Redefining GC-MS in the Laboratory with GC Orbitrap Michal Godula, - - PowerPoint PPT Presentation

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Redefining GC-MS in the Laboratory with GC Orbitrap Michal Godula, - - PowerPoint PPT Presentation

Jan 26, 2024 293 likes •671 views

Redefining GC-MS in the Laboratory with GC Orbitrap Michal Godula, Ph.D. Thermo Fisher Scientific michal.godula@thermofisher.com The world leader in serving science A New Chapter in GC-MS Thermo Scientific TM Q Exactive TM GC Hybrid

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The world leader in serving science

Michal Godula, Ph.D. Thermo Fisher Scientific michal.godula@thermofisher.com

Redefining GC-MS in the Laboratory with GC Orbitrap

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A New Chapter in GC-MS

Thermo ScientificTM Q ExactiveTM GC Hybrid Quadrupole-OrbitrapTM GC-MS/MS System

Offering Unprecedented Depth in GC-MS Analysis

  • Hugely powerful HR/AM performance for GC-MS
  • Uncompromised quantitative and qualitative capability
  • Reliable, robust and easy to operate
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Orbitrap Principles

Makarov A. Anal. Chem. 2000, 72, 1156-1162.

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Bringing GC and Orbitrap Technology Together

C-TRAP HCD Cell AQT Quadrupole Orbitrap Mass Analyzer Bent Flatapole ExtractaBrite™ Ion Source

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Breakthrough in GC-MS Performance

Unrivalled Resolving Power

  • Up to 120k @ m/z 200
  • Allows excellent selectivity in

complex matrix, maintaining confidence in detection and identification

  • Drives highly resolved data sets and

high capacity peak detection

  • Fast scanning Orbitrap for well

defined, reproducible GC peaks

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Breakthrough in GC-MS Performance

Chlorprophram

  • 10 ng/g

QuEChERS extract

  • f leek
  • Full scan with

resolving power of >30k (FWHM @ m/z 200) provides interference free detection

  • Excellent mass

accuracy for confident identification

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Pesticides screening

False negatives at 15,000 RP

Secure screening

  • 5 ppm identification

threshold

  • Higher resolving power

allows confident detection and identification

  • Mid- resolution of 15k

causes false negatives

  • Q Exactive GC

routinely operates at 60k (FWHM @ m/z 200)

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Breakthrough in GC-MS Performance

XIC of dieldrin in baby food (m/z 262.85642) 100ppb XIC of dieldrin in baby food (m/z 262.85642) 100ppb 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17

60,000 RP (m/z 200) 2.4 s peak

Fast acquisitions

  • Important for

accurate profiling of narrow GC peaks

  • Full scan with

resolving power of 60 (FWHM @ m/z 200) generates 17 scans

  • Fast enough for

GC!

17 scans

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Scan to-scan Scan to-scan XIC of dieldrin in baby food (m/z 262.85642) 100ppb XIC of dieldrin in baby food (m/z 262.85642) 100ppb

60,000 RP (m/z 200) 2.4 s peak

Breakthrough in GC-MS Performance

< 1ppm (internal) < 3ppm (external)

  • Scan-to-scan
  • Low level in matrix
  • Over full mass

range

  • Over full

concentration range

  • No need for

continuously calibrating in sequence

0.7 0.7 0.6 0.5 0.3 0.6 0.5 0.3 0.3 0.3 0.5 0.6 0.6 0.7 0.5

0.5 ppm RMS

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Breakthrough in GC-MS Performance

Mass accuracy over >6

  • rders
  • Hexachlorobenzene
  • 0.2 – 250,000 ng/mL
  • Mass accuracy within 1 ppm
  • Orbitrap spectral fidelity

independent of concentration

Over full concentration range Over full concentration range

Data courtesy of Hans Mol and Marc Tienstra, RIKILT

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Orbitrap GC-MS Family

Q Exactive GC system

Unprecedented Depth in Analysis RP 120,000 (FWHM @ m/z 200) EI/CI; Full-scan, Timed-SIM MS/MS capability

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Redefining Routine GC-MS RP 60,000 (FWHM @ m/z 200) EI/CI; Full-scan; Timed-SIM

