a new era in high speed analysis with dynamic mrm and … new... · page 1 a new era in high speed...
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A New Era in High Speed Analysis with Dynamic MRM and UHPLC
Tim Schlabach, Marketing
Anabel Fandino, Ph.D., R&D
PeteStone, Ph.D., Marketing
Agilent 1290 Infinity LC Attributes for MS - Infinitely Better for LC/MS
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1200bar @ 2mL/min for highest resolutionReduced Ion & Matrix Suppression
HT-Solution for up to 2,000 samples/dayComplete System Integration MassHunter
Less than 100 uL delay volumesFirst peaks can elute in < 6 seconds
High Pressures
LowDelay Vol.
High Throughput
Agilent 6000 Series LC/MS: Speed for UHPLC
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Fast MRMsfor UHPLC
40 MS Scans/sec10 MS/MS scans/sec
BestData
QualityFast Scans for UHPLC
Agilent 6460
Agilent 6530
1 ms dwell for MRM30 ms Pos/Neg Switching> 200 MRMs/ sec
Fast MRMs with 6400 Series QQQ – metabolism polarity switching in 30 millisecs plus 6 transitions (2 ISTDs)
Avg W1/2 = 1.3 sec 35 points across RSD [%] = 1.8
Avg W1/2 = 0.71 sec 15 points across RSD [%] = 2.9
Avg W1/2 = 1.3 sec 35 points across RSD [%] = 2.7
Avg W1/2 = 0.52 sec 11 points across RSD [%] = 6.5
Verapamil, 0.75 min
Buspirone, 0.40 min
Dextromethorphan, 0.53 min
Diclofenac, 1.04 min
Peaks widths with UHPLC only 2 seconds or less > ten uniform measurements across peak REQUIRED
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W = 2.1 sec
Cycle time < 115 ms
Avg. W1/2 = 0.72 sec
Verapamil, peak RSD 5%
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Dynamic MRM for 6400 Series Triple Quads monitors ion transitions only when compounds elute
With DMRM the number of concurrent MRMs are fewer than with Time Segments
Cycle Time:Concurrent MRMsDwell Time:
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Agilent 1290 Infinity LC + 6460 Triple Quad
9 replicates analyzed at 200 pg levelMean area %RSD = 3.2
Analysis of 600 Pesticide DMRM Transitions in 15 min.
17 min.
1290/6230 Accurate Mass TOF Pesticide Screening250 pesticides, 50 pg on column
3 min.
1.5 min.
Most peaks 1-2 seconds wide!
The Need for Speed in UHPLC/MS/MS
Cycle Times (MRM): <150 milliseconds
Dwell Times (MRM or DMRM): <= 2 millliseconds
Polarity Switching (MRM): < 50 milliseconds
Spectral scans: at 5 Hz or faster
PPM Mass Accuracy: at 5 Hz
20 K Resolution: at 5 Hz
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