LC/MS TOF, 6460 Triple Quad and 6530 QTOF CASU · 2016-08-30 · Pesticides Analysis with the New Agilent 6220 Screening, Confirmation and the New Agilent 6220 Quantitation LC/MS

Pesticides Analysis with the New Agilent 6220

Screening, Confirmation and Quantitationthe New Agilent 6220

LC/MS TOF, 6460 Triple Quad and 6530 QTOF

Quantitation

CASUDr. Jerry ZweigenbaumMarket Development SpecialistApril 14, 2009

Pesticide Analysis with New Agilent Jet Stream Technology

April 14, 2009Page 1

Agilent’s 35+ Years in Mass SpectrometryNearly 40,000 systems – Applied to Chromatographic Analysis

5930A 1971

5992A1976 5970B MSD

19865971 - 59751988 - present

6100 Single Quad2006, 2007

LC/MSD Series1997-2005

6000 Series2006-Present

4500, 7500 Series ICP/MS1994-Present

Ion Trap Series2000-Present

ESI-TOF Series2003-Present

6410 Triple Quad2006, 2007

6510, 6520 Q-TOF, Q2006, 2007



Cutting Edge Triple Quad & Q-TOF PerformanceASMS 2008 Introductions

6460 Triple Quad1pg – 1000:1 s/n

With… 6530 Accurate Mass QTOFWith…Agilent Jet Stream TechnologyUnmatched sensitivity for ultra trace analysis

6530 Accurate Mass QTOF

trace analysisUnique data mining tools for faster compound ID

1pg 50:1 s/n1pg – 50:1 s/n



New Agilent 6230 LC/MS TOFIntroduced at the MontreuxIntroduced at the MontreuxLC/MS Symposium this November



New 6230 TOF and 6530 QTOF Each have a 5 Stage Vacuum SystemEach have a 5 Stage Vacuum System

Collision – Free TOF Region

2 5 10 7 T

6230 TOF

1 2 3 4 5Stage 2.5 x 10-7 Torr

6530

3 421 5St

6530 QTOF

2 5 x 10-7 Torr3 421 5Stage 2.5 x 10 Torr



Agilent Jet Stream Technology Theory

Page 6 April 14, 2009


Agilent Jet Stream: Thermal Gradient Focussing

April 14, 2009


Page 7

Agilent Jet Stream Ion GenerationThermal Dynamics ViewyThis plot is a simulation showing the thermal profile of Agilent Jet Stream Technology

LC flow

Nebulizer N gas

Super-heated N2sheath gas

Thermal focusing produces the most efficient Nebulizer N2 gas

(near sonic velocity)

most efficient desolvation and ion generation

ibl !possible!

Patent Pending



How Does Agilent Jet Stream Work?

Page 9Pesticide Analysis with New Agilent

Jet Stream Technology

April 14, 2009

SCREENING PESTICIDES WITH LC/TOFLC/TOF



Pepper Extract – Power of MS Resolution and Mass Accuracy



Extracted Ion Chromatogram m/z 202.0433±0.002of Lettuce Extract

2x10

0 7

0.8

0.9

1+ESI TIC Scan Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d

1 1

of Lettuce Extract

0.3

0.4

0.5

0.6

0.7

0

0.1

0.2

-2x10

6 5

7

+ESI EIC(202.0433) Scan Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d Smooth

1.4411 1

3.5

4

4.5

5

5.5

6

6.5

0.725

Thiabendazole at 0.1 ppb

0

0.5

1

1.5

2

2.5

3

2.581

4.7683.323 6.146

Thiabendazole at 0.1 ppb

0

Counts (%) vs. Acquisition Time (min)0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5



Mass Spectrum of Chromatographic Peak for ThiabendazoleThiabendazole

5x10

2

2.1

2.2

+ESI Scan (2.501-2.660 min, 13 scans) Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d Subtract

390.1763

1 4

1.5

1.6

1.7

1.8

1.9

0.9

1

1.1

1.2

1.3

1.4

767.2738

193.0859

300 2019

0.3

0.4

0.5

0.6

0.7

0.8 300.2019

0

0.1

0.2722.4532234.5509

Counts vs. Mass-to-Charge (m/z)100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000



Spectrum of Thiabendazole at 0.1 ppb in Extracted Pepper

3x10

1.351.4

1.45

+ESI Scan (2.501-2.660 min, 13 scans) Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 ater.d Subtract

