dart-ms - ionsense€¦ · 14 1. objectives: • generate standard curve data for carbendazim in...

Elizabeth Crawford Applications Manager

[email protected]

2nd October 2012

126th AOAC Meeting – IonSense, Inc. Seminar Las Vegas, NV, USA

DART-MS: Strategies for Rapid Screening & Quantitation of Pesticides

Evolving Open Air Ionization

Venter, A.; Nefliu, M. Cooks, R.G., “Ambient Desorption Ionization

Mass Spectrometry”, Trends in Anal. Chem., 27, 284-290, 2008.

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DART-SVP Ionization Source

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Key DART Source Parameters • Source Parameters:

• Ionization gasses • N2 (Standby) • He (Run Mode)

• Temperature of ionization gas • Sample introduction speed

• Ions typically observed in DART–MS mass spectra: • [M+H]+, [M–H]–

• M+·, M–· (ionic compounds) • Use of dopants to promote

ionization: • [M+NH4]+ (ammonia) • [M+Cl]– (e.g. CH2Cl2) • [M+CF3COO]– (trifluoroacetic

acid)

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Figure courtesy of Prof. Jana Hajšlová ICT Prague, Czech Republic

DART-SVP: Experiments Overview

• Pesticide Screening • FDA swab screening method

• Quantitative Pesticide Analysis in Orange Juice • 1-D Transmission Experiment Module

• Fast liquid analysis – Carbendazim in orange juices

• Sample Concentration

• Stir-bar sorptive concentration followed by DART-MS

• X-Z scanner Experiment Module

• Higher throughput option for liquid sample analysis – up to 96 samples per run

• Mycotoxin Screening • HR-AM screening with DART-SVP

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Pesticide Screening and Quantitation

FDA Rapid Pesticide Screening Method

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Rapid Pesticide Screening

Spiking • Each piece of produce was weighed to determine the pesticide dosing volume based on the determined spiking levels.

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Transmission DART Using Polyurethane Swabs

• Concentrate trace amount of analytes on a sponge type swab for DART analysis.

• Scan up to 10 swabs in one run using the temperature ramp method with user defined sampling time and heater temperatures.

• The heated DART gas passes through the swab and into MS inlet.

Utility of High Resolution for Very Rapid Analysis of Complex Mixtures

Pesticide Mixture

Total Run Time 1.8 minutes

DART Heater Temperature Quickly Ramped from 95 – 350° C

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Data courtesy of Sara Robison DART-SVP coupled to Exactive at FDA FCC Cincinnati

Typical Results

• 114 of the 132 pesticides were detected (86%), (some were isobaric compounds)

• 88% of the analytes had a mass error of < 1 ppm, 97% of the analytes had a mass error of < 2 ppm

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S. E. Edison, L. A. Lin, L. Parrales. Food Add and Contam. Part A., 2011, 28: 1393-1404.

S. E. Edison, L. A. Lin, B. M. Gamble, J. Wong, K. Zhang. Rapid Commun. Mass Spectrom., 2011, 25, 127-129.

Data courtesy of Sara Robison

DART-SVP coupled to Exactive at FDA FCC Cincinnati

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1. Objectives:

• Generate standard curve data for carbendazim in orange juice ranging from 1 ppb – 2000 ppb.

• Screen for carbendazim in a variety of orange juices from the EU, India and the USA. – European maximum residue limit* (MRL) is set at 200 ppb for oranges and

carbendazim is not approved for use on citrus fruits in the US . The US FDA has set an action limit of 10 ppb.

2. Analysis Overview:

• No sample preparation – liquid from consumer juice beverages directly analyzed by transmission mode direct analysis in real time (DART) source coupled with Q Exactive.

• Positive ion mode - carbendazim targeted MS2 experiment. – m/z 192.07675 → 160.05032

* http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2011:152:0001:0021:EN:PDF

Quantitation - Carbendazim Unapproved Fungicide for Citrus Fruits in US

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3 µL of liquid sample pipetted onto

stainless steel mesh screen and allowed to completely dry

before DART analysis

Stainless steel mesh screens: Used as the sampling

surface for juice samples

Transmission DART Workflow

Juice is directly

applied to the mesh

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RT: 1.15 - 5.66

1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5

Time (min)

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RT: 4.00SN: 164

RT: 4.43SN: 162

RT: 4.84SN: 114RMS

RT: 3.54SN: 91RMS

RT: 3.16SN: 69RMSRT: 2.70

SN: 64RMS

RT: 1.48SN: 35RMS

RT: 1.97SN: 35RMS

RT: 2.39SN: 31RMS

3.96

4.09

4.934.82

NL: 5.18E4

m/z= 160.04952-160.05112 F: FTMS + p NSI Full ms2 [email protected] [50.00-215.00] MS 5,10,20ngmL_Carbendazim_NantucketNectOJ_140K_tMS2_02_20120514

