triple quadrupole gas chromatography-mass spectrometry/mass spectrometry re-imagined: increased...
DESCRIPTION
Key Learning Objectives: • Identify emerging triple quadrupole Gas Chromatography-Mass Spectrometry/Mass Spectrometry (GC-MS/MS) technology designed to address increasing regulatory demands and requirements • Explore potential time savings in sample prep, method development/transition, and data analysis • Demonstrate how to optimize the GC-MS/MS workflow from sample prep to sample analysis to automated data analysis Overview: Regulatory lab requirements continue to drive detection limits lower with an ever increasing list of compounds to analyze. These requirements also demand greater precision at these lower limits. Triple quadrupole GC-MS/MS is a viable option for enhanced analysis and increased productivity with an emphasis on simplicity. We discuss emerging trends and technologies designed to ensure that laboratories are well-equipped to address these increased demands with minimal investment in training and method development. Find out how you can adopt triple quadrupole GC-MS/MS technology in your laboratory using existing methods and source parameters in most instances while requiring less sample prep and enjoying the benefits of automated data analysis for increased simplicity and productivity. For more information: www.thermoscientific.com/tsq8000TRANSCRIPT
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The world leader in serving science
Increased Simplicity and Productivity
Triple Quadrupole GC-MS/MS Re-imagined
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Tough challenges faced in the laboratory
• High sample loads/short deadlines
• Keeping sample analysis costs down with more and more challenging LODs and matrices
• Integrating and maintaining new methods and technologies into production workflows to remain competitive
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What‘s required...
• Realization of the productivity advantages of high performance GC-MS/MS
• Minimizing the impact of adoption and implementation to current laboratory operations
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Triple Quadrupole GC-MS/MS is an essential part of a cost-effective, high productivity analytical method in today’s laboratory
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Many laboratories are already investing in and exploring GC-MS/MS as a tool to obtain a competitive edge in their analyses
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GC-MS/MS – What’s so special?
• Low detection limits
• Reduced sample preparation
• Consolidated analytical methods
• Faster, automated data processing
...it is a high selectivity technique...
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Selectivity in a method
McLafferty circa. 1980
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Method performance requirement
• Target compounds
• Matrices
• Sensitivity
Method performance requirement
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First sample prep..
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Method performance requirement
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...then instrument detection...
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Total method selectivity
Method performance requirement
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Analytical Benefits for Single Quadrupole GC-MS
• Robust• Run more samples between cleaning
• Sensitive Precision• Accurate and reproducible results at the lowest levels
• Unknown Analysis• Full scan for unknown library searches
• Alternating full scan/SIM for unknowns and low level analysis
• Flexibility• Switch quickly between dedicated EI and CI sources
• Easy to Use and Maintain
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Full Scan/SIM Methodology for Drinking Water
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Challenging Samples for Single Quadrupole GC-MS
• Matrix Challenges• Concentration
challenges• Difficult to prove
contamination
Gamma BHC
Methiocarb Mevinphos
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...what about GC-MS/MS?...
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Total method selectivity
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Method performance requirement
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Use GC-MS/MS to reduce clean-up...
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Use GC-MS/MS to consolidate methods...
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Method 1performance requirement
Method 2performance requirement
Method 3performance requirement
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Use GC-MS/MS to consolidate methods...
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Consolidated multi-residue method
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What will GC-MS/MS do for my lab?
High selectivity
• Possibility the reduce selectivity in sample preparation
• Reduced sample prep steps creates a more generic sample prep method – more compounds & matrices
• Consolidated GC-MS methods due to high performance – buffer against requirements
• Compressed chromatography possible
• Easy peak evaluation – auto-integrators
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Step 1: Ions are produced in the source
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Step 2: Ions are focused in the s-shaped prefilter
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Step 2: Ions focused by prefilter, removing neutrals
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Step 3: Ions are isolated by molecular weight in Q1
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Step 3: Ions are isolated by molecular weight in Q1
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Step 4: Ions further fragmented in collision cell
(With same mass as analyte)
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Step 4: Ions further fragmented in collision cell
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Step 5: Unique ions to analytes chosen in Q3
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Only unique fragments to analyte detected
Step 5: Unique ions to analytes chosen in Q3
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AutoSRM Overview
1) Precursor ion selection
2) Product ion selection
3) Collision energy optimization
SR
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Step 1 – Pick Your Precursor Ions
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Step 1 – Pick Your Precursor Ions
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Step 1 – Pick Your Precursor Ions
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Step 2 – Pick Your Product Ions
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Step 2 – Pick Your Product Ions
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Step 3 – Optimize Your Transitions
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Step 3 – Optimize Your Transitions
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Export from AutoSRM to Instrument Method
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Timed-SRM Method Overview
Acquisition Windows centered around retention time and Window
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Timed-SRM Method Overview
Acquisition windows allowed to overlap
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Timed-SRM Advantages
Segmented SRM
Timed SRM
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Timed-SRM Advantages
Acquisition Windows
Segmented SRM
Timed SRM
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Timed-SRM Advantages
• Removes wasted dwell time
• Allow higher overall dwell times
• Leads to higher sensitivity
Wasted Dwell Time
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Timed-SRM Advantages
• Peaks centered in acquisition window
• No peak elutes near acquisition break
• Allows for retention time shift (e.g. due to heavy matrix)
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Instrument Parameters
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Timed-SRM from Thermo Scientific TSQ 8000 GC-MS
Screenshot of a section of the analytical run showing the “acquisition map” automatically created by the TSQ™ 8000 System using t-SRM.
