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High‐performance liquid chromatographic determination of memantine hydrochloride in rat plasma using sensitive fluorometric derivatization
Author(s) -
Xie MeiFen,
Zhou Wei,
Tong XinYi,
Chen YiLe,
Cai Yi,
Li Yan,
Duan GengLi
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201000579
Subject(s) - chemistry , derivatization , chromatography , high performance liquid chromatography , memantine , detection limit , amantadine , hydrochloride , organic chemistry , nmda receptor , pharmacology , medicine , biochemistry , receptor
In this study, we investigated a simple, sensitive and reliable liquid chromatography‐fluorescence detection method for the determination of memantine hydrochloride in rat plasma which was based on derivatization with 9‐fluorenylmethyl chloroformate (FMOC‐Cl). For the first time, FMOC‐Cl was introduced into derivatization of memantine hydrochloride in rat plasma. The amino groups of memantine hydrochloride and amantadine hydrochloride (internal standard) were trapped with FMOC‐Cl to form memantine hydrochloride‐FMOC‐Cl and amantadine hydrochloride‐FMOC‐Cl compositions, which can be very compatible for LC‐FLD. Precipitation of plasma proteins by acetonitrile was followed by vortex mixing and centrifugation. Chromatographic separation was performed on a C 18 column (DIAMONSIL 150×4.6 mm, id 5 μm) with a mobile phase consisting of acetonitrile and water at a flow rate of 1.0 mL/min. The retention times of memantine hydrochloride‐FMOC‐Cl and amantadine hydrochloride‐FMOC‐Cl compositions were 23.69 and 40.27 min, respectively. Optimal conditions for the derivatization of memantine hydrochloride were also described. The limit of quantification (LOQ) was 25 ng/mL for memantine hydrochloride in plasma, the linear range was 0.025–5.0 μg/mL in plasma with a correlation coefficient ( r ) of 0.9999. The relative standard deviations (RSDs) of intra‐day and inter‐day assays were 4.46–12.19 and 5.23–11.50%, respectively. The validated method was successfully applied to the determination of memantine hydrochloride in rat plasma samples.