Novel Visible Spectrophotometric Methods for Estimation of Fenoverine in Pharmaceutical Formulations

 

A Sreelakshmi*1, G Devala Rao2 and G Sudhakara Sai Babu2

1Department of Biotechnology, Montessori Mahila Kalasala, Vijayawada-520 010, Andhra Pradesh, India.

2K.V.S. R. Siddhartha College of Pharmaceutical Sciences, Vijayawada-520 010- Andhra Pradesh, India.

*Corresponding Author E-mail:  sree_adusu@rediffmail.com

 

ABSTRACT:

Fenoverine is an antispasmodic drug. Two simple, sensitive and accurate spectrophotometric methods have been developed for the determination of fenoverine in pure state and in its pharmaceutical formulations. The developed Method A is based on the formation of picrate salt between picric acid and free base of fenoverine and it shows maximum absorption at λ max 410 nm and Linearity in the range of 10-50 μg/mL. Method B involves reaction between free base of fenoverine and chloranilic acid. The developed chromogen in Method B shows maximum absorption at λ max 525 nm and Linearity in the range of 100-500 μg/mL. The results obtained were statistically evaluated and were found to be accurate and reproducible.

 

KEYWORDS: Fenoverine, Spectrophotometric.

 


INTRODUCTION:

Fenoverine1 is an antispasmodic drug. Chemically it is 10-[4-(3, 4-dioxy methylene benzyl)-1-piperazinylacetyl] phenothiazine; 10-[(4-piperonyl-1-piperazinyl) acetyl] phenothiazine. Fenoverine acts by reducing the intracellular ca2+ concentration by decreasing the calcium gradient across the cell membrane, as well as decreasing the release of ca2+ from the intestinal pool more intensely than papaverine. This phenomenon has been observed in the intestinal smooth muscle and also in the genito-urinary tract. Thus it effectively synchronizes the strong, asynchronous spasmodic contractions, relieving the spasms and the associated symptoms2, 3.

 

Literature survey reveals that the pharmacodynamic profile of fenoverine on a smooth muscle4, formulation and evaluation of rapidly disintegrating fenoverine tablets5, few HPLC methods in pharmaceutical dosage forms and human plasma6, 7 were carried out. In the present investigation two simple, sensitive and accurate visible spectrophotometric methods have been developed for the estimation of fenoverine in pharmaceutical formulations   and in bulk drug. Method A shows λ max at 410 nm and Linearity in the range of 10-50 μg/mL. Method B exhibits λ max at 525 nm and Linearity in the range of 100-500 μg/mL.

 

Chemical structure of Fenoverine

 

EXPERIMENTAL

Spectral and absorbance measurements were made on systronics Double beam UV-Visible spectrophotometer model 2201 with 1cm matched quartz cells. Fenoverine was procured from a local pharmaceutical industry. All other reagents used were of analytical grade.

 

Reagents Preparation

For Method A, 400 mg of Picric acid was dissolved in 100 mL of chloroform.

For Method B, 100 mg of Chloranilic acid was dissolved in 20 mL of isopropyl alcohol, filtered and made upto 100 mL with chloroform.

 

Standard preparation

1 mg/ mL of pure Fenoverine standard solution prepared in chloroform was used for Method B and it was further diluted to get 100 µg/mL for Method A.

 

Sample preparation

Twenty tablets were taken and made into fine powder. The powder equivalent to average weight was taken and dissolved in 100 mL of chloroform. This was further diluted in respective concentrations required for Method A and used as such for Method B.


Table -1: Optical characteristics, regression data, Precision and accuracy of the proposed methods for Fenoverine

Parameter

Method A

Method B

λ max  (nm)

410

525

Beer’s law limits ( μg/mL)

10-50

100-500

Molar absorptivity (Lit.mole-1.cm-1)

8.226 x 103

6.61x 102

Detection limits ( µg/mL)

0.2352

5.223

Sandell’s Sensitivity ( μg/cm2/0.001 abs. unit)

0.0558

0.6944

Optimum photometric range

9-55

50-550

Regression equation (Y=a+bc) :

Slope (b)

 

0.0178

0.00145

 

Standard deviation of slope (Sb)

3.83x10-5

6.93 x 10-6

Intercept (a)

0.0012

-0.0018

Standard deviation of intercept (Sa)

0.00127

0.00223

Standard error of estimation (Se)

0.0012

0.0022

Correlation coefficient (r)

0.9999

0.9999

% Relative standard deviation*

14.812

0.4928

% Range of Error*

(confidence limits)

0.05 level

0.01 level

 

 

15.547

24.382

 

 

0.5712

0.8513

% Error in bulk samples**

0.21

0.34

* Average of six determinations, ** Average of three determinations

 

The values obtained for the determination of Fenoverine in several pharmaceutical formulations (tablets) and bulk drug by the proposed and reference methods were compared (Table 2).The results indicate that the proposed methods are simple, sensitive, accurate and reproducible and can be used for the routine determination of Fenoverine in bulk and pharmaceutical formulations.