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Calibration Curves
• All calibration curves correlation coefficients greater than 0.99
• Example calibration curve for Cyfluthrin, R2 = 0.9996
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Transitions for Pesticides from PAR ver 2
• SRM peaks at 4 ppb from Terbacil (left, 161.1 > 88.0, CE 15 V) and Alachlor(right, 188.1 > 130.1, CE 25 V)
• SRM peaks at 4 ppb from Tolylfluanid (left, 238.1 > 137.1, CE 15 V) and Pyridaben (right, 309.1 > 147.1, CE 15 V)
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The world leader in serving science
A Second Level of Selectivity
Structure and Mass Defect
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Thermo Scientific TSQ Quantum XLS Ultra – HyperQuad™ Technology Inside
Patented HyperQuad technology meets with GC/MS for the first time to create highest performing GC Triple on the market
6 mm hyperbolic precision quads allow excellent ion transmission at standard resolution as well as opportunity to use enhanced mass resolution to 0.1 Da peak width
Effective pre-cursor ion filtering with strongly reduced matrix interference
Improved signal/noise & quantitative precision
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U-SRM: Two Modes of Selectivity
• Ultra selective-Single Reaction
Monitoring (U-SRM)
• Offers a unique opportunity to increase selectivity using triple quadrupole.
• Combination• Increased Q1 mass resolution • MS/MS Structural based
selectivity
• For use when standard SRM does not provide enough selectivity
m/z
Increased mass resolution
Structural selectivity (MS/MS)
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Precursor Ion Selection Q1 (Standard SRM mode)
• At standard mass resolution precursor selection • Q1 = 0.7 Da FWHM• Higher probability that interfering
species are transmitted to the collision cell with the target compound mass
• These matrix interferents are often orders of magnitude higher than target compounds
• If they are not completely discriminated against in by CID then the resulting product ion detection can have a higher background noise present
Matrix components transmitted through Q1 during SRM
Q1= 218.9 m/z (0.7 Da res.)
Q1 Transmission Window
Q1= 0.7Da
219217
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Precursor Ion Selection Q1 (U-SRM mode)
HyperQuad does not allow matrix through through Q1 during U-SRM
• When operated in U-SRM mode, the TSQ Quantum XLS Ultra narrows the pre-cursor mass window to <0.2Da
• This increased resolution allows the HyperQuad to discriminate against common matrix component masses for targets showing a large enough
Δmass defect.
• This allows for better collision cell performance and robustness as well as reducing chemical noise
• Lindane carries a Δmass defect and can be isolated from matrix in this example
Q1= 0.1Da
219217 221
Q1= 218.86 m/z (0.1 Da res.)
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U-SRM: Effect of Increasing Q1 Resolution
• Real life effect of the lindane example…
• Lindane isomers in green tea
• As Q1 resolution is increased into the ultra range signal/noise ratio increases dramatically
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U-SRM in action
• PCB 28 in industrial soil sample at ca.100fg (ASE extraction with no clean-up)
• Continuum full scan spectrum shows background removed with HyperQuad at increased resolution
contocpSCAN003 #1594-1597 RT: 13.68-13.69 AV: 4 SM: 7B NL: 4.04E7T: + p EI Q1MS [200.000-300.000]
255.0 255.2 255.4 255.6 255.8 256.0 256.2 256.4 256.6 256.8 257.0 257.2
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XLSULTRA_NPV_OCP00016_110329102653 3/29/2011 10:26:53 AM
RT: 13.33 - 15.12 SM: 7G
13.4 13.6 13.8 14.0 14.2 14.4 14.6 14.8 15.0
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RT: 13.69
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14.4614.3014.64
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14.94 15.0214.09
14.05 14.8913.96
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TIC F: + c EI SRM ms2 255.960 [185.965-185.975] MS XLSULTRA_NPV_OCP00016_110329102653
SRM (Q1 0.7Da)
0.7 Da
256 m/z
Q1 precursor selection
2,4,4'-Trichlorobiphenyl
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U-SRM in action
• PCB 28 in industrial soil sample at ca.100 fg (ASE extraction with no clean-up)
• Continuum full scan spectrum shows background removed with HyperQuad at increased resolution.
