copy2016 Waters Corporation 40

Dr Sara Stead sara_steadwaterscom

18th March 2016

Rapid Evaporative Ionisation Mass Spectrometry (REIMS)

for Food amp Agri-Tech Applications

copy2016 Waters Corporation 41

Overview

Real time analysis for food testing

Food authenticity applications

ndash Fish amp meat speciation

ndash Botanical origin of honey

ndash Authenticity of PDO butter

ndash Phytochemical profiling in herbs

Agri-tech applications

ndash Phenotypic profiling

Summary amp future study

Acknowledgments

copy2016 Waters Corporation 42

Origins of REIMS

Rapid Evaporative Ionisation Mass Spectrometry

(REIMS) is an emerging technique that allows rapid

characterisation of biological tissues with no need for

sample preparation

The inventor Professor Zoltan Takats of Imperial College

London created the iKnife by connecting an

electrosurgical knife to a mass spectrometer

ndash Electrosurgical knives use an electrical current to rapidly

heat tissue cutting through it while minimising blood loss

ndash In doing so they vaporise the tissue creating smoke that is

normally sucked away by extraction systems but is actually a

rich source of biological information that can provide the

surgeon a real-time decision-making tool

copy2016 Waters Corporation 43

Sample

REIMS Instrument Configuration

iKnife Hand-held

sampling device(s)

Diathermy generator

Informatics system

Source

REIMS is an emerging technique that allows rapid characterisation of biological

tissues and food samples

Xevo G2-XS QTof amp SYNAPT G2 Si

Potential for automation ndash Tecan system

copy2016 Waters Corporation 44

How Does it Work

copy2016 Waters Corporation 45

REIMS for Food Research Workflow Chemometric Profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Different tissues

Different species

Different production

Identification of unknowns by similarity to database entries

copy2016 Waters Corporation 46

Direct Analysis for Flexible amp Rapid Screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Sample to report in seconds

Point-of-control analysis

copy2016 Waters Corporation 42

Origins of REIMS

Rapid Evaporative Ionisation Mass Spectrometry

(REIMS) is an emerging technique that allows rapid

characterisation of biological tissues with no need for

sample preparation

The inventor Professor Zoltan Takats of Imperial College

London created the iKnife by connecting an

electrosurgical knife to a mass spectrometer

ndash Electrosurgical knives use an electrical current to rapidly

heat tissue cutting through it while minimising blood loss

ndash In doing so they vaporise the tissue creating smoke that is

normally sucked away by extraction systems but is actually a

rich source of biological information that can provide the

surgeon a real-time decision-making tool

copy2016 Waters Corporation 43

Sample

REIMS Instrument Configuration

iKnife Hand-held

sampling device(s)

Diathermy generator

Informatics system

Source

REIMS is an emerging technique that allows rapid characterisation of biological

tissues and food samples

Xevo G2-XS QTof amp SYNAPT G2 Si

Potential for automation ndash Tecan system

copy2016 Waters Corporation 44

How Does it Work

copy2016 Waters Corporation 45

REIMS for Food Research Workflow Chemometric Profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Different tissues

Different species

Different production

Identification of unknowns by similarity to database entries

copy2016 Waters Corporation 46

Direct Analysis for Flexible amp Rapid Screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Sample to report in seconds

Point-of-control analysis

copy2016 Waters Corporation 43

Sample

REIMS Instrument Configuration

iKnife Hand-held

sampling device(s)

Diathermy generator

Informatics system

Source

REIMS is an emerging technique that allows rapid characterisation of biological

tissues and food samples

Xevo G2-XS QTof amp SYNAPT G2 Si

Potential for automation ndash Tecan system

copy2016 Waters Corporation 44

How Does it Work

copy2016 Waters Corporation 45

REIMS for Food Research Workflow Chemometric Profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Different tissues

Different species

Different production

Identification of unknowns by similarity to database entries

copy2016 Waters Corporation 46

Direct Analysis for Flexible amp Rapid Screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Sample to report in seconds

Point-of-control analysis

copy2016 Waters Corporation 44

How Does it Work

copy2016 Waters Corporation 45

REIMS for Food Research Workflow Chemometric Profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Different tissues

Different species

Different production

Identification of unknowns by similarity to database entries

copy2016 Waters Corporation 46

Direct Analysis for Flexible amp Rapid Screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Sample to report in seconds

Point-of-control analysis

copy2016 Waters Corporation 45

REIMS for Food Research Workflow Chemometric Profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Different tissues

