Author(s): B Prathap, Akalanka Dey, GHSrinivasa rao, P Johnson, P Arthanariswara
Chromatography, although primarily a separation tec hnique, is mostly employed in chemical analysis in which High8performance liquid chromatography (HPLC) is an extremely versatile technique where analytes are se parated by passage through a column packed with micrometer8sized particles. Now a day reversed8phase chromatography is the most commonly used separation technique in HPLC. The reasons for this include the simplicity, versatility, and s cope of the reversed8phase method as it is able to handle compounds of a diverse polarity and molecular mass. Reversed phase chromatography has found both analytical and prepar ative applications in the area of biochemical separation and purification. Molecules that possess some degree of hydrophobic character, such as proteins, peptides a nd nucleic acids, can be separated by reversed phase chromatography with excellent recove ry and resolution. This review covers the importance of RP8HPLC in analytical meth od development and their strategies along with brief knowledge of critical c hromatographic parameters need to be optimized for an efficient method development.