1. For Forensic Toxicology use Only
Quantitative Screening of 46 Illicit Drugs in Urine using Ultrahigh Resolution and Accurate Mass Exactive System
For Forensic Toxicology use Only
Research Use Only

2. For Forensic Toxicology use Only
Presentation overview
Exactive system introduction
Principle of operation
Method overview and salient features
Urine analysis results
Conclusions
Appendix
What’s different about this Orbitrap compared to the one in the LTQ Orbitrap?
Orbitrap itself is very similar but its mounting and wiring have been made more compact.
How do you get faster scan speed? Why is it faster than the LTQ Orbitrap?
We operate orbitrap mass analyzer at higher voltage which results in higher frequencies. In parallel, we have reduced overheads on ion transfer (as there is no LTQ) and thus we need less time for a spectrum and less time for inter-spectrum delay.
What’s the ultimate resolution?
100,000 at m/z 200 and 1 sec acquisition.
What’s the dynamic range
At least similar to that in LTQ Orbitrap, Full MS mode.
Pumping system?
There is a novel split-flow pump which allows to keep vacuum chamber smaller and make it from aluminium. Final vacuum is similar to that in a standard LTQ Orbitrap.
C Trap is this the same as the current LTQ Orbitrap
Yes, C-trap and ion optics are the same though their mounting is different (from a flange)
HCD Scan – how does this work and – what’s new?
Ions get accelerated through the C-trap into a quadrupolar collision cell where they fragment and get stored. After that, voltages are ramped and ions are transferred back to the C-trap from which they are injected into the orbitrap. There is no mass selection so only a narrow mass range of ions fragment with optimal sequence coverage, with lower m/z fragmenting excessively and higher m/z fragmenting only weakly.
Is the mass accuracy the same at the LTQ Orbitrap?  (based on work in Bremen, it was better than the LTQ Orbitrap)
Actually, we have a better layout and components of central electrode power supply so we expect mass accuracy at least as good as on LTQ Orbitrap XL.
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3. For Forensic Toxicology use Only
Exactive Benchtop LC-MS
Resolution
100,000 at 1 scan per second
10,000 at 10 scans per second
Mass accuracy
Sub ppm
Sensitivity
500 fg Buspirone with S/N >10:1
Dynamic range
>10,000 within a spectrum
Scan speed
Up to 10 scans per second
Mass range
m/z
Polarity switching
One positive and one negative scan < 1 second (25K Resolution)
What’s different about this Orbitrap compared to the one in the LTQ Orbitrap?
Orbitrap itself is very similar but its mounting and wiring have been made more compact.
How do you get faster scan speed? Why is it faster than the LTQ Orbitrap?
We operate orbitrap mass analyzer at higher voltage which results in higher frequencies. In parallel, we have reduced overheads on ion transfer (as there is no LTQ) and thus we need less time for a spectrum and less time for inter-spectrum delay.
What’s the ultimate resolution?
100,000 at m/z 200 and 1 sec acquisition.
What’s the dynamic range
At least similar to that in LTQ Orbitrap, Full MS mode.
Pumping system?
There is a novel split-flow pump which allows to keep vacuum chamber smaller and make it from aluminium. Final vacuum is similar to that in a standard LTQ Orbitrap.
C Trap is this the same as the current LTQ Orbitrap
Yes, C-trap and ion optics are the same though their mounting is different (from a flange)
HCD Scan – how does this work and – what’s new?
Ions get accelerated through the C-trap into a quadrupolar collision cell where they fragment and get stored. After that, voltages are ramped and ions are transferred back to the C-trap from which they are injected into the orbitrap. There is no mass selection so only a narrow mass range of ions fragment with optimal sequence coverage, with lower m/z fragmenting excessively and higher m/z fragmenting only weakly.
Is the mass accuracy the same at the LTQ Orbitrap?  (based on work in Bremen, it was better than the LTQ Orbitrap)
Actually, we have a better layout and components of central electrode power supply so we expect mass accuracy at least as good as on LTQ Orbitrap XL.
For Forensic Toxicology use Only 3

4. Orbitrap – Principle of Operationz φ
Hyper-logarithmic potential distribution:
“ideal Kingdon trap” r
Characteristic frequencies:
Frequency of rotation ωφ
Frequency of radial oscillations ωr
Frequency of axial oscillations ωz
Makarov A. Anal. Chem. 2000, 72,
For Forensic Toxicology use Only

5. Method overview and salient features
Negative urine sample
Dilute with 25% ACN
Spike 46 drugs at different concentrations
Spike isotopically labeled int standards
at 50 ng/mL
Inject 10 uL
Hypersil GOLD™ PFP
2.1 x 100 mm, 3 µm
Operated at res. 50,000 w/ Ext. Calib.
Mass accuracy better than 5 ppm
Note: LC/MS method details in appendix
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6. For Forensic Toxicology use Only
Results - ± 5 ppm at LOQs in urine B C D E F G H I J K L M N O P Q R S T U V W A
Note: Drug names and LOQs on slide 9 & 10; XIC – Extracted Ion Chromatogram
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7. For Forensic Toxicology use Only
Results - 5ppm at LOQs in urine. AA BB CC DD EE FF GG HH II JJ KK LL MM NN OO PP QQ RR SS TT UU VV WW
Note: Drug names and LOQs on slide 9 & 10; XIC – Extracted Ion Chromatogram
For Forensic Toxicology use Only

8. For Forensic Toxicology use Only
List of drugs and mass LOQ
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9. For Forensic Toxicology use Only
List of drugs monitored and mass LOQ
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10. For Forensic Toxicology use Only
Need for 50,000 resolution
Nicotine mass spectra at experimental (Exp) and different simulated (Sim) resolutions (FWHM)
Exp: 50,000
Sim: 50,000
Sim: 20,000
Sim: 10,000
Note: Experimental Nicotine at RT 1.40 min; exact mass =
For Forensic Toxicology use Only

11. For Forensic Toxicology use Only
Conclusions
The method is capable of easy confirmatory and quantitative analysis of 46 illicit drugs at LOQs of 0.5 – 5 ng/mL in urine.
Ultrahigh mass resolution of 50,000 FWHM provided the necessary selectivity to identify 46 drugs in urine in 9 minutes with minimal sample preparation
Analyte confirmation is achieved by utilizing better than 5 ppm mass accuracy with external mass calibration.
Good quantitative accuracy achieved by isotopically-labeled standard.
Nicotine example demonstrates the need for ≥50,000 resolution in addition to exact masses.
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12. For Forensic Toxicology use Only
Appendix
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13. For Forensic Toxicology use Only
LC/MS Method
HPLC system
Accela UHPLC™ Pump, PAL autosampler (CTC Analytics)
LC Column
2.1 x 100 mm, 3 µm Hypersil GOLD™ PFP
Injection Volume = 10 µL
Mobile Phase
(A) 5 mM NH4OOCH + 0.1% HCOOH, (B) ACN + 0.1% HCOOH
Flow Rate
0.35 mL/min
MS system
Exactive™ (Thermo Scientific)
Heated ESI; Polarity: Positive; Scan Range: m/z 110 – 800
Resolution: 50,000 FWHM;
External mass calibration
For Forensic Toxicology use Only