KIM706_Ders_6_Mass Spectrometry_111223.pdf

MassSpectrometry

1. KİM 706
KÜTLE SPEKTROMETRİSİ
11/12/2023
1

2. ELECTROSPRAY IONIZATION
(ESI)
2

3. Electrospray Ionization (ESI)
Atmospheric Pressure Chemical Ionization (APCI)
Atmospheric Pressure Photoionization (APPI)
Matrix Assisted Laser Desorption/Ionization (MALDI)
Desorption Electrospray Ionization (DESI)
Direct Analysis in Real Time (DART)
Liquid
phase
Solid
phase
3

4. ELECTROSPRAY
4

5. High voltage applied
to metal sheath (~4 kV)
Sample Inlet Nozzle
(Lower Voltage)
Charged droplets
+
+
+
+
+
+
+
+
++
+
+
+
+
++
+
+
+
++
+
++
+
++
+
+
+
+
+
+
+
+
+
+
+
++
+
++
+
+
+
MH
+
MH
3
+
MH
2
+
Pressure = 1 atm
Inner tube diam. = 100 um
Sample in solution
N
2
N2 gas
Partial
vacuum
Electrospray ionization:
Ion Sources make ions from sample molecules
(Ions are easier to detect than neutral molecules.)
5

6. ESI
6

7. ES SPECTRUM OF HORSE
MYOGLOBIN MW 16.951 A)
multiply charged ion distribution
from +12-24 shown at low
resolution B) the 17+ charge state
at a resolution of about 15000
showing the resolved isotope
peaks
7

8. 8

9. ESI spectra of disulfide-intact and
disulfide-reduced lysozyme
Konermann et al.
J. Am. Soc. Mass
Spectrom.
1998, 9, 1248-1254
9

10. 10

11. Nanoelectrospray
Flow rates 50 to 400 nL/min

12. Nano-Electrospray(nanoESI)
Gold-coated borosilicate glass

13. Features of NanoESI
Flow rates of 100-400nL/min (25-100 min analysis time)
Sample volumes down to 300 nL
Near 100 % sample utilization
Minimal instrument contamination
Zero sample cross contamination
Spray from 0% to 100% aqueous solvents

14. NanoESIvs. conventional ESIDroplets NanoES Conventional ES
Radius
Volume
Analyte molecules
per droplet
(at 1pmol/µL)
50 - 200nm
5·10
-13
- 4·10
-12
µL
0.3 - 2.5
1000 - 2000nm
4·10
-9
- 3·10
-8
µL
2.5·10
3
- 1.9·10
4

15. NanoESIvs. conventional ESI
Vialaret, Jérôme, et al. "Nano-flowvsstandard-flow: Whichis themoresuitableLC/MS methodforquantifyinghepcidin-25 in humanserum in
routineclinicalsettings?."Journalof ChromatographyB1086 (2018): 110-117.

16. 16