The decisive step from classical Liquid Chromatography (LC) into the high performance mode (HPLC) took place between 1965 and 1975 by the development of pressure stable columns packed with 5 to 10 μm reversed phase (RP) silica particles.In the following decade such columns were manufactured at a high reproducibility and with a substantial ruggedness which finally lead to the widespread and global application of HPLC in pharmaceutical and chemical analysis.Parallel to this achievements novel columns were developed for the fast, high resolution separation of biopolymers such as peptides, proteins, polynucleotides etc.The successful hyphenation of HPLC with mass spectrometry (MS) as a powerful separator and detector and the use of 3 to 5 μm particles as packings tremendously enhanced the performance of HPLC in terms of speed, resolution and sensitivity in the decade between 1990 and 2000.With the birth of the-omics approaches (proteomics, peptidomics, metabolomics etc) at the beginning of this century HPLC/MS expanded into the field of life science.