DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF METHOTREXATE, DEXAMETHASONE AND INDOMETHACIN

Authors

  • Narendra K. Lariya Department of Pharmaceutical Sciences, Dr. H. S. Gour University, Sagar, M. P., India
  • G. P. Agrawal Dr. H. S. Gour University

Keywords:

RP-HPLC, Methotrexate, Dexamethasone, Indomethacin, LOD, LOQ

Abstract

Objective: To develop a reverse-phase high performance liquid chromatographic method for simultaneous estimation of methotrexate (MTX), dexamethasone (DXM) and indomethacin (IND).

Methods: A simple, isocratic reverse-phase high performance liquid chromatographic (RP-HPLC) method has been developed for simultaneous estimation of methotrexate, dexamethasone and indomethacin using ibuprofen (IBP) as internal standard with photodiode array detection at 254 nm. The process was carried out on C18 column (5 μm, 250 mm X 4.6 mm) using methanol: orthophosphoric acid (1.67 % v/v) in 70: 30 ratio at a flow rate of 1.5 ml per minute.

Results: The retention time for MTX, DXM, IND and IBP were found to be 1.47, 4.05, 11.29 and 12.69 minutes, respectively. Calibration curves of the drugs were linear in the concentration range 1-500 μg/ml. The limit of detection and limit of quantitation was 3.3 ng and 10.9 ng for MTX, 0.3 and 0.9 ng for DXM and 2.1 and 6.7 ng for IND respectively. The intra-day precision varied from 0.2 to 1.6 % and inter-day precision ranged from 0.2 to 1.8 %. The intra-day accuracy ranged from 98.3 to 101.13 % while inter-day accuracy varied from 98.6 to 101.4 %.

Conclusion: The developed method is ideally suited for simultaneous estimation of the drugs.

Downloads

Download data is not yet available.

References

Khurana R, Berney SM. Clinical aspects of rheumatoid arthritis. Pathophysiol 2005;12:153-65.

Goodman and Gilman’s, The pharmacological basis of therapeutics. 10th edition. McGraw Hill Publication; 2001. p. 705-6, 710-2, 1399-404, 1438.

Martindale. The complete drug reference. 34th edition. Pharmaceutical Press: London, UK; 2005. p. 45-50, 568-73, 1097-9.

Hirai T, Matsumoto S, Ikuo K. Simultaneous analysis of several non-steroidal anti-inflammatory drugs in human urine by high-performance liquid chromatography with normal solid-phase extraction. J Chromatogr B 1997;692:375-88.

Novakova L, Matysova L, Havlikova L, Sohei P. Development and validation of HPLC method for determination of indomethacin and its two degradation products in topical gel. J Pharm Biomed Anal 2005;37:899-905.

Hess S, Teuber U, Ortwein J, Eger K. Profiling indomethacin impurities using high-performance liquid chromatography and nuclear magnetic resonance. Eur J Pharm Sci 2001;14:301-11.

Zhang Y, Zhang Z, Qi G, Sun Y, Wei Y, Ma H. Detection of indomethacin by high-performance liquid chromatography with in situ electrogenerated Mn(III) chemiluminescence detection. Anal Chim Acta 2007;582:229-34.

Al Za’abi MA, Deghhonzadeh GH, Nonis PLG, Charles BG. A rapid and sensitive microscale HPLC method for the determination of indomethacin in plasma of premature neonates with patent ductus arteriousus. J Chromatogr B 2006;830:364-7.

Cristofol C, Percz B, Pons M, Valladares JE, Marti G, Arboix M. Determination of indomethacin residues in poultry by high performance liquid chromatography. J Chromatogr B 1998;709:310-4.

Grippa E, Santini L, Castellano G, Gatto MT, Leone MG, Saso L. Simultaneous determination of hydrocortisone, dexamethasone, indomethacin, phenylbutazone and oxyphenbutazone in equine serum by high-performance liquid chromatography. J Chromatogr B 2000;738:17-25.

Iqbal MS, Shad MA, Ashraf MW, Bilal M, Saeed M. Development and Validation of an HPLC Method for the determination of Dexamethasone, Dexamethasone sodium phosphate and chloramphenicol in presence of each other in pharmaceutical preparations. Chromatographia 2006;3(4):64-72.

Yun-Kyoung S, Jeong-Sook P, Jin-Ki K, Chong-Kook K. HPLC determination of Dexamethasone in Human Plasma. J Liquid Chromatogra Related Technol 2004;27:2293-306.

Huetos O, Ramos M, Martín de Pozuelo M, Reuvers TBA, Andres MS. Determination of dexamethasone in feed by TLC and HPLC. Analyst 1999;124:1583-7.

Garcia CV, Breier AR, Steppe M, Schapoval EES, Oppe TP. Determination of dexamethasone acetate in cream by HPLC. J Pharm Biomed Anal 2003;31(3):597-600.

Kumar V, Mostafa S, Kayo MW, Goldberg EP, Derendorf H. Determination of dexamethasone in human plasma and its application to an in vitro release study from endovascular stents. Pharm 2006;61(11):908-11.

Skibinska L, Gregorczyk J, Jarmo-owicz A. HPLC determination of Methotrexte and its metabolites in blood plasma. Chem Anal 2005;50:551-6.

Sparreboom A, Looj WJ, Nooter K, Stoler G, Verweij J. Liquid chromatographic analysis and preliminary pharmacokinetics of methotrexate in cancer patients co-treated with docetaxel. J Chromatogr B 1999;735:111-9.

Merras ID, Mansilla AE, Gomez MJR. Determination of methotrexate, several pteridines, and creatinine in human urine, previous oxidation with potassium permanganate using HPLC with photometric and Xuorimetric serial detection. Anal Biochem 2005;346:283-92.

Li H, Luo W, Zeng Q, Lui Z, Luo H, Zhang Y. Method for the determination of blood methotrexate by high performance liquid chromatography with online post-column electrochemical oxidation and fluorescence detection. J Chromatogr B 2007;845:164-8.

Floridia L, Pictropaslo AM, Terazzani M, Rubino FM, Colombi A. High-performance liquid chromatography of methotrexate for environmental monitoring of surface contamination in hospital departments and assessment of occupational exposure. J Chromatogr B 1996;726:95-103.

McCrudden EA, Tett SE. Improved high–performance liquid chromatography determination of mehotrexate and its major metabolite in plasma using a poly(styrenevinylbenzene) column. J Chromatogr B 1999;721:87-92.

Shah VP, Midha KK, Findlay JWA, Hill HM, Hulse JD, McGilveray IJ, et al. Bioanalytical method validation--a revisit with a decade of progress. Pharm Res 2000;17:1551-7.

International Conference on Harmonization, (ICH) Q2A, Validation of Analytical Procedures, Definitions and Terminology, US FDA Federal Register; 1995. p. 60.

International Conference on Harmonization, (ICH) Q2B, Validation of Analytical Procedures, Methodology, US FDA Federal Register; 1997. p. 62.

Published

01-03-2015

How to Cite

Lariya, N. K., and G. P. Agrawal. “DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATION OF METHOTREXATE, DEXAMETHASONE AND INDOMETHACIN”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 7, no. 3, Mar. 2015, pp. 443-6, https://www.innovareacademics.in/journals/index.php/ijpps/article/view/5429.

Issue

Section

Original Article(s)