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Page 1 Agilent Pesticide TOTAL Solutions Vivek Dhyani Applications Chemist Agilent Technologies

Agilent Pesticide TOTAL Solutions · Agilent Pesticide TOTAL Solutions Vivek Dhyani Applications Chemist Agilent Technologies. Page 2 ... Ashitaba blank Area: 99 1 ppbSpiked Area:

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Page 1

Agilent Pesticide TOTAL Solutions

Vivek DhyaniApplications ChemistAgilent Technologies

Page 2

Business Issues and Challenges

• Global trade increases the number of pesticides to monitor (> several hundreds)– Typically 150-200 on a target list

• Regulation driving lower detection limits– 10 ppb or lower

• Speed: doing more analyses in a day– Fast chromatography and shorter cycle time

• Need for fast startup and long uptime– No method development

Faster Data Review and Reporting

Total Solution Offering

Screen for Unknowns

Page 3

TOTAL Solutions Outline

• Instrumentations and Agilent Portfolio

• GC and LC

• MS Analyzers Selection

• GC/MS

• QQQ (MS/MS)

• LC/Q-TOF

Page 4

Pesticide Analysis Instrumentations

Chromatography (Separation – the best friend of any detector)

• GC – volatile and semi-volatile non-polar compounds

• LC – polar or moderately polar or thermally labile compounds

Detectors

• GC (element selective, FPD, NPD, ECD) – high sensitivity, poor specificity

• LC (UV, FLD) – general purpose, limited sensitivity, poor specificity

• MS –

– SQ: high confidence from spectral confirmation, limited MDL

– QQQ: low detection limits of target compounds in dirty matrices (MRM)

– TOF/Q-TOF: always full spectrum and accurate mass for screening

– TOF-TOF, Q-Trap, …and others (generally not for ROUTINE use).

Page 5

Agilent Has the TOTAL Portfolio for Pesticide Analysis

(semi)-volatile or non-polar

(moderate)-polar or thermally labile

GCFPD/ECD

LCUV, FLD

GC/QDRS

LC/QLC/QQQMRM

GC/QQQMRM

Q-TOFTOF

GAS

LIQUID

Routine Analysis Identify Unknowns

phase

phase

Exact Mass Database Search

pesticides

Page 6

4.296 min.

4.064 min.

4.297 min.

Agilent Unique and Powerful Tools for Pesticide Analysis

GC/MS Retention Time Locked (RTL) Pesticide Libraries

– Pesticide and Endocrine Disruptors library with RT and spectra (926 entries)

– Japanese Positive List library with RT and spectra (430 entries)

Deconvolution Reporting Software

– Extract pesticide spectra from complex matrices

Capillary Flow Technology

– Flow splitting and backflushing

LC/MS METLIN Pesticide database

– with RT, formula, structure and exact mass (>1400 entries)

– To screen for non-target pesticides

Page 7

TOTAL Solutions Outline

• Instrumentations and Agilent Portfolio

• GC and LC

• MS Analyzers Selection

• GC/MS

• QQQ (MS/MS)

• LC/Q-TOF

Page 8

Almost All Pesticides Contain Heteroatoms (Oxygen is too common)

PNH

O

S OO

CH3

CH3

CH3

AcephateN

N N

NH NH

Cl

CH3

CH3 CH3

Atrazine

N+

N+

N

O-

O-

O

O

F

F

F

CH3

CH3

Trifluralin

Cl

Cl

Cl

Cl

Cl

Cl

Cl

Heptachlor

Element-selective GC detectors are commonly used fo r pesticide analysis for sensitivity and selectivity .

Page 9

Sort of Pesticide Instrument Injector column Detector Data system

Organo-Chlorine(CHM) and pyrethroid

GC

Agilent 7890ALS

HP-1 30m×0.25mm×0.25 µm

HP-17 30m×0.25mm×0.25 µm

ECD

ECDAgilent

ChemstationOrgano-

phosphorus(OP)

GC

Agilent 7890ALS

HP-1 30m×0.53mm×1.5 µm

HP-17 30m×0.53mm×1.0 µm

FPD

FPD

Carbamate

Agilent 1200 with

Pickering5200

ALS or manual injector

C8 4.6mm×250mm, 5 µm

C18 4.6mm×250mm, 5 µmFLD

Agilent Chemstation

ConfirmationGC-MS

Agilent 5975ALS HP-5MS 30m×0.32mm×0.5 µm MSD

Agilent Chemstation

Pesticide Residual – CDFA* Instrument Configuration

*California Department of Food and Agriculture

Page 10

2.1x80mm 600 bar RRHT ZORBAX EPA 555 Chlorinated phenoxy acid herbicides

Fast and high resolution

Page 11

TOTAL Solutions Outline

• Instrumentations and Agilent Portfolio

• GC and LC

• MS Analyzers Selection

• GC/MS

• QQQ (MS/MS)

• LC/Q-TOF

Page 12

Guiding Principles in MS Analyzer Selection

The correct choice of analyzer depends on whether one is “LOOKING for UNKNOWNS” or whether one is “CONFIRMING & MEASURING KNOWNS”.

