Thermo Scientific LCMS Solutions for Pesticide Analysis

Transcription

1 Thermo Scientific LCMS Solutions for Pesticide Analysis Dipankar Ghosh, Ph.D Director, Environmental & Food Safety - LSMS San Jose, CA April 20, The world leader in serving science

2 2 Routine Testing SOP Driven QQQ LC(GC)MS Methods

3 What else could be present? screening Unknown Emerging contaminants in environmental water Unknown contaminants in raw/processed food Food Authenticity and Fraud 3

4 Analytical Methodology Sample Preparation Chromatographic Separation Mass Spectrometry Detection Data Processing & Reporting 4

5 The Ideal Mass Analzyer? 5 Adapted from Alexander Makarov s presentation 18th IMSC Bremen

6 The Industry s Leading Portfolio of MS Solutions Exactives Non-targeted Analysis Tribrid Orbitrap HRAM Metabolomics Proteomics Bioanalysis Quantitative Food Safety Environmental Clinical/Toxicology Applied Markets Triple Quads Thermo Scientific LSMS Portfolio MS, MSn Research Markets Ion Traps Biomarker Discovery Proteomics Metabolism Qualitative Metabolomics PTM Analysis Lipidomics Targeted Analysis 6

7 QQQ : TSQ Endura & TSQ Quantiva LC-MS Systems Thermo Scientific TSQ Endura MS Extreme Quantitative Value Best-in-class sensitivity Unprecedented usability Ultimate robustness Thermo Scientific TSQ Quantiva MS Extreme Quantitative Performance Attogram sensitivity Unprecedented usability Exceptional robustness 7

8 Exactive Series Portfolio PERFORMANCE Thermo Scientific Exactive Plus (EMR) MS Orbitrap analyzer Mass Range m/z Mass Range EMR: ,000 Mass Accuracy: <1ppm Mass Resolution >140,000 Scan Speed up to 12 Hz Thermo Scientific Q Exactive Focus MS Orbitrap analyzer Mass Range m/z Mass Accuracy <1ppm Max. Mass Resolution >70,000 Scan speed up to 12Hz MS and 12 Hz MS/MS Polarity switching Top 2 ddms2 No Multiplexing Refined workflows that are optimal for routine laboratories Thermo Scientific Q Exactive MS & Q Exactive Plus MS Orbitrap analyzer Mass Range m/z Mass Accuracy <1ppm Max. Mass Resolution >140,000 Scan speed up to 12Hz MS and MS/MS Spectral Multiplexing AQT & AABG (Q Exactive Plus MS only) Optional Intact Protein Mode and Enhanced Resolution (280k) for Q Exactive Plus MS Thermo Scientific Q Exactive HF MS Ultra High Field Orbitrap analyzer Mass Range m/z Mass Accuracy <1ppm Max. Mass Resolution >240,000 Scan speed up to 18Hz MS and MS/MS Spectral Multiplexing AQT & AABG Optional Protein Mode VALUE 8

9 LC/MS Workflows for Environmental and Food Safety Tools to develop methods for known and emerging contaminants Dionex ASE Accelerated Solvent Extractor Thermo Scientific Dionex ASE Accelerated Solvent Extractor Thermo Scientific Dionex UltiMate 3000 RSLC systems Thermo Scientific EQuan MAX Plus online sample prep LCMS Thermo Scientific Exactive Plus and Thermo Scientific Q Exactive mass spectrometers: HRAM to identify, quantify and confirm TSQ Series mass spectrometers: triple stage quadrupole LC-MS/MS and GC- MS/MS for high sensitivity quantitation Custom software to meet the needs of your workflow: Thermo Scientific TraceFinder, Thermo Scientific SIEVE, Thermo Scientific Compound Discoverer, and mzcloud software Raw Sample Customized Reporting TSQ Quantiva MS TSQ Endura MS UltiMate 3000 RSLC Orbitrap Fusion MS EQuan MAX Plus System Orbitrap HR/AM 9

10 Environmental and Food Safety Workflows Workflow Quantitation Targeted Screening Non-Targeted and Unknown Screening Hardware TSQ HRMS (E+,QE) HRMS Series (E+,QE) HRMS Series (QE) Scan Modes Software SRM Full Scan PRM TSIM ddms 2 vdia Full Scan ddms 2 PRM, tsim, vdia TraceFinder SW Compound Discoverer SW m/zcloud Full Scan ddms 2 vdia SIEVE SW Thermo Scientific HRAM Spectral Library vdia is not available in the U.S. 10

