Chemical Analysis Business Operations Waters Corporation Milford MA

Transcription

1 The Detection and Identification of Unknown Contaminants During ToF Screening and Structural Elucidation for Pesticides in River Water Using an Integrated Software Approach Chemical Analysis Business Operations Waters Corporation Milford MA

2 Streamlined Screening Workflow Prepare samples Generic sample extraction Pre concentrate and clean-up Acquire comprehensive sample data High resolution chromatography and MS detection Simultaneous molecular ion and fragment ion data Locate and identify known components Maximise MS information Determine significance with quantification 4 Identify unknown peaks of interest Deconvolution of molecular ion and fragment ion data Comparing against known good sample or controls 5 Elucidate structures of unknowns Elemental composition determinations, online search Software based mass spectral interpretation 211 Waters Corporation COMPANY CONFIDENTIAL 2

3 Screening Enabling Technologies Oasis HLB SPE Efficient water extraction Xevo G2 QTof QuanTOF technology 2, resolution MS E enabled POSI±IVE workflow software Targeted screening using compound database ChromaLynx XS, MassFragment Locate and identify non-targeted unknown components

4 Rapid and Comprehensive data Rapid and Comprehensive data acquisition

5 Speed AND Resolution MS E with rapid chromatography Fastnet_431_water68 F Da e4 28 datapoints across spiroxamine peak Fastnet_431_water68 Fragments = 14 data points F Da e4 MS E = All the masses in ONE injection m/z = M =. ppm Fastnet_431_water F Da 9.4e4 Time Hundreds of Pesticides, <85 seconds m/z = M = 1.3 ppm Time Precursors = 14 data points

6 Speed AND Resolution MS E with rapid chromatography Fastnet_431_water68 F Da e4 Fastnet_131_water7 1 1: TOF MS ES Da 1.2e5 Ultra fast for simple matrices Medium speed for normal matrices Time Fastnet_631_water_15 1 1: TOF MS ES Da Time e4 Maximum resolution for the most complex matrices 3.42 Time

7 Comprehensive Data Acquisition - UPLC/MS E Simple, patented method for unbiased data acquisition Catalogs complex samples in a single analysis Comprehensive record of precursor and fragment ion data

8 Comprehensive Fragment ion High Speeds LE LE m1 sec EE D. EE MS E is a UNBIASED process All ions all the time Bateman et al, Anal Chem (22) Silva et al, MCP (25) *Patented technology EE EE 211 Waters Corporation COMPANY CONFIDENTIAL 8

9 Matrix Tolerance

10 In Spectrum Dynamic range - Matrix Tolerance 1 XIC m/z TIC m/z Time Malaoxon in sewage effluent,.2 µg/l Theoretical mass = m/z

11 Stability and Selectivity

12 Mass Stability Essential for High Quality Data Thiabendazole 2 ppb Scan No. Measured Mass M (mda) M (ppm) RMS = Excellent mass accuracy across peak m/z m/z 22.5

13 Targeted Screening Narrow XIC Windows mda 1 XIC fragment m/z mda mda mda Mass accuracy + stability ypermits use of narrow XICs XIC precursor m/z TIC mda T 1.3 mda.9 Time Fenuron m/z Thiabendazole High selectivity for precursors and fragments with MS E

14 Reducing False Positives Narrow XIC Windows Monuron m/z Fenazox m/z Cymoxamil m/z mda Mass extraction window 3. mda

15 S l ti it i i hi h l ti Selectivity gains using high resolution fragment ions

16 Comprehensive Data Acquisition - Resolve Isobaric Precursors with MS E 1 Desmetryn m/z Desmetryn Simetryn m/z Desmetryn m/z Simetryn m/z Time Simetryn or Desmetryn? m/z m/z Selective MS E fragment in standards

17 Screening Against a Target List of Screening Against a Target List of Compounds

18 Experimental 2 ml UK River Water Oaisis HLB Cartridge (2x concentration) ACQUITY UPLC System Runtime: 5 min Flow Rate:.6 ml/min Injection: 3. L Column: ACQUITY BEH C 18, 1.7 m, 2.1 x 5 mm (45 o C) Mobile Phase: A) Ammonium Acetate in Water. B) Ammonium Acetate in Methanol Time (min) Flow rate (ml/min) A B Curve Initial Waters Corporation COMPANY CONFIDENTIAL 18

19 Experimental Continued MS system: Xevo G2 QTof (Figure 1) Ionization mode: ESI positive Analyser: Resolution mode (>2K resolution) Scan time: 1.1 s Capillary voltage: 1. kv Sampling cone: 3. V Source temp: 12 o C Desolvation temp: 55 o C Mass range: m/z 5 1 MS E Low energy: 6. V MS E High energy: V LockSpray solution: Leucine enkephalin LockSpray masses: m/z and m/z Waters Corporation COMPANY CONFIDENTIAL 19

