A Review - Importance of RP-HPLC in Analytical Method Development

International Journal of Novel Trends in Pharmaceutical Sciences,2013,3,1,15-23.
Published:February 2013
Type:Review Article
Authors:
Author(s) affiliations:

B. Prathap1,*, Akalanka Dey2, G.H. Srinivasa Rao1, P. Johnson1 and P.  Arthanariswaran3

1Department of Pharmaceutical Analysis, Saastra College of Pharmaceutical Education and Research, Nellore - 524311, Andhra Pradesh, India.

2Faculty of Pharmacy, Annamalai University, Annamalai Nagar – 608002, Tamilnadu, India.

3Department of Pharmacognosy, Saastra College of Pharmaceutical Education and Research, Nellore – 524 311, Andhra Pradesh, India.

Abstract:

Chromatography, although primarily a separation technique, is mostly employed in chemical analysis in which High-performance liquid chromatography (HPLC) is an extremely versatile technique where analytes are separated by passage through a column packed with micrometer-sized particles. Now a day reversed-phase chromatography is the most commonly used separation technique in HPLC. The reasons for this include the simplicity, versatility, and scope of the reversed-phase method as it is able to handle compounds of a diverse polarity and molecular mass. Reversed phase chromatography has found both analytical and preparative applications in the area of biochemical separation and purification. Molecules that possess some degree of hydrophobic character, such as proteins, peptides and nucleic acids, can be separated by reversed phase chromatography with excellent recovery and resolution. This review covers the importance of RP-HPLC in analytical method development and their strategies along with brief knowledge of critical chromatographic parameters need to be optimized for an efficient method development.

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PRINCIPLE OF REVERSED PHASE CHROMATOGRAPHY WITH GRADIENT ELUTION