fast method development using autosrm and...
TRANSCRIPT
![Page 1: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/1.jpg)
1 The world leader in serving science
Inge de Dobbeleer
Fast Method Development using AutoSRM and autoSIM
![Page 2: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/2.jpg)
4
RT: 0.00 - 20.00 SM: 7G
0 2 4 6 8 10 12 14 16 18Time (min)
0
10
20
30
40
50
60
70
80
90
100
0
10
20
30
40
50
60
70
80
90
100
Rel
ativ
e A
bund
ance
RT: 8.23SN: 35
15.36
14.31
15.52
14.09 16.6718.8517.04
12.42 17.90 19.0113.229.33
12.2610.648.13 8.42 9.756.365.375.214.69RT: 8.23SN: 506
Pentachlorobenzene Masstrace m/z
250 in fullscan acquisition
S/N 35
Pentachlorobenzene m/z 250 in SIM
acquisition S/N 506
Short refresher on SIM benefit
Pesticides in soils
![Page 3: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/3.jpg)
5
Fullscan TIC
Dichlorvos in SIM @ m/z185
Dichlorvos in SRM 185>109
Dichlobenil in SIM @ m/z171
Dichlobenil in SRM 171>136
Refresher on MS/MS benefit
Pesticides in herbs
![Page 4: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/4.jpg)
6
Method development using the classical set up with segments or windows
RT: 7.61 - 29.04
8 10 12 14 16 18 20 22 24 26 28Time (min)
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
105
110
115
Rel
ativ
e Ab
unda
nce
15.88
9.3413.8912.06 14.01
19.83
13.24 14.6111.63 17.07
12.8511.149.84 15.59 18.128.60 16.96 23.4010.6919.90
16.27 18.1817.87 20.82 23.17 25.0321.24 23.61
18.67 25.45
22.0819.61 26.9627.32
23.0027.86
Segment this…… 150 pesticides
![Page 5: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/5.jpg)
7
Method development using the classical set up with segments or windows
RT: 12.95 - 17.25
13.0 13.5 14.0 14.5 15.0 15.5 16.0 16.5 17.0Time (min)
25
30
35
40
45
50
55
60
65
70
75
80
85
90
95
100
Relativ
e Abun
dance
15.88
13.89 14.01
13.24 14.6113.6517.07
14.2215.59
16.96
13.4515.07 16.27
16.6116.0014.83 15.3515.81
16.7315.25 15.44 16.44
14.49
14.45
16.1813.03
N1T M_
• In SIM we need 3 ions/comp or 126 ions/segment • In MS/MS we need two transitions or 82 ions/segment • Manual set up leads to errors • Matrix injections leads to RT shift, compounds fall out of the
window, so often methods are made with compounds in two segments
• Many ions/ segment= lower dwell times= lower S/N
42 compounds in a segment
![Page 6: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/6.jpg)
8
Modern method development for single and triple quads
Fullscan
• Library search • List of compounds and RT
develop
• autoSIM or • autoSRM
GO
• Link to software • Acquire and quantify • Update method
![Page 7: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/7.jpg)
9
AutoSIM
GC and AS method
Chromatography OK?
no
Yes. Automation can start
FS run + lib search Compound list
Or… Start automation here
Click on the compound; A mass intensity list and S/N is shown; Click on the desired m/z Create a working list
Attach working list to Instrument method; Quantitation software
![Page 8: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/8.jpg)
10
Productivity Features: t-SIM
Acquisition - Timed
Link the working list; the work list is in fact now a database
![Page 9: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/9.jpg)
11
AutoSRM
1) Precursor ion selection
2) Product ion selection
3) Collision energy optimization
![Page 10: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/10.jpg)
12
AutoSRM
• Automates the following: • Creation of fullscan, product ion scan and SRM methods • Creation of sample sequences • Creation of data layouts for analyzing results • Selection of precursor, product and collision energies
End result showing optimized transition
![Page 11: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/11.jpg)
13
Timed-SRM Method Overview
Acquisition - Timed
Link the working list; the work list is in fact now a database
Pre and post times are adjustable; So in case of shift due to heavy matrix the compound is still inside the window
![Page 12: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/12.jpg)
14
GC-MS and MS/MS Method Development Workflow
Develop missing compound/ SRM or SIM
Add to database
Multi-component method
Automatic SIM or MRM method creation
Self -customized Auto-optimized
RT Update
My target compound
list Auto SIM Auto SRM
Just ask us for existing databases
![Page 13: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/13.jpg)
15
Analysis of 300 Pesticides – Segmented vs. Timed
Segmented SRM Timed SRM Closest compound to segment break: 5 seconds
Can lead to compounds disappearing in case of RT shift
Closest compound to segment break: 15 seconds
Average number of simultaneous transitions: 55
Average number of simultaneous transitions: 15 (4X higher dwell times- better sensitivity)
![Page 14: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/14.jpg)
16
AutoSRM User Case
• Created and optimized > 250 transitions for > 80 compounds
• Minimal user interaction over 24 hours
![Page 15: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/15.jpg)
17
TSQ 8000 Timed-SRM User Case result
Terbacil
Alachlor
Tolylfluanid
Pyridaben
Tea Analysis: 4 pg on-column
![Page 16: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/16.jpg)
18
TSQ 8000 Timed-SRM User Case result
Spiked ketchup Analysis: 50 ppb Example diazinon
12.5 13.0 13.5 14.0 14.5 15Time (min)
RT: 13.64AA: 1555401SN: 80040
RT: 13.63AA: 1047822SN: 6301
Top: Timed SRM: - S/N 80040 - response: 1555401 Bottom: Segmented SRM: - S/N 6301 - response: 1047822
![Page 17: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/17.jpg)
19
TraceFinder Method Sync
• Links TraceFinder Method with instrument method
• Enables: • Compound based acquisition setup
• Automated update of acquisition windows
![Page 18: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/18.jpg)
21
Method Sync – Automated RT Update
…updates both TraceFinder Method and Timed-SRM Method
![Page 19: Fast Method Development using AutoSRM and autoSIMtools.thermofisher.com/content/sfs/brochures/Vendor-Semiar-3-ISCC... · Fast Method Development using AutoSRM and autoSIM . 4 . RT:](https://reader031.vdocuments.mx/reader031/viewer/2022022010/5afde9ee7f8b9a994d8e3b02/html5/thumbnails/19.jpg)
22
In Conclusion
Image courtesy of digital art FreeDigitalPhotos.net
• Auto SIM and auto SRM:
• Simplifies the method development • Speeds the method development up • Makes adding new compounds easy • Connects the instrument and quantifying method
• Timed SIM and SRM:
• Eliminates the need for cumbersome segmenting • Limits the number of ions per unit of time; • Thus ensuring optimal dwell time and increasing the signal
to noise