Different species

Different production

Identification of unknowns by similarity to database entries

copy2016 Waters Corporation 46

Direct Analysis for Flexible amp Rapid Screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Sample to report in seconds

Point-of-control analysis

copy2016 Waters Corporation 46

Direct Analysis for Flexible amp Rapid Screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Sample to report in seconds

Point-of-control analysis

copy2016 Waters Corporation 47

REIMS for Food Security

AGRI-TECHNOLOGY

PROTECTED STATUS

copy2016 Waters Corporation 48

Oceana Survey Undertaken in 2015

Only 6 of Northern red snapper fish collected were truly red snapper (Lutjanus campechanus)

84 of tuna sold in sushi restaurants was escolar (health effects)

Almost 50 of all grocery stores visited in Boston sold mislabelled fish

31 of cod in Boston was mislabelled

copy2016 Waters Corporation 49

Fish Speciation Study

Phylogentic tree

Gadidae ndash Cod Haddock Coley Whiting amp Pollock

Pangasiidae ndash River Cobbler amp Basa (Vietnamese Catfish)

Lophiida ndash Monkfish

Serranidae ndash Sea Bass

Salmo ndash Atlantic salmon

Pleuronectes ndash European plaice

copy2016 Waters Corporation 50

North Atlantic Cod (Gadus morhua) Gadidae genus

Glycerophospholipids PAs PEs PCs PIs

EPA Eicosapentaenoic

acid

DHA Docosahexaenoic

acid

Arachidonic acid

Hexadecanoic acid

Fatty acids Unsaturatedamp saturated

copy2016 Waters Corporation 51

Spot the Differences REIMS neg ion +IPA 200 microlmin + leu enk

Cod

Whiting

Monkfish

Leu enk [M-H]-

copy2016 Waters Corporation 52

PROTOTYPE Software - PoP Multi-Species Fish Model

Gadidae species Cod haddock whiting

coley

Atlantic Salmon

Salmo salar

Sea Bass Dicentrarchus

labrax

Basa Pangasius bocourti

Plaice Pleuronectes platessa

Monkfish Lophius piscatorius

PCA model LDA model

Real-time recogniser software - prototype

n=ge3

copy2016 Waters Corporation 53

Meat Speciation An Emerging Issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2016 Waters Corporation 54

Polyunsaturated Fatty Acid (PUFA) Content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2016 Waters Corporation 50

North Atlantic Cod (Gadus morhua) Gadidae genus

Glycerophospholipids PAs PEs PCs PIs

EPA Eicosapentaenoic

acid

DHA Docosahexaenoic

acid

Arachidonic acid

Hexadecanoic acid

Fatty acids Unsaturatedamp saturated

copy2016 Waters Corporation 51

Spot the Differences REIMS neg ion +IPA 200 microlmin + leu enk

Cod

Whiting

Monkfish

Leu enk [M-H]-

copy2016 Waters Corporation 52

PROTOTYPE Software - PoP Multi-Species Fish Model

Gadidae species Cod haddock whiting

coley

Atlantic Salmon

Salmo salar

Sea Bass Dicentrarchus

labrax

Basa Pangasius bocourti

Plaice Pleuronectes platessa

Monkfish Lophius piscatorius

PCA model LDA model

Real-time recogniser software - prototype

n=ge3

copy2016 Waters Corporation 53

Meat Speciation An Emerging Issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2016 Waters Corporation 54

Polyunsaturated Fatty Acid (PUFA) Content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2016 Waters Corporation 51

Spot the Differences REIMS neg ion +IPA 200 microlmin + leu enk

Cod

Whiting

Monkfish

Leu enk [M-H]-

copy2016 Waters Corporation 52

PROTOTYPE Software - PoP Multi-Species Fish Model

Gadidae species Cod haddock whiting

coley

Atlantic Salmon

Salmo salar

Sea Bass Dicentrarchus

labrax

Basa Pangasius bocourti

Plaice Pleuronectes platessa

Monkfish Lophius piscatorius

PCA model LDA model

Real-time recogniser software - prototype

n=ge3

copy2016 Waters Corporation 53

Meat Speciation An Emerging Issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2016 Waters Corporation 54

Polyunsaturated Fatty Acid (PUFA) Content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2016 Waters Corporation 52

