DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR PHENYTOIN SODIUM AND PHENOBARBITONE IN BULK AND PHARMACEUTICAL DOSAGE FORM
Keywords:Phenytoin Sodium, Phenobarbitone, RP-HPLC, Method Development, Validation
Objective: To develop an accurate, simple, rapid, precise, and linear RP-HPLC method for the simultaneous estimation of phenytoin sodium and phenobarbitone in tablet dosage form and validated as per ICH guidelines.
Methods: The method used was a reverse phase HPLC (RP-HPLC) method using Hypersil BDS C18, (250Ã—4.6 mm, 5 Î¼m) column, mobile phase comprising of methanol: phosphate buffer (pH 5.0) (50:50), flow rate of 1.0 ml/min and a detection wavelength of 215 nm using a UV detector.
Results: The retention time for phenytoin sodium and phenobarbitone was found to be 3.97 min and 6.90 min, respectively. The linearity of developed method was achieved in the range of 10-30 Î¼g/ml for phenytoin sodium and 3-9 Î¼g/ml for phenobarbitone. The detection (LOD) and quantitation (LOQ) limits were 1.44 and 4.36 Âµg/ml for phenytoin sodium and 0.40 and 1.35 Âµg/ml for phenobarbitone respectively.
Conclusion: A simple, accurate, precise, linear and rapid RP-HPLC method was developed for simultaneous quantitative estimation of phenytoin sodium and phenobarbitone in bulk and pharmaceutical formulation. The method was validated as per ICH guidelines. Hence, the method holds good for the routine analysis of phenytoin sodium and phenobarbitone in various pharmaceutical industries as well as in academics.
https://en.wikipedia.org/wiki/Phenytoin. [Last assessed on 15 Nov 2016]
http://monographs.iarc.fr/ENG/Monographs/vol66/mono66-13.pdf. [Last assessed on 15 Nov 2016]
http://www.drugbank.ca/drugs/DB00252. [Last assessed on 15 Nov 2016]
Soumya R, Sarayu P, Srikanth MS, Ramesh A, Keshava BS. Drug interaction induced phenytoin toxicity: a case report. Asian J Pharm Clin Res 2014;7 Suppl 1:1-2.
http://www.drugbank.ca/drugs/DB01174. [Last assessed on 15 Nov 2016]
https://en.wikipedia.org/wiki/Phenobarbital#Mechanism_of_action. [Last assessed on 15 Nov 2016]
http://monographs.iarc.fr/ENG/Monographs/vol79/mono79-11.pdf. [Last assessed on 15 Nov 2016]
http://www.drugs.com/drug-interactions/dilantin-with-phenobarbital-1863-1205-1846-0. html?professional=1. [Last assessed on 15 Nov 2016]
http://www.drugsupdate.com/generic/view/636/Phenobarbital-+-Phenytoin. [Last assessed on 15 Nov 2016]
Siew YT, Michael JR, Allan GAC, Siang Y, Seng NG. Development and validation of a stability-indicating isocratic reverse phase-liquid chromatography assay for determination of phenytoin in bulk and pharmaceutical formulations. Int J Pharm Pharm Sci 2015;7:258-63.
Varaprasad A, Sriram N, Godwin Isaac Blessing A, Jawahar M, Thangamuthu S. Method development and validation of phenytoin sodium in bulk and its pharmaceutical dosage form by RP-HPLC method. Int J Bio Pharm Res 2012;3:126-9.
Muralee K, Meghana N, Aniruddha VS, Ranjith R. Stability indicating RP-HPLC method validation for the assay of phenytoin sodium in phenytoin sodium capsules. J Chem Pharm Res 2015;7:230-6.
Muralee K, Meghana N, Aniruddha VS, Ranjith R. Stability indicating analytical method validation for determination of related substances by RP HPLC for phenytoin sodium in phenytoin sodium capsules. Int J PharmTech Res 2015;8:78-87.
Ravichandran S, Valliappan K, Ramanathan M. Validated RP-HPLC method for concurrent determination of phenytoin sodium and clopidogrel bisulphate in tablet dosage form. J Pharm Sci Res 2015;7:934-7.
Jeyaprakash MR, Sireesha V, Meyyanathan SN. Development and validation for the estimation of benzil impurity in phenytoin formulations by reverse phase HPLC. Asian J Pharm Anal Med Chem 2013;1:79-87.
Hisham H, Ayman AG, Hanna S. Development and validation of rapid stability indicating HPLC-determinations of antiepileptic drugs phenobarbital in suppositories and phenytoin in capsules as well as in urine sample. J Liq Chromatogr Relat Technol 2013;36:2292-306.
Thiyagu R, Rajendran SD, Satish G, Arulmani R, Karthik A. Simultaneous liquid chromatographic analysis of phenobarbital, phenytoin and carbamazepine in human serum an application to therapeutic drug monitoring. Newsletter-Pharmacologyonline 2010;1:512-21.
Serralheiro A, Alves G, Fortuna A, Rocha M, Falcao A. First HPLC-UV method for rapid and simultaneous quantification of phenobarbital, primidone, phenytoin, carbamazepine, carbamazepine-10,11-epoxide, 10,11-Trans-dihydroxy-10,11-Dihydrocarbamazepine, lamotrigine, oxcarbazepine and licarbazepine in human plasma. J Chrom B 2013;925:1-9.
Cristina S, Karin VL, Celia EO, Maria AB, Maria DP, Silvia RCJS. Micromethod for quantification of carbamazepine, phenobartital and phenytoin in human plasma by HPLC-UV detection for therapeutic drug monitoring application. Latt Am J Pharm 2008;27:485-91.
Ouakouak H, Ben MM, Ben CM, Abdelhamid Z. Phenobarbital analysis in biological matrix (Blood) by high performance liquid chromatography (HPLC). Int Lett Chem Phys Astron 2014;20:31-40.
International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human use. Validation of Analytical Procedures; Text and Methodology ICH Q2 (R1); 2005.
Tijare LK, Rangari NT, Mahajan UN. A review on bioanalytical method development and validation. Asian J Pharm Clin Res 2016:9:6-10.
Govindarao K, Sowjanya V, Naga VK. Development and validation of RP-HPLC method for simultaneous estimation of lamivudine and zidovudine in bulk. Int J Curr Pharm Res 2016;1:28-33.