Publication Ref No.

: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-7/SEP/012

ISSN 0974 9446

TO DEVELOP UV SPECTROPHOTOMETRIC METHOD FOR THE ESTIMATION OF SILDENAFIL CITRATE IN BULK AND TABLET Ashok Kumar1*,Gaurav Kumar Rajput2 Gaurav Srivastava3,Atul Kumar Singh4 Department of Biotechnology, HNBGU (A Central University), Srinagar- Garhwal-246174 (UK) INDIA 2 Ashok Kumar Department of Quality Control,Cooper Pharma Ltd. Selaqui, Dehradun-248002 (UK) INDIA 3 Uttarakhand Technical University,Suddhowala Dehradun-248007 (Uttarakhand) 4 H. N. B. Garhwal University Srinagar Garhwal-246174 (Uttarakhand), India Email: asokumr@gmail.com ABSTRACT A simple, sensitive, highly accurate UV spectrophotometric method has been developed for the determination of Sildenafil citrate in bulk and tablet dosage form. Solution of sildenafil citrate in methanol shows maximum absorbance at 291 nm. Beers law was obeyed in the concentration range of 5-30 g mL-1 correlation coefficients, detection and quantitation limits were also calculated. The proposed method has been applied successfully for the analysis of the drug in pure and in its tablets dosage forms. Result of percentage recovery and placebo interference showed that the method was not affected by the presence of common excipients. The percentages assay of sildenafil citrate in tablet was 100 mg of 99.87% and 50 mg of 99.84%. The method was validated by determining its sensitivity, accuracy and precision which proves suitability of the developed method for the routine estimation of Sildenafil in bulk and tablets. Key Words: Sildenafil citrate, UV spectroscopy, Estimation, Tablets.
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INTRODUCTION Sildenafil citrate (SC) is designated chemically as 1-[[3-(6, 7-dihydro-1-methyl-7-oxo-3-propyl1H-pyrazole [4, 3-d] pyromidine-5-yl)-4-

ethoxyphenyl] sulfonyl]-4 methylpiperazine citrate. The physiological mechanism of erection of the penis involves release of nitric oxide (NO) in the corpus cavernosum during sexual stimulation. Then, NO activates the enzyme

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ISSN 0974 9446

guanylate cyclase, which results in increased levels of cyclic guanosine monophosphate (cGMP), production of smooth muscle relaxation in the corpus cavernosum and inflow of blood. SC has no direct relaxant effect on isolated human corpus cavernosum, but enhances the effect of NO by inhibiting phosphodiesterase type 5 (PDES), which is responsible for degradation of cGMP in the corpus cavernosum (1, 2, and 3). When sexual stimulation causes local release of NO, inhibition of PDES by SC causes increased levels of cGMP in the corpus cavernosum, resulting in smooth muscle relaxation and inflow to the corpus cavernosum (2, 4, and 5).

Spectro Scan 80DV UV-Visible double beam spectrophotometer with 1 cm matched quartz cell (7). 2. Sildenafil stock solution: Standard stock solution was prepared by dissolving 70.25 mg of Sildenafil in 100 mL of methanol to get concentration of 500 g/mL solution (7). 3. Method development: Aliquots of stock solution were further diluted with methanol to get working solution of 5, 10, 15, 20, 25 and 30g mL-1. The working standards were scanned between 200-400 nm. The same max was used for the further measurement of the drug. 4. Procedure for calibration curve: Aliquots of stock solution were further diluted with Methanol to get working solution of 5, 10, 15, 20, 25 and 30 g mL-1. Finally, the prepared standards were measured after standing for 5.0 min at max as recorded in each case against a solvent blank similarly prepared. A calibration graph of the absorbance versus the concentration of the drug was plotted. Procedure for dosage forms: For analysis of commercial formulations, twenty tablets were taken and powdered. Tablet powder equivalent to 140.5 mg of sildenafil citrate was transferred to 100 mL volumetric flask and dissolved in methanol. Then the solution was sonicated for 15 min and filtered and it was for further diluted to get the required concentration. The absorbance of the prepared sample solution was measure against methanol blank at 2912 nm. A standard additions technique was also used to confirm the accuracy and precisions (7, 8).

Figure 1. Structure of Sildenafil Citrate. objective of the present work was to develop a simple, rapid, accurate and sensitive UV spectrophotometric method for the determination of sildenafil in pharmaceutical formulation. MATERIAL AND METHODS Pharmaceutical grade of sildenafil citrate was procured from Cooper Pharma Limited, Dehradun. All the chemicals were of analytical reagent grade of Merck (Germany) unless otherwise specified. Doubly distilled water was used to prepare all solutions. Freshly prepared solutions were always employed. Different brands of tablets of sildenafil were supplied from local stores. 1. Instrumentation:

