1. For Clinical Research and Forensic Toxicology Use Only
Exactive LC- MS for Quantitation in Clinical and Forensic Toxicology Research
For Clinical Research and Forensic Toxicology Use Only

2. For Clinical Research and Forensic Toxicology Use Only
Overview
Exactive As a Quantitative Analysis Tool
Clinical and Forensic Toxicology Research Methods
Drugs Supported in the Methods
Quantitation Workflow
What You Don’t Have To Do
Why it is Easy To Use
User Testimonials
Proof Statements and Support
Summary
Appendix
For Clinical Research and Forensic Toxicology Use Only

3. Why Consider Exactive For Quantitation
LOQ, Precision and Dynamic Range ~ Mid to High End QQQ
QQQ Assays Plus More
Easiest to Use
Large Panel of Drugs in
Single Assay
Ideal for Analytes
with
Poor Fragmentation
Sample Prep Requirement
Same As QQQ
Good Screening Capabilities
Unbeatable Price/Performance
For Clinical Research and Forensic Toxicology Use Only

4. Why Consider Exactive For Quantitation
Methods
Clinical Research
Forensic Tox Research
Immunosuppressant
Drugs in
Whole Blood
ETG/ETS
in Urine
Opioids
Benzodiazepines
in Urine
Vitamin D3
in Plasma
Opiates
in Urine
10 Steroid Panel
in Plasma
Drugs of Abuse Screening
For Clinical Research and Forensic Toxicology Use Only

5. Why Consider Exactive For Quantitation
Clinical Research
Forensic Research
Immunosuppressant Drugs
Serolimus
Tacrolimus
Everolimus
Cyclosporin A
ETG/ETS
Benzodiazepines
Lorazepam Diazepam
Nordiazepam Temazepam
Hydroxytriazolam Aminoclonazepam
Aminotriazepam Hydroxyalprazolam
Aminoflunitrazepam Desalkylflurazepam
Hydroxyethylflurazepam Oxazepam
Opioids
Norketamine
Ketamine
Nalbuphine
Norfentanyl
Fentanyl
Butorphanol
Alfentanil
Sufentanil
Vitamin D3
25 OH Vit D3
25 OH Vit D2
10 Steroid Panel
Aldosterone
Androstenedione
Corticosterone
Cortisol
Deoxycortisol
Dehydroepiandrosterone sulphate
Progesterone
17 alpha-hydroxyprogesterone
Testosterone
Opiates
Morphine, Hydromorphone, Oxymorphone
Codeine, Dihydrocodeine, Hydrocodone
Oxycodone, Meperidine, Normeperidine
6-Acetylmorphine
Drugs of Abuse Screening
For Clinical Research and Forensic Toxicology Use Only

6. Exactive Quantitation Workflow
What You Do
Sample Preparation
Urine Dilute n Shoot or
Turbo Flow
LLE
SPE
Data Acquisition LC - Exactive
Full Scan K resolution using HESI or APCI source
Mass Range m/z
Data Processing
Extract data for compounds and internal standards using better than 2ppm mass accuracy As
Easy It
Can
Get
Quantitate
Quantitate using peak area of extracted ion chromatograms for samples and calibrators
There is no more universal detection method for drugs-of-abuse or toxicants than electrospray LC-MS involving the simultaneous detection of positively and negatively charged ions. This is exactly the type of method used by ToxSpec where all ion source and analyzer parameters have been fully optimized for simultaneous positive and negative ion detection by mass spectrometry, for molecular weight determination, followed by data-dependent tandem mass spectrometry for fragment ion library identification. This methodology is summarized graphically in this slide. Each drug and toxicant in the ToxSpec library has it’s unique molecular ion and fragment ion spectrum; during the LC-MS run, ions are automatically chosen for tandem mass analysis based upon their relative intensities and molecular weights. The orthogonal metrics of relative retention time, molecular weight and unique ion fragmentation spectral fingerprint are more than enough to positively identify compounds of interest.
A +ve MS scan is performed, the m/z of ions detected over the background are compared against the expected m/z and Rt of the drugs in the method and tentative match recorded. MS2 scan is performed on up to 5 most intense ions and data recorded. A -ve MS scan is performed, the m/z of ions detected over the background are compared against the expected m/z and Rt of the drugs in the method and tentative match recorded. MS2 scan is performed on up to 2 most intense ions and data recorded.
Report Results
What You Don’t Have To Do (next slide)
For Clinical Research and Forensic Toxicology Use Only

7. Exactive Quantitation Workflow
What You Don’t Have To Do
Optimize SRM’s
Infusion or FIA optimization
Painstaking process of product ion
selection for best sensitivity and specificity
Collision gas optimization
Optimization of ion path lens voltages
Optimization of Timed SRM
For Clinical Research and Forensic Toxicology Use Only

8. Exactive Quantitation Workflow
Why it is Ease to Use
Easiest to operate
No SRM optimization
Quantitate large panels without
additional optimization considerations
Easiest to expand assay
Easiest to interpret data
Easiest to maintain
For Clinical Research and Forensic Toxicology Use Only

9. What Do Users Say
“The beauty of the Exactive MS is that it allows us to look for unknown metabolites at the same time that we quantitate known metabolites.  In our laboratory, we have been able to identify four different fentanyl metabolites with no need for UV detection, fraction collection with NMR, or MS/MS fragmentation patterns”
Uwe Christians, MD, PhD, Professor, Department of Anesthesiology, University of Colorado and iSiS Clinical Research & Development Lab Director
“We historically used triple quadrupole mass spectrometers but were very impressed with the Exactive demo. After acquiring the system, we had to do little to no method development. It’s flexibility in analyzing both dilute-and-shoot and SPE prepared samples and the ease of addition of new compounds to our drug panels helped us keep things simple. Overall, it turned out to be a wise choice for our Drugs of Abuse and Pain Management drug analyses”
Kent Johnson, Director
Fortes Lab, Portland Oregon, USA
For Clinical Research and Forensic Toxicology Use Only

10. Proof Statements and Support
Performance and Method Details (PPT files for each method)
Library of Compounds for Screening in ToxID
Assay Installation and Training
Customer Reference Sites
For Clinical Research and Forensic Toxicology Use Only

11. For Clinical Research and Forensic Toxicology Use Only
Summary
All Things Considered, Ultrahigh Resolution Exactive Bench Top LC/MS System Is a Very Efficient and Sensitive Alternative Quantitation Tool for Use in Clinical and Forensic Research Including Pain Management.
For Clinical Research and Forensic Toxicology Use Only

12. For Clinical Research and Forensic Toxicology Use Only
Appendix
For Clinical Research and Forensic Toxicology Use Only

13. Exactive Quantitation Workflow
Why is it easy to use
Easiest to operate
Full scan MS operation
No SRM optimization
Monitor m/z of interest at ultrahigh resolution and mass accuracies
Monitor large panels without additional optimization considerations
Just extract m/z of interest in any full scan MS acquisition
Easiest to expand assay
Full scan acquisition means all ions all the time
Easiest to interpret data
Just need to know the m/z of the compound of interest
Easiest to maintain
Less complex ion path, no collision cell and the necessary gas.
For Clinical Research and Forensic Toxicology Use Only