11 Typical LC-QqQ-MS/MS workflow QuEChERS LC-MS/MS SRM List Retention Time Product ions Ion ratios Identification Quantification Reporting 11

12 Increasing Productivity: Alternative to Optimization by Infusion Built-in QQQ methods Select compounds - TraceFinder Compound Database to create instrument method OR Select Pre-configured TSQ method with LC and SRM conditions included - Load and Go! OR Edit existing pre-configured method to create new 12

13 H-SRM : The story of Toltrazuril and Frankfurt! Toltrazuril is a triazinetrione derivative used in the prevention and treatment of coccidiosis of chicken, turkeys and pigs. Metabolism Two oxidation products, the sulfoxide and sulfone metabolites are obtained. TOL has been included in the Annex I of Council Regulation (ECC) No 2377/90 and maximum residue levels (MRLs) on the animal tissues have been established ( µg/kg) O H N O O O H N O O O H N O F 3 C S O N O N F 3 C S O N O N F 3 C S O O N O N Toltrazuril (TOL) Toltrazuril Sulfoxide (TOLX) Toltrazuril Sulfone (TOLS) 13

14 Toltrazuril and metabolites Frankfurt sausage SRM 0.7 FWHM H-SRM 0.2 FWHM TOLX TOLS TOL TOL-D 3 Relative Abundance m/z m/z m/z m/z m/z m/z m/z m/z Time (min) m/z m/z µg/kg m/z m/z m/z m/z m/z m/z Time (min) A. Martínez-Villalba et al., Anal. and Bioanal. Chem, 2010 Aug; 397(7):

15 Typical LC-HRMS workflow QuEChERS LC-HRMS Retention Time Exact Mass (± 5 ppm) Product ions Ion ratios Isotopic Pattern Library Matching Compound Inclusion List Identification Quantification Generic Instrument Method Screening Identification Reporting Reporting 15

16 Pesticides in Fruits and Vegetables by LC-HRMS QuEChERS Sample Treatment LC-HRMS Thermo Scientific Accucore aq column(13min) FS/ddMS 2 (Inclusion List) Thermo Scientific TraceFinder software Data Processing Data presented at EPRW

17 Mass Resolving Power & Mass Accuracy are they important? The best a QTOF is going to get at low mass Do you want to see 94% or 97 % of pesticides? Thermo Scientific Q Exactive Focus MS Data presented at 10th European Pesticide Residue Workshop (EPRW 2014) Courtesy Prof Amadeo R Fernandez Albar 17

18 YES - when you add sample complexity The best a QTOF is going to get at low mass Or worse!! Do you want to see 77% or 94 % of pesticides? Q Exactive Focus MS Data presented at 10th European Pesticide Residue Workshop (EPRW 2014) Courtesy Prof Amadeo R Fernandez Albar 18

19 Importance of Mass Accuracy on Unknown Screening m/z Number of Possible Candidates CHO CHON CHONS CHONSF Mass Accuracy (ppm) N. Cortés-Francisco, C. Flores, E. Moyano, J. Caixach. Anal. Bioanal. Chem (10):

20 Which HRAM Scan Mode is Right for Your Workflow? Precursor R=70,000 Leek extract spiked with 0.1 ppb imazalil Full Scan MS PRM/T-MS2 Product R=70, ppm Product Ion Matrix 20 M.M.Gomez-Ramos et al. J Chrom A, 1287, 2013, 24-37

21 Identification of Micropollutants and TPs by LC-HRMS Data Processing Thermo Scientific Hypersil GOLD aq column FS-ddMS2 (70K FWHM) Data Processing TraceFinder SW (Targeted Screening) Compound Discoverer SW Data Processing Sieve (Multivariate Data Analysis PCA) LC-HRMS Data presented at SETAC and ASMS

22 Workflow for suspect and non-targeted analysis Suspect Targeted Screening (TraceFinder SW 16:35 A Navigation through the Thermo Scientific Tracefinder Software Structure and Workflow Options; Frans Schoutsen, Thermo Fisher Scientific, Breda, Netherlands Search for Degradation Products (Compound Discoverer SW) Differential Analysis (Sieve SW) 22

