LR12-peptide quantitation in whole blood by RP-HPLC and intrinsic fluorescence detection: Validation and pharmacokinetic study
| Autor: | Sébastien Gibot, Lucie Jolly, Marianne Parent, Ariane Boudier, Mostafa Kouach, Pierre Leroy, Philippe Maincent, J.-F. Goossens, Igor Clarot, Marc Derive, Amir Boufenzer, Hafid Ait-Oufella |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Pharmacology Detection limit Bioanalysis Chromatography Chemistry 010401 analytical chemistry Clinical Biochemistry General Medicine Tandem mass spectrometry 01 natural sciences Biochemistry High-performance liquid chromatography 0104 chemical sciences Analytical Chemistry 03 medical and health sciences 030104 developmental biology Pharmacokinetics Drug Discovery Protein precipitation Solid phase extraction Molecular Biology Whole blood |
| Zdroj: | Biomedical Chromatography. 31:e3877 |
| ISSN: | 0269-3879 |
| Popis: | A simple, sensitive, selective and robust HPLC method based on intrinsic fluorescence detection was developed for the quantitation of a dodecapeptide (designated as LR12), inhibitor of Triggering Receptor Expressed on Myeloid cells-1 (TREM-1), in rat whole blood. Sample treatment was optimized using protein precipitation and solid phase extraction. Chromatographic separation was carried out in a gradient mode using a core-shell C18 column (150 × 4.6 mm, 3.6 µm) with mobile phases of acetonitrile and water containing trifluoroacetic acid at 1.0 mL/min. The method was validated using methodology described by the Food and Drugs Administration guidelines for bioanalytical methods. Linearity was demonstrated within the 50-500 ng/mL range and the lower limit of quantitation was 50 ng/mL. Finally, a preliminary pharmacokinetic study after intraperitoneal injection of LR12 in rats was conducted to evaluate both LR12 monomer and its corresponding disulfide dimer, the main product of degradation. Beyond the fact that this paper describes the first fully validated method for LR12 analysis in blood samples, the approach followed here to optimize pre-analytical steps could be beneficial to develop HPLC and/or MS methods for other pharmaceutical peptides. |
| Databáze: | OpenAIRE |
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