Visible Spectrophotometric Determination of Ticlopidine in Pharmaceutical Formulation

 

M Padmalatha¹, K Vanitha Prakash²*, and Eranna Dopadally³

¹Vijaya College of Pharmacy, Munganoor, Hayathnagar, Hyderabad

²SSJ College of Pharmacy, V.N. Pally, Gandipet, Hyderabad.

³Sultan-Ul-Uloom College of Pharmacy, Banjara Hills, Hyderabad.

*Corresponding Author E-mail: prakash.karnam@gmail.com

ABSTRACT:

Three simple, economical, precise, reliable and reproducible visible spectrophotometric methods (A, B and C) have been developed for the estimation of Ticlopidine in bulk as well as in Tablet formulation. The developed methods A, B and C are based on the formation of chloroform extractable complex of Ticlopidine with Bromocresol green (Method A), Bromophenol Blue (Method B), and Bromothymol Blue (Method C) which shows absorbance maxima at 430 nm, 449 nm and 427 nm respectively. The absorbance-concentration plot is linear over the range of 2.5-25 mcg/ml for Method A, and 2.5-20 mg/ml for Method B and 2.5-15 mcg/ml for Method C. The different experimental parameters affecting the development and stability were studied carefully and optimized. Results of analysis for all the methods were validated statistically and by recovery studies.

 

 

 

INTRODUCTION:

Ticlopidine ¹ with a chemical name 5-[(2-chlorophenyl) methyl]-6, 7-dihydro-4H-thieno [3, 2-c] pyridine. Ticlopidine is an effective inhibitor of platelet aggregation. The drug has been found to significantly reduce infarction size in acute myocardial infarcts and is an effective antithrombotic agent in arteriovenous fistulas, aorto-coronary bypass grafts, ischemic heart disease, venous thrombosis, and arteriosclerosis. Literature survey reveals many Chromatographic methods^2-5 for the determination of Ticlopidine in biological fluids and in tablets and very few Spectrophotometric methods were reported. Therefore the need for fast, low cost and selective Spectrophotometric method is obvious especially for routine Quality Control analysis of pharmaceutical formulation.

 

EXPERIMENTAL:

Instrument:

Elico double beam Ultraviolet-Visible double beam spectrophotometer SL-244 with 1 cm matched quartz cells was used for all spectral measurements.

 

Reagents:

All the chemicals used were of analytical reagent grade. Acid phthalte buffer pH 2.4 and

pH 3 were prepared as per I.P.

i)       Bromocresol green, BCG (0.1% w/v)-100 mg is weighed accurately and dissolved in 100 ml of acid phthalate buffer pH 2.4.

ii)      Bromophenol Blue, BPB (0.1% w/v)- 100 mg is weighed accurately and dissolved in 100 ml of acid phthalate buffer pH 3.0.

iii)     Bromothymol Blue, BTB (0.1% w/v)-100 mg is weighed accurately and dissolved in 100 ml of acid phthalate buffer pH 2.4.

iv)     Chloroform AR grade

 

Procedure:

Standard stock solution: A standard stock solution containing 1mg/ml was prepared by dissolving 100 mg of Ticlopidine in 100 ml of methanol. From this, a working standard solution containing 100 mcg/ml were prepared with methanol.

 

ASSAY PROCEDURE:

Method A: Aliquots of the drug solution of Ticlopidine 0.5-5.0 ml (100 mcg/ml) are taken and transferred into a series of 125 ml of separating funnel. To each funnel 2 ml of BCG reagent solution is added. Reaction mixture was shaken gently for 5 min.

Table: I - OPTICAL CHARACTERISTICS AND PRECISION DATA.

Parameters	Method A	Method B	Method C
l max (nm)	445	459	420
Beer’s law limits (mcg/ml)	5-50	5-50	5-40
Molar absorptivity (l/mol.cm)	1.85x104	1.052x104	1.938x103
Sand ell’s sensitivity (micrograms/cm2/0.001 absorbance unit)	0.5369	0.4921	0.2302
Regression Equation* (Y) Slope (m) Intercept (c)	0.004 0.0146	0.002 -0.0017	0.006 0.0609
Correlation Coefficient(r)	0.9999	0.9998	0.9989
Precision (%Relative Standard Deviation)	0.417	0.910	0.421
% range of error (confidence limits)* 0.05 level 0.01 level	0.972 1.524	0.313 0.492	0.246 0.386

*Y=mx+c, where X is the concentration in micrograms/ml and Y is absorbance unit.

 

Table II: ASSAY OF TICLOPIDINE IN TABLETS.

Sample No.	Labelled Amount (mg)	% Obtained* by proposed method			** % Recovery by the Proposed method
		Method A.	Method B	Method C	Method A	Method B	Method C
1	250	249.6	250.1	250.2	99.9	99.5	100.5
2	250	248.9	249.9	251.3	100.5	99.6	99.8

*Average of three determinations, ** After spiking the sample.

