Calibration peptides for HRM-MS™ (SWATH, DIA) for fully automated high-throughput data processing with Spectronaut™ software.
Hyper Reaction Monitoring Mass Spectrometry (HRM-MS™) is the next generation targeted proteomics technology invented at Biognosys that enables reproducible and precise quantification of 1000s of proteins in a single instrument run with data independent acquisition (DIA, SWATH). Spectronaut™ software in combination with the HRM Calibration Kit enables fully automated and accurate signal processing of HRM-MS™ data sets.
The HRM Calibration Kit, containing non-naturally occurring synthetic peptides in a pooled mix, serves to calibrate retention time and other parameters for the Spectronaut™ software enabling peak detection, scoring, mass calibration, noise reduction and other data analysis steps.
The use of the HRM Calibration Kit translates into better results:
The kit components are sufficient for approximately 1000 injections (or 100 samples) depending on sample volume and injection amount.
Using the HRM Calibration Kit requires just one step: Pipet the HRM Peptide Mix into your sample directly before measurement – everything else is handled by Spectronaut™ software. Spectronaut™ automatically detects the HRM peptides and reports successful calibration only if the HRM peptides are detected in the sample.
If your instrument is not on the list please contact us to get detailed information on the instruments we support.
See the Quick Reference Card for detailed information on how to use the HRM Calibration Kit.
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iRT is stands for indexed retention time. It is a dimensionless value that allows the transferring of retention time information of a peptide to different chromatographic set ups.What are iRT peptides?
iRT peptides are 11 non-naturally occurring chemically synthesized peptides that are used in the next generation proteomics workflows to accurately calculate the indexed retention time of peptides on a chromatographic set up.Why is it beneficial to add the iRT peptides to all my samples?
Apart from predicting the retention time the iRT peptides are used for mass calibration and QC monitoring of the LC-MS set up. In an HRM-MS experiment the iRT or HRM peptides are further used as anchor points for the data analysis.What is the difference between the iRT peptides and the HRM calibration peptides?
On top of the 11 peptides in the iRT kit the HRM calibration kit contains additional peptides. These are used in the older versions of Spectronaut. With the new algorithm of SN 9 the HRM peptides are no longer a requirement and can be replaced with the iRT peptides if desired. This has the advantage that only one set of peptides are required for customers using all three next-generation proteomics workflows offered by Biognosys.Do I really need to add the iRT peptides to all my runs because they make the per sample costs higher?
As of Spectronaut 9 the iRT peptides are not required in all cases for the samples used for spectral library generation. Yet, we strongly advise using them as in seldom cases spectral library generation can fail without them. For all DIA runs the peptides are required by the software. Adding the iRT peptides to the sample increase the per sample costs by less than 1% (~30cents/sample). Further, the dilution can be optimized on the newest generation of mass spectrometers to further reduce the per run cost additions.
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