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Acta Pharmaceutica

The Journal of Croatian Pharmaceutical Society

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Volume 61, Issue 4 (Dec 2011)


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

Mohammed Abonassif
  • Pharmaceutical Chemistry Department College of Pharmacy, King Saud University, P.O. Box 2457 Riyadh 11451, Saudi Arabia
/ Mohammed Hefnawy
  • Pharmaceutical Chemistry Department College of Pharmacy, King Saud University, P.O. Box 2457 Riyadh 11451, Saudi Arabia
/ Mohamed Kassem
  • Pharmaceutical Chemistry Department College of Pharmacy, King Saud University, P.O. Box 2457 Riyadh 11451, Saudi Arabia
/ Gamal Mostafa
  • Pharmaceutical Chemistry Department College of Pharmacy, King Saud University, P.O. Box 2457 Riyadh 11451, Saudi Arabia
Published Online: 2011-12-25 | DOI: https://doi.org/10.2478/V10007-011-0035-1

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

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.

Određivanje donepezil hidroklorida u humanoj plazmi i ljekovitim oblicima pomoću HPLC s detekcijom fluorescencije

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.

Keywords: donepezil hydrochloride; RP-HPLC; fluorescence detection; dosage form; plasma

Keywords: donepezil; RP-HPLC; dozirani oblik; plazma

  • H. Sugimoto, Y. Tsuchiya, H. Sungmi, K. Higurahi, N. Karibe, Y. Iimur, A. Sasaki, Y. Kawakami, T. Nakamura, S. Araki, Y. Yamanishi and K. Yamatssu, Novel piperidine derivatives. Synthesis and anti-acetylcholinesterase activity of 1-benzyl-4-[2-(N-benzoylamino)ethyl]piperidine derivatives, J. Med. Chem. 33 (1990) 1880-1887. [Crossref]

  • I. T. Lott, K. Osann, E. Dorn and L. Nelson, Down syndrome and Alzheimer disease: response to donepezil, Arch. Neurol. 59 (2002) 1133-1136.

  • S. Kafkala, S. Matthaiou, P. Alexaki, M. Abatzis, A. Bartzeliotis and M. Katsiabani, New gradient high-performance liquid chromatography method for determination of donepezil hydrochloride assay and impurities content in oral pharmaceutical formulation, J. Chromatogr. A 1189 (2008) 392-397; DOI: 10.1016/j.chroma.2007.12.015. [Crossref]

  • H. Pappa, R. Farrú, P. O. Vilanova, M. Palacios and M. T. Pizzorno, A new HPLC method to determine donepezil hydrochloride in tablets, J. Pharm. Biomed. Anal. 27 (2002) 177-182. [Crossref]

  • B. S. Sultry, S. Gananadhamu, D. Devadasu and K. Balaji, New RP-HPLC method for estimation of donepezil hydrochloride in pharmaceutical formulations, Int. J. Chem. Sci. 6 (2008) 1976-1983.

  • N. Y. Furukori, R. Furuya, T. Takahata and T. Tateishi, Determination of donepezil, an acetylcholinesterase inhibitor, in human plasma by high-performance liquid chromatography with ultraviolet absorbance detection, J. Chromatogr. B 768 (2002) 261-265.

  • M. A. Radwan, H. H. Abdine, B. T. Al-Quadeb, H. Y. Aboul-Enien and K. Nakashima, Stereoselective HPLC assay of donepezil enantiomers with UV detection and its application to pharmacokinetic in rats, J. Chromatogr. B. 830 (2006) 114-119; DOI: 10.1016/j.jchromb.2005.10.031. [Crossref]

  • J. S. Hiten, L. K. Mohan, P. Ankit, P. Shivkumar, S. Gunta, P. N. Chhagan, M. P. Dasharath and N. S. Bhanu, A rapid and specific approach for direct measurement of donepezil concentration in human plasma by LC-MS/MS employing solid-phase extraction, Biomed. Chromatogr. 23 (2009) 141-151, DOI: 10.1002/bmc.1095. [Crossref] [Web of Science]

  • P. E. Jeong, L. H. Won, J. H. Young, K. H. Yoon, L. M. Hyung, P. Eun-Seok, L. K. Choon and L. H. Suk, Hydrophilic interaction chromatography-tandem mass spectrometry of donepezil in human plasma: Application to a pharmacokinetic study of donepezil in volunteers, Arch. Pharm. Res. 31(2008) 1205-1211. [Web of Science]

  • Y. Hsin-Hua, Y. Yuan-Han, K. Ju-Yun and C. Su-Hwei, Sensitive analysis of donepezil in plasma by capillary electrophoresis combining on-column field-amplified sample stacking and its application of Alzheimer's disease, Electrophoresis 29 (2008) 3649-3657; DOI: 10.1002/elps.200800276. [Web of Science] [Crossref]

  • K. Nakashima, K. Itoh, M. Kono, M. N. Nakashima and M. Wada, Determination of donepezil hydrochloride in human and rat plasma, blood and brain microdialysates by HPLC with a short C30 column, J. Pharm. Biomed. Anal. 41 (2006) 201-206; DOI: 10.1016/j.jpba.2005.10.017. [Crossref]

  • G. M. Khaledi, Micelles as separation media in high-performance liquid chromatography and high-performance capillary electrophoresis: overview and perspective, J. Chromatog. A 780 (1997) 3-40.

  • V. Meyer, Practical High-Performance Liquid Chromatography, 3rd ed., Royal Society of Chemistry, London, 1998, pp. 14-16.

  • FDA Guidance for Industry: Bioanaytical Method Validation, US Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Biologics Evaluation and Research (CBER), Rockville (MD) 2001.

  • G. A. Shabir, Validation of high-performance liquid chromatography methods for pharmaceutical analysis, understanding the differences and similarities between validation requirements of the US Food and Drug Administration, the US Pharmacopeia and the International Conference on Harmonization, J. Chromatogr. A 987 (2003) 57-59; DOI: 10.1016/S0021-9673(02)01536-4. [Crossref]

  • International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use, ICH Harmonised Tripartite Guideline, Validation of Analytical Procedures: Text and Methodology Q2(R1), Complementary Guideline on Methododology dated 06 November 1996, incorporated in November 2005, London.

  • H. Ermer, Validation in pharmaceutical analysis. Part I. An integrated approach, J. Pharm. Biomed. Anal. 24 (2001) 755-767; DOI: 10.1016/S073-7085(00)00530-6. [Crossref]

  • J. N. Miller and J. C. Miller, Statistics and Chemometrics for Analytical Chemistry, 5th ed., Tottenham (UK), 2005.

  • Donepezil hydrochloride tablets, USP Pending Monograph Guideline, Version 2, USP Convention, Rockville (MD) 2009, USA http://www.usp.org

About the article

Published Online: 2011-12-25

Published in Print: 2011-12-01

Citation Information: Acta Pharmaceutica, ISSN (Online) 1846-9558, ISSN (Print) 1330-0075, DOI: https://doi.org/10.2478/V10007-011-0035-1. Export Citation

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