Varaprasad Bobbarala et al.

/International Journal of Chemical and Analytical Science 2010, 1(2),35-36

Research Article
Available online through
www.ijcas.info

RP-HPLC method development for the determination of assay of Donepezil hydrochloride

Useni Reddy Mallu1 K. Hussain Reddy 1, Varaprasad Bobbarala2*, Somasekhar Penumajji 3,
1Department of Chemistry, Sri Krishnadevaraya University, Anathapur-515003, A.P. India

*2Department of Botany, Andhra University, Visakhapatnam-530003, A.P. India.

3Vivimed labs Limited, 2nd, 4th Floor, Veeranag towers, Habsiguda, Hyderabad, A.P. India

Received on: 20-11-2009; Revised on: 16-12-2009; Accepted on:17-02-2010

ABSTRACT
RP-HPLC method was developed and validated by assessing linearity, accuracy, selectivity, limit of quantitation, and precision for the
determination of Donepezil hydrochloride. Donepezil was identified and quantitated on a C18 reversed phase column (3.9×150mm, 5.0µm),
using a mobile phase composed of acetate buffer-Acetonitrile (50:50v/v) delivered at a flow rate of 1.0mL/min, and with UV detection (λexcitation
= 230nm). The method was proven to be linear over a Donepezil hydrochloride concentration range of 12.5 to 75µg/mL with a mean correlation
coefficient of 0.9999.

Keywords: Donepezil hydrochloride, high performance liquid chromatography (HPLC), Acetonitrile, UV detection.

INTRODUCTION

Donepezil is a centrally acting reversible acetyl cholinest-
erase inhibitor1 its main therapeutic use is in the treatment of
Alzheimer’s disease, where it is used to increase cortical acetylcho-
line. Donepezil has been tested in other cognitive disorders including
Lewy body dementia 2 and vascular dementia 3 but it is not currently
approved for these indications. Donepezil has been found to improve
sleep apnea in Alzheimer’s patients 4. The studies found that speech
of austistic children who were mild to moderately affected appeared
to improve from the use of Donepezil5, 6 medication.
rations were performed using isocratic elution at ambient
temperature.The Waters HPLC system (Waters Corporation,
Milford, MA, USA) consisted of a model 2695 with UVdetector at
230nm and a computer running Waters Empower 3.2 software.The
mobile phase was composed of a mixture of buffer (1.0gm of Am-
monium acetate in to 1000ml of HPLC water, degas it with 0.45µ
filter) and Acetonitrile (50:50v/v) and degassed. A reversed phase
C18column (3.9x150mm, 5.0µ).The flow rate was set at 1.0mL/min
and the injection volume was 10µL.Donepezil hydrochloride stan-
dard solution was prepared to get known concentration of 50ppm
in diluent.Test solution was prepared by Weighing and crushing
of known quantity of 20 tablets accurately to get known concen-
tration of 50ppm in diluent.

Percentage (%) of Donepezil Hydrochloride=

At X WS X Avg. wt of tablet X P =
Figure 1: Structure of Donepezil Hydrochloride As X WT X Label Claim

Where in As is the Area of Donepezil Hydrochloride stan-

MATERIALS AND METHODS dard solution, At is the Area of Donepezil Hydrochloride test sample
solution, WS is the Donepezil Hydrochloride working standard weight
The chromatographic method was specifically developed for the taken for standard solution preparation in mg, WT is the Weight
determination of Donepezil hydrochloride. Chromatographic sepa-
taken for test solution preparation in mg and P is the Potency of
Donepezil Hydrochloride working standard.
*Corresponding author.
Dr. Varaprasad Bobbarala M.Sc., Ph.D. RESULTS AND DISCUSSION
# 36-92-248/23, Sreenivasa Nagar,
Kancharapalem, Visakhapatnam-530008,
Andhrapradesh, India. Representative chromatograms show that the peaks are well charac-
Tel.: + 91-9949129539 terized and meet acceptability criteria for peak separation.
E-mail:varaprasadphd@rediffmail.com

