ABSTRACT
This chapter focuses on such techniques using high performance liquid chromatography (HPLC) to separate complex peptide mixtures, followed by tandem mass spectrometry for sequence analysis. It discusses the vantage point of the human embryonic stem cells and proteomics techniques rely on mass spectrometry (MS) as a general detection technique that allows amino acid sequencing through analysis of peptide fragmentation. A strong point of electrospray ionization is its ease of utilization coupled to HPLC as a front end (LC/MS), a common separation method in proteomics, allowing real-time analysis of separations. Time-of flight (TOF) mass analyzers separate ions by the different times required to reach the detector. Ions are accelerated down a flight tube and ions with lower masses will have higher velocities and a shorter flight time. Based on the TOF, the instrument assigns a m/z to the various products detected following ion extraction.
