APGC-MS, ASAP-MS, and MAI-MS:
Expanding the Horizons of API-MS
Charles McEwen
University of the Sciences
DuPont VG-ZAB Double Focusing High Resolution Magnetic Sector MS
Atmospheric Pressure Chemical Ionization
Mass Spectrometry (1973)
Horning, E.C., Horning, M.G., Carroll, D.I., Dzidic, I., Stillwell, R.N.,
Anal. Chem., 1973, 45, 936-943
John B. Fenn ASMS Distinguished Contribution, 1992
Electrospray Ionization of Some Polypeptides and Small
Proteins C. K. Meng, M. Mann and J. B. Fenn, 36th ASMS Conf., 1988
The Electrospray Process; Utility Beyond Mass Spectrometry
Professor John FennVirginia Commonwealth University
April 23, 2002
E328/115
10:00 AM
Professor John Fenn is an entertaining speaker and promises to lead a
freewheeling discussion that briefly covers the history of electrospray
ionization and then moves to current and potential applications of this
technology. He will discuss uses such as electrospinning of polymers and
high-sensitivity collection and detection of airborne chemicals. Dr. Fenn has
had an enormous impact on science with his successful interface of
electrospray ionization with mass spectrometry and the discovery that
multiple charging allowed analysis of such large and nonvolatile compounds
as synthetic polymers and proteins. This technology has been especially
valuable to the biological sciences. These discoveries were made during
Professor Fenn's tenure at Yale where he was widely recognized for his
distinguished studies into the properties of supersonic free jets expanding
into vacuum.
John B. Fenn received a B.A. in chemistry from Berea College and a Ph.D. from Yale. After a
dozen years in industry he was appointed Director of Project SQUID, a Navy program of basic
and applied research in Jet Propulsion administered by Princeton University where he becameProfessor of Aerospace and Mechanical Sciences. He returned to Yale as Professor of Chemical
Engineering and Chemistry, remaining there as a Research Scientist after becoming Emeritus in
1987. In 1993 he moved to Virginia Commonwealth University as Research Professor. He hasserved as a visiting Professor at Trento University in Italy, the University of Tokyo, the Indian
Institute of Science at Bangalore, and the Chinese Academy of Science in Beijing and as a
distinguished lecturer at several other institutions. Author of one book and over a hundred papershe is sole or co-inventor on 19 patents. Much of his research has centered on the properties and
uses of supersonic free jets expanding into vacuum. Such jets can produce molecular beams with
much higher intensities and energies than can the classical effusion ovens they have replaced.
Their ability to cool molecules to ultra low temperatures, with or without condensation, hasrevolutionized molecular spectroscopy and made them versatile sources of clusters and van der
Waals molecules. In the scientific community, Fenn is best known for the first successful
interface of electrospray with mass spectrometry and the discovery that large nonvolatilecompounds such as polymers and proteins could be analyzed using this method. He has received
a number of awards for these inventions including the Thompson Medal and the Distinguished
Contribution to Mass Spectrometry award.
MS inlet HV
melting
Point tube
N2
LC Probe
WickSpray on a Waters Z-spray Source (2002)
wick
Method and apparatus for electrospray ionization
US 6297499 B1
ABSTRACT
Sample liquid is supplied to the ion source of an Electrospray
Ionization Mass Spectrometer (ESIMS) by capillarity induced
flow through a wick element comprising a permeable porous
aggregate of fibers or particles of material that is wetted by the
sample liquid. This method of liquid introduction eliminates the
need for pumps of pressurized gas to drive the flow. It also
makes possible the convenient extraction of a representative
sample from a stream of liquid flowing at any rate, no matter
how large.
This application claims benefit of Provisional Appln.
60/052,885 filed Jul. 17, 1997
Inventor: John B. Fenn
Wickspray Patent Issued to John Fenn (2001)
High Voltage
Power
Supply
Vented
Bottle
w/pouch
liner
Capillary
Wick
Level
Viewing
Window
Vents
Fragrance Dispersion WickSpray Device
Liquid dispersion device Patent number: 6871794
Abstract: A device that can be used for dispersing or dispensing a liquid is provided that comprises a
container, a nozzle device, a secondary wick, and a control device in which the container comprises, an
open end and a liquid; the nozzle comprises a primary wick, a first end being in fluid communication with
the liquid, and a second end extending through the open end of the container and having an opening; the
aperture has extended therethrough the primary wick; the secondary wick extends into proximity with the
primary wick; and the secondary wick is securely connected to a control device, which controls the distance
between the primary wick and the secondary wick.