Thermo ScientificTM ExactiveTM GC OrbitrapTM GC-MS System

Orbitrap GC-MS Family

Thermo ScientificTM Q ExactiveTM GC OrbitrapTM GC-MS/MS System

Unprecedented Depth in Analysis RP 120,000 (FWHM @ m/z 200) EI/CI; Full-scan, Timed-SIM MS/MS capability

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Insight into the Orbitrap GC-MS Systems

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Insight into the Orbitrap GC-MS Systems

Thermo FisherTM TRACETM 1310 GC system Unique modular injector and detector design Short cycle time

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Insight into the Orbitrap GC-MS Systems

Thermo Scientific™ ExtractaBrite™ ion source technology Routine grade robustness Patented RF lens Removable without breaking vacuum through VPI Vacuum-free column replacement through VPI TRACE 1310 GC system Unique modular injector and detector design Short cycle time

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Insight into the Orbitrap GC-MS Systems

Thermo Scientific™ ExtractaBrite™ ion source technology Routine grade robustness Patented RF lens Removable without breaking vacuum through VPI Vacuum-free column replacement through VPI Orbitrap mass analyzer Incredible HRAM performance Extended Dynamic Range TRACE 1310 GC system Unique modular injector and detector design Short cycle time

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Insight into the Orbitrap GC-MS Systems

Sensitivity Mass Accuracy Resolving Power

  • Maximum

selectivity

  • Fast enough for

GC

  • In full-scan
  • High selectivity
  • High spectral

fidelity

  • Excellent

coverage in sample profiling

  • “Triple quad

grade” quantitation in full-scan Dynamic Range

  • Every scan
  • All concentrations
  • In complex matrix
  • Across the mass

range

  • Everyday!

Up to 120,000 at m/z 200 < 1ppm ppt >6 orders

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Analytical Workflows Overview

Non- targeted full scan acquisition

Targeted quantitation

“the things you expect”

Targeted screening

“the things you may suspect”

Non-targeted screening

“the things you don’t expect”

i.e. Commonly used pesticides for a particular commodity i.e. Unusual, banned or alternate pesticides for particular commodity i.e. Other contaminants or compounds, could be known or unknown

Known unknown ID Unknown unknown ID

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  • 4 O-ring samples
  • A Red
  • B Brown
  • C White
  • D Black
  • Blank (control)

Pharmaceutical O-Rings Extractables Study

  • Solvents
  • Water
  • 5M NaCl
  • 50% Ethanol
  • 100% Ethanol
  • 1% PS-80
  • 0.5N NaOH
  • 0.1M Phosphoric Acid
  • 40 °C for 30 days
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Q Exactive GC MS Parameters Transfer line (°C): 280 Ionization type: EI Ion source(°C): 230 Electron energy (eV): 70 Acquisition mode: Full scan Mass range (Da): 50-650 Mass resolution (FWHM): 60k Lockmass (m/z): 207.03235

GC-MS Instrument Conditions

TRACE 1310 GC Parameters Injection Volume (mL): 1 Liner Single gooseneck Inlet (°C): 280 Carrier Gas, (mL/min): He, 1.2 Oven Temperature Program: Temperature 1 (oC): 40 Hold Time (min): 1 Temperature 2 (oC): 320 Rate (oC/min) 15 Hold Time (min): 10

Thermo Scientific™ Q Exactive™ GC Hybrid Quadrupole-Orbitrap GC-MS/MS Thermo Scientific™ Trace™ 1310 GC Thermo Scientific™ TraceFinder™

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TraceFinder Non-targeted Screening Overview

  • Sensitive and

selective peak detection

  • High resolution

spectral deconvolution

  • Clean spectrum
  • 1. Detect and refine
  • 2. Generate candidates
  • 3. Filter and identify
  • Search spectra against

spectral libraries

  • HRAM o low resolution

spectral libraries (NIST, Wiley…)

  • Candidates list

generated

  • High resolution

filtering of candidates

  • Putative

identifications made Automatic compound identification

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Example Results

Blank control Brown O-Ring

RT: 4.10 - 27.88 SM: 7B 6 8 10 12 14 16 18 20 22 24 26 Time (min) 10 20 30 40 50 60 70 80 90 100 10 20 30 40 50 60 70 80 90 100 Relative Abundance 5.66 4.78 21.86 22.45 21.02 23.25 24.83 20.64 26.44 20.03 19.59 7.65 19.16 18.56 6.66 8.84 17.88 10.33 11.65 16.62 12.80 13.82 17.48 5.67 24.62 21.86 24.39 20.80 20.37 25.02 26.04 5.10 15.42 5.70 11.10 19.19 NL: 6.86E8 TIC MS 03June15_0 08 NL: 2.54E9 TIC MS 03june15_01 1

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Step 1: Deconvolution of the Data

  • TraceFinder first performs an accurate mass deconvolution of the data.
  • Extracts all of the peaks in the chromatogram.
  • Provides a cleaned spectrum for library matching.
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Step 2: Quickly Isolate the Peaks of Interest

  • 2051 peaks were extracted from the brown O-ring sample

Heat Map indicates the peaks elevated in the sample.