• Dramatically improved detection using U-SRM
C:\XCALIBUR\...\contocpSCAN004 4/14/2011 5:27:58 PM
contocpSCAN004 #1469-1471 RT: 13.68-13.69 AV: 3 SM: 7B NL: 3.02E6T: + p EI Q1MS [200.000-300.000]
255.0 255.2 255.4 255.6 255.8 256.0 256.2 256.4 256.6 256.8 257.0 257.2
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contocpSCAN003 #1594-1597 RT: 13.68-13.69 AV: 4 SM: 7B NL: 4.04E7T: + p EI Q1MS [200.000-300.000]
255.0 255.2 255.4 255.6 255.8 256.0 256.2 256.4 256.6 256.8 257.0 257.2
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XLSULTRA_NPV_OCP00011_110329074139 3/29/2011 7:41:39 AM
RT: 13.33 - 15.12 SM: 7G
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RT: 13.69 NL: 3.21E3
TIC F: + c EI SRM ms2 255.960 [185.965-185.975] MS ICIS XLSULTRA_NPV_OCP00011_110329074139
XLSULTRA_NPV_OCP00016_110329102653 3/29/2011 10:26:53 AM
RT: 13.33 - 15.12 SM: 7G
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RT: 13.69
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14.4614.3014.64
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14.94 15.0214.09
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TIC F: + c EI SRM ms2 255.960 [185.965-185.975] MS XLSULTRA_NPV_OCP00016_110329102653
U-SRM (Q1 0.1Da)
Matrix interefent transmitted in standard mode (0.7Da) but rejected during U-SRM (0.1Da) precursor selection
PCB 28 Precursor mass 2 55.96 m/z selected
0.1
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0.7 Da
0.1 Da
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Environmental: Polychlorinated Biphenyls (PCBS)XLSULTRA_NPV_OCP00017_110329105955 3/29/2011 10:59:55 AM
RT: 13.17 - 16.75 SM: 7G
13.2 13.4 13.6 13.8 14.0 14.2 14.4 14.6 14.8 15.0 15.2 15.4 15.6 15.8 16.0 16.2 16.4 16.6
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RT: 14.04AA: 725579SN: 97
RT: 14.35AA: 37646SN: 280
RT: 14.78AA: 201199SN: 229
RT: 15.03AA: 140023SN: 185
RT: 14.38AA: 94563SN: 88
RT: 16.03AA: 24964SN: 94
RT: 16.03AA: 26376SN: 160
NL: 1.01E5
TIC F: + c EI SRM ms2 255.960 [185.965-185.975] MS XLSULTRA_NPV_OCP00017_110329105955
NL: 1.53E5
TIC F: + c EI SRM ms2 257.960 [185.965-185.975] MS ICIS XLSULTRA_NPV_OCP00017_110329105955
NL: 1.37E4
TIC F: + c EI SRM ms2 289.920 [219.935-219.945] MS ICIS XLSULTRA_NPV_OCP00017_110329105955
NL: 7.06E4
TIC F: + c EI SRM ms2 291.920 [219.935-219.945] MS ICIS XLSULTRA_NPV_OCP00017_110329105955
NL: 1.01E4
TIC F: + c EI SRM ms2 323.880 [253.905-253.915] MS ICIS XLSULTRA_NPV_OCP00017_110329105955
NL: 1.42E4
TIC F: + c EI SRM ms2 325.880 [255.905-255.915] MS ICIS XLSULTRA_NPV_OCP00017_110329105955
XLSULTRA_NPV_OCP00011_110329074139 3/29/2011 7:41:39 AM
RT: 13.17 - 16.75 SM: 7G
13.2 13.4 13.6 13.8 14.0 14.2 14.4 14.6 14.8 15.0 15.2 15.4 15.6 15.8 16.0 16.2 16.4 16.6
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RT: 13.69AA: 5479SN: 859
RT: 13.70AA: 5485SN: 862
RT: 14.37AA: 4122SN: 1454
RT: 14.36AA: 1393SN: 393
RT: 16.05AA: 2033SN: 352
RT: 16.04AA: 3693SN: 412
NL: 3.21E3
TIC F: + c EI SRM ms2 255.960 [185.965-185.975] MS ICIS XLSULTRA_NPV_OCP00011_110329074139
NL: 2.87E3
TIC F: + c EI SRM ms2 257.960 [185.965-185.975] MS ICIS XLSULTRA_NPV_OCP00011_110329074139