 

Then 10 ml of chloroform was added to each of them. The contents are shaken thoroughly for 5 min and allowed to stand, so as to separate the aqueous and chloroform layer. Colored chloroform layer was separated out and absorbance was measured at 430 nm against reagent blank. Calibration curve was prepared from absorbance values so obtained.

 

Fig 1: Absorption Spectrum of Ticlopidine with BCG, BPB and BTB

 

Method B: Aliquots of the drug solution of Ticlopidine 0.5-5.0 ml (100 mcg/ml) are taken and transferred into a series of 125 ml of separating funnel. To each funnel 5 ml of BPB reagent is added. Reaction mixture was shaken gently for 5 min. Then 10 ml of chloroform was added to each of them. The contents are shaken thoroughly for 5 min and allowed to stand, so as to separate the aqueous and chloroform layer.

Colored chloroform layer was separated out and absorbance was measured at 449 nm against reagent blank. Calibration curve was prepared from absorbance values so obtained.

 

Method C: Aliquots of the drug solution of Ticlopidine 0.5-4.0 ml (100 mcg/ml) are taken and transferred into a series of 125 ml of separating funnel. To each funnel 5 ml of BTB reagent is added. Reaction mixture was shaken gently for 5 min. Then 10 ml of chloroform was added to each of them. The contents are shaken thoroughly for 5 min and allowed to stand, so as to separate the aqueous and chloroform layer. Colored chloroform layer was separated out and absorbance was measured at 427 nm against reagent blank. Calibration curve was prepared from absorbance values so obtained.

 

Preparation of sample solution:

Tablets containing Ticlopidine were successfully analyzed by the proposed methods. Twenty tablets of Ticlopidine (Tyklid, 250 mg, Torrent Pharma) were accurately weighed and powdered. Tablet powder equivalent to 100 mg of Ticlopidine was dissolved in 50 ml of methanol and filtered and washed with methanol, the filtrate and washings were combined and the final volume was made to 100 ml with methanol. The solution was suitably diluted and analyzed as given under the assay procedure for bulk samples. The results are represented in Table II. None of the excipients usually employed in the formulation of tablets interfered in the analysis of Ticlopidine, by the proposed methods.

 

Recovery Studies:

To ensure the accuracy and reproducibility of the results obtained, adding known amounts of pure drug to the previously analysed formulated samples and these samples were reanalyzed by the proposed method performed recovery experiments. The percentage recoveries thus obtained were given in Table II.

RESULTS AND DISCUSSIONS:

In the present work three methods have been developed for the estimation of Ticlopidine from Tablet formulation. The developed Methods A, B and C are based on formation of chloroform extractable yellow colored ion-pair complexes with BCG, BPB and BTB respectively. The conditions required for the formation of colored complexes were optimized. Statistical analysis was carried out and the results of which were satisfactory. Relative Standard Deviation values were low that indicates the reproducibility of the proposed methods. Recovery studies were close to 100 % that indicates the accuracy and precision of the proposed methods.

 

The optical characteristics such as absorption maxima, Beer’s law limits, molar absorptivity and Sand ell’s sensitivity are presented in Table I.

 

The regression analysis using the method of least squares was made for slope (m), intercept (b) and correlation obtained from different concentrations and the results are summarized in Table I.

 

In conclusion, the proposed methods are simple, economical, sensitive, precise reliable and reproducible for the routine estimation of Ticlopidine in bulk as well as in tablet formulation.

 

ACKNOWLEDGEMENTS:

The authors are grateful to M/s Shilpa Medicare, Raichur, Karnataka State for the supply of Ticlopidine as a gift sample and to the Management, SSJ College of Pharmacy, Hyderabad, for providing the necessary facilities to carry out the research work.

REFERENCES:

1.       The Merck Index, XIII edition, 2001, Merck Research Laboratories, (Monograph No: 6316) page 6320.

2.       GU Jun-Stokes, Wang Peng, crystal Mei-Stokes, Gu, Meiling, Wang Peng,  Beijing Polytechnic University Journal (in English), 04 2002.

3.       Rona K, Ary K, Gachalyi B, Klebovich I., J Chromatogr B Biomed Sci Appl. 1997 June 6; 693(2):393-8.

4.       Furlan G, Malusa N, Strohmayer A, Klugmann FB, Decorti G, Candussio L, Klugmann S., Farmaco. 1996 Nov; 51(11):747-51.

5.       Dal Bo L, Verga F, Marzo A, Ambrosoli L, Poli A., J Chromatogr B Biomed Appl. 1995 Mar 24;665(2):404-9

 

 

Received on 11.02.2010        Modified on 08.03.2010

Accepted on 24.03.2010        © AJRC All right reserved