The correct choice of analyzer depends on the COMPLEXITY of the MATRIX.

Page 13

Basic Questions – Which MS Solution

Target analysis only?

Analysis of unknowns?

How much chemical noise from the matrix?

How much sample prep?

Scan or SIM or MS/MS

Scan MS with Quad, IT or TOF

Deconvolution or MS/MS

Backflush or MS/MS

Page 14

Representative Sample

Extraction

Pesticide Workflow: Screen , Confirm and Q uantify

Clean-up

LC/QQQ MRM – for known targets S

C

Q

QuEChERS

S C Q

C16 H19 N3 P ClC15 H25 O P S ClC18 H21 O P Cl

LC/QTOF or TOF Full Spectrum– for unknown compounds

S

Exact Mass Database Search

Molecular Formula Generation

Q C

Another injection for MS/MS (QQQ or QTOF)

GC/MS (PTV) SIM/Scan– for known and unknown Final Report

S

C

Q

Deconvolution

(+backflush)

3 1

3 1

3 1

GC/QQQ MRM – for known targets S

C

Q

Page 15

TOTAL Solutions Outline

• Instrumentations and Agilent Portfolio

• GC and LC

• MS Analyzers Selection

• GC/MS

• QQQ (MS/MS)

• LC/Q-TOF

Page 16

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00

5.00 10.00 15.00 20.00 25.00 30.00 35.00 40.00

TICs of Surface Water Extracts

How many pesticides are in these samples and how long does it take you to confirm?

Page 17

17 Surface Water TICs (Scan): Pesticide Analysis Using DRS with Pesticide Database**

*CDFA is the California Department of Food and Agriculture

CDFA* Agilent DRS

Targets Found (not counting

ISTD)37

Same 37+99 more

False Positives 1 0

Processing Time ~8 hrs 32 min

Saving 7.5 hoursDRS: Deconvolution Reporting Software

**App Note 5989-5076 has the list of all 926 compounds in the database.

September, 2008Page 18

Carbaryl

Peach

Scan at 5.615 min

Deconvoluted/extracted spectrum

Library spectrum

A component in the scan above.

The power of deconvolution is appreciated while comparing the top two spectra (raw scan and the compound spectrum hidden in the raw scan).

Application Note 5989-7670: Replacing Multiple 50-Minute GC and GC-MS/SIM Analyses with One 15-Minute Full-Scan GC-MS Analysis for Non-targeted Pesticides Screening and >10x Productivity Gain

Page 19

DRS A.04 Report after importing AMDIS results QEdit reviewed with manual integrations

Page 20

TOTAL Solutions Outline

• Instrumentations and Agilent Portfolio

• GC and LC

• MS Analyzers Selection

• GC/MS

• QQQ (MS/MS)

• LC/Q-TOF

Page 21

Analyzers for Trace Target Compounds

LC/QQQ MRM – for known targets S

C

Q

GC/QQQ MRM – for known targets S

C

Q

Who should use QQQ (MS/MS)?

Users doing Selected Ion Monitoring (SIM) for target compound analysis in traditional markets, needing additional sensitivity and selectivity with less sample prep , to meet more demanding analytical requirements.

MRM

Page 22

LC/QQQ - Multiple Reaction Monitoring (MRM) of 301 Compounds

5989-8614EN, includingcompound name, precursor ion and two product ions of all 301 compounds.

Agilent 1200 HPLC With 1.8 micron ZORBAX SB C-18 Column.

Page 23

301 Compounds (Page 1 of 4) - LC/QQQ3,4,5-Trimethacarb Benalaxyl Carbofuran Cycloate

3-Hydroxycarbofuran Bendiocarb Carboxin Cymoxanil

Acephate Benfuracarb Carfentrazone-ethyl Cyproconazole

Acetamiprid Bensulfuron-methyl Chlorbromuron Cyprodinil

Acetochlor Benzoximate Chlorfenvinphos Cyromazine

Acibenzolar-S-methyl Bifenox Chlorfluazuron Daminozide

Aclonifen Bitertanol Chloridazon Deethylatrazine

Alachlor Bromacil Chlorotoluron Deethylterbuthylazine

Aldicarb Bromoxynil Chloroxuron Deisopropylatrazine

Aldicarb Sulfone Bromuconazole 1 Chlorpropham Demeton-S-methyl-sulfone

Aldicarb Sulfoxide Bromuconazole 2 Chlorpyrifos methyl Desmedipham

Aldoxycarb Bupirimate Chlorsulfuron Diazinon

Ametryn Buprofezin Cinosulfuron Dichlofenthion

Aminocarb Butocarboxim Clethodim Dichlofluanid

Anilazine Butocarboxim-sulfoxide Clodinafop-propargyl Dichlorvos

Atrazine Buturon Clofentezine Diclobutrazol

Azamethiphos Butylate Clomazone Diethofencarb

Azinphos-ethyl Carbaryl Cloquintocet-mexyl Difeconazole-1

Azinphos-methyl Carbendazim Coumaphos Difeconazole-2

Azoxystrobin Carbetamide Cyanazine Difenoxuron

Page 24

GC/MS/MS of Aojiru (Kale): Cyfluthrin (4 isomers)

