BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF VARDENAFIL IN HUMAN PLASMA USING RP-HPLC

Dr.Kumaraswamy.Gandla; R.Lalitha; Dara Varun Kumar; P.VR Teja Shruthi

Dr.Kumaraswamy.Gandla Care College of Pharmacy, Oglapur (Vill),Damera,(Mdl),Warangal-Rural,(Dist)-506001.Telangana, India.
R.Lalitha Chaitanya College of Pharmacy Education and Research, Hanamkonda, Warangal, Telangana 506001.
Dara Varun Kumar Research Scholar, Career Point University, Kota, Rajasthan,-324005,India
P.VR Teja Shruthi Research Scholar, Career Point University, Kota, Rajasthan,-324005,India

Abstract

Purpose: A simple reverse phase liquid chromatographic analytical method has been developed and validated for estimation of Vardenafil in Human plasma. Methods: The separation was carried out on Eclipse XBD-C8 Plasma 5 µm ( 4.6× 150 mm) as Stationary phase, Mobile Phase: Acetonitrile: Aqueous solution containing 0.012M triethyl amine + 0.020M orthophosphoric acid. (40:60) Flow rate: 0.5 mL/min using uv detector at 225 & 285 nm. Results: The described HPLC method was linear over a concentration range of 100-3200 ng/mL. Sildenafil was used as a internal standard. The Vardenafil showed retention time 9.4 min & sildenafil showed retention time 3.7min respectively. The Accuracy and Precision, recovery, selectivity for Vardenafil was 100-3200 ng/mL respectively. The stability of thedrug spiked human plasma samples during three freeze thaw cycles were stable in plasma for about one month when stored at frozen state. Conclusions: It can be concluded that the developed bioanalytical method is capable of quantifying Vardenafil from spiked human plasma samples. The method meets the requirements of the USFDA Guidelines and can be applied to Bioavailability/ Bioequivalence studies of Vardenafil.

Keywords: Bioanalytical method, development, validation

Downloads

Download data is not yet available.

References

Kirthi, A., Shanmugam, R., Prathyusha, S.M. and Basha, J., A review on bioanalytical method development and validation by RP-HPLC. Journal of Global Trends in Pharmaceutical Sciences, 2014; 5(4): 2265-2271.

Bolton Sanford, “Pharmaceutical Statistics Practical and Clinical Applications”, Third Edition, Marcel Dekker Inc, 1984; 216-257, 265-315.

Braggio, S., Barnaby, R.J., Grossi, P. and Cugola, M., A strategy for validation of bioanalytical methods. Journal of pharmaceutical and biomedical analysis, 1996; 14(4): 375-388.

Snyder, L.R., Kirkland, J.J. and Glajch, J.L., Practical HPLC method development. John Wiley & Sons, 2012; 736: 77-84, 643-678.

Ramakrishna, N.V.S., Vishwottam, K.N., Puran, S., Koteshwara, M., Manoj, S., Santosh, M., Chidambara, J., Wishu, S. and Sumatha, B., Quantitation of Vardenafil in human plasma by liquid chromatography–tandem mass spectrometry with electrospray ionization. Journal of Chromatography B, 2004; 809(2): 243-249.

Shakya, A.K., Abu-Awwad, A.N., Arafat, T.A. and Melhim, M., Validated liquid chromatographic–ultraviolet method for the quantitation of Vardenafil in human plasma using liquid–liquid extraction. Journal of Chromatography B, 2007; 852(1-2):
403-408.

Karavadi, T. and Challa, B.R., Determination of Vardenafil in rat plasma by liquid chromatography tandam mass spectrometry: Application to a pharmacokinetic study. Der Pharmacia Lettre, 2012; 4: 1401-1413.

Nikolaou, P., Papoutsis, I., Athanaselis, S., Alevisopoulos, G., Khraiwesh, A., Pistos, C. and Spiliopoulou, C., Development and validation of a GC/MS method for the determination of Vardenafil in whole blood. Journal of pharmaceutical and biomedical analysis 2011; 56(3): 577-581.

Khan, Z.G., Patil, A.S. and Shirkhedkar, A.A., Estimation of Vardenafil using derivative spectrophotometry in bulk material and in pharmaceutical formulation. International Journal of Spectroscopy, 2014.

Evans, G., A handbook of bioanalysis and drug metabolism. CRC press, 2004; 8-69.

Tiwari, G. and Tiwari, R., Bioanalytical method validation: An updated review. Pharmaceutical methods, 2010; 1(1): 25.

Sonawane Lalit V. et al, “Bioanalytical Method Validation and Its Pharmaceutical Applications- A Review”, Pharmaceutica Analityca Acta, 2014; 05(03): 1-7.

Hartmann, C., Smeyers-Verbeke, J., Massart, D.L. and McDowall, R.D., Validation of bioanalytical chromatographic methods. Journal of pharmaceutical and biomedical analysis, 1998; 17(2): 193-218.

Heftmann, E. ed., Chromatography: Fundamentals and applications of chromatography and related differential migration methods-Part B: Applications. Elsevier, 2004.

Kelley, M. and DeSilva, B., Key elements of bioanalytical method validation for macromolecules. The AAPS journal, 2007; 9(2): E156-E163.

Nowatzke, W. and Woolf, E., Best practices during bioanalytical method validation for the characterization of assay reagents and the evaluation of analyte stability in assay standards, quality controls, and study samples. The AAPS journal, 2007; 9(2): E117-E122.

Liu, G., Snapp, H.M., Ji, Q.C. and Arnold, M.E., Strategy of accelerated method development for high-throughput bioanalytical assays using ultra high-performance liquid chromatography coupled with mass spectrometry. Analytical chemistry, 2009; 81(22): 9225-9232.