20 Screening against a Target List Non-selective sample extraction Non-Targeted (Unbiased) Data Acquisition Acquire single comprehensive dataset with independent precursor and fragment information Generate list of prospective compounds Quantify Targeted Data Analysis Compound databases nw-xic -RT - Mass error - Isotope patterns - Fragment ion data Quantitation of positives Benefits Utilise U-HPLC speed Precursors & Fragments in single injection Good selectivity it from UPLC & MS resolution, mass accuracy & stability, use of fragment ions Reduced false positives Required sensitivity for regulatory use & discovery of unknowns Works with complex matrices Screen & Quantify

21 Targeted Screening River Water MS E fragment ions 59 compounds targeted, 5 detected within tolerances MS E molecular ions Overview of all detected compounds in all samples

22 River Water Screening Codeine Detection M: +.4 mda M: mda M: +.1 mda M: -.1 mda MassFragment

23 Discovery & Identification of Discovery & Identification of Unknown Residues

24 Unknown Discovery and Identification Non-selective sample extraction Non-Targeted (Unbiased) Data Acquisition Acquire single comprehensive dataset with independent precursor and fragment information Non-Targeted Data Analysis Application Managers o Identify using spectral libraries o Compare o Detect t and identify transformation products Generate component list S List of detected known compounds Spectral deconvolution Apply application managers o Detect and identify markers of sample differentiation Elucidate structure of unknowns DISCOVERY o Apply structural elucidation tools o Confirm identity

25 Example Workflow Acquire single comprehensive dataset with independent precursor and fragment information DISCOVERY Spectral deconvolution Compare samples Use Acc Mass and isotope profile filters Non-Targeted Data Analysis Component List Detect Unknown Propose Elemental Composition 1 2 Search database for candidates Provide structure and fragment ion spectra to Massfragment Propose Candidate Structure Test Candidate Structure Confirm Candidate (Standard + MS/MS)

26 Non-Targeted Components Deconvoluting river water sample 1 deconvoluted non-targeted components Codeine detected in non-target mode

27 Non-Targeted Components Comparison with control sample 1 Unique components in each sample Unusual Chlorinated component 3 rd highest abundance

28 Non-Targeted Component Elucidating structure of unknown Unknown component RT 2.28min, m/z Elecomp and i-fit proposed C 6 H 13 O 4 PCl 3 as candidate

29 Non-Targeted Component Elucidating structure of unknown C 6 H 12 O 4 PCl 3 entered into Chemspider search ([M+H] + ) ChemSpider proposed a number of structures Tris(2-chloroethyl) Phosphate is a commonly used flame retardant and the top hit

30 Non-Targeted Component MassFragment and MS E MS E fragments m/z M = -.3 mda m/z M = -1.1 mda 4 Precursor m/z M = -.1 mda Unknown component in river water

31 Non-Targeted Component Isotope Model Comparison Isotope model based on proposed formula Original acquired mass spectrum ChemSpider result 211 Waters Corporation COMPANY CONFIDENTIAL 31

32 Non-Targeted Components Comparison with control sample 1 Another unusual compound not detected in the control

33 Non-Targeted Component Emerging g contaminants in river water Time Time Unknown component RT min, m/z Elecomp and i-fit ChemSpider proposed proposed C 15 H 13 N 2 O as Carbamazepine candidate 2

34 Non-Targeted Component MassFragment and MS E MS E fragments m/z M = -.3 Da MS/MS spectrum Precursor 4 1 m/z m/z M = +.3 Da m/z m/z Unknown component in river water Carbamazepine standard

35 Conclusions Compounds as diverse as the anticonvulsant mood stabilizing drug carbamazepine, the analgesic compound codeine and the flame retardant tris(2-chloroethyl) phosphate were discovered in the river water blank using this non-targeted structural elucidation workflow approach. The MS E functionality enables the acquisition of both low energy (precursor ion) and high energy (fragment ion) data in a single rapid screening run. MS E fragment ion data can be used to help with unequivocal identification of detected compounds. The compare functionality, elemental composition, and mass fragment strategies in the workflow provide powerful tools for screening, detection, and identification of true nontargeted species in environmental matrices. 211 Waters Corporation COMPANY CONFIDENTIAL 35

36 Many Thanks Riches E, 1 Morphet J, 1 Major H, 1 Keenan G, 2 Giela A, 2 Rosnack K, 3 1 Waters Corporation, Atlas Park, Simonsway, Manchester M22 5PP; eleanor_riches@waters.com 2 SASA, Roddinglaw Road, Edinburgh, Scotland EH12 9FJ; george.keenan@sasa.gsi.gov.uk 3 Waters Corporation, 34 Maple Street, Milford, MA, 1757; ken_rosnack@waters.com 211 Waters Corporation COMPANY CONFIDENTIAL 36

37 QUESTIONS? Chemical Analysis Business Operations Waters Corporation Milford MA