1. How Can Orbitrap Technology Help My Food Safety Analysis?
Larry Burchfield
Mass Spectrometry Product Manager
Latin America Region
August 27, 2012

2. EFS Target or Non-target analysis workflow
SAMPLE
Target Analysis
Non-Target Analysis
Target screening ?
Profiling
Fingerprinting
Authenticity
Quantitation
Quantitation
Food Safety
Environmental

3. Which LCMS Analyzer Do I Choose?
Pure Quantitative
Pure Qualitative
Determine structure
Detect & Quantify
Structural ID
Compound Confirmation
Reaction Monitoring
Process Monitoring
Metabolism
Proteomics
Metabolomics
Targeted screening
Targeted
& Unknowns screening
1 – >40 cpds
40 – 600 cpds
Multiple cpds
ORBITRAP
QTRAPs
Q-TOF
Triple Quads
Ion Traps
Exactive & Q-Exactive (Orbitrap)

4. Triple Quadrupoles for Target Quan : Traditional Workhorse and still 75% of EFS Instruments Sold in 2011
TSQ Quantum Access Max
TSQ Quantum Access
TSQ Quantum Ultra
TSQ Vantage
The following is what Thermo Scientific has to offer in the TSQ line. For the ultimate value we have the TSQ Quantum Access Max. For the utmost detection needed TSQ Vantage which was introduce in 2008.
Ultimate Value
Ultimate Performance

5. However, conventional thinking is evolving…
Food Safety: International Requirements and Innovative Analytical Solutions

6. EU Commission directive on HRAM
One monitored m/z at R=10K, gives 2 IPs
(but One QQQ SRM = 1.5 IP)
However, this R refers to Sector instruments
10% FWHM
ToFs/Orbis measure 50% FWHM
Thus, R = 10K (sector) corresponds to R= 50% FWHM (ToF/Orbi)
There`s still no specific clarity in EU directives about the use of HRAM (Orbi) for multi-residue work

7. EU - Q Exactive exceeds IP requirements (min = 4)
Score Max Possible 6.5 points   
EC Directive 96/23/EC for analytical methods and the interpretation of results

8. How does this translate to LCMS instrument selection in EFS labs today ?

9. Shift towards HRAM : Crossover Point
Official Food Control Authority of the Canton of Zurich

10. Shift towards HRAM : Crossover Point (cont.)

11. Quantitative Performance : Orbitrap vs TOF vs Triple Quad
Note: Orbi and TOF not MS/MS
Kaufmann et al, Rapid Comm Mass Spectrom, 2011, 25, 1-14

12. Presentation of Dr. Hans Mol

13. Visualizing the matrix…(TICs)
compound feed
honey
solvent
0.25 g matrix/ml extract
5 µl injection

14. Detection: selectivity
Pirimicarb
m/z
± 12 mDa
[0.012 Da/239*106]
What mass extraction window is
required for adequate selectivity
in single stage MS?
Pirimicarb 10 ng/g in horse feed
(R = 100,000 m/z 200)

15. Requirements for narrow mass extraction windows
1) high mass accuracy
intrinsic property of mass analyzer (after adequate mass calibration)
but: in real life analyte may co-elute with compounds with same
nominal mass. This may lead to wrong mass assignment
2) full separation of analyte from other compounds with same nominal mass
chromatographic resolution
mass spectrometric resolution

16. How much resolving power is required ?
XIC m/z ±5 ppm
mass profiles at rt = 3.69 min
Isopyrin, 25 ng/g in animal feed

17. Quantitative aspects: linearity
Solvent standards, R= 25,000; mass extr. window 5 ppm

18. Conclusions Detectability/selectivity: resolving power key parameter
needed for selectivity (narrow mass extraction windows)
higher degree of initial confirmation
more reliable quantification
minimum resolving power? => depends on rel. response analyte/matrix*
Identification: HCD (or IS-CID) fragmentation very useful for identification and
data generated suited for confirmation according to EU legislation
Quantitative aspects: suited for quantitative analysis
Single stage full scan hr/hma MS is a very promising option for comprehensive
screening, identification and determination of toxicants in food/feed
*M. Kellman, H. Muenster, P. Zomer, H. Mol,
J Am Soc Mass Spectrom. 20(8) (2009)

19. Q Exactive EFS Data
CDFA, Sacramento
US FDA, DC

20. Q Exactive for EFS Targeted Screening & GUS
High Confidence Confirmation Targeted screening and GUS
Goal
How Q Exactive addresses
the needs
Mode/s of
operation
Software
Data collected in HRAM MS and MS/MS
Software capable of identifying all
unique components in the
chromatographic data
Database searches for HRAM MS and
library searches with MS/MS spectra
Highest specificity with 140K resolution
Software that supports multiple
orthogonal ways of confirmation
including elemental composition
determination, isotope matching, DB
search, spectral library matching.
Xcalibur;
ExactFinder
Highest confidence in
Targeted screening
& GUS
Full Scan MS w AIF
Precursor ion selection w HCD MS/MS