Filed: May 1, 2003 Issued: March 29, 2005
Assignee: E. I. du Pont de Nemours and Company
Inventor: Charles Nehemiah McEwen
Liquid dispersion device Patent number: 6729552
Abstract: A device that can be used for dispersing a liquid is disclose. The device comprises a
container, a capillary device, and a housing. The container has an open end that is connected to the
capillary device and comprises a liquid. The capillary device comprises a substantially tubular member
having one end secured to the open end of the container and the opposing end extended therethrough a
substantially tubular capillary structure, which is coaxially aligned with the substantially tubular member. The
capillary structure is in fluid communication with the liquid in the container. The housing comprises a first
end having an opening attached thereon the container, a low voltage supplier attached to one wall, a high
voltage converter attached to another wall, a voltage contact and a counter electrode, optionally a
heat and/or lighting source, a wicking material, and further optionally electronics for voltage regulation. Also
disclosed is a process for dispensing liquid using the device.
Filed: April 22, 2003 Issued: May 4, 2004
Assignee: E. I. du Pont de Nemours and Company
Inventors: Charles Nehemiah McEwen, William J. Herron, Richard G. McKay
AP Ionization Technologies
• AP-GC/MS
• ASAP-MS
MS inlet HV
melting
Point tube
N2
LC Probe
WickSpray on a Waters Z-spray Source (2002)
wick
Gas Chromatography/Atmospheric
Pressure-Mass Spectrometry
Conversion of LC/MS instruments
to Dual LC/MS and GC/AP-MS
Ion source for a mass spectrometer US7642510
Korfmacher, et al. at the National Center for Toxicological Research in Jefferson,
Arkansas published on AP-GC/MS (negative ion) until 1990’s.
Product 1
1
19
3
20
21
4
23
22
9
24
19. LC interface probe
20. Nebulizer gas inlet
21. LC column connector
22. LC interface tube
23. LC transfer line
24. Metal sheath gas tube
25. GC purge gas tube and
heater assembly
5
6 8
16
7 25 18
Atmospheric Pressure GC/LC Ion Source Fig. 2
Mass Spec GC Inlet
LC Probe
Corona Needle
Waters QTof
GC with
autosampler
AP-GC/MS
Syagen Photoionization Source
APPI-GC/MS
Interface Probe
APGC/MS of 1ul Injection of 1ppm TOA 50:1 Split
GC column
N2 gas
in
Tee
Interface probe Ceramic tip
GC/AP-MS of Restek EPA 8270 Mixture
Air atmosphere
Nitrogen atmosphere
Air
Nitrogen purge
gcapms 8270 5 ml/min flow
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00 12.00 13.00Time0
100
%
gcapms8270fast TOF MS ES+ BPI
3.51e3
7.27
7.15
6.80
5.723.76
2.89
0.56
2.70
2.05
1.31
2.67
3.203.57
3.36
4.474.13
4.79
5.02
6.77
6.00
6.60
6.36
6.33
7.84
7.79
7.45
7.62
8.46
8.30
8.00
8.62
9.80
9.14
9.50
10.18
9.8811.99
11.66
10.90
11.07
13.4913.24
12.18
12.69
GCAPMS Base peak Chromatogram of Restek 8270 mixture
1ul injected/10:1 split
Positive ion, N2 Purge on:
73 of 76 compounds observed
gcapms 8270 5 ml/min flow
4.65 4.70 4.75 4.80 4.85 4.90 4.95 5.00 5.05 5.10Time0
