Development and Validation of Spectrophotometric Methods for Simultaneous Estimation of Indapamide and Amlodipine Besylate in Combined Dosage Form.

 

Mihir R. Patel*, Gaurav F. Patel, Jignesh R. Patel

Department of Quality Assurance, Shree Dhanvantary Pharmacy College, Kim (Surat), Gujarat, India 394110.

*Corresponding Author E-mail: patel.mihir57@gmail.com

 

ABSTRACT:

Two simple spectrophotometric methods have been developed for the simultaneous estimation of Indapamide and Amlodipine besylate in their combined dosage forms. In Dual wavelength Spectrophotometric method, two wavelengths on amlodipine besylate curve has been found out where it showed same absorbance at 256 nm and 360.0 nm. At 360.0 nm, Amlodipine besylate showed some absorbance while Indapamide showed zero absorbance. The method involved solving of an equation based on measurement of absorbances at two wavelengths 256 nm and 360.0 nm. In first order derivative spectrophotometric method in which, measurement of amplitudes at 237.4 nm (for IND) and 241 nm (for AML) in first derivative spectra. Developed methods were validated according to ICH guidelines. This methods obeyed Beer’s law in the concentration range of 3-18 μg/ml for IND and 10–60 μg/ml for AML for dual wavelength Spectrophotometric method and 1.5-9 μg/ml for IND and 5–30 μg/ml for AML for first order derivative method.The proposed methods were found to be simple, accurate and reproducible for the routine analysis of both drugs in combined tablet dosage forms.

 

KEY WORDS: Spectrophotometric, Indapamide, Amlodipine besylate, Simultaneous estimation, Dual wavelength spectrophotometric method, first order derivative spectrophotometric method.

 

 


INTRODUCTION:

Indapamide is a white to almost white powder and slightly soluble in water, freely soluble in methanol, sparingly soluble in ethanol, slightly soluble in 2-propanol. Chemically Indapamide is 4-chloro-N-(2-methyl-2,3-dihydro-1H-indol-1-yl)-3-sulfamoylbenzamide. Indapamide shows an alteration in vascular reactivity to angiotensin II and other agonists, suggesting the possibility of a direct vascular effect.

Indapamide

 

Amlodipine is a white crystalline powder which is slightly soluble in water, sparingly soluble in ethanol and freely soluble in methanol. Chemically Amlodipine besylate is 3-ethyl 5-methyl(4RS)-2-[(2-aminoethoxy)methyl]-4-(2-chlorophenyl)-6-methyl-1,4-dihydropyridine-3,5-dicarboxylate benzene sulphonate. Amlodipine is a dihydropyridine derivative with calcium antagonist activity. It is used in the management of hypertension. Amlodipine acts by inhibiting the transmembrane influx of calcium ions into vascular smooth muscle and cardiac muscle and also acts directly on vascular smooth muscle to cause a reduction in peripheral vascular resistance and reduction in blood pressure.

 

Amlodipine Besylate

 

The review of literature revealed that several methods are available for the determination of Indapamide1-7 and Amlodipine besylate8-18 individually. But, there is one analytical method has been reported yet for combination of these drugs19. The present research work aims to develop simple, sensitive, accurate and reproducible methods for simultaneous estimation of Indapamide and Amlodipine besylate in combined dosage form.

 

EXPERIMENTAL:

Instrument and material

All chemicals used were of analytical grade. Pure Indapamide was obtained from Supra Chemicals Pvt. Ltd., Thane, Navi Mumbai and Amodipine besylate from Prudence Pharmaceutical, Ankleshwar. The spectrophotometer used was Shimadzu UV Visible spectrophometer 2450 model with spectral bandwidth of 2 nm and wavelength accuracy (with automatic wavelength correction) of 0.5 nm. All the apparatus and instruments were calibrated and validated as per calibration and validation protocol specified before starting the experimental work.

 

Overlain spectra of SERIES A, B and mixture C

 

Linearity graph of Indapamide

 

Calibration curve for Indapamide

 

Linearity graph of Indapamide

 

Linearity graph of Amlodipine besylate

 

METHOD:

Standard preparation:

10 mg of IND and 10 mg of AML was accurately weighed and transferred in different 100 ml volumetric flask. Add sufficient amount of methanol to dissolve. Adjust the volume with methanol. It would give 100ppm IND and 100ppm AML stock solution.