20 ppb Spiked Area: 31007(No.2)

20 ppb Spiked Area: 30815(No.2)

Ashitaba blankArea: 0

1 ppb Spiked Area: 1211(No.2)

5 ppb Spiked Area: 8034(No.2)

20 pg on column

1 pg on column

5 pg on column

20 pg on column

Aojiru : Kale(dried power)

Page 25

GC/MS/MS of Ashitaba Tea:Chlorpyrifos

20 ppb Spiked Area: 88595

20 ppb Spiked Area: 86999

Ashitaba blankArea: 99

1 ppbSpiked Area: 4063

5 ppb Spiked Area: 24284

20 pg on column

1 pg on column

5 pg on column

20 pg on column

Ashitaba (Asian Herb)(dried power)

Sample Quantitation Results - Basil

Run 1 13.3

Run 2 12.2

Run 3 12.8

Run 4 12.5

Run 5 12.5

10 ppb Acephate

Run 1 5.4

Run 2 5.3

Run 3 5.2

Run 4 5.7

Run 5 5.2

5 ppb Quinalphos

Run 1 4.7

Run 2 5.7

Run 3 5.0

Run 4 5.1

Run 5 5.4

5 ppb Chlorfenapyr

Run 1 4.6

Run 2 4.8

Run 3 4.3

Run 4 4.9

Run 5 4.2

5 ppb Endosulfan-beta

Garlic : Single Quad SIM v.s. GC-QQQ MRM

5 ppb Deltamethrin

30 ppb Deltamethrin

GC-Q, SIM GC-QQQ, MRM

This demonstrates that GC-QQQ has much higher selectivity (sensitivity) than GC-Q

Garlic : Single Quad SIM v.s. GC-QQQ MRM

10 ppb Omethoate, MRM

Compound was confirmed easily

40 ppb Omethoate, SIM

Ion ratios were seriously distorted

GC-QQQ removes matrix interference and provides more reliable confirmation

Page 29

TOTAL Solutions Outline

• Instrumentations and Agilent Portfolio

• GC and LC

• MS Analyzers Selection

• GC/MS

• QQQ (MS/MS)

• LC/Q-TOF

Page 30

Business Issues and Challenges

• Global trade increases the number of pesticides to monitor (> several hundreds)– Typically 150-200 on a target list

• Regulation driving lower detection limits– 10 ppb or lower

• Speed: doing more analyses in a day– Fast chromatography and shorter cycle time

• Need for fast startup and long uptime– No method development

Faster Data Review and Reporting

Total Solution Offering

Screen for Unknowns

DRS and Q-TOF to

screen

GC- or LC-QQQ

Deconvolution integrated with

QEdit

• Databases and libraries available

• Universal MassHunter platform

• Backflushing

Page 31

Agilent has the TOTAL portfolio of target and non-target pesticide analyzers

Target Compound Non-target (unknown) Compound

GA

S P

hase

GC/QQQ (MRM)

• MRM method available for >200 compounds

• Backflushing

GC/MSD + Deconvolution

• The only company with deconvolution integrated into data analysis and quantitation (Easy and time saving)

• 2 Pesticide Libraries (for screening and confirmation)

• Backflushing to increase productivity

LIQU

ID P

hase

LC/QQQ (MRM)

• MRM method for >300 compounds

• Optimizer (>500 compounds)

LC/Q-TOF (or TOF) + Exact Mass database search

• Excellent mass accuracy (< 3ppm) to search against public and private databases provides confident results

• Molecular Formula Generator feature reduces data interpretation time

• NEW METLIN databases (1400 entries) for easy screening

Pesticide Analysis Most Popular Application using GCQQQ

• Pesticides in Traditional Chinese Medicine (TCM)

• Exceptional quantitative performance across a wide concentration range

• Exceptional precision for qualitative ion ratios

• Pesticides in carrot

• MS/MS succeeds where SIM has failed

Page 32

Excellent Result for Qualification in Real Sample - Cyanophos in TCM matrix, NOT solvent

•“ Inert” Qualifier Ion Ratio

0.1ppb(LOD)-1000ppb

Page 33

Excellent Result for Quantitation in Real Sample - Cyanophos in TCM matrix, NOT solvent

R^2=0.99940.2ppb(LOQ)-100ppb(9 Points)

•“ Stable” Quant AccuracyEven Up to 1000ppb (~104 linear Range)

Page 34

Build up your confidence in Quantitation Results - 1ppb Bromopropylate in TCM matrix, 10 times lower t han Uniform Limit*

RSD = 5.12%

*10 ppb is the uniform limit in Japan Positive List

Page 35

1ppb Cyanophos in TCM Matrix, 6 injection

RSD = 4.83%

Page 36

SIM vs MS/MS Comparison: Chlorpyrifos Not Confirmed in SIM Due to Matrix Interferences!

Chlorpyrifos (28 ppb) Easily Detected and Quantitated by GC/MS/MS – Incurred Carrot

Coun

ts

Resp

onse

s

Page 39 Sept. 2007