Selective Analysis of Dopamine Receptor Antagonist LE300 and its N-Methyl Metabolite in Mouse Sera at the Trace Level by HPLC–Fluorescence Detection
Abstract A highly selective, sensitive, and reliable high-performance liquid chromatography (HPLC) method was developed and validated for the simultaneous determination of a novel type of dopamine receptor antagonist LE300 and its N-methyl metabolite in mouse sera. LE300, its N-methyl metabolite, and verapamil (an internal standard) were detected using excitation and emission wavelengths of 275 and 340 nm, respectively. HPLC analysis using a deproteinization procedure was performed by injecting an aliquot of the supernatant into the chromatographic system. Chromatographic separation was achieved on a reversed-phase Spherisorb Cyano (CN) column with a mobile phase consisting of acetonitrile:50 mM phosphate buffer pH 3.5 (70:30, v/v) pumped at a flow rate of 1.0 mL $ min^{−1} $. Regression analyses showed excellent linearity (r = 0.999) for concentrations of LE300 ranging from 4 to 500 ng $ mL^{−1} $ and for concentrations of its N-methyl metabolite of 6–600 ng $ mL^{−1} $. The HPLC-FLD method had limits of detection of 1.6 ng $ mL^{−1} $ for LE300 and 2.4 ng $ mL^{−1} $ for its N-methyl metabolite in mouse sera. The precision results, expressed as the intraday and interday relative standard deviation (RSD) values, ranged from 0.65 to 2.85 % (repeatability) and from 0.37 to 2.62 % (intermediate precision) for LE300 and its N-methyl metabolite, respectively; these values are in line with ICH guidelines. The assay was successfully applied in a pharmacokinetic study. The mean values of Tmax and Cmax were 2 h and 25.03 ± 5.60 ng $ mL^{−1} $ for LE300 and 3 h and 19.92 ± 2.88 ng $ mL^{−1} $ for its N-methyl metabolite, respectively..
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2015 |
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| Erschienen: |
2015 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:78 |
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| Enthalten in: |
Chromatographia - 78(2015), 9-10 vom: 21. März, Seite 655-661 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Hefnawy, Mohamed [VerfasserIn] |
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| Links: |
Volltext [lizenzpflichtig] |
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| Themen: |
Fluorescence detection |
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| RVK: |
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| Anmerkungen: |
© Springer-Verlag Berlin Heidelberg 2015 |
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| doi: |
10.1007/s10337-015-2879-x |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
OLC2044361647 |
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| 520 | |a Abstract A highly selective, sensitive, and reliable high-performance liquid chromatography (HPLC) method was developed and validated for the simultaneous determination of a novel type of dopamine receptor antagonist LE300 and its N-methyl metabolite in mouse sera. LE300, its N-methyl metabolite, and verapamil (an internal standard) were detected using excitation and emission wavelengths of 275 and 340 nm, respectively. HPLC analysis using a deproteinization procedure was performed by injecting an aliquot of the supernatant into the chromatographic system. Chromatographic separation was achieved on a reversed-phase Spherisorb Cyano (CN) column with a mobile phase consisting of acetonitrile:50 mM phosphate buffer pH 3.5 (70:30, v/v) pumped at a flow rate of 1.0 mL $ min^{−1} $. Regression analyses showed excellent linearity (r = 0.999) for concentrations of LE300 ranging from 4 to 500 ng $ mL^{−1} $ and for concentrations of its N-methyl metabolite of 6–600 ng $ mL^{−1} $. The HPLC-FLD method had limits of detection of 1.6 ng $ mL^{−1} $ for LE300 and 2.4 ng $ mL^{−1} $ for its N-methyl metabolite in mouse sera. The precision results, expressed as the intraday and interday relative standard deviation (RSD) values, ranged from 0.65 to 2.85 % (repeatability) and from 0.37 to 2.62 % (intermediate precision) for LE300 and its N-methyl metabolite, respectively; these values are in line with ICH guidelines. The assay was successfully applied in a pharmacokinetic study. The mean values of Tmax and Cmax were 2 h and 25.03 ± 5.60 ng $ mL^{−1} $ for LE300 and 3 h and 19.92 ± 2.88 ng $ mL^{−1} $ for its N-methyl metabolite, respectively. | ||
| 650 | 4 | |a HPLC | |
| 650 | 4 | |a Fluorescence detection | |
| 650 | 4 | |a LE300 | |
| 650 | 4 | |a Trace analysis | |
| 650 | 4 | |a Pharmacokinetics | |
| 700 | 1 | |a Alanazi, Amer |4 aut | |
| 700 | 1 | |a Abounassif, Mohammed |4 aut | |
| 700 | 1 | |a Mohammed, Mostafa |4 aut | |
| 700 | 1 | |a Al-Swaidan, Ibrahim |4 aut | |
| 700 | 1 | |a Attia, Sabry |4 aut | |
| 700 | 1 | |a Mostafa, Gamal |4 aut | |
| 700 | 1 | |a El-Subbagh, Hussein |4 aut | |
| 700 | 1 | |a Lehmann, Jochen |4 aut | |
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