copy2015 Waters Corporation 1

Rapid Evaporative Ionisation Mass

Spectrometry (REIMS) for food amp agri-tech applications

copy2015 Waters Corporation 2

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

Summary amp future study

Acknowledgments

copy2015 Waters Corporation 3

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

copy2015 Waters Corporation 4

Sample

REIMS instrument configuration

iKnife Hand-held

sampling device(s)

Diathermy generator

Informatics system

Source

Xevo G2-XS QTof amp Synapt G2 Si

copy2015 Waters Corporation 5

How does it work

copy2015 Waters Corporation 6

REIMS for food research workflow chemometric profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Multivariate statistical analysis

Different tissues

Different species

Different production

Biomarker ID amp verification

Identification of unknowns by similarity to database entries

copy2015 Waters Corporation 7

Direct analysis for flexible amp rapid screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Point-of-control analysis

copy2015 Waters Corporation 2

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

Summary amp future study

Acknowledgments

copy2015 Waters Corporation 3

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

copy2015 Waters Corporation 4

Sample

REIMS instrument configuration

iKnife Hand-held

sampling device(s)

Diathermy generator

Informatics system

Source

Xevo G2-XS QTof amp Synapt G2 Si

copy2015 Waters Corporation 5

How does it work

copy2015 Waters Corporation 6

REIMS for food research workflow chemometric profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Multivariate statistical analysis

Different tissues

Different species

Different production

Biomarker ID amp verification

Identification of unknowns by similarity to database entries

copy2015 Waters Corporation 7

Direct analysis for flexible amp rapid screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Point-of-control analysis

copy2015 Waters Corporation 3

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

copy2015 Waters Corporation 4

Sample

REIMS instrument configuration

iKnife Hand-held

sampling device(s)

Diathermy generator

Informatics system

Source

Xevo G2-XS QTof amp Synapt G2 Si

copy2015 Waters Corporation 5

How does it work

copy2015 Waters Corporation 6

REIMS for food research workflow chemometric profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Multivariate statistical analysis

Different tissues

Different species

Different production

Biomarker ID amp verification

Identification of unknowns by similarity to database entries

copy2015 Waters Corporation 7

Direct analysis for flexible amp rapid screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Point-of-control analysis

copy2015 Waters Corporation 4

Sample

REIMS instrument configuration

iKnife Hand-held

sampling device(s)

Diathermy generator

Informatics system

Source

Xevo G2-XS QTof amp Synapt G2 Si

copy2015 Waters Corporation 5

How does it work

copy2015 Waters Corporation 6

REIMS for food research workflow chemometric profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Multivariate statistical analysis

Different tissues

Different species

Different production

Biomarker ID amp verification

Identification of unknowns by similarity to database entries

copy2015 Waters Corporation 7

Direct analysis for flexible amp rapid screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Point-of-control analysis

copy2015 Waters Corporation 5

How does it work

copy2015 Waters Corporation 6

REIMS for food research workflow chemometric profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Multivariate statistical analysis

Different tissues

Different species

Different production

Biomarker ID amp verification

Identification of unknowns by similarity to database entries

copy2015 Waters Corporation 7

Direct analysis for flexible amp rapid screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Point-of-control analysis

copy2015 Waters Corporation 6

REIMS for food research workflow chemometric profiling

iKnife and Xevo

G2-XS QTof

Model Generation

Multivariate statistical analysis

Different tissues

Different species

Different production

Biomarker ID amp verification

Identification of unknowns by similarity to database entries

copy2015 Waters Corporation 7

Direct analysis for flexible amp rapid screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Point-of-control analysis

copy2015 Waters Corporation 7

Direct analysis for flexible amp rapid screening

Sub-sampling Determination

(5s)

Data analysis (2s)

Report

Point-of-control analysis

copy2015 Waters Corporation 8

REIMS for Food Security

AGRI-TECHNOLOGY

PROTECTED STATUS

copy2015 Waters Corporation 9

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

copy2015 Waters Corporation 10

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

copy2015 Waters Corporation 11

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

Cod

Whiting

Monkfish

Leu enk [M-H]-

copy2015 Waters Corporation 12

White fish database

Proof-of-concept

Species Atlantic cod - Gadus morhua Haddock - Melanogrammus aeglefinus Atlantic pollock - Pollachius pollachius Coley - Pollachius Virens Whiting (lunar whiting) - Merlangius merlangus

Authentic samples obtained from the UK seafish industry amp verified by rtPCR

copy2015 Waters Corporation 11

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

Cod

Whiting

Monkfish

Leu enk [M-H]-

copy2015 Waters Corporation 12

White fish database

Proof-of-concept

Species Atlantic cod - Gadus morhua Haddock - Melanogrammus aeglefinus Atlantic pollock - Pollachius pollachius Coley - Pollachius Virens Whiting (lunar whiting) - Merlangius merlangus

Authentic samples obtained from the UK seafish industry amp verified by rtPCR

copy2015 Waters Corporation 12

White fish database

Proof-of-concept

Species Atlantic cod - Gadus morhua Haddock - Melanogrammus aeglefinus Atlantic pollock - Pollachius pollachius Coley - Pollachius Virens Whiting (lunar whiting) - Merlangius merlangus

Authentic samples obtained from the UK seafish industry amp verified by rtPCR

copy2015 Waters Corporation 13

Preliminary multi-species model Raw fish muscle

Group Number of spectra Number of passes Number of failures Number of outliers Correct Classification Rate

G 2268 2133 135 0 9405

Total 2268 2133 135 0 9405

Species Cod Coley Haddock Pollock Whiting Total

Cod 638 11 46 0 0 695

Coley 1 260 0 0 7 268

Haddock 51 2 512 0 1 566

Pollock 0 0 0 373 0 373

Whiting 0 16 0 0 350 366

Cross validation results

REIMS negative ionisation acquired in sensitivity mode on the Xevo G2 XS Qtof using monopolar knife sampling Spectral data were acquired between 200-1200 mz 200 biological samples amp gt2000 spectra

PCA

Coley Pollock

Whiting

Haddock Cod

PCALDA

copy2015 Waters Corporation 14

PROTOTYPE real-time recogniser software

Question Which species of fish

are present

copy2015 Waters Corporation 15

Meat Speciation an Emerging issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2015 Waters Corporation 16

Polyunsaturated fatty acid (PUFA) content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2015 Waters Corporation 14

PROTOTYPE real-time recogniser software

Question Which species of fish

are present

copy2015 Waters Corporation 15

Meat Speciation an Emerging issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2015 Waters Corporation 16

Polyunsaturated fatty acid (PUFA) content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2015 Waters Corporation 15

Meat Speciation an Emerging issue in India (October 2015)

Bos taurus vs Bison bonasus

copy2015 Waters Corporation 16

Polyunsaturated fatty acid (PUFA) content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2015 Waters Corporation 16

Polyunsaturated fatty acid (PUFA) content

Ca x16

Arachidonic acid Ω6

C20H32O2

Eicosapentaenoic acid Ω3

C20H30O2

Ca x7

Good ratio of Ω36 acids

copy2015 Waters Corporation 17

PROTOTYPE real-time recogniser software

copy2015 Waters Corporation 18

Food Fraud amp authenticity applications

copy2015 Waters Corporation 19

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

copy2015 Waters Corporation 20

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

copy2015 Waters Corporation 21

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)

copy2015 Waters Corporation 21

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)