Lcms Basics

LCMS Basics explained with several animations to help understand the theory behind MS. All images are cited that are not my own. If you have any questions please feel free to reach out to me.

1. Long Island University | April 6 th , 2018 LC-MS/MS Basics Ben Kuzma

2. Long Island University | April 6 th , 2018 Objectives Introduce basic MS concepts Explain the fundamentals of MS Demonstrate the benefits of different systems Method Development using concepts

3. Long Island University | April 6 th , 2018 Key Terms Parent Ion (Precursor ion) Daughter Ion (fragment ion) MS/MS MRM http:// www.waters.com / webassets / cms /library/docs/720000807en.pdf

4. Long Island University | April 6 th , 2018 Overview https:// www.idex-hs.com /education-and-tools/educational-materials/ hplc -center

5. Long Island University | April 6 th , 2018 Overview https:// www.idex-hs.com /education-and-tools/educational-materials/ hplc -center

6. Long Island University | April 6 th , 2018 Why use LCMS? Selectivity Lidocaine and Prilocaine Elute with 0.1 minutes of each other Speed (UHPLC) HPLC Run time – 5-20 minutes UHPLC Run time 3 minutes Sensitivity HPLC LLOQ - 10 ng/ml LCMS/MS – 400 pg /ml (0.4ng/ml) 25 fold difference

7. Long Island University | April 6 th , 2018 Fundamentals of LCMS How does it work? Mass to charge ratio (m/z) Ionization Methods Electrospray ionization (ESI) Atmospheric Pressure Chemical ionization (APCI) Atmospheric pressure photoionization (APPI)

8. Long Island University | April 6 th , 2018 Fundamentals of LCMS http:// www.ecs.umass.edu /eve/background/methods/chemical/ Openlit /Chromacademy%20LCMS%20Intro.pdf

9. Long Island University | April 6 th , 2018 Ionization Methods Electrospray ionization (ESI) Limited fragmentation http:// www.particlesciences.com /news/technical-briefs/2009/mass-spectrometry- bioanalysis.html

10. Long Island University | April 6 th , 2018 Ionization Methods Atmospheric Pressure Chemical ionization (APCI) http:// www.particlesciences.com /news/technical-briefs/2009/mass-spectrometry- bioanalysis.html

11. Long Island University | April 6 th , 2018 Ionization Methods Atmospheric pressure photoionization (APPI) http:// ccc.chem.pitt.edu / wipf /Agilent%20LC-MS%20primer.pdf

12. Long Island University | April 6 th , 2018 Type of Mass Detectors Quadrupole Ion Trap Time of Flight Magnetic Sector Triple Quad = 2 Mass analyzers

13. Long Island University | April 6 th , 2018 Quadrupole Uses electric field to separate ions of interest (volts) If the m/z ratio is “correct” then it will not collide with the poles and it will continue on to the next part of the analyzer. Gold standard for quantitation ( QqQ )

14. Long Island University | April 6 th , 2018 Ion Trap Trap the ions of interest in a field then release them m/z intensity

15. Long Island University | April 6 th , 2018 Time of Flight (TOF) Fast Scan rate Metabolites Structural elucidation m/z intensity F Ion Source Acceleration Time - of - Flight Detector

16. Long Island University | April 6 th , 2018 LC-MS/MS Gold standard for quantitation http://what-when- how.com /proteomics/quadrupole-mass-analyzers-theoretical-and-practical-considerations-proteomics/

17. Long Island University | April 6 th , 2018 Application of Fundamentals

18. Long Island University | April 6 th , 2018 Ionization type ESI Positive Mode Negative Mode APCI Waters LCMS Prep Guide

19. Long Island University | April 6 th , 2018 MS Scan Mass Range of 120 – 600 m/z

20. Long Island University | April 6 th , 2018 MS Scan

21. Long Island University | April 6 th , 2018 ESI+ 120-600 M/z

22. Long Island University | April 6 th , 2018 Daughter Ions Argon Gas Also called MSMS Mode

23. Long Island University | April 6 th , 2018 Daughter Ions

24. Long Island University | April 6 th , 2018 Daughter Ions

25. Long Island University | April 6 th , 2018 Daughter Ions Argon Gas

26. Long Island University | April 6 th , 2018 10 eV 15 eV 20 eV 25 eV 30 eV

27. Long Island University | April 6 th , 2018 Daughter Ions Argon Gas

28. Long Island University | April 6 th , 2018 Development of MRM MW +H Transition (MSMS) Time transitions are analyzed Voltage applied during ionization Energy of Argon gas for fragments

29. Long Island University | April 6 th , 2018 Final Product of MRM

30. Questions?

31. Resources http://ccc.chem.pitt.edu/wipf/Waters%20LC-MS%20primer.pdf http://www.ecs.umass.edu/eve/background/methods/chemical/Openlit/Chromacademy%20LCMS%20Intro.pdf http://www.rsc.org/images/AMC%20LCMS%20Guide_tcm18-240030.pdf