Determination of Memantine in Plasma and Vitreous Humour by HPLC with Precolumn Derivatization and Fluorescence Detection

A new HPLC procedure with precolumn derivatization and rimantadine as the internal standard for determining memantine, a candidate agent for the treatment of glaucoma in plasma and vitreous humour, has been developed and validated. Precolumn derivatization was performed with 9-fluorenylmethyl-chloro...

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Bibliographic Details
Published in:Journal of chromatographic science 2011-11, Vol.49 (10), p.745-752
Main Authors: Puente, Belen, Garcia, Maria Angeles, Hernandez, Esther, Bregante, Miguel Angel, Perez, Susana, Pablo, Luis, Prieto, Esther
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Chromatography, High Pressure Liquid - methods
Drug Stability
Eye
Fluorenes - chemistry
Hexanes - chemistry
Hydrogen-Ion Concentration
Linear Models
Medical sciences
Memantine - analysis
Memantine - blood
Memantine - chemistry
Memantine - pharmacokinetics
Pharmacology. Drug treatments
Rabbits
Reproducibility of Results
Rimantadine - analysis
Rimantadine - blood
Sensitivity and Specificity
Spectrometry, Fluorescence - methods
Vitreous Body - chemistry
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Summary:A new HPLC procedure with precolumn derivatization and rimantadine as the internal standard for determining memantine, a candidate agent for the treatment of glaucoma in plasma and vitreous humour, has been developed and validated. Precolumn derivatization was performed with 9-fluorenylmethyl-chloroformate-chloride (FMOC-CI) as the derivatization reagent and followed by a liquid-liquid extraction with n-hexane. Optimal conditions for derivatization were an FMOC-CI concentration of 1.5 mM, a reaction time of 20 min, the temperature at 30°C, the borate buffer pH 8.5, and a borate buffer-acetonitrile ratio of 1:1. The derivatives were analyzed by isocratic HPLC with the fluorescence detector λex 260 nm λem 315 nm on a Novapack C18 reversed-phase column with a mobile phase of acetonitrile-water (73:27, v/v), 40°C, and a flow rate of 1.2 mL/min. The linear range was 10-1000 ng/mL with a quantification limit of ~ 10 ng/mL for both types of samples. This analytical method may be suitable for using in ocular availability studies.
ISSN:0021-9665
1945-239X
DOI:10.1093/chrsci/49.10.745