Exploring the use of mass spectrometry data for differential proteomics
Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility...
Transcript of Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility...
![Page 1: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/1.jpg)
Differential Mobility Spectrometry (DMS)
An orthogonal separation mechanism based upon charge-to-mass and
molecular shape
Lecture 2, Page 84
![Page 2: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/2.jpg)
Hardware Overview
SelexIon – Planar DMS
Lecture 2, Page 85
![Page 3: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/3.jpg)
Mass Spec Vacuum draws ions and gas through planar DMS electrodes
(5 millisecond flight time)
2.3 torr
2.8 L/min
1 mm
3 cm
760 torr
Lecture 2, Page 86
![Page 4: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/4.jpg)
The physics and chemistry of the separation
principle.
Specificity from nuclear properties. •Radioactivity •High Mass Resolution.
• Specificity from molecular properties. • NMR • Collision-Induced Dissociation - MSn
• Specificity from chemical properties. • Chemical interactions of analyte with environment
• HPLC, CZE, immunochemsity
• Ion Molecule Chemistry (ICP-MS Dynamic Reaction Cell)
• Ion Molecule Chemistry Mobility
Sources of Selectivity
Lecture 2, Page 87
![Page 5: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/5.jpg)
Chemical Enhanced Separations Driven by Mobility Changes Due to Clustering / Declustering.
Low field
clustering-
mobility decreases
+
HV=3kV
Schneider BB, Covey TR, Coy SL, Krylov EV, and Nazarov EG, “Chemical Effects in the Separation Process of a Differential Mobility/Mass
Spectrometer System”, Anal. Chem., 2010, 82, 1867-1880.
High field
declustering;
mobility increases
L V=0.3 kV
F ~ 1 MHz, T= 10-6 s
Lecture 2, Page 88
![Page 6: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/6.jpg)
Ionograms of 3 compounds with/without clustering modifiers
Dianabol
Benoxinate
Clenbuterol
Nitrogen
transport gas
Nitrogen with
44% helium
transport gas
Nitrogen with
1.5% isopropanol
transport gas
Increased mobility with helium
favors hard sphere behavior
….peaks shift to higher CV
and narrow slightly.
Lecture 2, Page 89
![Page 7: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/7.jpg)
Different effects with different modifiers
Ketamine (blue) and sulfadiazine (red)
Ethylacetate
Acetone
Acetonitrile
Methanol
Isopropanol
m/z 238.1 m/z 251.1
Lecture 2, Page 90
![Page 8: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/8.jpg)
Lecture 2, Page 8
Effects of Different Modifiers-Gas phase LC?
1.0
0.8
0.6
0.4
0.2
0.0
Norm
aliz
ed S
ignal
-50 -40 -30 -20 -10 0
CV (V)
Isopropanol Separation
1.0
0.8
0.6
0.4
0.2
0.0
Norm
aliz
ed S
ignal
-50 -40 -30 -20 -10 0
CV (V)
Acetone Separation
1.0
0.8
0.6
0.4
0.2
0.0
Norm
aliz
ed S
ignal
-50 -40 -30 -20 -10 0
CV (V)
Acetonitrile Separation
Red: Ephedrine Black: Acyclovir Green: Norfentanyl Blue: Clenbuterol Purple: Imipramine Brown: Diazepam Pink: Quinoxifen
Lecture 2, Page 91
![Page 9: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/9.jpg)
Applications where Chromatography is
employed and integrated with DMS.
Lecture 2, Page 92
![Page 10: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/10.jpg)
Testosterone Female #3 Plasma Sample: LLE
DMS OFF/
289/109
DMS OFF/
289/97
DMS ON/
289/109 DMS ON/
289/97
Interference
Ion ratio is Peak of interest Interference
Peak of interest
Lecture 2, Page 93
![Page 11: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/11.jpg)
Testosterone Female #3 Plasma Sample: PPT
DMS OFF/
289/109
DMS OFF/
289/97
DMS ON/
289/109
DMS ON/
289/97
Peak of interest
Peak of interest @
RT 3.37 min
Lecture 2, Page 94
![Page 12: Differential Mobility Spectrometry (DMS) › wp-content › ... · Differential Mobility Spectrometry (DMS) An orthogonal separation mechanism based upon charge-to-mass and molecular](https://reader036.fdocuments.net/reader036/viewer/2022062603/5f031ed47e708231d407a348/html5/thumbnails/12.jpg)
Separation Science References
• Basics of HPLC – http://www.waters.com/waters/nav.htm?cid=100490
64&locale=en_US
• Basics of nano LC – http://www.nanoseparations.com/
• Basics of CE – http://matematicas.udea.edu.co/~carlopez/cnq533/C
E_Basics.pdf
• Basics of IC – http://www.melvynbecerra.cl/images/ICPractica.pdf
Lecture 2, Page 95