International Journal of Chemical and Analytical Science Vol.1.Issue 2.February 2010 35-36
 Varaprasad Bobbarala et al. /International Journal of Chemical and Analytical Science 2010, 1(2),35-36

Table.1: Linearity of Donepezil

0.16
Linearity Concentration Area

Donepezil-Std - 3.754
0.14 Level (ppm)
0.12

0.10
1 12.5 358269
2 25 796392
AU

0.08
3 37.5 1194635
0.06 4 50 1605070
0.04 5 62.5 2050335
0.02
6 75 2435453
0.00 Correlation co-efficient 0.9999
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00
Minutes

Donepezil Linearity

Chromatogram.1: Donepezil hydrochloride standard

Chromatogram.1 shows the retention time 3.7min corre-

sponds to Donepezil Hydrochloride. Chromatogram.2 represents the
system suitability of the method. Five replicated injections of the
sample shows that the area of Donepezil is consistent in all the injec-
tions and passes the system suitability (%RSD) of not more than 2%.
0.000
AU

-0.005

-0.010

Graph1: Linearity graph of Donepezil

3.754

0.10
AU

0.00
3.734

0.10 The system suitability of the method shows that the per-
AU

0.00 centage Relative standard deviation is less than 2% and the Linearity
over a concentration range of 12.5ppm to 75ppm with a correlation
3.752

0.10
AU

co-efficient of 0.9999 indicates that developed HPLC method is very

0.00
effective and it can be used for analysis of the pharmaceutical dosage
3.770

0.10
form of Donepezil Hydrochloride. Based on the System suitability
AU

0.00 and the Linearity conformances this method can be used for analysis.
3.746

0.10
AU

0.00
REFERENCE
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00
Minutes
1. Birks J, Harvey RJ, Donepezil for dementia due to Alzheimer’s dis-
ease. Cochrane Database Syst Rev 2006 (1): CD001190. doi:10.1002/
Chromatogram.2: System suitability 14651858.CD001190.pub2. PMID 16437430
2. Rojas-Fernandez CH, Successful use of donepezil for the treatment of
Chromatogram.3 shows that method is linear in different dementia with Lewy bodies. Ann Pharmacother, 2001, 35 (2): 202–
concentrations from 12.5ppm to 75ppm of Donepezil with a Correla- 205.
3. Malouf R, Birks J, Donepezil for vascular cognitive impairment.
tion coefficient of 0.9999. This result indicates that the method is Cochrane Database Syst Rev, 2004 (1): CD004395. doi:10.1002/
suitable for analysis. 14651858.CD004395.pub2. PMID 14974068.
0.24 4. Moraes W, Poyares D, Sukys-Claudino L, Guilleminault C, Tufik S,
Donepezil - 3.733

0.22 Donepezil improves obstructive sleep apnea in Alzheimer disease: a

0.20 double-blind, placebo-controlled study, Chest 2008, 133 (3): 677–83.
0.18 5. Alzheimer’s Drug Shows Promise as Treatment for Autism-Arehart-
0.16 Treichel, Psychiatric News (pn.psychiatryonline .org), 2001:11-16.
0.14 6. http://pn.psychiatryonline.org/cgi/content/full/36/22/16-a.
0.12 7. Michael G. Chez, Thomas M. Buchanan, Marina Becker, Jami Kessler,
AU

0.10
Mary C. Aimonovitch, Susan R. Mrazek Autism and Epilepsy,
0.08
Donepezil hydrochloride: a double-blind study in autistic children
0.06
Journal of Pediatric Neurology 2003; 1(2): 83-88.
0.04

0.02

0.00 Source of support: Nil, Conflict of interest: None Declared

-0.02
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00
Minutes

Chromatogram.3: Linearity
International Journal of Chemical and Analytical Science Vol.1.Issue 2.February 2010 35-36