Development and Validation of RP-HPLC Method for Simultaneous Estimation of Albendazole and Praziqantel in Tablet Dosage Form

 

Kumaraswamy. Gandla1*, R. Lalitha1, Sadhana.Bommakanti1, R. Suthakaran1, K. Pallavi2

1Department of Pharmaceutical Analysis, Teegala Ram Reddy College of Pharmacy,

Meerpet, Hyderabad- 505172, Telangana

2SSJ College of Pharmacy, Vattinagulpalli, Hyderabad, Telangana.

*Corresponding Author E-mail: kumaraswamy.gandla@gmail.com

 

ABSTRACT:

A Simple, specific and sensitive an isocratic simultaneous Estimation by RP-HPLC analytical Method were developed and validated for the quantification Albendazole and Praziqantel. Quantification was achieved by using the mobile phase Phosphate Buffer pH 4.0 :Acetonitrile and Methanol in the ratio of 20:70:10 Inertsil ODS,C-18,250×4.6mm ID, 5µm Particle size was used as stationary phase. The flow rate was 1.0 ml/min. Measurements were made at a wavelength of 238nm. The average retention times for Albendazole and Praziqantel was found to be 2.713 and 4.770min. The proposed method was validated for selectivity, precision, linearity and accuracy. The assay methods were found to be linear from 60-180µg/ml and5-15µg/ml for Albendazole and Praziqantel respectively. All validation parameters were within the acceptable range. The developed methods were successfully applied to estimate the amount of Albendazole and Praziqantel.

 

KEYWORDS: Albendazole and Praziqantel, RP-HPLC method, isocratic elution Tablet dosage forms.

 

 

INTRODUCTION:

Albendazole1-3 is an orally administered broad-spectrum anthelmintic. Chemically, it is methyl 5-(propylthio)-2-benzimidazolecarbamate. Its molecular formula is C12H15N3O2S. Its molecular weight is 265.34. It has the following chemical structure (fig 01.)

 

Figure 1: Chemical structure of Albendazole

 

Albendazole is a white to off-white powder. It is soluble in dimethylsulfoxide, strong acids, and strong bases. It is slightly soluble in methanol, chloroform, ethyl acetate, and acetonitrile. Albendazole is practically insoluble in water. Each white to off-white, film-coated tablet contains 200 mg of albendazole

 

Praziquantel is a trematodicide4-5 provided in tablet form for the oral treatment of schistosome infections and infections due to liver fluke. praziquantel is 2-(cyclohexylcarbonyl)-1, 2,3,6,7, 11b-hexahydro-4H-pyrazino [2, 1-a] isoquinolin-4-one with the molecular formula; C19H24N2O2. The structural formula is as follows (fig.02).

 

Figure 2: Chemical structure of Praziqantel

Praziquantel is a white to nearly white crystalline powder of bitter taste. The compound is stable under normal conditions and melts at 136-140°C with decomposition. The active substance is hygroscopic. Praziquantel is easily soluble in chloroform and dimethylsulfoxide, soluble in ethanol and very slightly soluble in water.

 

Literature survey6-13 revealed that a various analytical methods have been reported for the determination of Albendazole and Praziqantel in pure drug, pharmaceutical dosage forms and in biological samples using liquid chromatography either in single or in combined forms. Confirmation of the applicability of the developed method was validated according to the International Conference on Harmonization (ICH) 14-16 for the simultaneous determination of Albendazole And Praziqantel  in bulk and in tablet dosage form.

 

MATERIALS AND METHODS:

Reagents and chemicals

Methanol HPLC grade was procured from E. Merck Ltd, Mumbai. Methanol, Acetonitrile. Fine chemicals, Hyderabad. Water HPLC grade was prepared using Millipore purification system. Albendazole and Praziqantel reference standards procured from Chandra pharma labs, Hyderabad.

 

Instrumentation

The HPLC system consists of water Empower -2690/5 having photodiode array detector system, which was connected with the help of Empower 3000 software for data integration and processing. Inertsil ODS‐3V (250 X 4.6 mm) 5μ column was used for the analysis.

 

METHOD 

The RP-HPLC Method of Albendazole and Praziqantel were achieved by isocratic elution technique with PDA Detector. Albendazole and Praziqantel were determined at 245nm   respectively with the concentration range of 60.-180 mg/ml and 5.0-15mg/ml for both Albendazole and Praziqantel respectively.fig.03 and04 and Table 1.  For analysis of tablet formulation the tablet powder equivalent to 25 mg was taken, dissolved in 25 ml volumetric flask and made up to 25ml with Methanol. The solution was sonicated for 15min, centrifuged at 100 rpm for 15 min and filtered through Whatmann filter paper No.41. From clear solution, further dilutions were made to get 10 mg/ml of Albendazole and Praziqantel theoretically.  