23 Q Exactive MS: HRAM Target Screening & Quantitation of Pesticides GB National Food Standard method was created in China to specifically analyze 450 pesticides in fruits and vegetables using LC-MS/MS To specifically focus on this issue we have created a turnkey (acquisition + processing) Thermo Scientific Q Exactive MS method that can Screen and Quantitate 500 pesticides PARTNERS: FSANZ Health Canada IFIC Foundation JIFSAN NCFPD USDA IICA FDA Advisory Council CFIA Food Communications Compass, Japan CFSA VAAS STEPRI 23

24 500+ Pesticides Covering All Major Groups 24

25 HRAM Library: Targeted Screening & Quantitation Full MS / dd-ms 2 Inclusion list activated: 500 pesticides Quantitation &Confirmation: Fragment ion matching MS/MS library matching Search by MS/MS spectrum Q Exactive Focus MS 25

26 Boscalid at 0.5ppb (Beet Extract) R^2>0.998, Quan peak, Fragment ion matching and Spectra Library Scoring 26

27 Trifloxystrobin at 0.1ppb (Beet Extract) R^2>0.99, Quan Peak, fragment ion matching and Spectra Library Scoring 27

28 Industry Leading HRAM MS/MS Spectral Library Comprehensive HRAM Library and compound database created on Thermo Scientific Q Exactive MS at R 140,000 Searchable in TraceFinder software Consists of: EFS (Pesticides, Mycotoxins, Veterinary Drugs, Environmental Contaminates, PFCs) Clin/Tox (Drugs of Abuse, Therapeutic Drugs, Poisons, Performance Enhancing Drugs) The new spectra library will include the following: 3 ramped 20, 30, 40 ev and 2 step collision 40 with 50% and 70 with 50% Contains RTs and RRTs using the same group of IS for both EFS + Clin/Tox EFS Compounds: EFS MS/MS spectra: (5~6 spectra/compound) 28

29 HRAM Ametryn Example - Different CEs Advantage HCD 20 HCD 30 HCD 40 HCD 50 HCD Stepped 40 HCD Stepped 70 29

30 Quantified and Confirmed Chromatogram Fragment matching Spectra matching Calibration curve Quantified and Confirmed with Built-in HRAM Spectra/Isotope/Fragments DB Imidacloprid at 0.64 ppb standard curve 30

31 mzcloud Novel highly curated MS n fragmentation library for next generation compound identification TM mzcloud is a unique approach to mass spectral fragmentation libraries Combining MS n, multiple fragmentation techniques, multiple energy levels, and a high level of curation Providing informative metadata 31

32 mzcloud - Constantly Growing! 2,469 compounds with 78,813 spectra- 32 spectra per compound Compounds currently available on-line and those in que 32

33 FOOD: Status of Legislation in Different Regions Japan European Union Well developed methods and standards Well developed infrastructure Food safety relatively under control Well developed methods and standards Well developed infrastructure Food safety relatively under control No prescribed methods: must meet LOD s, LOQs China USA Developing methods Developing infrastructure Food safety control being developed. Developed methods and standards Prescribed methods by FDA and ISO, but no requirement to follow them Lack of testing resources 33

34 Overview of Identification Criteria LC-MS/MS 34

35 Environmental: Summary of Global Regulations & Enforcement Country or Region Regulation Enforcement European Commission sets directive, member states have 8 years to implement Labs can use any method, but must prove validation. Customer use ISO/DIN/AFNOR etc. EU Directive 98;/83/EC - Drinking water Directive 86/278/EEC metals Directive 2013/39/EU: SURFACE waters and BIOTA The new Directive 2008/50/EC of the European Parliament and of the Council of 21 May 2008 on ambient air quality US EPA Office of Ground Water and Drinking Water: Maximum Contaminant Limits (MCLs ) for drinking water under Safe Water Drinking Act (SWDA) EPA Office of Research and Development: Methods and MCL s for compliance to the clean water act. EPA RCRA (formerly Office of Solid Waste): Solid Wastes Labs must use approved methods; EPA, ASTM, Standard Methods EPA Regional Offices: 10 regions in the US. NELAC: Separate body which is responsible for compliance in some states. Customers are most concerned about enforcement bodies! Enforcement bodies often don t know what the EPA has approved! Rely on you for information 35

36 Moving Towards a Safer Way of Monitoring the Global Food & Environmental Chain Standardization of Performance Criteria yes! Standardize global methods? Standardize global technologies? Sample prep Chromatography Detection Mass Spectrometry analyzer Reporting Your thoughts 36

37 Thank You! 37