rapid detection of pesticides in fruit juice without sample preparation - waters corporation food...

©2015 Waters Corporation 1

Rapid Detection of Pesticides in Fruit Juice

using tandem quadrupole LC/MS/MS without

the need for Sample Preparation


Presentation Overview

Overview of pesticide residue analysis

Experimental

– System Solution

Results

– Multi-Residue Story

o Sample preparation comparison

• Both scenarios working well…

• One method is better than the other…

o Robustness and repeatability data

Conclusion


Food Contaminant Testing What are the challenges?

Changing Legislation

Demanding Performance

Multitude of Food Types

Variety of Compounds


Ultra Trace Analysis An Analogy

Whole product Representative sample Ultra trace detail


Ultra trace analysis What are the challenges?

Challenging matrix/analyte combinations

Low LOQs

• Low target levels

Amount of sample

• Mass/volume possible to extract

Matrix effects

• Complex matrices

Reproducibility

• Trace levels can affect statistics


What impact does this have on the laboratory?

Demand for Flexible methods

– Many analytes

– Many matrices

– Simple set-up / modification

Method performance

– High sensitivity

– Quantitative accuracy

Data management

– Automated processing

– Reporting

– Archive and back-up


Pesticide Residue Analysis

Recent trend for pesticide residue analysis is to run multi-analyte methods

Many labs now running QuEChERS (or variations of…)

Experimental Questions

– Is it possible to use dilute-and-shoot for pesticide residue analysis?

– How does QuEChERS compare with dilute-and-shoot?

– Are we able to reduce to sample prep time even further for the detection of pesticides in food and beverage products?

– Can we meet the regulatory detection limits required?

– Can we produce repeatable data with a dilute-and-shoot approach for pesticide residues in fruit juices?


• Automatically sets up instrument,

• Optimises MRMs and performs system suitability

Setup & Optimize Complete Analytical System

• Generates optimised methods, time scheduled MRMs

Generate / Select Methods to Run

• Methods generated automatically by QuanPedia,

• Clear flagging when data is out of regulatory tolerance

• Reduced risk of errors

Review Data & Prepare Reports

• Real time QC, Intelligent ‘on the fly’ decision making

Check QC & Calibration Standards

• Long term system monitoring, Lab management tool

Track System & Method Performance / Trends

QUAN Workflow


Experimental Information

Multi-Residue Method


Pesticide Residues Targeted Screening Workflow


Sample Preparation Two methods employed

Measure out 10 mL of fruit juice (spiked at 10ng/mL)+ 15mL of 1%

acetic acid in Acetonitrile into DisQuE tube 1

Centrifuge and transfer 1 mL of the extract into DisQuE tube 2

Centrifuge Dilute 10 times with water

(~ 15x dilution)

Dilute

and

Shoot

Measure out 10 mL of fruit juice (spiked at 10ng/mL)