 

Figure 3: Calibration Graph of Albendazole

 

Figure 4: Calibration Graph of Praziqantel

 

Chromatographic conditions

The contents of the mobile phase were Methanol and Acetonitrile the ratio of 50:50. These were filtered through 0.45μ membrane filter and degassed by sonication before use. The flow rate of mobile phase was optimized to 1.0 ml / min. The run time was set at 8 min and column temperature was maintained at ambient. The volume of injection was 20 μl, and the eluent was detected at 245nm. Each of standard and test preparations was injected into the column and the responses recorded (Fig.05 and Fig.06.).

 

Figure 5: Typical chromatograms for recovery studies

 

For recovery studies, to the reanalyzed formulation, solutions of raw material containing different concentrations were added and the amount of drug recovered was calculated. The procedure was repeated as per the analysis of formulation. The amount of drug recovered was calculated by using slope and intercept values from the calibration graph. Finally the method was validated as per ICH guide lines for precision, accuracy, specificity, linearity, reproducibility, limit of detection and limit of quantification.

 

Figure 6: Typical chromatogram of Albendazole and Praziqantel

 

RESULTS AND DISCUSSION:

A simple, selective, rapid and precise validated RP-HPLC Method for Simultaneous Estimation of Albendazole and Praziqantel in bulk material and in pharmaceutical formulation has been developed and validated. The correlation coefficient was found to be 0.9997 and 0.9998 for Albendazole and Praziqantel respectively. In this method the % purity of Albendazole and Praziqantel were found to be 101.25 ± 1.074 and 100.19 ± 1.031 respectively. The recovery studies range is.99.98-100.01% and 99.94–100.03 % for Albendazole and Praziqantel, respectively. The Intraday and Inter day analysis carried out for precision. The ruggedness study was performed. In this method the % purity were found to be 100.25 ± 1.0054 and 101.49 ± 1.9305 for Albendazole and Praziqantel, respectively. The recovery studies range is 99.98-100.01% and 99.94 – 100.03 %. Table 2 and 3.The Intraday and Inter day analysis carried out for precision. The ruggedness study was performed. The method was validated for statistical analysis

 

Table 1: Linearity results

Preparations

Volume from standard stock  transferred in ml

Volume made up in ml

(with mobile phase)

Concentration of solution

(µg /ml)

ALBENDAZOLE

PRAZIQUANTEL 

Preparation 1

0.5

50

60

5.0

Preparation 2

0.75

50

90

7.5

Preparation 3

1.0

50

120

10.0

Preparation 4

1.25

50

150

12.5

Preparation 5

1.5

50

180

15

 

Table 2: Recovery Studies of Albendazole.

Recovery level

Accuracy ALBENDAZOLE

Amount taken(mcg/ml)

Area

Average area

Amount recovered(mcg/ml)

%Recovery

 

75%

90

2947.566

2932.667

 

89.20

 

99.11

 

90

2902.955

90

2947.480

 

100%

120

3571.503

3601.162

 

120.76

 

100.63

 

120

3666.372

120

3565.612

125%

150

4317.878

4389.011

 

152.24

 

101.49

 

150

4323.066

150

4526.088

 

Table 3.Accuracy of Albendazole

Recovery level

Amount taken(mcg/ml)

Area

Average area

Amount recovered (mcg/ml)

%

Recovery

 

 

75%

90

2947.566

2932.667

 

89.20

 

99.11

 

90

2902.955

90

2947.480


100%

120

3571.503

3601.162

 

120.76

 

100.63

 

120

3666.372

120

3565.612

 

125%

150

4317.878

4389.011

 

152.24

 

101.49

 

150

4323.066

150

4526.088

 

 

CONCLUSION:

The developed RP-HPLC method was validated and the statistical validation was performed with the simplicity and ease of operation ensures that the validated method can successfully used for routine Analysis of Albendazole and Praziqantel in bulk and tablet dosage formulation.

 

ACKNOWLEDGEMENTS:

The authors would like to thank MS. Chandra Labs, for providing the gift samples of Albendazole and Praziqantel for the project work.

 

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Received on 24.06.2015          Accepted on 28.07.2015        

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Ana. 5(3): July- Sept. 2015; Page 115-118

DOI: 10.5958/2231-5675.2015.00018.6