• Narottam Pal Department of Pharmaceutical Analysis, Bhaskar Pharmacy College, Yenkapally, Moinabad, Hyderabad, India, 50075
  • Avanapu Srinivasa Rao Department of Pharmaceutical Analysis, Bhaskar Pharmacy College, Yenkapally, Moinabad, Hyderabad, India, 50075
  • Pigilli Ravikumar Bio-analytical Department, Aizant Drug Research Solution Pvt. Ltd, Hyderabad, India, 50014


Objective: To develop a new method and validate the same for the determination of Febuxostat (FBS) in human plasma by liquid chromatography–mass spectrometry (LCMS).

Methods: The present method utilized reversed-phase high-performance liquid chromatography with tandem mass spectroscopy. Febuxostat D9 (FBS D9) was used as internal standard (IS). The analyte and internal standard were separated from human plasma by using solid phase extraction method. Zorbax Eclipse XDB, C8, 100 mm x 4.6 mm, 3.5 µm column was used and HPLC grade acetonitrile, 5 millimolar (mM) ammonium format (80: 20, v/v) as mobile phase, detected by mass spectrometry operating in positive ion and multiple reaction monitoring modes.

Results: The parent and production transitions for FBS and internal standard were at m/z 317.1→261.0 and 326.1→262.0 respectively. The method was validated for system suitability, specificity, carryover effect, linearity, precision, accuracy, matrix effect, sensitivity and stability. The linearity range was from 20.131 ng/ml to10015. 534 ng/ml with a correlation coefficient of 0.999. Precision results (%CV) across six quality control samples were within the limit. The percentage recovery of FBS and internal standard from matrix samples was found to be 76.57% and 75.03% respectively.

Conclusion: Present study describes new LC-MS method for the quantification of FBS in a pharmaceutical formulation. According to validation results, it was found to be a simple, sensitive, accurate and precise method and also free from any kind of interference. Therefore the proposed analytical method can be used for routine analysis for the estimation of FBS in its formulation.

Keywords: Febuxostat, Febuxostat D9, Liquid chromatography, Mass spectrometry


Download data is not yet available.


1. Laurence L Brunton, John S LGZO, Keith L Parker. Goodman and Gilman's The Pharmacological Basis of therapeutics. 11th ed. New York: McGraw-Hill; 2006. p. 706-11.
2. Roger Walker, Cate Whittlesea. Clinical pharmacy and therapeutics. 5th ed. Churchill Livingstone; 2012. p. 848-57.
3. Richard Finkel, Michelle A Clark, Luigi X Cubeddu. Lippincott’s Illustrated Reviews: Pharmacology. 5th ed. Wolter kluwer (India Pvt. Ltd.); 2013. p. 515-7.
4. HP Rang, MM Dale, JM Ritter, RJ Flower. Rang and dale pharmacology. 6th ed. Churchill Livingstone; 2008. p. 238-9.
5. Grewa HK, Martinez JR, Espinoza LR. Febuxostat: drug review and update. Expert Opin Drug Metab Toxicol 2014;10:747-58.
6. Diaz-Torné C, Perez-Herrero N, Perez-Ruiz F. New medications in development for the treatment of hyperuricemia of gout. Curr Opin Rheumatol 2015;27:164-9.
7. Baker JF, Schumacher HR. Update on gout and hyperuricemia. Int J Clin Pract 2010;64:371-7.
8. Chou G. An update on the treatment options for gout and calcium pyrophosphate deposition. Expert Opin Pharmacother 2005;6:2443-53.
9. Paramdeep Bagga, Mohd Salman, HH Siddiqui, Abdul M Ansari, Tariq Mehmood, Kuldeep Singh. A simple UV spectrophotometric method for the determination of Febuxostat in bulk and pharmaceutical formulation. Int J Pharm Sci Res 2011;2:2655-9.
10. Sowjanya G, Srinivas L, Rajasri A. New validated UV spectrophotometric methods for the determination of Febuxostat in bulk and formulation. CIBTech J Pharm Sci 2014;3:7-13.
11. Mukthinuthalapati MA, Bandaru SP, Bukkapatnam V, Mohapatro C. Development and validation of a stability-indicating RP-HPLC method for the determination of Febuxostat (a xanthine oxidase inhibitor). J Chromatogr Sci 2013;51:931-8.
12. Jyothirmai B, Tata Santosh, Syama Sundar B. Development and validation of an RP-HPLC method for the determination of Febuxostat in human plasma. Indo Am J Pharm Res 2014;4:1040-8.
13. Sudhir S Muvvala, Venkata Nadh Ratnakaram, Rama Rao Nadendla. A validated RP-HPLC method for the estimation of Febuxostat in bulk drugs. Int J PharmTech Res 2012;4:1358-66.
14. BRC Sekhar Reddy, Nallagatla Vijaya Bhaskar Rao. A stability indicating RP-HPLC method development for the determination of Febuxostat in tablet dosage form. Caribbean J Sci Technol 2013;1:228-37.
15. K Nageswara Rao, S Ganapaty, A Lakshmana Rao. Development and validation of RP-HPLC method for estimation of Febuxostat in bulk and tablet dosage form. Int J Res Pharm Chem 2012;2:1104-8.
16. Monita Gide, Pankaj Sharma, Ravindra Saudagar, Birendra Shrivastava. Method development and validation for determination of febuxostat from spiked human plasma using RP-HPLC with UV detection. Chromatogr Res Int 2014;1-6.
17. Ojikumar Lukram, Shivaji Parmar, Amit Hande. Determination of febuxostat in human plasma using ultra-performance liquid chromatography tandem mass spectrometry. Drug Test Anal 2013;5:492–9.
18. Yingli Wu, Zhengsheng Mao, Youping Liu, Xin Wang, Xin Di. Simultaneous determination of febuxostat and its three active metabolites in human plasma by liquid chromatography–tandem mass spectrometry and its application to a pharmacokinetic study in Chinese healthy volunteers. J Pharm Biomed Anal 2015;114:216–21.
19. Babu Rao Chandu, Kanchanamala Kanala, Nagiat T Hwisa, Prakash Katakam, Mukkanti Khagga. Bioequivalence and pharmacokinetic study of Febuxostat in human plasma by using LC-MS/MS with liquid-liquid extraction method. Springer Plus 2013;2:194.
428 Views | 846 Downloads
How to Cite
Pal, N., A. S. Rao, and P. Ravikumar. “NEW METHOD DEVELOPMENT AND VALIDATION FOR THE DETERMINATION OF FEBUXOSTAT IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY–MASS SPECTROMETRY”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 8, no. 9, Sept. 2016, pp. 61-70, doi:10.22159/ijpps.2016v8i9.11968.
Original Article(s)