Dilute 10 and 100 times with water

Filter and Inject


UPLC system ACQUITY UPLC System

Column: ACQUITY UPLC BEH C18 2.1 x 100 mm, 1.7 m

Column temp 40 C

Mobile phase A Water + 0.1% Formic acid

Mobile phase B Methanol + 0.1% Formic acid

Flow Rate 0.45 mL/min

Inject. volume 10 L

Gradient

ACQUITY UPLC & Xevo TQ-S Conditions

Min % A %B

0.00 95 5

0.25 95 5

7.75 0 100

8.50 0 100

8.51 95 5

10.00 95 5


ACQUITY UPLC & Xevo TQ-S Conditions

MS System XEVO TQ-S

Ionization mode ESI Positive

Capillary voltage 3.0 kV

Cone voltage 30.0 V

Source temp 150 C

Desolvation temp 500 C

Desolvation gas 1000 L/hr

Cone gas 50 L/hr

Routine targeted quantification

with information rich qualitative data

Specifically designed for sensitivity AND robustness




IntelliStart


Quanpedia




TargetLynx


QCMonitor


TrendPlot

Waters QUAN Workflow


Quanpedia

A central data base for

quantitative LC/MS methods

Based on compound name

A tool to aid MS method creation

– Automatically creates data

acquisition methods

– Automatically creates data

processing methods


Quanpedia

Quanpedia – manage compounds and methods


Quanpedia


Quanpedia improves data quality

Manual MS method

1 MRM per compound

6.6 % RSD

8 data points

across peak

Quanpedia MS method

1 MRM per compound

2.4 % RSD

12 data points

across peak

Quinoxyfen @ 1ppb

Manual average = 23 % RSD

Quanpedia average = 8.6 % RSD

80 pesticides spiked,

acquired using 402 compound MS method


90 Multi-Analytes Screened

Acaricide Monocrotophos Herbicide Atraton Herbicide Aziprotryne

Acaricide/Insecticide Methamidophos Herbicide Prometryn Herbicide Propaquizafop

Acaricide/Insecticide Dimethoate Herbicide MCPA Herbicide Isoproturon

Acaricide/Insecticide Dichlorvos Herbicide Isoxaben Herbicide Terbuthylazine

Acaricide/Insecticide Methiocarb Herbicide Asulam Herbicide Tebutam

Acaricide/Insecticide Parathion Herbicide Dimethachlor Herbicide Thidiazuron

Acaricide/Insecticide EPN Herbicide Picloram Herbicide Diuron

acaricides/insecticides Phosmet Herbicide Isocarbamide Herbicide Siduron

Breakdown product 2,6-Dichlorbenzamid Herbicide Tebuthiuron Herbicide Hexazinone

Fungicide Oxadixyl Herbicide Diphenamid Herbicide Propanil

Fungicide Hexaconazole Herbicide Quizalofop-ethyl Herbicide Clodinafop-propargyl

Fungicide Carbendazim Herbicide Cyanazine Herbicide Dimethenamid

Fungicide Difenoconazole Herbicide Prometon herbicides Haloxyfop

Fungicide Pyracarbolid Herbicide Naptalam herbicides Propisochlor

Fungicide Propiconazole Herbicide Napropamide herbicides Butachlor

Fungicide Thiabendazole Herbicide Rimsulfuron herbicides 2,4-DB

Fungicide Flutriafol Herbicide Propyzamide herbicides MCPB

Fungicide Imazalil Herbicide Atrazine Insecticide Cyromazine

Fungicide Boscalid Herbicide Terbutryn Insecticide Dicrotophos

Fungicide Fenamidone Herbicide Desmetryn Insecticide Thiodicarb

Fungicide Tebuconazole Herbicide Ametryn Insecticide Fenpropathrin

Fungicide Benomyl Herbicide Simazine Insecticide Methomyl

Fungicide Azoxystrobin Herbicide Metazachlor Insecticide Acephate

Fungicide/Herbicide Ethoxyquin Herbicide Alachlor Insecticide Imidacloprid

Fungicide Tridemorph Herbicide Mecoprop Insecticide Trichlorfon

Fungicide Octhilinone (2-Octyl-4-isothiazoline-3-one) Herbicide Propachlor Insecticide Phoxim

Insecticide Phorate Insecticide Propoxur insecticides Tetramethrin

Insecticide Pymetrozine Insecticide Diflubenzuron insecticides Fenitrothion

Insecticide Pirimicarb Insecticide Buprofezin insecticides Allethrin

Insecticide Diazinon Insecticide Deltamethrin insecticides Rotenone

Insecticide Carbofuran Insecticide/Acaricide Azinphos-methyl Metabolite Aldicarb sulfone

Insecticide Parathion-methyl Insecticide/Acaricide Aldicarb Metabolite Atrazine-desethyl

Insecticide Carbaryl Insecticide/Nematicide Oxamyl Metabolite Aldicarb sulfoxide

Insecticide Fenthion Insecticide/Nematicide Phosphamidon Metabolite Atrazine-desisopropyl

