Comparison between a linear ion trap and a triple quadruple MS in the sensitive detection of large peptides at femtomole amounts on column

摘要

In addition to the importance of sample preparation and extract separation, MS detection is a key factor in the sensitive quantification of large undigested peptides. In this article, a linear ion trap MS (LIT-MS) and a triple quadrupole MS (TQ-MS) have been compared in the detection of large peptides at subnanomolar concentrations. Natural brain natriuretic peptide, C-peptide, substance P and D-Junk-inhibitor peptide, a full D-amino acid therapeutic peptide, were chosen. They were detected by ESI and simultaneous MS~1 and MS~2 acquisitions. With direct peptide infusion, MS~2 spectra revealed that fragmentation was peptide dependent, milder on the LIT-MS and required high collision energies on the TQMS to obtain high-intensity product ions. Peptide adsorption on surfaces was overcome and peptide dilutions ranging from 0.1 to 25 nM were injected onto an ultra high-pressure LC system with a 1mm id analytical column and coupled with the MS instruments. No difference was observed between the two instruments when recording in LC-MS ~1 acquisitions. However, in LC-MS~2 acquisitions, a better sensitivity in the detection of large peptides was observed with the LIT-MS. Indeed, with the three longer peptides, the typical fragmentation in the TQ-MS resulted in a dramatic loss of sensitivity (≥ 10x). & 2010 Wiley-VCH Verlag GmbH & Co. KGaA.