PROTOTYPE Software - PoP Multi-Species Fish Model

Gadidae species Cod haddock whiting

coley

Atlantic Salmon

Salmo salar

Sea Bass Dicentrarchus

labrax

Basa Pangasius bocourti

Plaice Pleuronectes platessa

Monkfish Lophius piscatorius

PCA model LDA model

Real-time recogniser software - prototype

n=ge3

copy2016 Waters Corporation 53

Meat Speciation An Emerging Issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2016 Waters Corporation 54

Polyunsaturated Fatty Acid (PUFA) Content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2016 Waters Corporation 53

Meat Speciation An Emerging Issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2016 Waters Corporation 54

Polyunsaturated Fatty Acid (PUFA) Content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2016 Waters Corporation 54

Polyunsaturated Fatty Acid (PUFA) Content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2016 Waters Corporation 55

Multi-Species Muscle Tissue ndash MVA Models

2 biological replicates of each species 10 technical replicates per sample

PCA model

Non-ruminants

Ruminants

Buffalo Bovine

Porcine

Ovine

Gallus gallus

copy2016 Waters Corporation 56

PROTOTYPE Real-Time Recogniser Software

copy2016 Waters Corporation 56

PROTOTYPE Real-Time Recogniser Software

copy2016 Waters Corporation 57

Food Fraud amp Authenticity Applications

copy2016 Waters Corporation 58

REIMS Spectra

Kaempferol

Apigenin

Oleanolic acid ursolic acid

Herbs amp spices

PDO status Belgian butter

Botanical origin of honey

Essential Ω6 fatty acid

Ω9

Galangin Myricetin

Geographical origin of coffee

Caffeine Tanzania

Peru

Kenyan

Colombian

copy2016 Waters Corporation 59

Herbicide Selectivity in Cereals

and Competing Weeds

Collaboration with Prof Rob Edwards

University of Newcastle

School of Agriculture Food amp Rural Development

copy2016 Waters Corporation 60

Black-Grass vs Rye Grass Spectral Comparison

(50 to 1200 mz 6-8 combined scans)

Rye grass

B-G Pel

B-G Roth

B-G Oxford

B-G Nots

Black-grass grouped vs Rye grass EZInfo OPLS-DA scores amp S-plot

-10

-09

-08

-07

-06

-05

-04

-03

-02

-01

00

01

02

03

04

05

06

07

08

09

10

-014 -013 -012 -011 -010 -009 -008 -007 -006 -005 -004 -003 -002 -001 000 001 002 003 004 005 006 007 008 009 010 011 012 013 014 015 016 017 018 019 020 021 022

p(corr)[1] (C

orrela

tio

n)

p[1] (Loadings)

S-Plot (Black grass = -1 Lollium = 1)

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EZInfo controlled by Progenesis QI 3 - Black grass vs lolliumusp (M9 OPLS-DA) - 2015-12-11 171517 (UTC+0)

copy2016 Waters Corporation 61

Summary amp Future Prospects

REIMS for food authenticity

ndash Direct analysis technique results possible in seconds

o New ionisation technique

ndash Simplicity of operation amp potential for automation

ndash Point-of-control analysishellip bringing the analysis to the sample

o RTR software commercialisation June 2016

Chemometric profiling

ndash Holistic profiling of the sample for QC

ndash Detection of the ldquounexpectedrdquo for authenticity

o Potential to identify amp verify unique product markers

Liquid sampling REIMS

ndash Alcoholic beverages fruit juices milk amp edible oils

Implementation of ion mobility into the workflow to enhance power to differentiate sample types amp model generation (CCS)

copy2016 Waters Corporation 62

Acknowledgements

Prof Zoltan Takats (Imperial College London)

Dr Simon Hird Dr Jonathan Williams Dr Emrys Jones Dr Julia Balog

Dr Steve Pringle Dr Mike Morris Dr Jackson Pope (Waters Manchester

and Budapest)

Prof Chris Elliott Dr Olivier Chevallier Connor Black (Queens

University Belfast)

Prof Rob Edwards Dr Melissa Brazier-Hicks Dr Catherine Tetard-Jones

More Fish

copy2016 Waters Corporation 62

Acknowledgements

Prof Zoltan Takats (Imperial College London)

Dr Simon Hird Dr Jonathan Williams Dr Emrys Jones Dr Julia Balog

Dr Steve Pringle Dr Mike Morris Dr Jackson Pope (Waters Manchester

and Budapest)

Prof Chris Elliott Dr Olivier Chevallier Connor Black (Queens

University Belfast)

Prof Rob Edwards Dr Melissa Brazier-Hicks Dr Catherine Tetard-Jones

More Fish