

Review On: RP-HPLC
Abstract
Pharmaceutical analysis plays a vital role in determining the qualitative and quantitative composition of materials, essential for understanding sample characteristics. Analytical chemistry divides into two branches qualitative, which identifies components, and quantitative, which measures them. In recent years, high-performance liquid chromatography (HPLC) has gained prominence for its ability to separate, identify, and quantify various components in biological fluids, pharmaceutical formulations, and drug samples. Notably, reversed-phase chromatography (RP-HPLC) is widely used due to its efficacy in analyzing molecules with hydrophobic properties, such as proteins, peptides, and nucleic acids.
RP-HPLC method development involves optimizing chromatographic parameters, selecting appropriate stationary and mobile phases, and adjusting conditions to achieve accurate separation of analytes. Validation of RP-HPLC methods is crucial, ensuring reliability, consistency, and compliance with International Council for Harmonisation (ICH) guidelines. Key validation parameters include accuracy, precision, specificity, detection and quantitation limits, linearity, robustness, and ruggedness.
The importance of RP-HPLC method validation extends to quality control in pharmaceuticals, minimizing product failures and production costs while enhancing productivity and quality. With well-established protocols, validated methods improve reliability across laboratories, ensuring robust and reproducible results. Overall, RP-HPLC and its validation are instrumental in maintaining high standards in pharmaceutical analysis, enhancing product quality, and achieving efficient and accurate separation and measurement of pharmaceutical compounds.
References
High performance liquid chromatography [Internet]. 2009 [Accessed 2009 Jan 20]. Available from: en. wikipedia.org/wiki/file:Agilent1200HPLC.jpg.html.
Renlund S, Erlandsson I, Hellman U, Silberring J, Wernstedt C, Lindström L, Nyberg F: Micropurification and amino acid sequence of -casomorphin-8 in milk from a woman with postpartum psychosis Peptides. 1993;14 :1125-1132.
Adeyeye, E.; Kapil, V.; Lobo, M.D. Hypertension. Medicine (Baltimore). 2022, 50, 399–407. doi:10.1016/j.mpmed. 2022.04.002.
Rimoldi, S.F.; Messerli, F.H.; Chavez, P.; Stefanini, G.G.; Scherrer, U. Efficacy and Safety of Calcium Channel Blocker/Diuretics Combination Therapy in Hypertensive Patients: A Meta-Analysis. J. Clin. Hypertens. 2015, 17, 193–199. doi:10.1111/jch.12462
James, P.A.; Oparil, S.; Carter, B.L.; Cushman, W.C.; Dennison-Himmelfarb, C.; Handler, J.; Lackland, D.T.; LeFevre, M.L.; MacKenzie, T.D.; Ogedegbe, O.; Smith, S.C.; Svetkey, L.P.; Taler, S.J.; Townsend, R.R.; Wright, J.T.; Narva, A.S.; Ortiz, E. 2014 Evidence-Based Guideline for the Management of High Blood Pressure in Adults: Report from the Panel Members Appointed to the Eighth Joint National Committee (JNC 8). Jama. 2014, 311 (5), 507–520. doi:10.1001/jama.2013. 284427.
Kaushal C, Srivastava B. A Process of Method Development: A Chromatographic Approach, J Chem Pharm Res, 2010; 2(2): 519-545.
Snyder, L.R., J.J. Kirkland and J.L. Glajch, 1997. Practical HPLC Method Development, 2 Edition, Wiley-Interscience, New York, pp: 41-43.
MS. Charde, AS. Welankiwar, J.Kumar. METHOD DEVELOPMENT BY LIQUID CHROMATOGRAPHY WITH VALIDATION. International Journal of Pharmaceutical Chemistry. (2014) 04(02): 57-61.DOI: https://doi. org/10.22270/ajprd.v8i4.656.
Melander, W.R. and C. Horvath, 1980. "High Performance Liquid Chromatography, Advances and Perspectives" (C. Horvath, ed.), Academic Press, New York, pp: 113-119
S.Ravisankar. Text book of pharmaceutical analysis. Rx Publications. 3, 2.2- 2.5, 13.1, 17.14- 17.18 & 18.2-18.6.
Refbacks
- There are currently no refbacks.