202.0431(M+H)+

Pepper

11.05

1.11.15

1.21.25

1.3

201 1091

Resolving power= 15,000

0.650.7

0.750.8

0.850.9

0.95201.1091

0 30.35

0.40.45

0.50.55

0.60.65

204.1217

00.05

0.10.15

0.20.25

0.3

201 201 2 201 4 201 6 201 8 202 202 2 202 4 202 6 202 8 203 203 2 203 4 203 6 203 8 204 204 2 204 4 204 6 204 8 205 205 2 205 4 205 6Counts vs. Mass-to-Charge (m/z)

201 201.2 201.4 201.6 201.8 202 202.2 202.4 202.6 202.8 203 203.2 203.4 203.6 203.8 204 204.2 204.4 204.6 204.8 205 205.2 205.4 205.6



Calculated Formula of Thiabendazole at 0.1 ppb in Extracted PepperExtracted Pepper



Sensitivity of New Agilent 6230 vs 6220C d 6230/6220Compound 6230/6220

Thiabendazole 3.0Methomyl 8 7Methomyl 8.7Deisopropylatrazine 3.9Imazopry 3.6Imdaocloprid 1 9Imdaocloprid 1.9Imazalil 2.8Monuron 2.0Atrazine 3.2Procloraz 2.1Molinate 2.1Acetachlor 1.5Diazonin 3.5

Average= 3.2



Average 3.2

Carbofuran at 0.8 ppb

+ EIC(222.1125) Scan Pest_MX_20_08ppb_03.d

Cou

nts 3x10

4.75

54.790

3.25

3.5

3.75

4

4.25

4.5

Collected at 0.8 s per scan6 i t th k

1.75

2

2.25

2.5

2.75

3 6 points across the peak

0.25

0.5

0.75

1

1.25

1.5

Acquisition Time (min)4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1

-0.5

-0.25

0



Carbofuran at 0.16 ppb


Cou

nts 3x10

1.6

1.7

1.2

1.3

1.4

1.5

0.8

0.9

1

1.1

0.4

0.5

0.6

0.7

Acquisition Time (min)4.2 4.3 4.4 4.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1

0.2

0.3



Calibration of Carbofuran

Carbofuran - 5 Levels, 4 Levels Used, 5 Points, 4 Points Used, 0 QCs

espo

nses 6x10

22.1

y = 15310.9890 * x - 7233.1887R^2 = 0.99988175

Re

1 31.41.51.61.71.81.9

2

0.60.70.80.9

11.11.21.3

-0.10

0.10.20.30.40.5

Concentration (ng/ml)-10 -5 0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 105 110

-0.3-0.2



Atrazine at 0.16 ppb (0.8 pg on-column)


Cou

nts 3x10

3.2

3.45.142

2.2

2.4

2.6

2.8

3

1.2

1.4

1.6

1.8

2

0.2

0.4

0.6

0.8

1

Acquisition Time (min)4.5 4.6 4.7 4.8 4.9 5 5.1 5.2 5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1 6.2 6.3 6.4

-0.2

0



Atrazine 0.16 ppb to 100

Atrazine - 6 Levels, 5 Levels Used, 6 Points, 5 Points Used, 0 QCs

espo

nses 6x10

7

y = 68689.2636 * x + 17563.2362R^2 = 0.99965838

Re

5

5.5

6

6.5

2 5

3

3.5

4

4.5

0.5

1

1.5

2

2.5

Concentration (ng/ml)-5 0 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 10

0



Lettuce Extract with EIC for Atrazine at 0.1 ppb2x10

0.7

0.8

0.9

1+ESI TIC Scan Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d

1 1

0.2

0.3

0.4

0.5

0.6

0

0.1

0

-1x10

1 1

1.2

1.3+ESI EIC(216.1010) Scan Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d

0.6721 1

0 5

0.6

0.7

0.8

0.9

1

1.1

3.217

5.470

3 522

0

0.1

0.2

0.3

0.4

0.5 3.522

0.911

6.9152.236 8.0024.6494.171 10.481 11.2501.626

0 5 1 1 5 2 2 5 3 3 5 4 4 5 5 5 5 6 6 5 7 7 5 8 8 5 9 9 5 10 10 5 11 11 5



Counts (%) vs. Acquisition Time (min)0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5

Spectrum of Atrazine at 0.5 pg on-column

3x10

6 5

7

7.5+ESI Scan (5.457-5.497 min, 4 scans) Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d