NL: 2.74E3
TIC F: + c EI SRM ms2 289.920 [219.935-219.945] MS ICIS XLSULTRA_NPV_OCP00011_110329074139
NL: 1.02E3
TIC F: + c EI SRM ms2 291.920 [219.935-219.945] MS ICIS XLSULTRA_NPV_OCP00011_110329074139
NL: 1.54E3
TIC F: + c EI SRM ms2 323.880 [253.905-253.915] MS ICIS XLSULTRA_NPV_OCP00011_110329074139
NL: 2.53E3
TIC F: + c EI SRM ms2 325.880 [255.905-255.915] MS ICIS XLSULTRA_NPV_OCP00011_110329074139
Tri, Tetra, Penta PCBs 100 fg in Contaminated land (industrial soil) sample
Standard SRM ( Q1=0.7 Da)
TSQ Quantum XLS Ultra U-SRM
( Q1=0.1Da )
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Environmental: Pesticides
Endrin 1pg in contaminated land sample ran both in SRM (Q1 0.7 amu) and U-SRM (Q1 0.1 amu)
U-SRM
SRM
C:\Xcalibur\...\XLSULTRA_NPV_0104OCP048 4/2/2011 7:39:50 PM
RT: 16.93 - 17.25 SM: 3G
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NL: 1.68E5
m/z= 190.43-191.43 F: + c EI SRM ms2 262.910 [190.925-190.935, 192.925-192.935] MS XLSULTRA_NPV_0104OCP048
NL: 1.83E5
m/z= 192.43-193.43 F: + c EI SRM ms2 262.910 [190.925-190.935, 192.925-192.935] MS XLSULTRA_NPV_0104OCP048
NL: 1.25E4
m/z= 190.43-191.43 F: + c EI SRM ms2 262.910 [190.925-190.935, 192.925-192.935] MS ICIS xlsultra_npv_0104ocp031
NL: 1.86E4
m/z= 192.43-193.43 F: + c EI SRM ms2 262.910 [190.925-190.935, 192.925-192.935] MS ICIS xlsultra_npv_0104ocp031
XLSULTRA_NPV_OCP00015_110329095349 3/29/2011 9:53:49 AM
RT: 15.48 - 17.07 SM: 7G
15.5 15.6 15.7 15.8 15.9 16.0 16.1 16.2 16.3 16.4 16.5 16.6 16.7 16.8 16.9 17.0
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RT: 16.56AA: 28533SN: 31RMS
RT: 15.95AA: 29627SN: 31RMS
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NL: 1.74E4
m/z= 175.47-176.47 F: + c EI SRM ms2 245.950 [175.965-175.975] MS ICIS XLSULTRA_NPV_OCP00015_110329095349
NL: 7.78E3
m/z= 245.45-246.45 F: + c EI SRM ms2 317.940 [245.945-245.955] MS XLSULTRA_NPV_OCP00015_110329095349
XLSULTRA_NPV_OCP00013_110329084745 3/29/2011 8:47:45 AM
RT: 15.48 - 17.07 SM: 7G
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RT: 16.56AA: 12784SN: 708RMS
RT: 15.95AA: 7444SN: 412RMS
RT: 16.86AA: 380SN: 22RMS
RT: 16.67AA: 198SN: 19RMS
RT: 16.52AA: 129SN: 14RMS
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NL: 6.83E3
m/z= 175.47-176.47 F: + c EI SRM ms2 245.950 [175.965-175.975] MS ICIS XLSULTRA_NPV_OCP00013_110329084745
NL: 1.16E3
m/z= 245.45-246.45 F: + c EI SRM ms2 317.940 [245.945-245.955] MS XLSULTRA_NPV_OCP00013_110329084745
U-SRM
SRM
o,p-DDE & p,p-DDE 100fg in contaminated land sample ran both in SRM (Q1 0.7 amu) and U-SRM (Q1 0.1 amu)
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• New technology• Sample through-put
• Time savings
• Lower detection limits
• Triple Quadrupole GC-MS/MS provides selectivity with flexibility
• Advanced Triple Quadrupole GC-MS/MS can deliver two modes of selectivity• Increased resolution
• Structural selectivity through MS/MS
Conclusion
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Thank You for Your Attention!
Questions?
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