Validation of Oxolamine Citrate by derivative UV Spectrophotometric Method in Pharmaceutical Dosage Form

 

Rele Rajan V.*

Central Research Laboratory, D.G. Ruparel College, Matunga, Mumbai 400016.

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

 

ABSTRACT:

A simple and precise third order derivative method have been developed and validated for the estimation of oxolamine citrate in bulk and its tablet formulation. Oxolamine citrate was estimated at 254.6 nm for the third 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 0.9995 for third order derivative method. The method was tested and validated for various parameters according to ICH guidelines. The precision expressed as relative standard deviation, which was as 2.0487 % for the above method. The proposed method was successfully applied for the determination of oxolamine citrate 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.

 

KEYWORDS: Oxolamine citrate, Third order derivative spectroscopy.

 

 

 

INTRODUCTION:

In this communication the present work proposes UV spectrophotometric method for assay of oxolamine citrate from bulk drug and pharmaceutical formulation.  It’s chemical name is 5- (2 -[diethyl amino] ethyl ) 3-phenyl-1,2,4 oxadiazole citrate. Oxolamine is an anti-inflammatory drug. This drug is in Chemical Abstracts Service Registry Number [1].

 

 

Drug is not official in any pharmacopeia. Literature survey reveals liquid chromatography methods [2-5]  and spectrophotometric [6-9] and non aqueous titration[10] for assay of this drug. Rapid, simple and reliable UV spectrophotometric methods are developed for the determination of oxolamine citrate. These methods can be used for the routine analysis and research organization. In the proposed work optimization and validation of these methods are reported. The structure of oxolamine citrate is shown in Fig.1.

 

Fig. 1: Chemical structure of oxolamine citrate

 

MATERIALS AND METHODS:

Instrument and reagents

1)     Shimadzu UV-spectrophotometer, model 1800 (Shimadzu, Japan) with spectral band width of 0.5 nm with automatic wavelength corrections by using a pair of 10 mm quartz cells. All spectral measurements were done by using UV-Probe 2.42 software.

2)     Shimadzu analytical balance (0.01 mg) was used

 

Reagents and materials

Reference standard of oxolamine citrate was obtained from reputed firm with certificate analysis.

 

Preparation of standard drug solution

100 mg standard oxolamine citrate was weighed accurately and transferred to a 100 ml volumetric flask and sonicated with 30 ml of distilled water for 15 minutes. The volume was made up to the mark with distilled water to give a stock solution of concentration 1000 μg /ml. From this solution, 10 ml of solution was pipetted out and transferred into 100 ml volumetric flask. The volume was made up to mark with distilled water to give a working standard solution of concentration 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 oxolamine citrate was weighed and transferred in 100 ml of volumetric flask. A 30 ml of distilled water was added and sonicated for 15 minutes and filtered. The filtrate and washing were diluted up to the mark with distilled water to give concentration as 100 μg /ml. Such solution was used for analysis.

 

Experimental

For the selection of analytical wavelength, 10 μg/ml solution of oxolamine citrate was scanned in the spectrum mode from 400 nm to 200 nm by using distilled water as blank. The third order derivative spectrum was obtained by using derivative mode by UV probe 2.42 software. From the spectrum, the absorbance of the derivative spectrum was measured at 254.6nm (Fig. 2).

                                                                                                                                         

Fig.2. Third order derivative overlay spectrum of oxolamine citrate (1- 14 μg/ml) showing absorbance at 254.6 nm

 

 

Linearity:
Into series of 10 ml graduated flask, varying amount of sample solutions of oxolamine citrate were pipetted out and volume was adjusted with distilled water. Solutions were scanned between 400 nm to 200 nm in spectrum mode. The third order derivative spectra were obtained by using derivative mode. Amplitudes of the resulting solutions were measured at between 254.6nm by using distilled water as blank. The calibration curve was prepared in the concentration range of 1 to 14 μg/ml (Fig. 3).

 

 

Fig. 3. Calibration curve for oxolamine citrate at 254.6nm by third order derivative  Spectroscopy

 

 

Results of the analysis are given in table 1.

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

Parameter

Values

Detection Wavelength (nm)

254.6

Beer Law Limits (µg/ml)                             

1-14

Correlation coefficient(r2)                                      

0.9995

Regression equation

(y=b+ac)

 

Slope (a)                                                                 

0.0002

Intercept (b)                                                            

0.00001

 

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).

 

Table 2: Results of recovery of oxolamine citrate for third order derivative method

Amount  of sample added in

(μg/ ml)

Amount of standard added in( μg/ ml)

Total amount recovered in μg/ ml

Percentage recovery(%)

Standard deviation

Percentage of relative standard deviation (%) C.O.V.)

2

0

1.904762

95.2381

0.251976

13.22876

2

2

3.952381

98.80952

0.125988

3.187652

2

4

6.047619

100.7937

0.299912

4.959172

2

6

7.952381

99.40476

0.230022

2.89249

 

Precision

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

Table 3: Precision- method precision

Experiment no.

Weight of oxolamine citrate taken in mg.

