Skip to main content

ESTIMATION OF DRONEDARONE HYDROCHLORIDE BY SPECTROPHOTOMETRIC METHOD IN PHARMACEUTICAL FORMULATION

 

Clinical courses

About Authors:
Kashyap K. Bhatt, Emanual Michael Patelia*, Ishani Amin
Department of Pharmaceutical Chemistry and Analysis,
Indukaka Ipcowala College of Pharmacy,
New Vallabh Vidyanagar – 388121, Gujarat, India.
*ricky.emanual@gmail.com

Summary
The UV- spectrophotometric method was developed for estimation of DRO in its dosage form. UV- spectrophotometric method was developed using methanol as solvent. Detection Wavelength was found to be 289nm. Detection wavelength was 289nm. In UV- spectrophotometric linear range was found to be 6 – 16 μg/ml & mean recoveries were found to be 99.83-100.04%. This spectroscopic method is economic, sensitive, and less time consuming than other chromatographic procedures. It is a user-friendly and importance tool for analysis of combined tablet dosage forms.

REFERENCE ID: PHARMATUTOR-ART-1630

1 Introduction
DRO (DRO;N-(2-Butyl-3-{4-[3-(dibutylamino)propoxy]benzoyl}-1-benzofuran-5- yl) methanesulfonamide hydrochloride; Figure 1a) is a widely used for theparoxysmal or persistent atrial fibrillation or atrial flutter[1-2].


Figure 1 Structure of Dronedarone


Literature survey revealed that various analytical methods like spectrophotometric [3], HPLC [4] and LC-MS in plasma [5] have been reported for the determination of DRO and either individually or combination with some other drugs, but no UV method was reported for estimation estimation of DRO in dosage forms. The review of literature prompted us to develop an accurate, selective and precise estimation method for the estimation of dosage forms.

2 Experimental

2.1 Chemicals and Materials
DRO were procured from Cadila pharmaceuticals, Ahmedabad. Methanol (HPLC Grade), Acetonitrile (HPLC Grade), Water (HPLC Grade), Hydrochloric acid (A. R. Grade), Potassium di-hydrogen Phosphate (A. R. Grade), Sodium hydroxide (A. R. Grade), Hydrogen peroxide (A. R. Grade) and Ortho phosphoric acid (A. R. Grade) were used as solvents to prepare the mobile phase. Dronedarone equivalent to 400 mg of DRO (Proprietary Name: MULTAQ) tablets were procured from local market.

2.2 Selection of analytical wavelength
The solution of DRO was prepared in Methanol at a concentration of 10 μg /ml. It was scanned in the wavelength range of 200-400 nm. Data were recorded at an interval of 1 nm. Analytical wavelength of 289 nm was selected for determination of DRO. This wavelength can be employed for the determination of DRO without any interference from the other excipients in its formulations.

2.3 Sample Preparation
Twenty tablets were weighed and powdered, powder equivalent to 400 mg of DRO was transferred in to a 10 ml volumetric flask. Methanol (5 ml) was added to it and sonicated for 20 min. The solution was filtered through Whattman filter paper No. 41 and the volume was adjusted up to the mark with Methanol. This solution is expected to contain 10 mg DRO.

2.4 Method Validation
The developed method was validated for linearity and range, specificity, accuracy, precision, Limit of detection, Limit of quantitation, robustness and solution stability as per ICH guidelines.

2.4.1 Linearity and Range
The linearity was determined at six levels over the range of 6 – 16 μg/ml. Take absorbances of above linearity solution preparations at each concentration. Calculate mean absorbance at each concentration and plot a graph of mean absorbance (y-axis) versus concentration (x-axis). (n=6)

Calculate and record value of correlation co-efficient (r), y-intercept & slope of regression line.

2.4.3 Accuracy (% Recovery)
The accuracy of the method was determined by calculating recoveries of DRO by method of standard additions. Known amount of DRO (0%,50%, 100%, 150%) were added to a pre quantified sample solution, and the amount of DRO was estimated by measuring the peak areas and by fitting these values to the straight-line equation of calibration curve.