100
%
gcapms8270fast TOF MS ES+ BPI909
4.79
4.70
5.02
4.92
5.09
Peak widths half height < 2.5 sec
GCAPMS Resolution on Restek 8270 Misture
Accurate Mass Measurement of APGC/MS
Peaks Using GC Bleed as Reference Mass
• Meas. Mass Calc. Mass mDa err Formula
• 113.0928 113.0966 -3.8 C7 H13 O
• 122.0593 122.0606 -1.2 C7 H8 N O
• 242.2821 242.2848 -2.6 C16 H36 N
R2=0.99941 SD=1.67453
GCAPMS of 2-Methylbenzophenone Calibration Curve
from 1ng to 5pg injected using Qtof I Mass Spectrometer
Acquisition m/z 100-400
0 200 400 600 800 1000
0
20
40
60
80
100
120
140
Are
a
PPB Methylbenzophenone
PPB Area
1000 121
500 57
250 29
50 5.9
25 3.5
10 2.1
5 1.9
Perfume Sample Anti-Counterfeit Analysis
UV Diode Array
LC/MS Electrospray
GC/APMS
2.50 5.00 7.50 10.00 12.50 15.00 17.50 20.00 22.50 25.00 27.50 30.00 32.50 35.00 37.50 40.00 Time 0
100
%
-6
100
%
K25-574A 3: Diode Array TIC
6.53e7
10.9
7.80
1.99 0.12
22.82 18.10
17.12 11.75
14.82
22.29
21.10
24.3 30.45 30.90
K25-574A 1: TOF MS ES+ TIC
1.23e4 17.16
11.0
16.21
12.88
24.4
22.37 17.58
19.91 22.92
29.55 29.82 30.99
APGCMS Perfume4 30m DB1
4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 Time 2
100
%
apciperfume4_cv32 Sm (Mn, 1x1) TOF MS ES+ BPI
1.10e3 845
822
783
461
587 692
660
638 747
711
1174
878
928
1110
1019
995
971 1341
13.3
Waters APGC-MS
Thanks to
Peter Hancock
Waters GC-APMS Technology
APGC – How does it work?
Mass Analyser GC Oven
Corona discharge at needle creates plasma at atmospheric pressure
N2 make-up gas delivered through heated transfer line
N2 meets GC eluent flow at transfer line tip
Molecules are ionised after GC elution and directed to the mass analyser
APGC Source Enclosure
…familiar processes
N2+●
N2 e-
2e-
2N2
N4+● M+●
M
M+●
M
Charge Transfer
“Dry” source conditions
Favoured by relatively non-polar compounds
Corona discharge needle
…familiar processes
Corona discharge needle N2
+●
N4+●
H2O
H2O+●
H2O
H3O+●
+OH●
MH+
M
Proton Transfer
Modified source conditions, e.g. with water or methanol
Favoured by relatively polar compounds
m/z280 281 282 283 284 285 286 287 288 289 290 291 292
%
0
100
%
0
100
ANAPGC200309TEST001 (0.026) Is (1.00,0.10) C6Cl6 TOF MS AP+ 3.40e12283.8102
281.8131
285.8073
287.8043
ANAPGC200309TEST001 905 (7.830) Cm (903:907-885:901) TOF MS AP+ 6.35e3283.8097
281.8131
285.8077
287.8047
m/z20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320
%
0
100
ANAPGC200309TEST001 905 (7.830) Cm (903:907-887:900) TOF MS AP+ 6.36e3283.8097
281.8131
285.8077
287.8047
(mainlib) Benzene, hexachloro-
20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 3200
50
100
2435
47
53
71
83
95
107
118
130
142
153165
177
190
214
237
249
284
Cl
Cl
Cl
Cl
Cl
Cl
Monoisotopic Mass, Odd and Even Electron Ions
600 formula(e) evaluated with 64 results within limits (all results (up to 1000) for each mass)
Elements Used:
C: 0-20 H: 0-30 N: 0-4 O: 0-10 Cl: 0-6
Minimum: -1.5
Maximum: 100 10 50
Mass Calc. Mass mDa PPM DBE i-FIT Formula
