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Validation of UV spectrophotometric method for determination of atenolol

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Research Article

Validation of UV Spectrophotometric Method for

Determination of Atenolol
SHOAEB SYED ∗, MUBASHIR MOHAMMED
Research Scholor Y B Chavan College of Pharmacy, Dr. Rafiq Zakaria Campus Rauza bagh, Aurangabad 431 001, India.

ABSTRACT
A rapid and sensitive UV spectrophotometric method for routine evaluation of atenolol in tablets was developed
and validated as per ICH guidelines. UV spectrophotometric method was performed at 226.6 nm, and samples
were prepared with a solution of phosphate buffer pH 6.8. The linearity demonstrated a correlation coefficient
of 0.9988, various validation parameters like accuracy, precision, LOD, LOQ, recovery study, range were
determined The proposed method was simple, rapid, precise, accurate and sensitive, and can be used for the
routine analysis of atenolol.

Key word: Atenolol, UV spectrophotometry, validation.

INTRODUCTION official methods and with other methods published. The

UV spectrophotometric method is simpler than the oth-
Atenolol, designated chemically as (RS)-4-(2hidroxy-3- ers studied because it does not need derivative and
isopropylaminopropoxy) phenylacetamide, is commer- chemometric assistance. Moreover, this method can be
cially available as a racemic mixture (Fig. 1), it is found used in dissolution studies because it uses its own dis-
in the form of tablets, oral solution, and sterile solution solution medium as diluent.
for injectable. Atenolol (ATN) is a β1 selective (cardiose-
lective) β-adrenergic receptor-blocking agent without
membrane-stabilizing or intrinsic sympathomimetic
(partial agonist) activities [1]. ATN is also used to treat
myocardial infarction (heart attack), arrhythmias
(rhythm disorders), angina (chest pains), and disorders
arising from decreased circulation and vascular con-
striction, including migraine [2] In the European Phar-
macopoeia [3], the described method for atenolol quan- Figure 1: Structure of atenolol
tification uses technique of titration with perchloric acid.
In the British Pharmacopoeia [4] the indicated methods MATERIALS AND METHODS
use spectrophotometric in the ultraviolet at 275 nm. The
UV spectrophotometric method is very simple, rapid, Instrumentation
economical, and it allows the determination in pharma- An UV-Visible double beam spectrophotometer (Varian
ceuticals with enough reliability. For the UV spectropho- Cary 100) with 10 mm matched quartz cells was used
tometric method, the survey of literature revealed very for Spectrophotometric method. All weighing were done
complex methods, using bands of the visible which on electronic balance (Model Shimadzu AUW-220D).
have used complexometry [5], derivative or chemomet- Spectroscopic grade methanol was used throughout the
ric assistance and interpolation on the calibration curve study. Ultrasonicator (Model 5.5 150H) was used for
[5]. The aim of this work was the development and fully sample solution preparation.
validation of a new UV spectrophotometric method,
which can be more economical and simpler than the Reagents and chemicals

∗Address for correspondance

Shoaeb Syed, Research Scholor Y B Chavan College of Pharmacy, Dr. Rafiq Zakaria Campus Rauza bagh, Au-
rangabad 431 001, India.
E-mail address: shoaibmohammad87@yahoo.com, Phone: +91-9730572605
Received: 12/11/2013, Revised: 30/11/2013, Accepted: 10/12/2013

International Journal of Pharmaceutical Research | January-March 2014 | Volume 6 | Issue 1 | 25