21. Q Exactive Quan capabilities for EFS
Quantitation Performance Similar
to High End QqQ
Goal
How Q Exactive addresses
the needs
Mode/s of
operation
Software
Unit mass resolution precursor ion
selection like QqQ
Full scan HRAM MS/MS with stable
fragment ion ratio
Higher specificity than QqQ due to
HRAM fragmment ions
Ability to confirm with many fragment
ions seen in full scan MS/MS
Four to five orders of dynamic range
UHPLC compatible
Targeted MS/MS similar
to SRM; Data
dependent MS/MS
where quant done using
precursor ion in full scan
MS and confirmation
using HRAM fragment
ions in MS/MS; Full
scan MS; HRAM SIM
Quant capability of high-end QqQ
ExactFinder or
Xcalibur

22. Q Exactive TM Hardware Innovations
Hardware Demonstration

23. Q Exactive RSLC
< 1.5 m

24. California Dept of Food & Ag
Sample : 60+ pesticides in crop extracts
In neat
In Green Bell Peppers
In avocado
Calibration levels vary depending on analytes
500 ppb – 1 ppt
Mass tolerance set at 5ppm
Sample injection size = 5uL
Column = Hypersil Gold aQ 100 x 2.1mm 1.9u

25. Q Exactive Workflows for EFS :
Full Scan :
Screening (target and non-target)
Quan
Full scan Targeted Data Dependent MS/MS :
Full scan screening
Targeted precursor ion selection
DD MS/MS of precursor ions
UHPLC

26. Full Scan Targeted DD MS/MS
Quadrupole scans from a mass range of
Sends the ions to Ctrap, compress and send into orbitrap for analysis
From a targeted list (Data Dependent MS2
Quadrupole selects the ion of interest from list
Pass it to Ctrap
Then pass it to HCD for fragmentation
Then sends data packet back to Ctrap and injects to Orbitrap to analyze for MS2
Full Scan Data Dependent (MS2) (TopN) – Ions are passed from a selected mass range, then triggered of a targeted listed (precursor) to collect ms/ms data for confirmation – result is 2 scans (FS, MS2) – This type of experiment is Quan followed by Confirmation. You can also run this experiment with “No Targeted” and than instrument will trigger of most intense ion in the spectrum and give you MS2 scan (great for metabolism)

27. CDFA samples (targeted list)
Parent selection list of 62 pesticide analyzed by CDFA of Sacramento
Method transferred
from TSQ Ultra

28. Q Exactive Full Scan Quan (Azoxystrobin at 1.5ppb)
Direct comparison of neat vs matrix

29. Q Exactive Quantitation Full Scan Targeted DD MS/MS (Azoxystrobin at 1
Q Exactive Quantitation Full Scan Targeted DD MS/MS (Azoxystrobin at 1.5ppb)
Direct comparison of neat vs matrix
Quan IP = 2

30. Quanfirmation Boscalid 1.5ppb
HRAM IP = 2
Boscalid
Diuron
Azoxystrobin
Methomyl
Indoxacarb
Spinosad D
Number of scans across a peak doing MS/MS
Quantitation
Confirmation
Boscalid – MS/MS Accurate Mass Fragmentation
HR MS(n) IP = 2.0

31. Quanfirmation Azoxystrobin 1.5ppb
Quantitation
Number of scans across a peak doing MS/MS
Diuron
Azoxystrobin
Boscalid
Methomyl
Spinosad D
Indoxacarb
Azoxystrobin
MH+ =
Confirmation

32. Q Exactive Quan performance :
Repeat 3x injections at LOD/LOQ in Matrix (Green Bell Pepper)
0.1 – 1.1 ppb
data represented from FS/ Targeted DD MS2 QE

33. Comparison of CDFA pesticides in matrix : Q Exactive vs TSQ Vantage LOD/LOQ (ppb)

34. UHPLC compatibility : 60+ pesticides in 2 mins, 4s wide peaks, Full scan
Accela Q Exactive
R = 70,000
All compound elute in 2mins, with peak width of 0.04mins = 9scans under the peak

35. Target Screen Method
Select database, Identify with mz and confirm with isotope, library match

36. Unknown Screen Method
Select database, Identify with mz and confirm with isotope, library match

37. Library/Database Searching Capabilities- Spectral Library Included
List selected based on customer feedback and from the major worldwide regulated lists
Over 1000 compounds
Over 6000 spectra
HCD
CID
Positive
Negative
Multiple CE
Pesticides, hormones, POPs, PPCP
Created on LTQ Orbitrap Velos
HRAM library courtesy : Eric Genin, Les Ulis

38. Q Exactive Quanfirmation
High performance HRAM Quantitation and Confirmation bench top LCMS system, capable of :
Multi-residue quan performance similar to mid-high end Triples
Ideal for targeted and general unknown screening
Highest confidence confirmation with R = 140K, and MS/MS
UHPLC compatible

39. History and Advances in Orbitrap Technology
Salón EBRO 11:00 -12:00 Tuesday, 28 August
History and Advances in Orbitrap Technology
Larry A. Burchfield
Latin America Manager
for
Chromatography and
Mass Spectrometry

40. The End
Thank You!
See y’all tomorrow…