281.8131 281.8131 0 0 4 14 C6 Cl6
281.8217 -8.6 -30.5 -0.5 93.9 C2 H2 N O2 Cl6
281.8091 4 14.2 0 108.4 C N2 O2 Cl6
281.8203 -7.2 -25.5 0 124 N4 O Cl6
281.8342 -21.1 -74.9 -1 125.1 C3 H4 O2 Cl6
281.7979 15.2 53.9 0 126.2 C2 O3 Cl6
Spectral characteristics
Theoretical isotope pattern
APGC data
APGC Spectrum
NIST EI Spectrum
EI
Faster chromatography at higher flows
Linearity
HCB 22fg to 10,000fg
MRM on Xevo TQ-S
APGC-MS Examples
(mainlib) .lambda.-Cyhalothrin
10 40 70 100 130 160 190 220 250 280 310 340 370 400 430 4600
50
100
1527
41
51
65
77
91115
141
152
181
197
208
225241 262 313 349 368 404
449
Cl
FF
F
O
O
N
O
Time5.00 6.00 7.00 8.00 9.00 10.00 11.00
%
0
100
5.00 6.00 7.00 8.00 9.00 10.00 11.00
%
0
100
ANAPGC270409TEST005 TOF MS AP+ 450.107 0.03Da
897Area, Height
11.1129.566
895
11.029.499305
ANAPGC270409TEST005 TOF MS AP+ 423.095 0.03Da
395Area, Height
11.1112.728
394
11.023.716108
10.523.114
90
Time4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00
%
0
100
4.00 5.00 6.00 7.00 8.00 9.00 10.00 11.00
%
0
100
ANAPGC270409TEST010 TOF MS AP+ 450.103 0.03Da
156Area, Height
11.135.278156
11.051.952
666.06
0.99732
ANAPGC270409TEST010 TOF MS AP+ 423.097 0.05Da
69.0Area, Height
11.132.156
69
50 pg/µL standard
m/z20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460
%
0
100450.1071
423.0949
225.0285
-cyhalothrin in persimmon
10 ppb in persimmon
NIST EI spectrum
APGC spectrum
Deltamethrin in nectarine
Background
• Aerosol pharmaceutical products must be tested to ensure
the quality and safety of the final product
TIC of EI GC/MS on Single
Quad
1 5 .6 01 5 .7 01 5 .8 01 5 .9 01 6 .0 01 6 .1 01 6 .2 01 6 .3 01 6 .4 01 6 .5 01 6 .6 01 6 .7 0
5 0 0 0 0
1 0 0 0 0 0
1 5 0 0 0 0
2 0 0 0 0 0
2 5 0 0 0 0
3 0 0 0 0 0
3 5 0 0 0 0
4 0 0 0 0 0
4 5 0 0 0 0
5 0 0 0 0 0
5 5 0 0 0 0
T im e -->
A b u n d a n c e
T IC : 1 2 0 6 1 0 0 0 7 .D \ d a ta .m s
1 6 .0 3 21 6 .0 9 3
1 6 .2 2 2
1 6 .3 5 2
EI Spectrum of
Peak at 16.03mins
2 0 4 0 6 0 8 01 0 01 2 01 4 01 6 01 8 02 0 02 2 02 4 02 6 02 8 03 0 03 2 03 4 03 6 03 8 04 0 04 2 00
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
6 0 0
7 0 0
8 0 0
9 0 0
1 0 0 0
1 1 0 0
1 2 0 0
m / z -->
A b u n d a n c e
S c a n 1 7 1 9 (1 6 .0 3 4 m in ): 1 2 0 6 1 0 0 0 7 .D \ d a ta .m s (-1 7 1 5 ) (-)5 6
2 2 0
1 0 57 9
3 83 2 53 4 2 4 3 11 9 3 2 8 2
1 3 3
2 5 11 6 3
No library match due to low response and poor spectral quality
CI GC/MS on Single Quad
1 5 .0 0 1 5 .5 0 1 6 .0 0 1 6 .5 0 1 7 .0 0 1 7 .5 0
3 9 5 0 0 0
4 0 0 0 0 0
4 0 5 0 0 0
4 1 0 0 0 0
4 1 5 0 0 0
4 2 0 0 0 0
4 2 5 0 0 0
4 3 0 0 0 0
4 3 5 0 0 0
4 4 0 0 0 0
T im e -->
A b u n d a n c e
T IC : 1 2 1 6 1 0 0 0 9 .D \ d a ta .m s
1 6 .2 1 0
No peak detected
APGC High and Low Energy
Chromatograms
High energy
Low energy
Unaligned data
APGC High/Low Energy Spectra Unknown
Impurity 15.75mins
High energy
Low energy
Summary and Conclusions
Complementary to EI allowing selection of molecular ion or
fragmentation.
