Add time:08/17/2019         Source:sciencedirect.com

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-μm Capcell Pak C18 UG120 column (250×4.6 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 μg/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.

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