This Application Note demonstrates how Intelligent Sample Loading automates and optimizes sample enrichment on the HPLC-Chip, and also shows how Intelligent Sample Loading increases the number of peptides detected in a protein digest test mixture.
The document describes an application using the Agilent HPLC-Chip/MS system for automated sample enrichment. It highlights that conventional nanocolumn LC-MS systems require time-consuming manual adjustment of valve event timetables when injection volumes change, which can degrade efficiency.
Intelligent Sample Loading (ISL) is presented as a solution that decouples the sample loading process from analysis. The system automatically calculates the required loading time based on actual flow rates and capillary delay volumes, eliminating the need for manual re-optimization. This feature reduces data file sizes and increases the number of peptides detected in protein digest mixtures.
| Enrichment column | 40 nL, 5 um ZORBAX 300SB-C18 |
|---|---|
| Analytical column | 75 pm x 43 mm, 5 pm ZORBAX 300SB-C18 |
| Injection flush volume (IFV) | 5.926 pL |
It automates and optimizes sample loading by automatically calculating the required time to effectively load the sample, eliminating the need for manual re-adjustment when injection volumes change.
It reduces data file size because MS data acquisition coincides with the switching from enrichment to analysis mode instead of starting at the beginning of the autosampler sample loading cycle.