High-performance liquid chromatographic determination of memantine hydrochloride in rat plasma using sensitive fluorometric derivatization.
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.
Department of Pharmaceutical Analysis, School of Pharmacy, Fudan University, Shanghai, PR China.
Mei-Fen
MF
Xie
Wei
W
Zhou
Xin-Yi
XY
Tong
Yi-Le
YL
Chen
Yi
Y
Cai
Yan
Y
Li
Geng-Li
GL
Duan
D000544:Alzheimer Disease / Q000097:blood; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015259:Dopamine Agents / Q000097:blood* / Q000737:chemistry; D005449:Fluorenes / Q000737:chemistry; D006801:Humans; D008559:Memantine / Q000097:blood* / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar
D015259:Dopamine Agents; D005449:Fluorenes; C054007:1-(9-fluorenyl)methyl chloroformate; D008559:Memantine