Content of oxolamine citrate found in mg.

1

10

10.000

2

10

10.666

3

10

10.000

4

10

9.333

5

10

10.000

6

10

10.666

7

10

10..000

Standard deviation

%R.S.D.

0.5039

4.9453

Inter-day and intra-day precision

An accurately weighed quantity of tablets powder equivalent to 10 mg of oxolamine citrate was transferred to 100 ml of volumetric flask. A 30 ml of distilled water was added and sonicated for 15 minutes and filtered. The filtrate and washing were diluted up to the mark with distilled water to give concentration as 100 μg /ml. Such solution was used for analysis.

 

Solution was scanned between 400 nm to 200 nm in spectrum mode. The third order derivative spectrum was obtained by using derivative mode. Amplitude of the resulting solution was measured at 254.6nm by using distilled water as blank. The amplitude of final solution was read after 0 hr., 3 hrs. and 6 hrs. in 10 mm cell at 254.6nm for third order derivative. Similarly the amplitude of the same solution was read on 1st, 2nd and 5th day. The amount of   oxolamine citrate was estimated by comparison with standard at 254.6nm for third order derivative, table 4.

 

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

Sr. No.

Parameters

third order derivative method

1

Intra day precision  (N=3)

amount found  ± % R.S.D.

 98.57%±

2.8927

2

Inter day precision 

( N=3)amount found ± % R.S.D.

 98.56%±

2.261

3

Ruggedness analyst to analyst ± % R.S.D.

 97.26%±

2.256

 

Ruggedness

The ruggedness of the method is defined as degree of reproducibility of results obtained by analysis of oxolamine citrate sample under variety of normal test conditions such as different laboratories, different analysts and different lots of reagents. Quantitative determination of oxolamine citrate 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.

 

RESULTS AND DISCUSSION:

The third order derivative UV-spectroscopic method is useful for routine analysis of oxolamine citrate 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 procedure [11]. Oxolamine citrate has the absorbance maxima at 254.6nm. 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, 3. The percentage recovery value indicates non interference from excipients used in formulation. The reproducibility and accuracy of the methods were found to be good, which was evidenced by low standard deviation.

 

CONCLUSION:

The most striking features of method 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 and in vitro of oxolamine citrate in pharmaceutical formulation.

 

ACKNOWLEDGEMENT:

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

 

REFERENCES:

1.     Chemical Abstract Service Register Number 1949-20-8.

2.     Ozan Pirol, Morat Sukuroglu, Tancel Ozden. Simultaneous determination of paracetamol, phenylephirne hydrochloride, oxolamine citrate and chlorpheniramine maleate by HPLC in pharmaceutical dosage form. J Chemistry,8(3); 2011:1275- 1279 .

3.     R.V. Rele, S.P. Patil. Reverse phase high pressure liquid Chromatographic Technique for determination of oxolamine citrate from pharmaceutical formulation. Research J. Pharm. and Tech 4(5) ; 2011: 787-789.

4.     Rele R.V. , Validation Method of Oxolamine Citrate From Bulk Drug And Pharmaceutical Formulation by High Performance Liquid Chromatography Method. International Journal of Pharmtech Research, 7(4); 2014: 549-553.

5.     Rele R.V.,Sawant S.A. High performance liquid chromatography for validation method of oxolamine citrate from bulk drug and pharmaceutical formulation, Journal  of  Chemical and Pharmaceutical  Research,7(1); 2015: 865-869, .

6.     Rele R.V. , Sawant S.A. UV spectrophotometric method for estimation of oxolamine citrate from bulk drug and pharmaceutical formulation. Der Pharma Chemica, 4(6); 2012: 2389- 2392.

7.     Rajan V. Rele, Swapnil A. Sawant, Sachin S. Patil and Pankaj J. Gurav, Spectrophotometric estimation of oxolamine citrate in bulk and pharmaceutical dosage form by first order derivative and area under curve methods. Journal of Chemical and Pharmaceutical Research, 5(5); 2013: 12-17.

8.     Rajan V. Rele, Spectrophotometric Estimation of Oxolamine citrate in Bulk and Pharmaceutical Dosage Form by Second Order Derivative Method. Research J. Pharm. and Tech.,7(10); 2014: 1150-1152.

9.     Rele R.V., Sawant S.A, Dissolution Study of Oxolamine Citrate by UV Spectrophotometric Method in Pharmaceutical Dosage Form, Journal of Chemical and Pharmaceutical Research, 8(7); 2015:108-112.

10.  Rele R.V., Patil S.S. A validated non-aqueous titration method for the quantative determination of oxolamine citrate from pharmaceutical preparation, American  Journal of Pharm-Tech Research, 3(1);2013: 445-449.

11.  ICH, Q2 (R1) Validation of Analytical Procedures: text and methodology. 2005, 1-13.

 

 

 

 

Received on 04.08.2016         Modified on 16.08.2016

Accepted on 04.10.2016         © AJRC All right reserved

Asian J. Research Chem. 2016; 9(10):475-478.

DOI: 10.5958/0974-4150.2016.00071.7