 Syed & Mohammed / Validation of UV spectrophotometric method for determination of atenolol
Pure drug sample of atenolol was kindly supplied asa
gift sample by WOCKHARDT LIMITED Waluj Aurangabad.
Tablet formulation manufactured by Zydus cadilla In-
dustries was purchased from local market containing
ATEN 50 Containing atenolol 50 mg per tablet.
Preparation of standard stock solution
Standard drug solution of Atenolol was prepared by dis-
solving 100 mg in phosphate buffer pH 6.8 and the vol-
ume was made up to 100 ml to obtain stock solution of
1000 µg/ml concentration. Ultrasonication was done to
obtain a clear solution.
Determination of analytical wavelength
From the standard stock solution, 0.1 ml was pipette
out into 10 ml volumetric flask. The volume was made
up to 10 ml with phosphate buffer solution of pH 6.8.
The resulting solution containing 10µg/ml was scanned
between 200 and 400 nm.
Preparation of Calibration curve of Atenolol
in PBS pH 6.8
Aliquots of 0.1 to 0.6 ml portion of stock solutions were
transferred to a series of 10 ml volumetric flasks, and
volume made up to the mark with phosphate buffer pH
6.8. The serial dilution of the range of 10, 20, 30, 40,
50, and 60 µg/ml was prepared. The absorbance was
measured at λmax 226.6 nm.
UV Method Validation
Linearity and range
The linearity of the response of the drug was verified at
10 to 60 µg/ml concentrations. The calibration graphs
were obtained by plotting the absorbance versus the
concentration data and were treated by linear regres-
sion analysis. The equation of the calibration curve for
atenolol obtained. The linearity range of calibration
curve and correlation coefficient (r2) of determination
was obtained.
Precision
The accuracy of the method was determined by recov-
ery experiments. Each solution was recorded in tripli-
cate and the percentage recovery was calculated. The
precision of the method was demonstrated by intra-day
and inter-day variation studies. In intra-day studies,
three repeated measurements of standard and sample
solutions were made in a day and percentage RSD were
calculated. In inter-day studies, three repeated meas-
urements of standard and sample solutions were made
on three consecutive days and percentage RSD were
calculated.
Limit of detection (LOD) and Limit of quan-
tification (LOQ):
LOD and LOQ were calculated by the equations;
LOD = 3.3σ / S and LOQ = 10σ / S
26 | International Journal of Pharmaceutical Research | January-March 2014 | Volume 6 | Issue 1
Where, S is slope of the calibration curve and σ is the
residual standard deviation
Recovery Study
Accuracy of the method was studied by recovery exper-
iments. The recovery was performed at three levels,
80,100 and 120% of atenolol standard concentration.
The recovery samples were prepared in a before men-
tioned procedure. Three samples were prepared for
each recovery level. The solutions were then analyzed
and the percentage recoveries were calculated from the
calibration curve.
RESULTS AND DISCUSSION
Precision
Precision of method was evaluated for atenolol. The
reproducibility (inter-day precision) of the method and
repeatability (intra-day) was evaluated in the same la-
boratory. The relative standard deviation (RSD) values
obtained were 0.1973 and 0.2569 respectively (Table
1).
Table 1: Precision determination
Intra-Day Inter-Day
Sample no.
Precision* Precision*
1 99.98 98.71
2 99.85 98.52
3 99.65 98.65
Mean ± SD 99.68 ± 0.0026 98.62 ± 0.0020
%RSD 0.2569 0.1973
*mean values of three determination
Accuracy (Recovery Study)
Accuracy of the method was studied by recovery exper-
iments. The recovery was performed at three levels,
80,100 and 120% of atenolol standard concentration.
Three samples were prepared for each recovery level.
The solutions were then analyzed and the percentage
recoveries were calculated from the calibration curve.
The recovery value for atenolol is 99.21±0.275 and RSD
is 0.321which is less than 2, which shows that the
method has good reproducibility (Table 1).
Limit of detection (LOD) and Limit of quan-
tification (LOQ)
Limit of detection and Limit of quantification was found
to be 0.2586 and 0.7788 respectively.
Specificity
Specificity is the ability of the method to accurately
measure the analyte response in the presence of all
potential sample components (excipients). The results
were compared with the analysis of a standard atenolol
 Syed & Mohammed / Validation of UV spectrophotometric method for determination of atenolol

and tablet formulations. Excipients of the solid dosage

form did not interfere with the analyte (Table 2). ACKNOWLEDGEMENT
CONCLUSION We are thankful to Padmashree Mrs Fatema Rafiq Za-
karia for providing infrastructure and facilities for carry-
UV method was validated for atenolol as per ICH guide- ing out the work. We are also thankful to WOCKHARDT
lines. The proposed UV spectrophotometric method is LIMITED Waluj Aurangabad for providing atenolol as gift
simple, sensitive, accurate, precise and cost effective sample.
and hence can be used for the routine analysis of
atenolol in bulk, tablet formulations and for dissolution REFERNCES
studies.
1. The Indian Pharmacopoeia Commission. Indian
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Absorption maxima
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Standard regression
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Figure 3: Calibration curve of Atenolol

International Journal of Pharmaceutical Research | January-March 2014 | Volume 6 | Issue 1 | 27

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