cysteine); C027717:2,3-naphthalenedicarboxaldehyde