HPLC is a form of column chromatography that pumps a sample mixture or analyte in a solvent at high pressure through a column with chromatographic packing material. The different components in the mixture pass through the column and the constituents of a sample can be identified by comparison of retention times with those of known standards run under the same conditions. The term reversed-phase describes the chromatography mode that is just the opposite of normal phase, namely the use of a polar mobile phase and a non-polar [hydrophobic] stationary phase. Today, because it is more reproducible and has broad applicability, reversed-phase chromatography is used for approximately 75% of all HPLC methods.

Applications

• Separation of large molecular weight materials that have very low volatility Analysis of biologically important aromatic amines
• Determination of pharmaceutical products such as antibiotics, sedatives, steroids and analgesics
• Determination of artificial sweeteners, antioxidants, aflatoxins, amino acids, proteins, carbohydrates, lipids and vitamins