By utilizing StepWave Technology within the SYNAPT G2-S platform, comprehensive untargeted identification and quantification of compounds can be carried out at the lowest levels in complex matrices while minimizing MS method development. A Step Change in High-Resolution Quantitative Performance Combining Novel StepWave, QuanTof, and MS E Technology in the SYNAPT G2-S System GOAL To successfully quantify compounds while acquiring high resolution full scan spectral data to allow unequivocal identification. BACKGROUND Routine quantitation has traditionally been reserved for tandem quadrupole instruments where the high level of selectivity enables scientists to detect very low levels while still providing a large dynamic range. ToF platforms, while providing the scientist with comprehensive amounts of data for untargeted quantitation, have often been limited not only in their detection levels when compared to tandem quadrupoles, but also in the range of concentrations that can be accurately determined in a single batch. Both ToF platforms and tandem quadrupole instruments are limited to detecting whatever ions can be efficiently introduced via the source. Simply allowing more ions into the machine is never a guarantee that detection limits will be lowered. The StepWave™ device allows significantly more ions to be introduced while utilizing a robust mechanism for elimination of neutral components and gas stream from the rest of the instrument. QuanTof Technology also provides a wide dynamic range to maximize both quantitative and qualitative performance with high resolution MS. THE SOLUTION A SYNAPT ® G2 coupled with an ACQUITY UPLC ® System was used to chromatograph and detect Sulfadimethoxine. Calibration standards had been prepared by spiking separate solutions with increasing amounts of Sulfadimethoxine. These were then injected and analyzed in sequence. The entire sequence was repeated using the same solutions on a SYNAPT G2-S System, featuring high-sensitivity StepWave ion optics. Figure 1. Increase in limit of quantitation by over 10X with StepWave Technology with over four orders of linearity provided by the high resolution QuanTof analyzer. Conc 0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0 50.0 Response 0 10000 20000 30000 40000 50000 ULOQ of 500 pg on column LLOQ of 25 fg on column 4.3 orders of linearity Residuals < 20% R 2 = 0.99 SYNAPT G2-S (with StepWave) Conc 0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 Response 0 2000 4000 6000 8000 10000 LOQ of 250 fg on column SYNAPT G2 (no StepWave)
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By utilizing StepWave Technology within the SYNAPT G2-S platform, comprehensive untargeted identification and quantification of compounds can be carried out at the lowest levels in complex matrices while minimizing MS method development.
A Step Change in High-Resolution Quantitative Performance Combining Novel StepWave, QuanTof, and MSE Technology in the SYNAPT G2-S System
GOA L
To successfully quantify compounds while
acquiring high resolution full scan spectral
data to allow unequivocal identification.
BAc kG rOu n d
Routine quantitation has traditionally been
reserved for tandem quadrupole instruments
where the high level of selectivity enables
scientists to detect very low levels while
still providing a large dynamic range. ToF
platforms, while providing the scientist with
comprehensive amounts of data for untargeted
quantitation, have often been limited not only
in their detection levels when compared to
tandem quadrupoles, but also in the range
of concentrations that can be accurately
determined in a single batch.
Both ToF platforms and tandem quadrupole
instruments are limited to detecting whatever
ions can be efficiently introduced via the
source. Simply allowing more ions into the
machine is never a guarantee that detection
limits will be lowered.
The StepWave™ device allows significantly
more ions to be introduced while utilizing a
robust mechanism for elimination of neutral
components and gas stream from the rest of the
instrument. QuanTof Technology also provides
a wide dynamic range to maximize both
quantitative and qualitative performance
with high resolution MS.
T h e sO Lu T iO n
A SYNAPT® G2 coupled with an ACQUITY UPLC® System was used to
chromatograph and detect Sulfadimethoxine. Calibration standards had
been prepared by spiking separate solutions with increasing amounts of
Sulfadimethoxine. These were then injected and analyzed in sequence.
The entire sequence was repeated using the same solutions on a
SYNAPT G2-S System, featuring high-sensitivity
StepWave ion optics.
