Validated RP-HPLC Method for the Quantitation of Candesartan in Bulk and Pharmaceutical Dosage Forms.

 

Syeda Kulsum1, G. Vidya Sagar2 and M. Padmalatha1

1Department of Pharmaceutical Analysis, Vijaya College of Pharmacy, Munaganoor (V), Hyathnagar (M), Hyderabad, India-501505.                2KSKV Kachchh University, Bhuj, Kutch, Gujarat, India.

*Corresponding Author E-mail: syedakulsum@gmail.com

 

ABSTRACT:

A simple, specific, accurate, precise and sensitive Reverse Phase High Performance Liquid Chromatographic method has been developed for the quantitation of candesartan in both pure and pharmaceutical dosage forms. An Inertsil ODS-3V C-18, 5 um column having 250 x 4.6 mm internal diameter in isocratic mode with mobile phase containing ortho phosphoric acid: Acetonitrile (20:80v/v) and adjust the pH to 3.2 by using 0.03M potassium hydrogen phosphate buffer. The flow rate was 1.0 ml / min and the effluents were monitored at 210 nm. The retention time was 8.057min. The linearity was in the range of 50-160g / ml. This method was validated for linearity, precision, limit of detection, limit of quantitation and accuracy. Statistical analysis proves that the method is reproducible and selective for the estimation of the said drug.

 

KEYWORDS: RP-HPLC, candesartan, Validation,mobile phase,inertsil column.

 

 


 

INTRODUCTION:

Candesartan , a angiotensin-receptor blocker (ARB), is used alone or with other antihypertensive agents to treat hypertension. Candesartan competes with angiotensin II for binding at the AT1 receptor subtype.(1,2)

 

The chemical name of Candesartan cilexetil is((±)-1-hydroxyethyl-2-ethoxy- 1-[p-(o-1H-tetrazol-5-ylphenyl)benzyl]-7-benzimidazolecarboxylate cyclohe-xyl carbonate(fig.1). Candesartan cilexetil is metabolised completely by esterases in the intestinal wall during absorption to the active candesartan moieity.[3,4] Several analytical methods that have been reported for the estimation of candesartan in biological fluids or pharmaceutical formulations which includes spectrofluorimetry,[5] derivative spectrophotometry,[6] Q- Analysis spectrophotometry,[7] and HPTLC[8] methods. The objective of the work was to develop simple, accurate, precise and economic RP-HPLC method with lesser run time to estimate the candesartan in bulk and pharmaceutical dosage forms.

 

MATERIALS AND METHODS:

The liquid chromatographic system consisted of following components : A Shimadzu HPLC model containing LC-20AT (VP Series) Pump, variable wavelength PDA detector and Hamilton syringe (50 µL).

 

Chromatographic analysis was performed using empower software on a Inertsil ODS-3V C-18 (250x4.6mm, 5µm) column. The mobile phase consisting of ortho phosphoric acid:Acetonitrile (20:80 v/v) and adjust the pH to 3.2 by using potassium hydrogen phosphate buffer. The optimized chromatographic conditions are summarized in Table.1. The standard solution of candesartan was prepared by dissolving 135mg in 100ml of acetonitrile which is used as diluent. 5ml of above solution was made up to 50ml with diluent. The mobile phase and the solution were soinicated for 10min and filtered using whatman filter paper No.1. The various dilutions of candesartan in the concentration of 50-160g/ml were prepared. The solutions were injected using a 20 µl fixed loop in to the chromatographic system at the flow rate of 1.0ml/min and the effluents were monitored at 210 nm, chromatograms were recorded. The candesartan was eluted at 8.057min as shown in Fig: 2 the method was extended for determination of candesartan in pharmaceutical dosage form. The pharmaceutical dosage form containing 4mg strength was taken.

 

 

Table 1: Optimized Chromatographic conditions for the proposed method

Parameters

Optimized condition

Column

Inertsil ODS-3V C-18 (250 x 4.6 mm, 5)

Mobile phase

0.03M potassium hydrogen phosphate in

 

Water pH 3.2 with ortho phosphoric acid: Acetonitrile (20:80).

