Determination of donepezil hydrochloride in human plasma and pharmaceutical formulations by HPLC with fluorescence detection

• Published in: Acta pharmaceutica (Zagreb, Croatia) Vol. 61; no. 4; pp. 403 - 413
• Main Authors: Abonassif, Mohammed, Hefnawy, Mohammed, Kassem, Mohamed, Mostafa, Gamal
• Format: Journal Article
• Language: English
• Published: Croatia Versita 01.12.2011; De Gruyter Poland
• Subjects: Chemistry, Pharmaceutical - methods; Chromatography, High Pressure Liquid - methods; Chromatography, Liquid - methods; donepezil; donepezil hydrochloride; dosage form; dozirani oblik; fluorescence detection; Humans; Indans - blood; Indans - chemistry; Male; Micelles; Piperidines - blood; Piperidines - chemistry; plasma; plazma; RP-HPLC; Spectrometry, Fluorescence
• ISSN: 1330-0075; 1846-9558
• DOI: 10.2478/V10007-011-0035-1

A sensitive, isocratic reversed-phase high performance liquid chromatographic method involving fluorescence detection was developed for the determination of donepezil hydrochloride in tablets and in human plasma. Pindolol was used as an internal standard. Good chromatographic separation was achieved by using an analytical column C18. The system operated at room temperature using a mobile phase consisting of methanol, phosphate buffer (0.02 mol L-1) and triethyl amine (pH 3.5) (55: 45: 0.5, V/V/V) at a flow rate 0.9 mL-1 min. The analyte and internal standard were extracted from human plasma via liquid-liquid extraction. The proposed method was validated for sensitivity, selectivity, linearity, accuracy and precision. The calibration curve was linear over the range of 5-2000 ng mL-1 of donepezil with detection limit of 1.5 ng mL-1. Intra- and inter-day relative standard deviations were less than 2.5 %. The method was found to be suitable for quality control of donepezil hydrochloride in bulk drug as well as in human plasma. Ovaj rad opisuje HPLC metodu određivanja donepezil hidroklorida (DP) u tabletama i u ljudskoj plazmi u nano području. Postavljena je osjetljiva metoda izokratične HPLC s fluorescencijskom detekcijom. Kao unutarnji standard upotrebljen je pindolol. Dobro kromatografsko odjeljivanje postignuto je primjenom analitičke kolone C18. Radna temperatura bila je sobna, a kao mobilna faza upotrebljena je smjesa metanola, fosfatnog pufera (0,02 mol L-1) i trietilamina (pH 3,5) (55: 45: 0.5, V/V/V). Analit i unutarnji standard su ekstrahirani iz ljudske plazme ekstrakcijom tekuće-tekuće. Predložena metoda je validirana s obzirom na selektivnost, područje linearnosti, ispravnost i preciznost. Kalibracijska funkcija bila je linearna u području od 5-2000 ng mL-1 donepezila, a granica detekcije iznosila je 2 ng mL-1. Relativna standardna devijacija za repetabilnost i intermedijarnu preciznost bila je manja od 2,5 %. Metoda je primjenljliva u kontroli kvalitete ljekovitih formulacija s DP-om i u praćenju DP-a u ljudskoj plazmi.