Quantification over wide concentration range
Faster separations and higher sensitivity the EI GC/MS
Positive/negative ion switching
Detection of compounds that are not observed in LC/MS
Does not require analyte solubility
API source with ASAP® probe, GC/MS as well as LC/MS
extends the range of analytes addressed by one instrument
Product 2
ASAP-MS
Rapid technique for the analysis of solids,
liquids, polymers, and tissue samples
ASAP®-MS
MS inlet ASAP Probe
discharge
needle
melting
Point tube
N2
heated gas
LC Probe
Atmospheric Solids Analysis Probe (ASAP®)
Simple
Fast
Sensitive
Versatile
McEwen, C. N.; McKay, R. G.; Larsen, B. S., Anal. Chem. 2005, 77, 7826-7831.
DART® Cocaine on Dollar Bill
ASAP Cocaine on a Dollar Bill
Cocaine
C17H22O4N
-PhCOOH
C10H16O2N
Phathalate
C8H5O3
C24H39O4
150 200 250 300 350 400 450 500
m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100 R
ela
tive A
bundance
149.023
C 8 H 5 O 3
304.154
C 17 H 22 O 4 N 1
182.117
C 10 H 16 O 2 N 1
391.284
C 24 H 39 O 4
167.034
C 8 H 7 O 4 367.269
C 18 H 39 O 7
425.311
C 22 H 41 O 4 N 4 279.159
C 16 H 23 O 4 483.352
C 25 H 47 O 5 N 4
353.305
C 22 H 41 O 3
117.090
C 6 H 13 O 2
135.116
C 10 H 15
ASAP
• Solids probe replacement
• Accurate mass of mixtures
• Quantification
• Analysis of drugs (or explosives)
• Analysis of biological tissue
• Analysis of polymer additives
• Analysis of food residues
• Analysis of everything volatile
EPA8270_060524143111 #4 RT: 0.06 AV: 1 NL: 6.03E7
T: FTMS + p APCI corona Full ms [ 50.00-1000.00]
95 100 105 110 115 120 125 130 135 140 145 150 155 160 165 170
m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Re
lative
Ab
un
da
nce
131.12C6 H15 O1 N2
109.06
C7 H9 O1
139.11C9 H15 O1
94.06
C6 H8 N1
143.09
C11 H11
128.03C9 H4 O1
123.08
C8 H11 O1155.09C12 H11
153.07C12 H9
163.04C9 H7 O3107.05
C7 H7 O1170.10
C12 H12 N1
121.06C8 H9 O1
149.02
C8 H5 O3
135.08
C9 H11 O1
96.05
C1 H8 O3 N2
166.08C13 H10
105.07
C8 H9
145.06C10 H9 O1
111.04
C6 H7 O2
115.05C9 H7
119.09C9 H11
1
2
3
4
5
6
7
8
9
1. Aminobenzene
2. Phenol
3. Methylphenol
4. Dimethylphenol
5. Nitroso-di-propylamine
6. Isophorone
7. Methylnapthalene
8. Acenaphthylene
9. Diphenylamine
ASAP of Restek EPA 8270 Megamix
US 301
75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600 625 650 675 700m/z0
100
%
cnmtest 145 (2.694) Cm (140:147-(126:137+151:163)) TOF MS ES+1.53e485.0
84.0
127.0
97.0
99.0
115.0
145.1
289.1
271.1
163.1
253.1
241.1229.1
180.1 211.1
325.1
307.1
326.2
US 301
410 420 430 440 450 460 470 480 490 500 510 520 530 540 550 560 570 580 590 600 610 620 630 640 650 660 670 680 690m/z0
100
%
cnmtest 145 (2.694) Cm (140:147-(126:137+151:163)) TOF MS ES+ 264537.5
416.2
415.2
408.2
433.2
431.4
417.4
451.2434.2
446.2
536.5
469.2460.2
472.2
538.5
539.5
545.6591.6577.6
575.6
561.6
619.6617.6
