SMASH 2008 Recap:Introducing Citation, Your Hard-
Working Lab Assistant
Dean Olson, Ph.D.Director of Customer Support
Protasis Corporation
Citation:Push-Button Sample Loader
• Chromatography-grade Fluidics• Easy-to-use, Web-based Software• Built-in Instrument Controller• Integral Degassing• Field-serviceable Valves• Small-footprint benchtop packaging• Enjoy the reassurance of feedback
control, auto-calibration, and system self-maintenance in applications where the robot is not required
• Completely upgradeable to full robotic automation
Chromatography grade pumps
Field-serviceable valves
Integrated feedback control
sensors
Degas, blanket, sparge
On-board instrument controller
Protasis One-Minute™ NMR Lite Software
The size of a plant metabolome is unknown it is estimated at a few thousand constituents
Plant metabolomics
LC-NMR 10-6 mol
LC-UV 10-9 mol
GC-MS 10-12 mol
LC-MS 10-15 mol
LC-LIF 10-19 mol
CE-LIF 10-22 mol
Sumner et al. Phytochemistry 2003, 62,
817-836.
Rapid, Reproducible and Non-Destructive
Low Sensitivity
NMR Metabolomics
High Sensitivity
Instrumentation variability
LC-MS: problem in RT repeatability
MS Metabolomics
100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000m/z0
100
%
0
100
%
291.72
277.71
265.66
113.43
159.46133.48 175.45 223.57
305.74
309.70339.60
577.63353.73
397.77
381.76 531.80431.71 441.78623.61593.62
823.81807.83
739.60
291.71
277.70
265.66
113.43
223.60
305.73
319.76339.63
353.73397.75
381.75 441.76 577.69531.77 609.68 805.80647.65
100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000m/z0
100
%
0
100
%
291.72
277.71
265.66
113.43
159.46133.48 175.45 223.57
305.74
309.70339.60
577.63353.73
397.77
381.76 531.80431.71 441.78623.61593.62
823.81807.83
739.60
291.71
277.70
265.66
113.43
223.60
305.73
319.76339.63
353.73397.75
381.75 441.76 577.69531.77 609.68 805.80647.65
CONTROL
WOUNDED
PCA of total crude extract MS spectra
WA
CA
WB
CB
WA
CA
WB
CB
spiked
spiked
Total crude extract TOF MS spectra
RANKED LIST OF INDUCED IONS
MATLABm/z coord t1209 0.092353338 0.088507308 0.087165226 0.086949352 0.086062237 0.079287446 0.078662307 0.077829256 0.076834263 0.076811194 0.076683272 0.075681250 0.075639238 0.07556
Total crude extract TOF MS spectra
CONTROL
WOUNDED
C12
H17
O3
209.1256
209.1256
Data Courtesty of Data Courtesty of Jean-Luc Wolfender, Jean-Luc Wolfender, University of Geneva, University of Geneva,
SwitzerlandSwitzerland
UPLC-TOF-ES-MS of A. thaliana
11:06:54wt wound conc NEG new col 150 mmm aper 1 30 v
Time5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00
%
0
100
5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00
%
0
100
5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00
%
0
100
060316_wt_w_conc_neg_1_ap30 1: TOF MS ES- 209.12 10PPM
5.41e326.37209
060316_wt_w_conc_neg_1_ap30 1: TOF MS ES- 209
5.44e326.37209
1.15210 22.63
20910.76371
1.30;436 12.76338
31.46309
28.31501
060316_wt_w_conc_neg_1_ap30 1: TOF MS ES- BPI
3.84e419.14577
14.3673911.24
492
8.72285
1.26404
1.85436
6.263385.07
500
9.66447
13.68431
17.26593
16.47431
26.88545
26.51517
23.74579
19.69498
28.00689
33.8129130.83
26329.26527
37.1881935.94
809 38.75311
11:06:54wt wound conc NEG new col 150 mmm aper 1 30 v
Time5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00
%
0
100
5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00
%
0
100
5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00
%
0
100
060316_wt_w_conc_neg_1_ap30 1: TOF MS ES- 209.12 10PPM
5.41e326.37209
060316_wt_w_conc_neg_1_ap30 1: TOF MS ES- 209
5.44e326.37209
1.15210 22.63
20910.76371
1.30;436 12.76338
31.46309
28.31501
060316_wt_w_conc_neg_1_ap30 1: TOF MS ES- BPI
3.84e419.14577
14.3673911.24
492
8.72285
1.26404
1.85436
6.263385.07
500
9.66447
13.68431
17.26593
16.47431
26.88545
26.51517
23.74579
19.69498
28.00689
33.8129130.83
26329.26527
37.1881935.94
809 38.75311
Induced by wounding
m/z 209.12 (+/- 10 ppm)
m/z 209.12 (+/- 0.5 Da)
CAP-NMRCAP-NMR
Multiple collection Multiple collection triggered by MStriggered by MS
Data Courtesty of Jean-Luc Wolfender, Data Courtesty of Jean-Luc Wolfender, University of Geneva, SwitzerlandUniversity of Geneva, Switzerland
CAP-NMR of ion m/z 209
5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5Chemical Shift (ppm)
CAP-NMRca 1 gNT=5000 12
10 9 7 3,2,8 11,5 4 12
Standard 50 g
COOH
O 1187
5
43
2
19 10
Data Courtesty of Jean-Luc Wolfender, Data Courtesty of Jean-Luc Wolfender, University of Geneva, SwitzerlandUniversity of Geneva, Switzerland
Further example details can be found in…
A LC-MS approach is presented for the isolation of minor key plant biomarkers, in view of their characterization by NMR at the microgram scale …using ultra-performance liquid chromatography-time-of-flight mass spectrometry (UPLC-TOF-MS) … then transferred to semi-preparative LC conditions with MS detection … and subsequent characterization by capillary NMR (CapNMR).
Hot Off the Press: Natural Products Workflow
Herein we present a method of extract evaluation using HPLC profiling with biological evaluation16 followed by capillary probe NMR spectroscopy/ESMS/UV combined with NMR database (AntiMarin) evaluation, which reduces the crude extract requirement for dereplication to submilligram quantities.17
Full RoboticAutomation
Automated Push-Button Discrete Sample Loadingvs.
You pick
Top Related