2.4.4 Method Precision (Repeatability)
Standard solutions of DRO (6, 8, 10, 12, 14 and 16 μg/ml) were prepared and spectrums were recorded. Absorbance was measured at 289 nm using methanol as a blank. The absorbances of the same concentration solution were measured six times and %RSD was calculated.

2.4.5 Intermediate Precision (Reproducibility)
Variation of results of three different concentrations(6, 10 and 16 μg/ml) within the same day (intra- day) & variation of results between different days (inter- day) were analyzed. Intra-day precision was determined by analyzing DRO for three times in the same day. Inter-day precision was determined by analyzing DRO daily for three days.

2.4.6 Limits of Detection (LOD) and Limits of quantitation (LOQ)
The limit of detection (LOD) is defined as the lowest concentration of an analyte that can reliably be differentiated from background levels. Limit of quantification (LOQ) of an individual analytical procedure is the lowest amount of analyte that can be quantitatively determined with suitable precision and accuracy. LOD and LOQ were calculated using following equation as per ICH guidelines.LOD = 3.3 ×σ /S; LOQ = 10 ×σ /S; Where sis the standard deviation of y-intercepts of regression lines and S is the slope of the calibration curve.

2.4.7 Solution Stability
The sample preparations were analyzed by UV at regular intervals for 24 hrs as per test procedure

3 Results and Discussion

3.1 Selection of analytical wavelength
The solution of DRO was prepared in Methanol at a concentration of 10 μg /ml. It was scanned in the wavelength range of 200-400 nm. Data were recorded at an interval of 1 nm. Analytical wavelength of 289 nm was selected for determination of DRO Analytical wavelength of 289 nm was selected for determination of DRO.

Figure 2: Overlain Spectrum of DRO (6 – 16 μg/ml) in Methanol

3.2 Validation of the Method

3.2.1 Linearity
The linearity was determined at six levels over the range of 6 – 16 μg/ml. Calibration data at 289 nm for DRO. Overlay spectra is shown in (figure 3). Calibration curve for DRO was plotted between absorbance and concentration(Figure 5.8). The following equation for straight line was obtained for DRO. Linear equation for DRO , y = 0.0379x + 0.0137. The developed Spectrophotometry method was validated. The linear range, correlation coefficient, detection limit and standard deviation for DRO by Spectrophotometry method.

Figure 3:-Calibration plot of DRO by the developed UV method.

3.2.2 Specificity
The specificity study was carried out to check the interference from the excipients used in the formulations by preparing synthetic mixture containing both the drugs and excipients. The chromatogram showed peaks for the drug without any interfering peak and the recoveries of the drug were above 99%.

Figure 4 UV-Spectral of DRO (200-400 nm).

Table 1

Peak purity correlation results of DRO in two formulations at peak start, middle and end using CAMAG TLC SCANNER III

Sample

Correlation of center and

slope spectra

r (s, m)

r (m, e)

DRO

0.991

0.995

DRO formulation

0.996

0.991

3.2.3 Accuracy
Accuracy was determined by calculating the recovery. The method was found to be accurate with % recovery 99.86 – 100.44 for DRO (Table 2).

Table 2 Results from accuracy study.

Level

Amount

Taken (µg/ml)

Amount

Added (µg/ml)

Amount

Found (µg/ml)

%

Recovery

 

 

Mean

recovery

%RSD

0

6

0

5.97

99.5

99.83 %

0.78

6

0

6.01

100.16

6

0

5.99

99.83

50 %

6

3

8.99

99.89

100.44 %

0.89

6

3

9.01

100.11

6

3

9.12

101.33

100 %

6

6

12.11

100.90

100.43 %

0.87

6

6

11.92

99.33

6

6

12.13

101.08

150 %

6

9

15.01

100.06

99.86

0.68

6

9

14.98

99.86

6

9

14.95

99.66

3.2.4 Precision

a) Repeatability
The % RSD < 2 for DRO which indicate that the method is precise.

b) Intra and inter day precision
Variation of results within the same day (intra- day), variation of results between days (inter- day) were analyzed.. For intra-day (n=3) % RSD was found to be 0.43 – 0.64 and % RSD for inter-day (n=3) was 0.32 – 0.99 for DRO. The % RSD < 2 for DRO which indicate that the method is precise. (Table 3).

