HPLC METHOD DEVELOPMENT AND VALIDATION: SIMULTANEOUS DETERMINATION OF ACTIVE INGREDIENTS IN COUGH AND COLD PHARMACEUTICALS
Objective: This study aimed to develope a simple reversed-phase high performance chromatographic method for simultaneous determination of pseudoephdrine HCI, pheniramine maleate, acetaminophen, guaifenisin, pyrilamine maleate, chlorpheniramine maleate, triprolidine HCI, dextromethorphan HBr, diphenhydramine HCI in cough and cold pharmaceuticals.
Methods: The separation of these compounds was achieved within 37.9 min on a Nucleodur gravity C18 column (250 x 4.0 mm, 5Î¼m). The chromatographic separation of these compounds performed in a single run by using isocratic mobile phase consisting of methanol:buffer mixture (38:62, v/v) at room temperature, with flow rate of 0.75 mL.min-1.
An ultraviolet absorption at 210 nm was monitored. 2,4,6-trimethoxybenzaldehyde was used as an internal standard (ISTD). The selectivity, linearity of calibration, accuracy, intraday and interday precision and forced degradation studies were examined as parts of the method validation.
Results: The concentrationâ€“response relationship was linear over a concentration range of 0.2-250 Âµg.mL-1 for acetaminophen, 0.5â€“250 Âµg.mL-1 for pseudoephdrine HCI and pheniramine maleate, 1â€“250 Âµg.mL-1 for guaifenisin, 2.5-250 Âµg.mL-1 for chlorpheniramine maleate and triprolidine HCI, 5-250 Âµg.mL-1 for pyrilamine maleate and diphenhydramine HCI, 10-20 Âµg.mL-1 for dextromethorphan HBr with correlation coefficients better than 0.9993. The relative standard deviations of the intraday and interday were all less than 4%. Conclusion: The proposed liquid chromatographic method was successfully applied for the routine analysis of these compounds in different cough and cold pharmaceutical preparations such as syrups and tablets.
2. Dong YM, Chen XF, Chen YL, Chen XG, Hu ZD. Separation and determination of pseudoephedrine, dextromethorphan, diphenhydramine and chlorpheniramine in cold medicines by nonaqueous capillary electrophoresis. J Pharm Biomed 2005;39(1-2):285-9.
3. Gasco Lopez AI, IzquierdoHornillos R, Jiminez A. Development and validation of a high-performance liquid chromatography method for the determination of cold relief ingredients in chewing gum. J Chromatogr A 1997;775(1-2):179-85.
4. Galli V, Barbas C. High-performance liquid chromatographic analysis of dextromethorphan, guaifenesin and benzoate in a cough syrup for stability testing. J Chromatogr A 2004;1048(2):207-11.
5. Marin A, Garcia E, Garcia A, Barbas C. Validation of a HPLC quantification of acetaminophen, phenylephrine and chlorpheniramine in pharmaceutical formulations: capsules and sachets. J Pharm Biomed 2002;29(4):701-14.
6. Shabir GA, Arain SA. Determination of paracetamol, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, and propyl parahydroxybenzoate in medicinal suspension and tablets by rplc/uv-dad. J Liq Chromatogr R T 2011;34(9):719-29.
7. Kompany-Zareh M, Mirzaei S. Spectrophotometric resolution of ternary mixtures of pseudoephedrine hydrochloride, dextromethorphan hydrobromide, and sodium benzoate in syrups using wavelength selection by net analyte signals calculated with hybrid linear analysis. Anal Chim Acta 2004;526(1):83-94.
8. Bolser DC, Hey JA, Chapman RW. Influence of central antitussive drugs on the cough motor pattern. J Appl Physiol 1999;86(3):1017-24.
9. Grattan TJ, Marshall AE, Higgins KS, Morice AH. The effect of Ä±nhaled and oral dextromethorphan on citric-acid Ä±nduced cough in man. Brit J Clin Pharm 1995;39(3):261-3.
