Innovations in fish fraud detection using rapid evaporative ionisation mass spectrometry (REIMS)
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1 Innovations in fish fraud detection using rapid evaporative ionisation mass spectrometry (REIMS) Connor Black Institute for Global Food Security June 2017
2 Outline Introduction Ambient mass spectrometry REIMS background & setup Fish speciation study using REIMS Chemometric fish models Validation Comparison with PCR Other aspects of fish fraud Conclusions
3 Ambient mass spectrometry (AMS) First conceptualised in 1998 with paper spray mass spectrometry (PS-MS) Spray or jet ionisation Electric discharge ambient ionisation Ambient gas-, heat- or laser assisted desorption/ionisation Desorption electrospray ionisation (DESI) Desorption atmospheric pressure photo ionisation (DAPPI) Desorption electrospray/metastable-induced ionisation (DEMI)* Atmospheric pressure glow discharge desorption ionisation (APGDDI) Desorption atmospheric pressure chemical ionisation (DAPCI) Direct analysis in real time (DART) Atmospheric solid analysis probe (ASAP) Electrospray-assisted laser desorption ionisation (ELDI) Extractive electrospray ionisation (EESI) First publication in 2004 using desorption electrospray ionisationmass spectrometry (DESI-MS) Desorption sonic-spray ionisation (DESSI) / Easy ambient sonic-spray ionisation (EASI) Electrode-assisted desorption electrospray ionisation (EADSI) Electrostatic spray ionisation (ESTASI) Jet desorption electrospray ionisation (JeDI) Dielectric barrier discharge ionisation (DBDI) Desorption corona beam ionisation (DCBI) Helium atmospheric pressure glow discharge ionisation (HAPGDI) Liquid sampling-atmospheric pressure glow discharge (LS-APGD) High voltage-assisted laser desorption ionisation (HALDI) Infrared laser ablation metastableinduced chemical ionisation (IR-LAMICI)* Laser ablation electrospray ionisation (LAESI) Laser desorption spray post-ionisation (LDSPI) Minimal to no sample preparation and fast assay running times Quantitative analysis an issue Liquid extraction surface analysis (LESA) Low temperature plasma (LTP) Laser spray ionisation (LSI) Paper spray (PS) Transmission mode desorption electrospray ionisation (TM-DESI) Microwave induced plasma desorption ionisation (MIPDI) Plasma assisted desorption ionisation (PADI) Matrix assisted laser desorption electrospray ionisation (MALDESI) Rapid evaporative ionisation mass spectrometry (REIMS) C. Black, O.P. Chevallier, C.T. Elliott; Trends in Analytical Chemistry. 82 (2016) 268.
4 Rapid evaporative ionisation mass spectrometry (REIMS)
5 Creation of fish database 478 samples Cod Coley - 51 Haddock Whiting - 50 Pollock - 50 Stability varying freeze/thawing cycle (n=32) Repeatability intra day repeatability Reproducibility inter day operators, different cutting methods
6 Stored at -80 o C prior to analysis Prototype Medimass REIMS source operated in negative-ion and sensitivity mode, with each sample cut 10 times Lockmass solution of Leucine Enkephalin (2ng / µl) in isopropanol (IPA) REIMS setup
7 Raw data
8 A prototype Waters software (OMB model building) Principal component analysis (PCA) & linear discriminant analysis (LDA) Phospholipid region m/z Spectrum lockmassed to m/z & binning of spectrum per sample Data handling
9 WHITING _RH_S387_ (0.283) Cm (16:21) Lunar Whiting TOF MS ES- 2.27e6 POLLOCK _RH_S233_ (0.283) Cm (15:19) Pollock TOF MS ES- 1.10e6 % % m/z m/z COD _RH_S3_ (0.283) Cm (16:19) TOF MS ES- 3.33e6 HADDOCK _RH_S178_ (0.283) Cm (16:21) Haddock TOF MS ES- 4.81e % % m/z m/z
10 Chemometric models Fish speciation m/z Cod Coley Haddock Pollock Whiting PCA LDA 5 clear clusters, each one representative of a fish species
11 Internal 20% cross validation Standard deviation set at 5 Samples outside standard deviation values assigned as outliers Correct classification rate of 99.37% (including outliers) Cod Coley Haddock Pollock Whiting Outlier Total Correct Classification Rate Cod % Coley % Haddock % Pollock % Whiting % Total %
12 Cod Coley Haddock Pollock Whiting Species markers Pollock markers OPLS-DA R 2 = 0.829; Q 2 = 0.809
13 Validation 99 samples Cod - 22 Coley - 20 Haddock - 20 Whiting - 17 Pollock - 20
14 Cod Coley Haddock Pollock Whiting Outlier Total Correct Classification Rate Cod % Coley % Haddock % Pollock % Whiting % Total % Standard deviation value of % correct classification Accurate fish identification achievable near-instantaneously
15 Further analysis of speciation model(s) 6 suspect haddock samples in the model building process All clustered with cod samples PCR testing identified 99% similarity to cod & NOT HADDOCK Time comparison of 30 mins 24 hours Tested seabass (n=6) & seabream (n=8) All identified as outliers using recognition software Statistical check = 100% correct classification
16 Catch method Speciation, geographic origin, chain of custody, catch method, fishery, illegal fishing (IUU), processed raw material authenticity Haddock Trawl Haddock Line PCA Generated chemometric models of line v trawl caught haddock (1 spectrum per sample) LDA
17 Mediterranean sea Croatia Italy Greece Spain Geographic origin - seabream PCA LDA
18 Croatia Italy - Sardinia Italy Tuscany (Orbetello) Spain Geographic origin - seabass PCA LDA
19 Conclusions Fish speciation very achievable using REIMS technology Validation of fish models with external set of samples provided a 98.99% classification Instantaneous results obtained (1-2 seconds delay) Important time comparisons with conventional techniques (PCR) Markers for each species of fish identifiable Potential for catch method and geographic origin to be determined
20 Acknowledgements Olivier Chevallier Julia Balog Zoltan Takats Maria Riina Simon Haughey Sara Stead Francesca Martucci Christopher Elliott Steven Pringle Pier Acutis
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