Simultaneous determination of glycyl-L-histidyl-L-lysine and its metabolite, L-histidyl-L-lysine, in rat plasma by high-performance liquid chromatography with post-column derivatization.
A selective and sensitive high-performance liquid chromatographic (HPLC) method was developed for the determination of glycyl-L-histidyl-L-lysine (GHK), a liver-cell growth factor isolated from human plasma, and its metabolite, L-histidyl-L-lysine (HK), in rat plasma. Both high selectivity and sensitivity were achieved by the use of solid-phase extraction with a Bond-Elut Certify cartridge, ion-pair chromatography with 1-pentanesulfonate on a 5-microm Capcell Pak C18 UG120 column (250x4.6 mm I.D.) with a guard column, and by post-column derivatization with o-phthalaldehyde (OPA). GHK and HK were extracted from 0.1 ml of rat plasma after addition of o-phenanthroline to protect against degradation. The limit of detection for GHK and HK were 50 and 15 ng/ml, respectively, and the calibration curves were linear in the range 0.1-5.0 microg/ml. The developed method was applied to the pharmacokinetic study of GHK after a single dose was administered intravenously to rats. GHK was rapidly degraded to HK, which was eliminated rapidly.
Endo
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T
Pharmacological Laboratories, Kissei Pharmaceutical Co., Ltd., Minamiazumi, Nagano, Japan.
Miyagi
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M
Ujiie
A
A
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004151:Dipeptides / Q000097:blood*; D004355:Drug Stability; D006133:Growth Substances / Q000097:blood* / Q000493:pharmacokinetics; D008297:Male; D009842:Oligopeptides / Q000097:blood* / Q000493:pharmacokinetics; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D009764:o-Phthalaldehyde
D004151:Dipeptides; D006133:Growth Substances; D009842:Oligopeptides; C020905:glycyl-histidyl-lysine; D009764:o-Phthalaldehyde; C035809:glycyllysine