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RESULTS AND DISCUSSION The absorption spectrum of sildenafil was measured in the range 200400 nm against the blank methanol [Figure 1]. The working standards were scanned between 200-400 nm which showed the maximum absorbance at 291 nm [Figure 2]. The curve for 10 ppm methanol and 10 ppm HCl was given in figure 3 and 4. The standard solution show maximum absorbance at max for each three systems as recorded in Table 1. And the method was validated by studying the following parameters as ICH guide lines for method validation (9). ( Please Refer Table No. 04 at bottom) The precision of the method was investigated with respect to repeatability. For intra-day precision, standard solution of fixed concentration was analyzed at various time interval and %RSD was noted (limit %RSD<2.0%). And the day-to-day precision was studied by taken the absorbance of the same concentration of standard solution at various days and the %RSD was calculated (%RSD<2.0%) as shown in Table 2.( Please Refer Table No. 02 at bottom) The specificity of the method was conducted to prove that the free from determined interference of solvent and commonly used tablet excipients. This is evidenced by the lack of absorbance at the specified max for the excipients in the placebo and blank solutions. The applicability of the proposed method for the assay of sildenafil in tablet formulation was examined by analyzing formulations and the results were tabulated in Table 2. The results obtained were good agreement with the label claims. The results were reproducible with low %RSD values. The results of analysis of the commercial tablets and the recovery study of drug suggested that there is no interference

from any excipients (such as starch, lactose, titanium dioxide, and magnesium stearate) which are commonly present in tablets. CONCLUSION A method for the determination of sildenafil in the bulk drug and tablet formulation has been developed. From the spectrum of sildenafil citrate as shown in Figure 2, it was found that the maximum absorbance is at about 291 nm in methanol. A good linear relationship (0.9998) was observed between the concentration ranges of 5-30 g/mL. The assay of sildenafil tablet was found to be 100 mg of 99.87% and 50 mg of 99.84%. The high percentage recovery indicates the high accuracy of the method. This demonstrates that the developed spectroscopic method is simple, accurate and reproducible. Thus the developed method can be easily used for the routine quality control of sildenafil in bulk and tablet dosage form. ACKNOWLEDGEMENTS The authors are thankful to Cooper Pharma Limited (Labs), Dehradun for providing gift sample of sildenafil. Authors are also grateful to Mrs. K.P. Rathoure for technical support. REFERENCES:

1. Boolell M, Allen MJ, Ballard SA, GepiAttee S, Muirhead GJ, Naylor AM, Osterloh IH, Gingell C (1996). Sildenafil: an orally active type 5 cyclic GMP-specific phosphodiesterase inhibitor for the treatment of penile erectile dysfunction. Int J Impot Res 8 (2): 4752. 2. Dinesh, N. D., Nagaraja, P., Made Gowda, N. N. and Rangappa, K. S. (2002). Extractive spectrophotometric methods for the assay of sildenafil citrate (Viagra) in pure form

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and in pharmaceutical formulations. Talanta 57: 757-764. 3. Nicholas K. Terrett, Andrew S. Bell, David Brown and Peter Ellis. (1996). Sildenafil (Viagra), a potent and selective inhibitor of Type 5 cGMP phosphodiesterase with utility for the treatment of male erectile dysfunction. Bioorg Med Chem Lett 6: 18191824. 4. Webb, D.J.; Freestone, S.; Allen, M.J.; Muirhead, G.J. (1999), Sildenafil citrate and blood-pressure-lowering drugs: results of drug interaction studies with an organic nitrate and a calcium antagonist. Am. J. Cardiol. 83 (5A): 21C28C. 5. Jean-Paul Richalet, Pierre Gratadour, Paul Robach, Isabelle Pham, Michle Dchaux, Aude Joncquiert-Latarjet, Pascal Mollard, Julien Brugniaux and Jrmy Cornolo. (2005), Sildenafil inhibits altitude-induced hypoxemia

and pulmonary hypertension. Am. J. Respir. Crit. Care Med. 171 (3): 27581. 6. K. Harikrishna, B. S. Nagaralli and J. Seetharamappa (2008). Extractive Spectrophotometric Determination of Sildenafil Citrate (Viagra) in Pure and Pharmaceutical Formulations. J. Food & Drug Anal, 16 (1): 1117. 7. Vogel, A. I. (1969). A text book of quantitave in-organic anlysis. 3rd ed. p. 35. ELBS and Longman. 8. Analytical Methods Committee. (1987), Recommendations for the definition, estimation and use of the detection limit. Analyst 112: 199204 9. ICH draft Guidelines on Validation of Analytical Procedures (1995); Definitions and Terminology, Federal Register, 60, IFPMA, Switzerland: 1260.

TABLES AND FIGURES: Table 1: Parameters for determination of sildenafil citrate against methanol Parameters A max (nm) Beers law limit (g mL-1) Correlation coefficient (r) Values 291 2 5 30 0.9999

Table 2: Results of Assay and Precision Studies Sample Label Amoun (%)* claim, t mg/tab found, mg /tab 100 99.87 99.87 mg 0.1890 0.1890 50 mg 49.92 99.84 0.1958 0.1986 C.V.* Precision** Interday Repeatabili Intraday ty 0.326 0.511 0.701

Sildenafil citrate Sildenafil citrate

0.1514

0.1689

0.312

0.507

0.731

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ISSN 0974 9446

Figure 1. Standard plot of Sildenafil Citrate.

0.9 0.8 0.7 Absorbance 0.6 0.5 0.4 0.3 0.2 0.1 0 5 10 15 20 25 Concentration ppm 30 0.137 0.274 0.411 0.548

Standard Curve
0.685

0.822

Figure 2. UV spectra of Sildenafil Citrate in Methanol 20 ppm.

Figure 3: UV Spectra for Sildenafil Citrate in Methanol 10 ppm.

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Figure 4. UV spectra of Sildenafil Citrate in HCl 10 ppm.

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