10. Manap RA, Wright CE, Gregory A, Rostami-Hodjegan A, Meller ST, Kelm GR, et al. The antitussive effect of dextromethorphan in relation to CYP2D6 activity. Brit J Clin Pharm 1999;48(3):382-7.
11. Matthys H, Bleicher B, Bleicher U. Dextromethorphan and codeine-objective assessment of antitussive activity in patients with chronic cough. J Int Med Res 1983;11(2):92-100.
12. Convention TUSP, United States Pharmacopoeia, The National Formulary. Rockville; 2002.
13. Goodman G, The Pharmacological Basis of Therapeutics. New York: McGraw-Hill;1996.
14. Ravisankar S, Vasudevan M, Gandhimathi M, Suresh B. Reversed-phase HPLC method for the estimation of acetaminophen, ibuprofen and chlorzoxazone in formulations. Talanta 1998;46(6):1577-81.
15. Celma C, Allue JA, Prunonosa J, Peraire C, Obach R. Simultaneous determination of paracetamol and chlorpheniramine in human plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr A 2000;870(1-2):77-86.
16. Vasudevan M, Ravisankar S, Sathiyanarayanan A, Chandan RS. Simultaneous estimation of phenylpropanolamine HCl, guaiphenesin and diphenylpyraline HCl in syrups by LC. J Pharm Biomed 2000;24(1):25-31.
17. Franeta JT, Agbaba D, Eric S, Pavkov S, Aleksic M, Vladimirov S. HPLC assay of acetylsalicylic acid, paracetamol, caffeine and phenobarbital in tablets. Farmaco 2002;57(9):709-13.
18. Hood DJ, Cheung HY. A chromatographic method for rapid and simultaneous analysis of codeine phosphate, ephedrine HCl and chlorpheniramine maleate in cough-cold syrup formulation. J Pharm Biomed 2003;30(5):1595-601.
19. Qi ML, Wang P, Zhou L, Gu JL, Fu RN. Simultaneous determination of acetaminophen, dextromethorphen hydrobromide and pseudoephedrine hydrochloride in a new drug formulation for cold treatment by HPLC. Chromatographia 2003;57(3-4):139-42.
20. El-Gindy A, Emara S, Mostafa A. Application and validation of chemometrics-assisted spectrophotometry and liquid chromatography for the simultaneous determination of six-component pharmaceuticals. J Pharm Biomed 2006;41(2):421-30.
21. Palabiyik IM, Onur F. The simultaneous determination of phenylephrine hydrochloride, paracetamol, chlorpheniramine maleate and dextromethorphan hydrobromide in pharmaceutical preparations. Chromatographia 2007;66:S93-S6.
22. Manassra A, Khamis M, El-Dakiky M, Abdel-Qader Z, Al-Rimawi F. Simultaneous HPLC analysis of pseudophedrine hydrochloride, codeine phosphate, and triprolidine hydrochloride in liquid dosage forms. J Pharm Biomed 2010;51(4):991-3.
23. Louhaichi MR, Jebali S, Loueslati MH, Adhoum N, Monser L. Simultaneous determination of pseudoephdrine, pheniramine, guaifenisin, pyrilamine, chlorpheniramine and dextromethorphan in cough and cold medicines by high performance liquid chromatography. Talanta 2009;78(3):991-7.
24. Rowe RC, Sheskey PJ, Cook WG, Fenton ME. Handbook of Pharmaceutical Excipients. UK: Pharmaceutical Press; 2009.
25. Mutton I, Boughtflower B, Taylor N, Brooke D. The design and use of a simple System Suitability Test Mix for generic reverse phase high performance liquid chromatography-mass spectrometry systems and the implications for automated system monitoring using global software tracking. J Chromatogr A 2011;1218(23):3711-17.
26. Zakeri-Milani P, Barzegar-Jalali M, Tajerzadeh H, Azarmi Y, Valizadeh H. Simultaneous determination of naproxen, ketoprofen and phenol red in samples from rat intestinal permeability studies: HPLC method development and validation. J Pharm Biomed 2005;39(3-4):624-30.
27. ICH Steering Committee,Validation of Analytical Procedures:Text and Methodology Q2(R1); 2005. p. 1-17.