evaluated reference ms/ms spectra libraries current and future nist programs
TRANSCRIPT
![Page 1: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/1.jpg)
Evaluated Reference MS/MS Spectra Libraries
Current and Future NIST Programs
![Page 2: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/2.jpg)
NIST/EPA/NIH Library of EI Mass Spectra
Search Program/Algorithms
Archive ProgramComputer Assisted Evaluation
![Page 3: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/3.jpg)
NIST MS/MS Library
• June 2005 - 5,191 Spectra– 1,920 Precursor ions (incl. 292 Anions)
• From NIST, Contributors and literature• Ion trap, Triple Quad, …• New data structure/software
– Energy and precursor ion variations– Evaluation/Search logic– Range of Energies (collision cell)
![Page 4: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/4.jpg)
Archive Program – MS/MS Mode
![Page 5: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/5.jpg)
Archive Program + MS Interpreter
![Page 6: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/6.jpg)
NIST MS Search Program – MS/MS Library
MW /
Formula /
Name /
Conditions
Levels
![Page 7: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/7.jpg)
NIST Library of Peptide Ion Fragmentation Spectra
• Peptide sequences are used to infer proteins – from protein digests
• Derived from ‘Shotgun’ proteomics– Identifications originate from sequence-based search
method
• Human, Yeast, Selected Proteins, …– 90,000 Consensus spectra
• Ion trap and qtof spectra (tof/tof soon) of ESI and MALDI generated ions
![Page 8: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/8.jpg)
Processing Pipeline
• From Raw data to Identified Proteins– Complex series of data transformations– Peptide sequences are identified by matching
m/z values against all possible sequences
• Consensus spectra extracted from pipeline– New QA/QC methods
• Library re-inserted to pipeline for faster, more reliable, more sensitive IDs
![Page 9: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/9.jpg)
Input list
Matching peptide and probability scores
Reference spectrum and annotation
Query MS/MSHead to tail sample and reference spectra comparison
![Page 10: Evaluated Reference MS/MS Spectra Libraries Current and Future NIST Programs](https://reader035.vdocuments.us/reader035/viewer/2022072010/56649db95503460f94aa9362/html5/thumbnails/10.jpg)
Fragmentation depends on energy/time
• FT-ICR– Lowest energies (SORI, IRMPD, SID, …)
• Ion Trap– Allows multiple stages of fragmentation
• Collision cell (Low E)– Dissociation increases with energy
• PSD – Low energy, limited energy/time
• Collision cell (High E)– Single collision, wide E distribution