What is LC-MS (liquid chromatography–mass spectrometry)?
LC-MS couples liquid chromatography with a mass spectrometer so that each separated component can be identified by its mass; it is the standard method for confirming a peptide's identity.
How LC-MS works
The sample is first separated by liquid chromatography. As components leave the column, they are ionized, most often by electrospray ionization, and passed into a mass spectrometer, which measures their mass-to-charge ratio (m/z).
Electrospray gives a large peptide several charges at once, so one molecule appears as a series of ions rather than a single peak at its full molecular weight. Identity is confirmed when the observed ions match the values calculated from the expected molecular weight.
Identity on KPV certificates
KPV certificates from ILS Laboratories confirm identity by HPLC retention-time comparison with a reference standard (HPLC-RTM) rather than by LC-MS. The method is named next to the result on every lot record.
Limits
LC-MS confirms molecular weight. Two peptides with the same amino acids in a different order have the same mass, so sequence order needs fragmentation (tandem MS) or other methods. Signal intensity also depends on how easily each species ionizes, so LC-MS is not used for the purity figure.