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Top 10 Differential Peaks in Brown O-ring

Peaks list can also be sorted by fold difference compared with control to isolate the differential peaks that could be low or high intensity.

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26 Hits from NIST are sorted based on:

  • Spectral matching
  • High Resolution Filtering

(HRF) score. Step 3: Identify the Compound – Searching NIST 14

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Step 3: Identify the Compound – Searching NIST 14

Combined SI and HRF values give an overall score (%) to quickly and confidently identify the compound. Eliminates other hits that would be valid if only SI used.

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Step 4: Fragments Can Be Explained With < 1ppm Mass Accuracy

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NEW: VeV Electron Ionization Technology

  • Top: spectrum acquired with

classic 70eV

  • Bottom: spectrum acuquired with

low 12eV, showing enhancement

  • f molecular ion

The key benefits of VeV are: Fully automated for optimum performance: VeV set-up is very simple and easy with fast, fully automated tuning. Increased confidence in identification: VeV promotes molecular ion and diagnostic high mass signals, important information for compound identification and confirmation.

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NEW: HRAM Library for Metabolomics Database

  • High resolution, high mass accuracy

spectra acquired at 60,000 RP (m/z 200)

  • Spectra refined and curated with

elemental composition of each EI fragment verified

  • Kovats retention index for each entry
  • PubChem ID for unique metabolites
  • Can be used in combination with existing

unit mass libraries

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Q Exactive GC: Compound Discovery and Identification

Remove entire ion source or change to CI source in under 2 minutes without venting …

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Q Exactive GC: Compound Discovery and Identification

Fragmentation in HCD Cell

  • Thermo ScientificTM Mass FrontierTM software can be used for sub-structural interpretation
  • Sub ppm accurate mass allows for higher confidence in fragment IDs
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EI & PCI Spectra for Peak at 15.17 mins.

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Number of Proposed Formulae for m/z 324.13541

5 10 15 20 25 30 35 1

1 ppm 3 ppm 5 ppm 20 ppm 10 ppm

  • No. proposed formulae

4 1 6 12 25

Top hit = C20H20O4

Elements used: C (1-30), H (1-60, N (1-5), O (1-5), P (1-2), S (1-2)

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MS/MS m/z 325.14 to Support Proposed Formula

_ T: FTMS + p CI Full ms2 325.14@hcd10.00 [50.00-350.00] 60 80 100 120 140 160 180 200 220 240 260 280 300 m/z 5 10 15 20 25 30 35 40 45 50 55 60 65 70 75 80 85 90 95 100 Relative Abundance 223.07545 C 15 H11 O 2 = 223.07536 0.42975 ppm 251.10672 C 17 H15 O 2 = 251.10666 0.26844 ppm 195.08049 C 14 H11 O = 195.08044 0.24907 ppm 105.03358 C 7 H5 O = 105.03349 0.83676 ppm 145.02854 C 9 H5 O 2 = 145.02841 0.92080 ppm 279.10166 C 18 H15 O 3 = 279.10157 0.30906 ppm 117.07012 77.03894 161.90226

>13 fragments can be explained based on C20H20O4

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Conclusion

  • Thermo Scientific Q Exactive and Exactive GC systems are easy-to-use,

dedicated HRAM GC-MS platforms that provides highly sensitive, routine grade performance in both targeted and untargeted screening experiments

  • Thanks to the superior resolving power, accurate mass measurements and

extended linear dynamic range, the Orbtrap GC-MS systems provide a greater confidence in the discovery, identification and quantification of compounds for the ultimate sample analysis workflow

Find out more about our GC Orbitrap GC-MS and GC-MS/MS http://info1.thermoscientific.com/OrbitrapGCMS

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Thank you for your attention!