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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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Total method selectivity

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Method performance requirement

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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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Total method selectivity

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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

M C

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Wo

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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

221

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219

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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0.7 Da

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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

14.54

14.4614.3014.64

14.68

14.7214.4014.16

14.7713.73 14.24

14.94 15.0214.09

14.05 14.8913.96

13.94

13.81

13.56

NL: 1.05E5

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

m/z

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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

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 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

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

14.54

14.4614.3014.64

14.68

14.7214.4014.16

14.7713.73 14.24

14.94 15.0214.09

14.05 14.8913.96

13.94

13.81

13.56

NL: 1.05E5

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

Da

0.7

Da

SRM (Q1 0.7Da)

0.7 Da

0.1 Da

56

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

16.95 17.00 17.05 17.10 17.15 17.20

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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

16.59

16.56

16.71

16.89

16.8416.6516.7715.95

16.30 16.5016.3816.23 16.43

16.0615.85 16.0815.77

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

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: 12784SN: 708RMS

RT: 15.95AA: 7444SN: 412RMS

RT: 16.86AA: 380SN: 22RMS

RT: 16.67AA: 198SN: 19RMS

RT: 16.52AA: 129SN: 14RMS

16.56

15.95

16.53 16.6216.4116.11 16.2315.85 16.76 16.7916.3616.03 16.91

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)

58

• 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

59

Thank You for Your Attention!

Questions?

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