RT: 1.15 - 5.66

1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5

Time (min)

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RT: 4.00SN: 164

RT: 4.43SN: 162

RT: 4.84SN: 114RMS

RT: 3.54SN: 91RMS

RT: 3.16SN: 69RMSRT: 2.70

SN: 64RMS

RT: 1.48SN: 35RMS

RT: 1.97SN: 35RMS

RT: 2.39SN: 31RMS

3.96

4.09

4.934.82

NL: 5.18E4

m/z= 160.04952-160.05112 F: FTMS + p NSI Full ms2 [email protected] [50.00-215.00] MS 5,10,20ngmL_Carbendazim_NantucketNectOJ_140K_tMS2_02_20120514

20 ppb ; 7 % CV

10 ppb ; 16 % CV

5 ppb ; 9 % CV

16 16 16

m/z 192.07675 → 160.05032

XIC generated with +/- 5 ppm window around 160.05032

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160.0 160.5 161.0 161.5 162.0

m/z

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161.05390

162.05725

160.05039

161.05955

NL:1.06E5

C 8 H 6 O N 3: C 8 H 6 O 1 N 3

p (gss, s /p:8) Chrg 1R: 140000 Res .Pwr . @FWHM

NL:9.65E3

5,10,20ngmL_Carbendazim_NantucketNectOJ_140K_tMS2_02_20120514#342-366 RT: 3.10-3.32 AV: 25 F: FTMS + p NSI Full ms2 [email protected] [50.00-215.00]

158 159 160 161 162 163 164 165

m/z

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160.05054

161.05390

162.05725

160.05039

161.05955

165.04798

163.04483 164.07048

NL:1.06E5

C 8 H 6 O N 3: C 8 H 6 O 1 N 3


NL:9.65E3


158 159 160 161 162 163 164 165

m/z

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160.05054

161.05390

162.05725

160.05039

161.05955

165.04798

163.04483 164.07048

NL:1.06E5

C 8 H 6 O N 3: C 8 H 6 O 1 N 3


NL:9.65E3


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Carbendazim: Major Fragment

10 ppb Carbendazim spiked in orange juice

Carbendazim fragment [C8H6N3O]+ spectral simulation

Carbendazim product ion δ mass accuracy: -0.9 ppm

Interference ion at m/z 161.05955

161.040 161.045 161.050 161.055 161.060 161.065

m/z

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161.05390

161.04757

161.05682

161.05955

161.05366

NL:9.14E3

C 8 H 6 O N 3: C 8 H 6 O 1 N 3


NL:3.46E3


A+1 isotope completely resolved from + 37 ppm interference at m/z 161.05955 (140,000 FWHM)

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y = 24684x - 412957 R² = 0.9987

0.00E+00

1.00E+07

2.00E+07

3.00E+07

4.00E+07

5.00E+07

6.00E+07

0 200 400 600 800 1000 1200 1400 1600 1800 2000

Ave

rage

Pe

ak A

rea

Concentration (ppb)

Carbendazim Curve in OJ: Q Exactive tMS2

0.00E+00

2.00E+06

4.00E+06

6.00E+06

8.00E+06

1.00E+07

1.20E+07

1.40E+07

0 100 200 300 400 500

n = 3 per conc.

18 18 Concentration Range: 1 ppb – 2,000 ppb

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0

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USA OJ 1 NL OJ 1 PL OJ 1 PL OJ 2 DE OJ 1 IT OJ 1 IN OJ 1 IN OJ 2 IN OJ Bev 1 IN OJ Bev 2

Me

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Le

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pb

Orange Juice Consumer Beverages

Screening for Carbendazim in OJ: Q Exactive tMS2 m/z 192.07675 → 160.05032

19 n = 3; except n = 9 for German OJ

ND

ND = Not Detected

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1. Objectives:

• Generate standard curve data for carbendazim in orange juice ranging from 1 ppb – 5000 ppb.

• Screen for carbendazim in a variety of orange juices from the EU (2 countries) and the USA. – European maximum residue limit* (MRL) is set at 200 ppb for oranges and

carbendazim is not approved for use on citrus fruits in the US . The US FDA has set an action limit of 10 ppb.

2. Analysis Overview:

• No sample preparation – liquid from consumer juice beverages directly analyzed by transmission mode direct analysis in real time (DART) source coupled with triple quad.

• Positive ion mode; carbendazim transition m/z 192.2 → 160.2.