593.6 621.6 681.6
AP Solids Probe of Tomato
Lycopene, m/z 537
537
m/z 400 - 700
US 301
75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500 525 550 575 600 625 650 675 700m/z0
100
%
cnmtest 262 (4.959) Cm (261:263) TOF MS ES+ 2.00e3112.1
102.1
84.1
86.1
130.1
337.4
132.1
166.1
133.1
147.1
165.1
257.3
239.3
182.1191.2
225.3
309.4
285.3
267.3
295.4310.4
326.4
365.5338.4
341.4
354.5
369.4
395.4
386.5
565.7
431.4400.5537.5
435.6
463.5538.5
582.7
583.7625.7
610.7
US 301
410 420 430 440 450 460 470 480 490 500 510 520 530 540 550 560 570 580 590 600 610 620 630 640 650 660 670 680 690m/z0
100
%
cnmtest 262 (4.959) Cm (261:263) TOF MS ES+ 357565.7
431.4
400.5
414.5
412.3
411.5415.5
425.5
537.5
435.6
463.5
451.4437.5
442.5 451.5
481.6498.6
490.5512.6
501.5 523.5
513.6 535.6
538.5551.5
538.6
554.7
555.6
582.7
566.7
570.6
579.6
583.7
625.7
610.7593.7
607.7
594.7613.6 626.7
653.8649.8638.7
666.8 694.8682.8
AP Solids Probe of Spinach Leaf
Apo-carotenal, 431
Beta-carotene, 537
Canthaxanthin, 565
Astaxanthin?
m/z 400 - 700
ASAP of Nonwoven Fiber
1
2
3
4
5
6
1. Erucamide
2. Fragment of 4
3. Irganox 1076
4. Irgafos 168
5. Irgafos 168 oxidized
6. Irganox 3114
:
300 350 400 450 500 550 600 650 700 750 800
m/z
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Rela
tive A
bundance
646.451 C 42 H 63 O 3 P 1
530.469 C 35 H 62 O 3
441.291 C 28 H 42 O 2 P 1
338.341 C 22 H 44 O 1 N 1
783.517 C 48 H 69 O 6 N 3
+
+
-
- +
+
+ +
+ ASAP
Probe
AP
CI
Pro
be
HV
Mass
Analyzer
N2
+
Crude Oil Sample by ASAP MS
PTEY = 150407+1.13976e+007*X R 2̂ = 0.9891 W: 1/X
0 5 10 15 20 25
0
50000000
100000000
150000000
200000000
250000000
300000000
Are
a
DZY = -496002+1.27123e+007*X R 2̂ = 0.9894 W: Equal
0 5 10 15 20 25
0
50000000
100000000
150000000
200000000
250000000
300000000
Are
a
MLTY = -499440+680790*X R^2 = 0.9911 W: Equal
0 5 10 15 20 25
0
5000000
10000000
15000000
Are
a
PTE
1 ng/mL
MLT
1 ng/mL
DZ
1 ng/mL
OEt
OEt
Pr-iMe
O P
SN
N
Diazinon (DZ)
EtO OEt
OMe
OMe
CH
S
CH 2C
P
C
O
S
O
Malathion (MLT)
OEt
OEt
O 2 N
PO
S
Parathion-ethyl (PTE)
Waters ASAP® Probe
Conclusion
• Direct analysis of vaporizable compounds
• Liquids, solids, and materials
• Minimal modification of ESI/APCI source
• Rapid switching from ASAP to ESI/APCI
• Simple to use and highly sensitive
Future Product?
MAI
Matrix-assisted ionization
Vacuum (MAIV), 2013
• Solid Matrix
• No Laser
• No Heat
• No voltage
• All MAIV matrices sublime in vacuum
eliminating instrument contamination
Inutan, E. D.; Trimpin, S., Mol. and Cell. Proteomics 2013, 12, 792-796
Ubiquitin Mass Spectrum using the
SYNAPT G2 MALDI Source
m/z 700 800 900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900
%
0
100 1071.51
952.56
857.41
779.56
714.77
1224.44
1428.33
1713.98 +5
+6
+7
+8
+9
+10
+11
+12
No laser and only 5 V extraction!