3.2.5 Limits of detection (LOD) and Limits of Quantification (LOQ)
Under the experimental conditions used, the lowest amount of drug that could be detected (LOD) for DRO was found to be 0.19 μg/ml. The limit of quantification(LOQ) for DRO was found to be 1.19 μg/ml, with an RSD <2%.

Table 3 Summary of validation parameters of developed UV method.

Parameters

DRO

Range

6-16 μg/ml

Accuracy(%)

99.86 – 100.04

Precision (%RSD)

Intra-day (n=3)

0.43-0.64

Inter-day (n=3)

0.32-0.89

Instrument precision  (%RSD)

0.67

Specificity

Specific

Solvent suitability

Solvent suitable for 24 hrs.

3.2.6 Solution stability
The sample preparations were analyzed by UV system at regular intervals for 24 hrs as per test procedure. The method is also rugged as there was no change in absorbance up to 24 hours of preparation of solution in Methanol.

3.4 Method Application
The proposed, developed and validated method was successfully applied to analysis of DRO in their marketed formulation. There was no interference of excipients commonly found in tablets as described in specificity studies. The assay results obtained were satisfactory, accurate, and precise as indicated by the good recovery and acceptable standard deviation values (Table 4). The good performance of the method indicates that it can be used for the determination of DRO in pharmaceutical formulation.

Table 4:- Results from analysis of Dronederone in the tablet dosage form

Component

Amount taken

in (μg/ml)

Amount Found (μg/ml)

Amount found

(%)

DRO

10

9.95

99.5

Mean ± SD

1198.1 ± 3.37

99.50± 0.26

% RSD

0.18

0.22

n = number of determinations

4 Conclusion
This developed and validated method for analysis of DRO in pharmaceutical preparations is very rapid, accurate, and precise. The method was successfully applied for determination of DRO in its pharmaceutical tablet formulation. Moreover it has advantages of short run time and the possibility of analysis of a large number of samples, both of which significantly reduce the analysis time per sample. Hence this method can be conveniently used for routine quality control analysis of DRO in its pharmaceutical formulation.

5 Acknowledgment
The authors are thankful to Cadila Pharmaceuticals Ltd., Ahmedabad for providing gift sample of DRO. The authors are very thankful to Sophisticated Instrumentation Center for Applied Research & Testing (SICART), Vallabh Vidyanagar, India), for providing necessary facilities to carryout research work. The authors are also thank full to indukaka ipcowala college of pharmacy (IICP) for providing laboratories facilties.

Reference
1.    chemspider.com/imageview.aspx
2.    rxlist.com/multaq.drug.htm
3.    Patel A, Akhtar J, Sharma C , “Spectrophotometric Estimation Of Dronedarone In Pure Drug And Pharmaceutical Formulation” Asian J. Of Bio. and Pharm. Res. 2012 ,2 , 2231-2560.
4.    Bolderman RW, Hermans JJ and Maessen JG, “Determination Of Class III Arrrhythmic Drugs Dronedarone And Amiodarone And Their Principal Metabolites In Plasma And Myocarium By High Performance Liquid Chromatography and UV Detection” J.Chrom B Anlyt Technol Biomed Life Sci. 2009,pp 1727-31.
5.    Cen X, Shilei Y, Dafang Z, Xiaojian D and Xiaoyan C, “Simultaneous Determination Of Dronedarone And It’s Active Metabolite Debutyldronedarone In Human Plasma By Liquid Chromatography- Tandem Mass Spectrophotometry : Application To A Pharmacokinetic Study” J. Of Chrom B. 2011,879,28, 3071-3075.

NOW YOU CAN ALSO PUBLISH YOUR ARTICLE ONLINE.

SUBMIT YOUR ARTICLE/PROJECT AT articles@pharmatutor.org

Subscribe to Pharmatutor Alerts by Email

FIND OUT MORE ARTICLES AT OUR DATABASE