UV Spectrophotometric Estimation of Carvedilol Hydrochloride by Second Order Derivative Methods in Bulk and Pharmaceutical Dosage Form

 

Rajan V. Rele *, Prathamesh P. Tiwatane

Central research laboratory, D.G. Ruparel College, Matunga, Mumbai 400016.

 

ABSTRACT:

Simple and precise UV spectrophotometric methods by second order derivative have been developed and validated for the estimation of carvedilol hydrochloride in bulk and its tablet formulation. The standard and sample solutions of carvedilol hydrochloride were prepared in absolute alcohol. Carvedilol hydrochloride was estimated at 247.5 nm for the second order derivative UV-spectrophotometric method. Beer’s law was obeyed in the concentration range of 1 to 14 μg / ml with coefficient of correlation value 0.9999 for second order derivative method. These methods were tested and validated for various parameters according to ICH guidelines. The precision expressed as relative standard deviation were of 0.03243 % for the above method respectively. The proposed methods were successfully applied for the determination of carvedilol hydrochloride in pharmaceutical formulation. Results of the analysis were validated statistically and were found to be satisfactory. The proposed methods are simple, easy to apply, low-cost and require relatively inexpensive instruments.

 

 

 

INTRODUCTION:

Its chemical name is   (±)-1-(isopropyl amino)-3-[p-(2-methoxyethyl)phenoxy]-2-propanol hydrochloride. Carvedilol hydrochloride is official in USP¹, EP². Literature survey reveals the Spectrophotometric^3-8 HPLC^9-16, UPLC¹⁷ methods for the estimation of carvedilol hydrochloride. Simple, rapid and reliable UV spectrophotometric methods are developed for the determination of carvedilol hydrochloride. These methods can be used for the routine analysis. In the proposed methods optimization and validation of this method are reported.

 

Structure of carvedilol 

MATERIAL AND METHODS:

Shimadzu UV-1800 was used with 10 mm matched quartz cell to measure absorbance of solution. A Shimadzu analytical balance with 0.01 mg was used.

 

Chemical and reagents:

Reference standard of carvedilol hydrochloride was obtained from reputed firm with certificate analysis.  All spectral absorbance measurements were made on Shimadzu UV-1800 with 10 mm matched cell.

 

Preparation of standard solution:

About 10 mg of standard carvedilol hydrochloride was weighed accurately and transferred in 100 ml of volumetric flask. About 30 ml of absolute alcohol was added and sonicated for 15 minutes. The volume was adjusted up to the mark with absolute alcohol to give concentration as 100 μg /ml.

 

Estimation from Tablets:

Twenty tablets were weighed accurately and average weight of each tablet was determined. Powder equivalent to 10 mg of carvedilol hydrochloride was weighed and transferred in 100 ml of volumetric flask. A 30 ml of absolute alcohol was added and sonicated for 15 minutes and filtered. The filtrate and washing were diluted up to the mark with absolute alcohol to give concentration as 100 μg /ml. Such solution was used for analysis.

 

Fig. 2. Overlay spectra of second  order derivative spectrum of carvedilol hydrochloride (2-14 μg/ml) showing  absorbance at  247.5 nm

 

Experimental:

Method: Second Order Derivative Method:

For the selection of analytical wavelength, 10 μg /ml solution of carvedilol hydrochloride was scanned in the spectrum mode from 300 nm to 200 nm by using absolute alcohol as blank. The second order derivative spectrum was obtained by using derivative mode by UV probe 2.42 software. From the spectrum, the amplitude of the derivative spectrum was measured at 247.5 nm (Fig. 2).

 

 

Into series of 10 ml graduated flask, varying amount of standard solutions of carvedilol hydrochloride was pipette out and volume was adjusted with absolute alcohol as solvent. Solutions were scanned between 300 nm to 200 nm in spectrum mode.

 

Fig. 3. Calibration curve for carvedilol hydrochloride at 247.5 nm by second order derivative   Spectroscopy

 

The second order derivative spectra were obtained by using derivative mode. Amplitudes of the resulting solutions were measured at between 247.5 nm by using absolute alcohol as blank. The calibration curve was prepared in the concentration range of 1 to 14 μg/ml.(Fig. 3)

 

Results of analysis are given in table 1.

 

Table 1: Values of results of optical and regression of drug

Parameter	Values
Detection Wavelength (nm)	247.5
Beer Law Limits (µg/ml)	1-14
Correlation coefficient(r2)	0.9999
Regression equation (y=b+ac)
Slope (a)	0.0045
Intercept (b)	0.0006

 

Validation:

Accuracy:

Accuracy of the proposed methods was carried as on the basis of recovery studies. It is performed by the standard addition method. Recovery studies were performed by adding standard drug at different levels to the pre-analyzed tablets powder solution and the proposed method was followed. From the amount of the drug estimated, the percentage recovery was calculated. The results of the analysis are shown in table 2.

 

Precision:

The method precision was established by carrying out the analysis of homogenous powder blend of tablets. The assay was carried out of drug by using proposed analytical method in six replicates. The values of relative standard deviation lie well within the limits indicated the sample repeatability of the method. The results obtained are tabulated in table 3.