N
N H
N
N
C l
100 200 300 m/z
Full MS Scan
270.08
272.08
250.19
192.07 144.11
0
%
100
360.34
327.15 [M+H]+
4.32e4
MAI-MSE: Untargeted MS/MS of Drug Spiked Human Urine
Woodall, D.W., Wang, B., Inutan, E.D., Narayan, S.B., Trimpin, S. Anal. Chem., 2015, 87, 4667-
4679.
Clozapine (MW 326)
227.05
N
N H
N
N
C l 192.07
270.08
100 200 300 m/z
Full MS Scan
270.08
272.08
250.19
192.07 144.11
0
%
100
360.34
327.15 [M+H]+
4.32e4
B
%
300200100
100192.07
270.08
227.05
0
m/z
MSE Product Ion Spectra
100
0 100 200 300
192.07
227.05
270.08
m/z
%
MAI-MSE: Untargeted MS/MS of Drug Spiked Human Urine
Woodall, D.W., Wang, B., Inutan, E.D., Narayan, S.B., Trimpin, S. Anal. Chem., 2015, 87, 4667-
4679.
270.09(3.66) 227.05(3.66) 192.08(3.66)
300
100
200
400
m/z
Drift time (ms) 5.0 4.0 3.0 2.0
Clozapine (MW 326) CID in Transfer Collision Cell
ESI
source
enclosure
Overridden
Z-Spray
source
ESI source
enclosure
and skimmer
cones
removed
Operating
temperature
used
INSTRUMENTATION Waters ACQUITY QDa
Detector Mass Spectrometer
Inlet
aperture
Fast Polarity Switching using Waters QDa,
MAI of Angiotensin I
[M-2H]2-
[M+2H]2+
[M+3H]3+
m/z 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700
%
0
100
m/z 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700
%
0
100 4.08e5 202.9588
141.9816
126.9913
142.9810
167.0691
217.1569 647.03
289.0624 218.1568
268.0585 301.0654 335.6765
316.3699 364.1879
576.6978 393.1011 473.2426
1.37e7 649.12
433.154
325.0662 129.0050 198.2893
132.5229 175.2030
272.0847 245.0887 338.2089
395.0921 591.5055 502.8628 445.8398
513.3965 561.3858 640.6224
668.0519 680.7145
Negative
Positive
Mass Analyzer
X-Y-Z Stage Movable Carriage
1 µL Syringe
Loaded Syringe is introduced to MS Inlet
0.3 µL matrix/ analyte
deposited and dried on
needle
MSTM platform
Syringe Method (1uL)
• 1ul syringe
• 10mg/ml 3-NBN solution
• Inlet temperature : 70 C
• 0.2ul of analyte was pulled into the syringe
and ejected out
• 0.2ul of 3-NBN solution was withdrew into
syringe and pushed out to make droplet at
a tip of needle
• Rinse with water/methanol between
samples.
69
zero
de
ad
vo
lum
e s
yrin
ge
Prototype MAI Platform for Waters Instruments
APGC Source Housing
Cover Removed
1 µL Syringe Holder
MH2+2
MH3+3
MH+
Substance P (1 pmol/µL) MAI Needle Method Waters Xevo Alpha
MH2+2
MH+
MH3+3
Conclusion
• Matrix-assisted ionization (MAI) is sensitive,
easy to use, and robust
• Provides a rapid means of analyzing volatile,
nonvolatile, low- and high-mass compounds
Acknowledgements
Barbara Larsen, DuPont: Rich McKay, M&M: Sarah Trimpin, WSU
McEwen lab: Shuba Chakrabarty, Vince Pagnotti,
Milan Pophristic , Christian Reynolds
Khoa Hoang , Dan Woodall
Jessica DeLeeuw Zachary Devereaux
MSTM
solutions
NSF STTR Phase I Grant
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