Flow rate

1.0 ml / min

Injection volume

20 l

Detection

210 nm

Temperature

Ambient

Retention time

8.057 min

 

20 tablets of candesartan (containing 4 mg) were weighed and powdered in glass mortar and the powder equivalent to 250mg of candesartan was transferred into 100ml volumetric flask  and acetonitrile was used as diluent to make up the volume to 100ml. 5ml of above solution was made up to 50ml with diluent. Flask was kept for Ultrasonication for 10 min and the solution was filtered through whatman filter paper No.1. From this solution various dilutions were made with the mobile phase, which were analysed. The concentration of the drug in tablet sample solution was calculated by comparing the peak area of standard. The proposed method was validated as per the ICH guidelines.[9,10]

 

Fig.1 : Chemical Structure of candesartan

 

Fig.2: Typical RP-HPLC Chromatogram of candesartan by the proposed method.

 

RESULTS AND DISCUSSION:

A suitability test was applied to representative chromatograms for various parameters. The results obtained were within acceptable limits (Table 2). Thus, the system meets suitable criteria. The calibration curve was obtained for a series of concentration in the range of 50-160g/ml and it was found to be linear. The precision was measured in terms of repeatability, which was determined by sufficient number of aliquots of a homogenous sample. The % RSD was found and lying with in 2. This showed that the precision of the method was satisfactory. The accuracy of the method was inferred by establishing the precision and linearity studies of the standard. The % RSD was less than 2.0. This showed that the recoveries of candesartan by the proposed methods are satisfactory. The % RSD values were calculated from precision study was less than 2.0. Limit of detection (LOD) and Limit of quantiation (LOQ) were determined by the proposed methods. The results of validation parameters are summarized in Table 3. The results of recovery studies obtained by the proposed method were validated by statistical evaluation and are given in Table 4.

 

Table 2: System Suitability Test Parameters for the proposed method.

Parameters

Values

Required limits

Retention time

8.057

RSD1%

Theoretical plates

10792.26

N > 2000

Tailing factor

1.17

T  2

 

Table 3: Summary of Validation Parameters for the proposed method

Parameters

Values

Limit of detection (g/ml)

0.1358

Limit of quantitation (g/ml)

0.4074

*Precision (% RSD)

 

Intra-day precision

0.40

Inter-day precision

0.46

 

Table 4: Assay Results of candesartan tablets using proposed method

Brand used

Labelled amount (mg)

Amount found (mg)

% Recovery

Tablet(candesar)

4

3.969

99.22

 

REFERENCES:

1.        Chen X et al.Therapeutic Target Database. Nucleic Acids  Res;  2002;30(1):412-5.

2.        Cervenka L et al.Renal responses of the nonclipped kidney of two- kidney/one-clip Goldblatt hypertensive rats to type 1 angiotensin  II receptor blockade with candesartan. J Am Soc  Nephrol;  1999 ;10 (11):197-201.

3.        Pfeffer M et al. Effects of candesartan on mortality and morbidity in patients with chronic heart failure. CHARM-Overall programme; 2003; 362(9): 759–66.

4.        Julius S et al. Feasibility of treating prehypertension with an angiotensin- receptor blocker.New England journal of medicine; 2006; 354   (16):345.

5.        Amir Alhaj Sakur et al. Determination of candesartan cilexitil in tablets by spectrofluorimetry. Int J pharm Sci; 2010; 4(1):12.

6.        Naseem Ahmad  et al. Determination of candesartan cilexetil in tablet dosage forms and dissolution testing samples by first derivative UV spectrophotometric method. Analytical letters; 2009; 42(14): 2232-2243.

7.        Patel Jignesh et al. Q-Analysis spectrometric method for determination f candesartan cilexetil and hydrochlorothiazide in tablet dosage form. J chem. Pharm Res; 2010; 2(3): 10-14.

8.        Bipin H et al. HPTLC-Densitometric analysis of candesartan cilexetil and hydrochlorothiazide in tablets. Planar chromatography; 2008; 21(3): 173-176.

9.        Robert A. Nash and Alfred H. Wachter, pharmaceutical process validation, James swarbrick, North Carolina, An international 3rd edition, Revised and Expanded, Volume 129, Marcel Dekker Inc.,New York,507-522.

10.     Green J M: A practical guide to analytical method validation,anal chem. News Feat 305A/309A(May 1,1996). 

 

 

 

 

Received on 03.03.2011          Modified on 18.03.2011

Accepted on 21.03.2011         © RJPT All right reserved

Research J. Pharm. and Tech. 4(6): June 2011; Page 997-998