 

Method I: Dual wavelength spectrophotometry16

The solutions of standard IND and AML were prepared in the range of 3 to 18 μg/ml and 10 to 60 μg/ml respectively in methanol. AML were found to be given adequate absorbance at 360 nm. Thereafter, individual spectra were recorded for AML at 360 nm was found and by using cursor function same absorbance was found at 256 nm.

 

IND showed no interference at 360 nm and showed zero absorbance. So 360 nm was selected for determination of AML in the mixture. 256 nm was selected for estimation of IND in mixture and difference between 256nm and 360nm was taken for estimation of IND

 

Method II: First derivative spectrophotometry11

0.6 ml of standard stock solution of IND (100 μg/ml) and 2 ml of standard stock solution of AML (100 μg/ml) were pipette out into two separate 10 ml volumetric flask and volume was adjusted to the mark with methanol to get 6 μg/ml of IND and 20 μg/ml of AML. Each solution was scanned between 200-400 nm against methanol as a reagent blank. The first order derivative spectra of each solution were obtained. The zero crossing points were found to be 241 nm and 237.4 nm for IND and AML respectively. The solutions of standard AML and IND were prepared in the range of 5 to 30 μg/ml and 1.5 to 9 μg/ml respectively. Calibration graph for AML and IND are shown below.

 

Overlain first order spectra of IND (3 μg/ml)

and (2) AML (10 μg/ml)

 

Overlain first order calibration spectra of IND (1.5-9μg/ml) and AML (5-30 μg/ml)

 

Calibration curve of standard IND at 237.4 nm by first order derivative spectrophotometry

 

Calibration curve of standard AML at 241 nm by first order derivative spectrophotometry

 


Table 1

Sr.No

Parameters

Result for IND

Result for AML

1.

Linearity Range

3-18 μg/ml

10-60 μg/ml

2.

Correlation coefficient(n=6)

0.99969

0.99962

3.

Precision (C.V. (%))

 

 

1. Repeatability (n=6)

0.7290

0.4834

2. Intraday precision (n=3)

0.430

0.608

3. Interday precision (n=3)

0.656

0.651

4.

Robustness (n=3)

0.408-0.542 %

0.425-0.568 %

5.

 Ruggedness (n=3)

0.541-0.717 %

0.423-0.712 %

6.

Assay (% found)(n=6)

99.32

101.34

7.

Accuracy (% Recovery)

100.0905

99.7870

8.

LOD (μg/ml)

0.136

0.475

9.

LOQ (μg/ml)

0.413

1.440

 

Table 2

Sr.No

Parameters

Result for IND

Result for AML

1.

Linearity Range

1.5-9 μg/ml

5-30 μg/ml

2.

Correlation coefficient(n=6)

0.99983

0.99966

3.

Precision (C.V. (%))

 

 

1. Repeatability (n=6)

0.8481

0.3548

2. Intraday precision (n=3)

0.454

0.471

3. Interday precision (n=3)

0.606

0.628

4.

Robustness (n=3)

0.408-0.542 %

0.425-0.568 %

5.

 Ruggedness (n=3)

0.541-0.717 %

0.423-0.712 %

6.

Assay (% Found)

99.72

100.28

7.

Accuracy (%recovery)

99.782

99.812

8.

LOD (μg/ml)

0.116

0.306

9.

LOQ (μg/ml)

0.351

0.927

 


Assay of Combined dosage form by both Method

20 tablets were powdered and an amount equivalent to 1.5 mg IND and 5 mg AML was weighed and dissolved in 50 ml methanol. Solutions were filtered using whatmann filter paper. Appropriate dilutions were prepared in methanol taking suitable aliquots of the clear filtrates and subjected to analysis.

 

RESULT AND DISCUSSION:

Developed spectrophotometric methods for the simultaneous were validated according to ICH guidelines20 and data complying with the standards were obtained. The results of validation parameters for all the three methods are reported in Table 1 and Table 2.

 

CONCLUSION:

The validated Spectrophotometric method employed here proved to be simple, fast, accurate, and precise and robust thus can be used for routine analysis of Indapamide and Amlodipine besylate in combined dosage form.

 

ACKNOWLEDGEMENT:

The authors are also thankful to Supra Chemicals, Thane and Prudence Pharma, Ankleshwar for providing gift samples of drugs.

 

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Received on 04.06.2012       Modified on 29.07.2012

Accepted on 12.08.2012      © RJPT All right reserved

Research J. Pharm. and Tech. 5(9): September 2012; Page 1213-1217