216.1010(M+H)+

5

5.5

6

6.5

3

3.5

4

4.5

217.1169

218.0985

1

1.5

2

2.5

0

0.5

Counts vs. Mass-to-Charge (m/z)215.6 215.8 216 216.2 216.4 216.6 216.8 217 217.2 217.4 217.6 217.8 218 218.2 218.4





Spectrum of Atrazine at 0.1 ppb in Lettuce after subtraction before and after peaksubtraction before and after peak

3x10

6

8

+ESI Scan (5.470-5.484 min, 2 scans) Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d Subtract (2)

216.1011(M+H)+

218.0983

0

2

4217.1230

3x10

4

+ESI Scan (5.417-5.431 min, 2 scans) Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d

0

1

2

3

4

03x10

1

1.5

+ESI Scan (5.523-5.537 min, 2 scans) Frag=120.0V Pest MX_20 Lettuce_01 ppb in 1_3 water.d

0

0.5

Counts vs. Mass-to-Charge (m/z)215.6 215.8 216 216.2 216.4 216.6 216.8 217 217.2 217.4 217.6 217.8 218 218.2 218.4 218.6 218.8



Pesticide Screening Workflow

• Simple Extraction QuEChERS or SPE

Analysis by RRHT LC/MS TOF• Analysis by RRHT LC/MS TOF

• Screening of database containing large list of pesticides

• Validated pesticides updated with retention times

• Confirmation of positives by another method (LC/QQQ?)



Result of a Database screen using Accurate Mass(10 ppb each pesticide in Pepper Matrix)(10 ppb each pesticide in Pepper Matrix)



Personal Compound Database – Structures and ability to update retention times semi-automaticallyability to update retention times semi automatically



Summary

• Designed for high sensitivity• High MS resolutionHigh MS resolution• Mass Accuracy better than 2 ppm• High dynamic range (in spectrum)High dynamic range (in spectrum)• Excellent for Screening

•New Personal Compound DatabaseNew Personal Compound Database •First offering 1600 pesticides•Customizable•Customizable



Break Number 1

For questions, at break q ,please dial 1 on your phone, or type onto the chat screen at any time yduring the presentation.

Page 30 Pesticide Analysis with New Agilent Jet Stream Technology

April 14, 2009

Confirmation of Pesticides with Confirmation of Pesticides with LC/Triple QuadLC/Triple QuadLC/Triple QuadLC/Triple Quad

Page 31 Pesticide Analysis with New Agilent Jet Stream Technology

April 14, 2009

New 6460 Triple Quad: World’s Most Sensitive QQQwith Agilent Jet Stream Technologywith Agilent Jet Stream Technology


Jet Stream TechnologyApril 14, 2009

Sensitivity of 6460 in Apple Matrix

Apple matrixApple matrixApple matrix Apple matrix BlankBlank

5ppt spiked in 5ppt spiked in apple matrix, apple matrix, 5uL injection5uL injection

Page 33


April 14, 2009

Agilent Jet Stream increased Sensitivity for Pesticides by about 5 Fold

600000

700000

800000

oxamyl

Pesticides by about 5 Fold

Agilent Jet Stream

0

100000

200000

300000

400000

500000

avg.

are

a re

spon

se

200 500 760

ESI 200 and 325 Cjet stream

oxamyl: signal enhancement 5.1 at SGT 500 C

Agilent Jet Stream

100 250 380

Sheath gas temperature (C)

400000

500000

600000

pons

e

imazalil

Agilent Jet Stream

0

100000

200000

300000

avg.

are

a re

sp

200 500 760

Sheath gas temperature (C)

ESI 200 and 325 Cjet stream

imazalil: signal enhancement 8.1 at SGT 760 C

g J S

100 250 380

Summary: Pesticides evaluated 11

Mean Jet Stream enhancement 5.8



ea Jet St ea e a ce e t 5 8

April 14, 2009

Fast MRMs & 100 msec polarity switching on 6460

4x10

4.5

5

+ MRM (226.20000 -> 170.09961) tz012908_BE_Athena_040.d Noise (RMS) = 1.95; SNR (5.892min) = 28717.7

5.8921

11 triazine herbicides + 5 sulfa drugs at 10 pg on-column

2

2.5

3

3.5

4 5 msec dwells

0

0.5

1

1.5

6.599

Counts vs. Acquisition Time (min)0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8 8 8.2 8.4 8.6 8.8 9 9.2 9.4 9.6 9.8

