Translate the Amino Acid Sequence of the Peptide by Mass Spectrometry Analysis
Liquid Chromatography-Mass Spectrometry (LC-MS) is a widely used analytical technique for determining the amino acid sequences of peptides. By integrating liquid chromatography (LC) with mass spectrometry (MS), LC-MS enables comprehensive structural characterization of peptides with high sensitivity, accuracy, and resolution.
Fundamental Principles of Liquid Chromatography-Mass Spectrometry
The principle of LC-MS involves two key stages: chromatographic separation followed by mass spectrometric detection. First, the peptide sample is separated using liquid chromatography before being introduced into the mass spectrometer for ionization and analysis. During the chromatographic phase, peptides pass through a column packed with a stationary phase (typically a chemically bonded material). Differences in peptide amino acid sequences lead to variations in their interactions with the stationary phase, enabling chromatographic separation. In the mass spectrometry stage, peptide ions are separated and detected based on their mass-to-charge ratio (m/z) under the influence of an electric field. Common mass spectrometry techniques include Time-of-Flight Mass Spectrometry (TOF-MS), Ion Trap Mass Spectrometry (IT-MS), and Quadrupole Mass Spectrometry (Q-MS).
Peptide Sequencing via LC-MS
In peptide sequencing, LC-MS is often coupled with tandem mass spectrometry (MS/MS). In an MS/MS experiment, selected peptide ions undergo collision-induced dissociation (CID) after the first mass spectrometric stage, generating a series of fragment ions. These fragment ions are then analyzed in a second mass spectrometric stage. By interpreting the mass-to-charge ratios and intensities of the fragment ions, the original peptide sequence can be reconstructed.
LC-MS serves as a powerful tool for high-precision peptide sequencing and has been extensively applied in diverse fields, including biomedicine, food science, and environmental science. Moreover, advancements in mass spectrometric techniques, such as Electrospray Ionization (ESI) and Matrix-Assisted Laser Desorption/Ionization (MALDI), are expected to further enhance the analytical performance of LC-MS.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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