* http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2011:152:0001:0021:EN:PDF

Quantitation - Carbendazim Unapproved Fungicide for Citrus Fruits in US

API 4000 QTRAP with DART-SVP

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MRM Settings • Scan Parameters:

• Positive Ion Mode

• Resolution: Unit

• Fragmentation:

carbendazim major transition monitored m/z 192.2 → 160.2

• DP 56 V

• CE 27 V

• Dwell Time: 100 ms

DART-SVP Settings • DART Source:


• Heater Temperature: 250° C

• Motorized Linear Rail:

• Sample Introduction Speed: 0.5 mm/s

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22

0

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USA Apple USA FruitPunch

USA OJ 1 USA OJ 2 USA OJ 3 France OJ 1 Germany OJ1

Germany OJ2

Me

asu

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Le

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pb

Fruit Juice Consumer Beverage

Carbendazim Levels: API 4000 QTRAP MRM m/z 192.1 → 160.1

22 n = 8; except n = 16 for German OJs

ND ND ND ND ND

ND = Not Detected

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Gerstel Twister Spin Bar: Sample Concentration

Twister was spun overnight in orange juice with

50 ppb level of 10 pesticides

Higher Log Kow = greater uptake onto the Twister sorptive surface. 24

10 Pesticide Mix [Exact Mass + H] + Log Kow

Carbaryl 202.08626 2.4

Metalaxyl 280.15433 1.7

Spiroxamine 298.27406 Unknown

Fenhexamid 302.07091 3.5

Diazinon 305.10833 3.8

Pyriproxyfen 322.14377 5.6

Pyridaben 365.14489 5.5

Dimethomorph 388.13101 2.7

Azoxystrobin 404.12410 2.5

Difenoconazole 406.07197 4.2

RT: 0.00 - 8.20

0 1 2 3 4 5 6 7 8

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1005.03

5.60 5.756.75 7.47 7.502.75 4.333.33 3.990.79

5.035.60 5.75

6.75 7.47 7.502.75 4.333.33 3.990.79

5.00 5.035.60

5.85 6.65 7.27 7.47

4.99 5.055.595.58 6.75 7.135.85 7.43

4.99 5.055.59

5.85 7.04 7.27 7.50

5.055.035.59 5.85 7.216.74 7.31

5.055.00 5.087.275.60 7.367.13

5.087.315.03 5.59 7.477.045.77

NL: 4.23E4

m/z= 202.08512-202.08714 MS 120424-01_Twister19&20_OSCranLt_Blank&50ppbSpin

NL: 4.23E4


NL: 2.20E5


NL: 6.78E4


NL: 1.88E6


NL: 2.62E6


NL: 9.00E5


NL: 1.71E5


10 Pesticides Concentrated onto Twister: 50 ppb in orange juice spun overnight → DART/MS

Exactive XICs generated with +/- 5 ppm m/z window.

Twister Blank Spun in Blank OJ

Twister Spun in Spiked OJ

50 ppb level mixture of pesticides were well concentrated onto the Twister and were then directly desorbed from the Twister sorptive material for direct exact mass analysis with DART.

120424-01_Twister19&20_OSCranLt_Blank&50ppbSpin #598-654 RT: 4.99-5.41 AV: 57 SB: 116 0.46-1.46 NL: 1.34E6T: FTMS {1,1} + p NSI Full ms [100.00-500.00]

200 220 240 260 280 300 320 340 360 380 400 420

m/z

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322.14321

305.10788

365.14431

406.07125

280.15399 388.13052217.17969

372.09673

298.27375

234.20614 355.33151

338.34136

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Direct Twister DART-MS Analysis

10 Pesticide Mix (50 ppb) in orange juice DART Heater 325° C; linear rail movement 0.2 mm/s

• There is a very good concentration effect on the Gerstel Twister for direct screening for low levels (50 ppb) of pesticides with ambient ionization DART source.

• Long lasting signal from the Twister experiments indicates that a large amount of material is collected on the Twister surface • A temperature ramp of the DART heater would yield thermal

separation and long lasting signal would be beneficial for MS/MS experiments.

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Summary

Haunschmidt, M. et al. Determination of organic UV filters in water by stir bar sorptive extraction and direct analysis in real-time mass spectrometry. Anal. Bioanal. Chem., 2010, 397(1): 269-275.

MS Instrumentation

API 5500 QTRAP Settings

• MRM Parameters for Pesticide A:


• Q1: 382.1 ; 382.1

• Q3: 342 ; 314

• Dwell Time: 150 ms/transition

• DP: 16 ; 16

• EP: 10 ; 10

• CE: 31 ; 35

• CXP: 28 ; 24

DART-SVP ion source with X-Z Scanner Experiment Module for Transmission Mode DART-MS/MS

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1. Spot 5 µL aliquots of liquid sample onto the mesh surface

2. Let sample spots dry

4. Set up the desired X-Z scanner method in the DART software to run up to 96 samples/run

3. Insert the mesh plate into the slot on the X-Z scanner module

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Mycotoxin Screening

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USDA Mycotoxin Screening

• Aflatoxin B1 (AFB1) Detection off of Corn Kernels

Major area for future work…

Questions?

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dart-ms - ionsense€¦ · 14 1. objectives: • generate standard curve data for carbendazim in...

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