Figure 1. Increase in limit of quantitation by over 10X with StepWave Technology with over four orders of linearity provided by the high resolution QuanTof analyzer.
Conc0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0 40.0 45.0 50.0
Resp
onse
0
10000
20000
30000
40000
50000
ULOQ of 500 pg on columnLLOQ of 25 fg on column4.3 orders of linearityResiduals < 20%R2 = 0.99
SYNAPT G2-S (with StepWave)
Conc0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100
Resp
onse
0
2000
4000
6000
8000
10000 LOQ of 250 fg on column
SYNAPT G2 (no StepWave)
Waters corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com
scientists while still allowing the capacity to collect high resolution molecular and
fragment ion spectral data for hundreds of compounds. High resolution ToF data
reduces the need for method development as there is no requirement for MRM
discovery or optimization, which can be a labor-intensive process for applications
where many analytes are targeted. Unbiased full scan data also enables archived
data to be interrogated at any point in the future for new compounds of interest.
Waters, SYNAPT, ACQUITY UPLC, and UPLC are registered trademarks of Waters Corporation. T he Science of What’s Possible, MassFragment, and StepWave are trademarks of Waters Corporation. All other trademarks are the property of their respective owners.
©2011 Waters Corporation. Produced in the U.S.A.May 2011 720003963EN LB-PDF
The data was collected in UPLC®/MSE mode. This allows
both intact molecular ion and fragment ion information
to be collected for all molecules in the sample. (No prior
knowledge of the samples was required.)
The data shown in Figure 2 demonstrates that the
use of the StepWave in the SYNAPT G2-S instrument
resulted in a six-fold increase in sensitivity (signal-to-
noise) over that obtained with a SYNAPT G2 System.
Figure 1 shows that the quantitative performance
standard, i.e. linear dynamic range and reproducibility,
was not adversely affected by the use of StepWave
Technology. In fact the StepWave enabled a 10x
improvement in the limit of quantitation. The SYNAPT
G2 achieved an LOQ of 250 femtograms and was
linear up to 100 picograms demonstrating over 3.5
orders of dynamic range. The SYNAPT G2-S System
with StepWave achieved a 10x lower detection limit,
25 femtograms, and more than 4 orders of linear
dynamic range.
Due to the ‘data independent’ MSE acquisition
mode employed in this analysis, it is possible to
unequivocally identify the components in the sample
using high resolution molecular and fragment ion
spectral data from every delectable component.
For example, Figure 3 shows the high and low energy
spectra obtained for one of the Sulfadimethoxine
peak at 2.5 pg on column injections.
suMMA rY
The data collected demonstrates excellent sensitivity
and linearity together with high mass resolution and
exact mass accuracy. The StepWave successfully
enables the SYNAPT G2-S platform to meet the
quantitative performance required by today’s
Figure 3. Exact mass molecular ion (lower spectrum) and fragment ion (upper spectrum) data from the UPLC/MSE analysis enabling rapid, unequivocal structural identification of Sulfadimethoxine with MassFragment™ Software.
m/z100 120 140 160 180 200 220 240 260 280 300 320 340 360
%
0
1001.12e4156.0776
127.0507
311.0808
245.1049218.0229
176.0325 287.1771 364.2942329.3606
-1.1mDa
-0.6mDa
-0.6mDa
+0.2mDa
m/z100 120 140 160 180 200 220 240 260 280 300 320 340 360
%
0
1008.42e4311.0816
209.3962123.8988 179.3056 245.1001 280.3845 349.0429
Figure 2. Increase in signal:noise with the combined ability of StepWave to maximize ion sampling efficiency while eliminating neutral contaminants.
Time0.00 0.50 1.00 1.50 2.00
%
0
100
0.00 0.50 1.00 1.50 2.00
%
0
1000.38
0.38
0.23 1.521.390.72
0.61 1.161.00 2.232.051.712.01
Signal to noise
SYNAPT G2-S (with StepWave) 600:1SYNAPT G2 (no StepWave) 100:1Increase X6
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