Chromatography Techniques

Hyphenated HPLC Methods

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The hyphenated technique is developed from the coupling of a separation technique and an on-line spectroscopic detection technology. Several remarkable improvements in hyphenated analytical methods over the last two decades have significantly broadened their applications in the analysis of biomaterials, especially natural products, pre-isolation analyses of crude extracts or fraction from various natural sources, isolation and detection of natural products, chemical fingerprinting, and metabolomics. Techniques like HPLC coupled to NMR (Nuclear Magnetic Resonance) or electrospray ionization tandem mass spectrometry (ESI-MS-MS) have been proven to be extremely powerful tools in natural product analysis, as they aid in the fast screening of crude natural product extracts or fractions for detailed information about metabolic profiles, with minimum quantity of material. The application of various hyphenated techniques even allows the discovery of new molecules, including complete and conclusive structure elucidation, and relative configurations as compared to time-consuming and costly isolation and purification processes. Hyphenated HPLC techniques include HPLC-MS, HPLC-ESI-MS, HPLC-IC-MS, HPLC-NMR-MS, HPLC-DAD, HPLC-CE-MS, HPLC-UV, Coupling LC and MALDI-TOF. MALDI (Matrix Assisted Laser Desorption Ionization) is a very sensitive technique for determining the mass of proteins, polymers and peptides. MALDI basically used in protein identification. MALDI sample preparation is fast and easy and therefore a primary choice in proteomics.