Overview of Rapid Evaporative Ionisation Mass Spectrometry (REIMS) and it's application to food authenticity studies including fish speciation.

1. ©2015 Waters Corporation 1
Rapid Evaporative Ionisation Mass
Spectrometry (REIMS) for food & agri-
tech applications

2. ©2015 Waters Corporation 2
Overview
 Real time analysis for food testing
 Food authenticity applications
– Fish & meat speciation
– Botanical origin of honey
– Authenticity of PDO butter
– Phytochemical profiling in herbs
 Summary & future study
 Acknowledgments

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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
– Electrosurgical knives use an electrical current to
rapidly heat tissue, cutting through it while
minimising blood loss
– 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

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Sample
REIMS instrument configuration
iKnife
Hand-held
sampling device(s)
Diathermy
generator
Informatics system
Source
Xevo G2-XS QTof
& Synapt G2 Si

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How does it work?

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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 &
verification
Identification of unknowns by
similarity to database entries

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Direct analysis for flexible & rapid
screening
Sub-sampling
Determination
(5s)
Data analysis
(2s)
Report
 Point-of-control analysis

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REIMS for Food Security
AGRI-TECHNOLOGY
PROTECTED STATUS

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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

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North Atlantic Cod (Gadus morhua)
Gadidae genus
Glycerophospholipids
PAs
PEs
PCs
PIs
EPA
Eicosapentaenoic
acid
DHA
Docosahexaenoic
acid
Arachidonic acid
Hexadecanoic
acid
Fatty acids
Unsaturated&
saturated

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Spot the differences?
REIMS neg ion +IPA 200 µl/min + leu enk
Cod
Whiting
Monkfish
Leu enk
[M-H]-

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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 & verified by rtPCR

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Preliminary multi-species model
Raw fish muscle
Group Numberofspectra Numberofpasses Numberoffailures Numberofoutliers CorrectClassificationRate
G 2268 2133 135 0 94.05%
Total 2268 2133 135 0 94.05%
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 m/z
 200 biological samples & >2000 spectra
PCA
ColeyPollock
Whiting
Haddock
Cod
PCA/LDA

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PROTOTYPE real-time recogniser
software
Question
Which species of fish
are present?

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Meat Speciation an Emerging issue in
India (October 2015)
Bos taurus vs. Bison bonasus

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Polyunsaturated fatty acid (PUFA)
content
Ca. x1.6
Arachidonic acid
Ω6
C20H32O2
Eicosapentaenoic acid
Ω3
C20H30O2
Ca. x7
 Good ratio of Ω3:6 acids

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PROTOTYPE real-time recogniser
software

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Food Fraud & authenticity applications

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REIMS spectra
Kaempferol
Apigenin
Oleanolic acid /
ursolic acid
Herbs & 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

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Summary & future prospects
 REIMS for food authenticity?
– Direct analysis technique results possible in seconds
o New ionisation technique
– Simplicity of operation & potential for automation
– Point-of-control analysis… bringing the analysis to the sample
o RTR software commercialisation June 2016
 Chemometric profiling
– Holistic profiling of the sample for QC
– Detection of the “unexpected” for authenticity
o Potential to identify & verify unique product markers
 Liquid sampling REIMS
– Alcoholic beverages; fruit juices; milk & edible oils

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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)