Table 2: Results of recovery of carvedilol hydrochloride

Amount of Sample Added in (µg/ml)	Amount of Standard Added in (µg/ml)	Total amount recovered	Percentage recovery (%)	Standard deviation	Percentage of relative standard deviation (C.O.V.)
2	0	2.0215	100.6144	0.02098	1.0429
2	2	3.9969	99.9232	0.03384	0.84666
2	4	5.9354	98.9247	0.02777	0.46775
2	6	7.8279	98.3871	0.03511	0.44617
				Mean=0.0557	Mean=1.130

Table 3: Precision- method precision

Experiment no.	Carvedilol hydrochloride taken in μg/ ml	Values
1	10	9.9784
2	10	9.9784
3	10	9.9569
4	10	10.0215
5	10	9.9784
6	10	10.0430
	Standard deviation	0.03237
	%RSD	0.03243

 

Inter-Day and Intra-Day Precision:

An accurately weighed quantity of tablets powder equivalent to 10 mg of carvedilol hydrochloride was transferred to 100 ml of volumetric flask. A 30 ml of absolute alcohol was added and sonicated for 15 minutes and filtered. The filtrate and washing were diluted up to the mark with absolute alcohol to give concentration as 100 μg /ml. Such solution was used for analysis. Solution was scanned between 300 nm to 200 nm in spectrum mode. The second order derivative spectrum was obtained by using derivative mode. Amplitude of the resulting solution was measured at 247.5 nm by using absolute alcohol as blank. The amplitude of final solution was read after 0 hr., 3 hrs. and 6 hrs. in 10 mm cell  at 233.7 nm for first order derivative (method A). Similarly the amplitude of the same solution was read on 1^st, 2^nd and 5^th day. The amount of  carvedilol hydrochloride was estimated by comparison with standard at 247.5  nm for second order derivative, table  4.

 

Table 4: Summary of validation parameter for intra-day and inter-day

Sr. no.	Parameters	First order derivative method
(A)	Intra-day precision ( n=3) Amount found ± % RSD	99.846% 0.7008
(B)	Inter-day precision ( n=3) Amount found ± % RSD	98.387% 0.4461
(c)	Ruggedness  Analyst to analyst( n= 3) %RSD	0.7006

 

Limit of Detection (LOD) and Limit of Quantification (LOQ):

The limit of detection (LOD) is defined as the lowest concentration of an analyte that an analytical process can reliably differentiate from back-ground levels. In this study, LOD and LOQ were based on the standard deviation of the response and the slope of the corresponding curve using the following equations-

LOD = 3.3 σ/S    and   LOQ = 10 σ/S

 

Where σ   is the standard deviation of the signal to noise ratio of the sample and S is the slope of the related calibrations graphs. The limit of quantification (LOQ) is defined as the lowest concentration of the standard curve that can be measured with an acceptable accuracy, precision and variability .The values of LOD and LOQ are given in table 5.

 

Table 5: Values of results of LOD and LOQ

Parameters	Values
Limit of Detection (μg/ml)	0.07152
Limit of Quantification (μg/ml)	0.2167

 

 

Ruggedness:

The ruggedness of the method is defined as degree of reproducibility of results obtained by analysis of carvedilol hydrochloride sample under variety of normal test conditions such as different laboratories, different analysts and different lots of reagents. Quantitative determination of carvedilol hydrochloride was conducted spectrophotometrically on one laboratory. It was again tested in another laboratory using different instrument by different analyst. The assays obtained in two different laboratories were well in agreement.  It proved ruggedness of the proposed methods.

 

RESULT AND DISCUSSION:

The second order derivative and area under curve UV-spectroscopic methods are useful for routine analysis of carvedilol hydrochloride in bulk drug and formulation. The derivative spectroscopy method applied has the advantage that it locates hidden peak in the normal spectrum. It eliminates the interference caused by the excipients and the degradation products present, if any, in the formulation. The method was validated according to International Conference on Harmonization guidelines for validation of analytical procedures. Carvedilol hydrochloride has the absorbance maxima at 247.5 nm. The polynomial regression data for the calibration plots showed good linear relationship in the concentration range of 1 to 14 μg/ml and given in table1. Recovery studies were carried out by adding the pure drug to the previously analyzed tablet powder sample and shown in table 2. The percentage recovery value indicates non interference from excipients used in formulation. The reproducibility and accuracy of the method were found to be good, which was evidenced by low standard deviation.

 

CONCLUSION:

The most striking features of two methods are its simplicity and rapidity, not requiring tedious sample solutions preparations which are needed for other instrumental methods.  From the results obtained it can be concluded that the proposed methods are fully validated and found to be simple, sensitive, accurate, precise, reproducible, rugged and robust and relatively inexpensive. So, the developed methods can be easily applied for the routine quality control analysis of carvedilol hydrochloride in pharmaceutical formulation.

 

ACKNOWLEDGMENT:

Authors express sincere thanks to the Principal, Dr. Tushar M. Desai of D. G. Ruparel college.

 

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Received on 04.11.2014          Modified on 12.11.2014

Accepted on 17.11.2014         © RJPT All right reserved