 

 

TABLE 2: Assay and recovery of Fenoverine in dosage forms

Method

Pharmaceutical

Formulation

Labeled  Amount

(mg)

Proposed Method

% recovery by Proposed

methods** ± S.D

Amount found* (mg) ± S.D

t (value)

F (value)

A

Brand-1

100

98.96 ± 0.015

0.617

1.874

100.2 ± 0.54

B

Brand-1

100

99.95 ± 0.008

0.401

2.638

99.92 ± 1.04

* Average ± Standard deviation of six determinations, the t and F values refer to comparison of the proposed method with reference method.

Theoretical values at 95 % confidence limits t = 2.571 and F = 5.05, ** Average of five determinations.

 

 


Procedure for estimation:

Method A: Aliquots of standard solution containing from 1.0 to 5.0 mL (10-50 µg/ml) were transferred into a series of 10 mL volumetric flasks. Now the volume in all flasks is made upto 1 mL with chloroform, followed by addition of 1 mL of picric acid reagent and made upto volume with chloroform. The absorbance of the yellow colored species was measured at 410 nm against the reagent blank. The amount of fenoverine present in the sample solution was computed from the standard calibration curve.

 

Method B:  Aliquots of standard solution containing from 0.5-2.5 mL (100-500 µg/mL) were transferred into a series of 10 mL volumetric flasks. Now the volume in all the flasks is made upto 1 mL with chloroform, followed by addition of 1 mL of chloranilic acid reagent and made upto volume with chloroform. The absorbance of the purple colored species was measured at 525 nm against the reagent blank. The amount of fenoverine present in the sample solution was computed from the standard calibration curve.

 

RESULTS AND DISCUSSION:

The developed Method A is based on the quantitative yellow colored complex formation of drug with picric acid reagent. Picric acid in chloroform solution will have very little or no color. When chloroform solution of drug is added to this picric acid reagent it develops intense yellow color. The yellow colored salt formed between fenoverine and picric acid can be attributed to presence of cyclic tertiary nitrogen in fenoverine.

 

The proposed Method B is based on the drug which possesses cyclic tertiary nitrogen of aliphatic nature and function as electron donor and participates in charge transfer interaction with substituted quinones (chloranilic acid). The colored species formed due to the formation of radical cation.

 

The interference studies revealed that the common excipients usually present in the dosage forms do not interfere in the proposed method.

 

The optical characteristics and validation parameters were given in Table 1. To evaluate the accuracy and reproducibility of the method, known amounts of the pure drug was added to the previously analyzed pharmaceutical formulations and the mixture were reanalyzed by the proposed methods and the recoveries (average of six determinations) were given in Table 2.

 

ACKNOWLEDGEMENTS:

The authors are grateful to Montessori Mahila kalasala, Vijayawada for their continuous support and encouragement and for providing the necessary facilities.

REFERENCES:

(1)     The Merck Index-An Encyclopedia for Chemicals, Drugs and Biologicals, Merck and Co., 12th Edn., p. 676 (1996).

(2)     Gonella J, et al. Fenoverine: a novel synchronizer of smooth muscle motility by interference with cellular calcium flow. Curr Med Res Opin 1987; 10: 427-435.

(3)     Mironneau J, et al. Fenoverine inhibition of calcium channel currents in single smooth muscle cells from rat portal vein and myometrium. Br J Pharmacol 1991; 104: 65-70.

(4)     De Santis D, Marrazzo R, Losasso C, et al. Pharmacodynamic profile of fenoverine, a novel modulator of smooth muscle motility. Drugs Exp Clin Res 1989; 15: 37-42.

(5)     Sunil Kumar Battu; Michael A. Repka; Soumyajit Majumdar; Rao Y. Madhusudan. Formulation and Evaluation of Rapidly Disintegrating Fenoverine Tablets. Drug Development and Industrial Pharmacy 2007; 33(11): 1225 – 1232.

(6)     Suresh Bandari., Ramesh Gannu., K.V.S. Naidu., Madhusudan Rao Yamsani. Determination of Fenoverine in Human Serum. Journal of Liquid Chromatography & Related Technologies 2008; 31(14): 2101-2112.

(7)     Oliver Yoa-Putlu, Pu-Hsiang Chen, Yaw-Ju Fang, Hung-Shang Tang, Li-Heng Pao, Kin-Man Kwok, Ming-Lu King. Determination of Fenoverine , a modulator of smooth muscle motility, in capsule and in human plasma. Journal of Pharmaceutical Sciences 2006; 81 (1): 91-93.

 

 

 

 

Received on 14.10.2009       Modified on 11.12.2009

Accepted on 16.01.2010      © RJPT All right reserved

Research J. Pharm. and Tech. 3(2): April- June 2010; Page 449-451