Insecticide Acetamiprid Insecticide/Nematicide Dichlofenthion Metabolite Carbofuran-3-hydroxy

Insecticide Sulfotep Insecticide Phenothrin


Results and Discussion


Developing a Suitable Method

Analysis of food samples can be challenging – need to

develop a sample preparation that can:

o Reduce matrix effects (e.g. peak distortion, ion

suppression…)

o Extract relevant compounds

o Provide a robust, repeatable method

Method development to investigate different

extraction techniques can be time intensive

Useful to have tools that support this process and help

improve lab efficiency

LC-MS/MS Analysis

Modify analytical method

LC-MS/MS Analysis

Modify sample

preparation


Spiked Orange Juice without RADAR


On of the ways to determine sample prep efficiency…

What is RADAR?

– Rapid electronics allow instrument to switch between MRM & full scan

– Added information can be gained from full scan data in routine analyses

• Intelligent decision making

• Improves method development

• Monitor matrix components

• Detect un-targeted adulterants

How do I set it up on the instrument?

- Parallel MRM with full scan data acquisition


Apple juice Grape juice Orange juice

Q Q

Q

100X 100X 100X

10X 10X

10X

Juice Samples – Matrix Contribution QuEChERS versus Dilute-and-Shoot


Sample prep comparison When both options work…

0.7 ng/mL

1 ng/mL

0.1 ng/mL

Atraton @ 10 ng/mL

10x 10x

100X 100X 100X

10x

Quechers Quechers Quechers


Sample prep comparison When QuEChERS works…

Rotenone @ 10 ng/mL


Sample prep comparison When Dilute & Shoot Doesn’t work…

Rotenone @ 10 ng/mL

Sample Diluted 10x


Q - diluted Q - diluted Q - diluted

Sample prep comparison When Dilute-and-Shoot works…

Q

10X

Q

10X

Q

10X

Poor peak shape Shift in RT




IntelliStart


Quanpedia




TargetLynx


QCMonitor


TrendPlot



TargetLynx Reporting Designed for food safety customers


Linearity and Repeatability

Compound name: Atraton

Correlation coefficient: r = 0.997944, r^2 = 0.995892

Calibration curve: 22116.2 * x + 949.701

Response type: External Std, Area

Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

ng/mL0 10 20 30 40 50 60 70 80 90 100

Re

sp

on

se

0

500000

1000000

1500000


Dilute-and-Shoot Calibration Curve

Compound name: Atraton

Correlation coefficient: r = 0.998102, r̂ 2 = 0.996207

Calibration curve: 0.0161973 * x + 0.00307921

Response type: Internal Std ( Ref 16 ), Area * ( IS Conc. / IS Area )

Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

ng/mL0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320

Resp

onse

0.00

1.00

2.00

3.00

4.00

5.00




IntelliStart


Quanpedia




TargetLynx


QCMonitor


TrendPlot



Compound Repeatability & Robustness Dilute-and-Shoot Approach


Pressure Repeatability Dilute-and-Shoot Approach

1st injection

100th injection

155th injection


Conclusions

Variety of different fruit juice samples have been analysed

Direct comparison of QuEChERS and dilute-and-shoot for pesticide

residue screening

Analysis was performed using a targeted multi-analyte screening

solution:

o Setting up the method – IntelliStart, Quanpedia

o Sample preparation- DisQUE and ‘dilute-and-shoot’

o Reliable chromatography - ACQUITY UPLC & ACQUITY BEH C18 columns

o Robust and Repeatable results - Xevo TQ-S

o Matrix monitoring tools - RADAR

o Review of quantitative data –TargetLynx, TrendPlot

rapid detection of pesticides in fruit juice without sample preparation - waters corporation food...

Science

waters corporation

residue method

ultra trace analysis

optimised methods

flexible methods

sample prep time

multianalyte methods

complete analytical