4x10

3

3.5

4

4.5

5

+ MRM (226.20000 -> 170.09961) tz012908_BE_Athena_039.d

5.9021 1

20 msec dwells

0

0.5

1

1.5

2

2.5

6.609

02 04 0 6 0 8 1 1 2 1 4 1 6 1 8 2 2 2 2 4 26 28 3 32 34 3 6 3 8 4 4 2 4 4 4 6 4 8 5 5 2 5 4 5 6 58 6 62 64 66 6 8 7 7 2 7 4 7 6 7 8 8 8 2 8 4 8 6 88 9 92 94 96 9 8



Counts vs. Acquisition Time (min)0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2 4.4 4.6 4.8 5 5.2 5.4 5.6 5.8 6 6.2 6.4 6.6 6.8 7 7.2 7.4 7.6 7.8 8 8.2 8.4 8.6 8.8 9 9.2 9.4 9.6 9.8

April 14, 2009

Quantitation and Confirmation of Diazinon



Screening and ConfirmationScreening and Confirmation on a LC/MS QTOFQ





Sensitivity – Agilent 6530 vs 6520

65206520

T t M t i 5 b ik d 10 l i j tiT t M t i 5 b ik d 10 l i j tiTomato Matrix: 5ppb spiked 10ul injectionTomato Matrix: 5ppb spiked 10ul injection

6530 with Agilent Jet Stream6530 with Agilent Jet Stream

Tomato Matrix: 5ppb spiked 10ul injectionTomato Matrix: 5ppb spiked 10ul injection

Compounds 6530(Integration Area) 6520 (Integration Area)

1 Simazine 1.74e5 5.07e4

2 Prometon 1.15e6 1.80e5

3 Atrazine 4 82e4 1 78e43 Atrazine 4.82e4 1.78e4

4 Ametryn 8.59e5 1.25e5

5 Propazine 1.19e5 5.12e4

6 Prometryn 1.11e6 1.52e5

Page 39

7 Terbutryn 1.22e6 1.30e5

Sensitivity – 6530 vs 6520

Tomato Matrix (5ppb level)Tomato Matrix (5ppb level)

April 14, 2009


Page 40

Workflow for QTOF Pesticide Analysis

• Extract sample

Run in Single MS Mode• Run in Single MS Mode

• Database Search for compound ID

• Targeted MS/MS for found Compounds

• Confirm and Quantify

April 14, 2009


Page 41

Screen with QTOF – Single MS of Fruit ExtractExtract



DataBase Search of 600 Pesticides

Compound Table

Name RT Mass Abund. Formula Tgt MassDiff

(ppm) DB Formula DB Diff (ppm) Hits (DB)

Cpd 527: Famoxadone 0.9 374.12628 2696 C22H18N2O4 374.12666 -1.01 C22H18N2O4 1.01 1

Cpd 335: SPIROXAMINE 11 297.26687 2710 C18H35NO2 297.26678 0.31 C18H35NO2 -0.31 1

Cpd 375: Tebuconazole 14 307.14498 28381 C16H22ClN3O 307.14514 -0.54 C16H22ClN3O 0.54 1

Cpd 491: TEBUFENOZIDE 15 352.21569 4345 C22H28N2O2 352.21508 1.74 C22H28N2O2 -1.74 1

Page 43April 14, 2009


Identification by Accurate Mass FormulaGenerationGeneration

Page 44

April 14, 2009


Accurate Mass MS/MS for Confirmation

m/z Formula Abund% Difference (ppm) Loss Mass Loss Formula70 03997 4 9770.03997 4.97

100.1122 C6H14N 16.91 -1.27 198.16198 C12H22O2

144.13817 C8H18NO 78.12 0.8 154.13577 C10H18O

OCH3 CH3

NH+

CH3

CH3

O

CH2+

O

O

NCH3

CH3

CH3 CH3

CH3CH3N

CH2

CH3

Page 45

April 14, 2009


Wrap-up E-Seminar Questions

Thank you for attending Agilent e-Seminars. Our y g ge-Seminar schedule is expanding every week.

Please check our website frequently at:Please check our website frequently at: www.agilent.com

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Page 46Pesticide Analysis with New Agilent Jet

Stream TechnologyApril 14, 2009

Documents

LC/MS TOF, 6460 Triple Quad and 6530 QTOF CASU · 2016-08-30 · Pesticides Analysis with the New Agilent 6220 Screening, Confirmation and the New Agilent 6220 Quantitation LC/MS