Evaluation accutof dart for postmortem toxicology screening peter r stout
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Evaluation AccuTOF DART for Postmortem Toxicology Screening Peter R. Stout. RTI International is a trade name of Research Triangle Institute. NIJ Project. Grant No. 2006-DN-BX-K014 Opinions are those of the authors and not necessarily the U.S. Department of Justice. Objective.

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Evaluation accutof dart for postmortem toxicology screening peter r stout l.jpg

Evaluation AccuTOFDART for Postmortem Toxicology ScreeningPeter R. Stout

RTI International is a trade name of Research Triangle Institute


Nij project l.jpg
NIJ Project

  • Grant No. 2006-DN-BX-K014

  • Opinions are those of the authors and not necessarily the U.S. Department of Justice


Objective l.jpg
Objective

  • Evaluate the Jeol Ltd. AccuTOF DART system as a novel application for use in postmortem toxicology laboratories

    • Qualitative

    • Screening methodology

  • Specifically postmortem

    • Urine

    • Blood and tissue


Presentation goals l.jpg
Presentation goals

  • Equipment set up

  • Discuss standards run

  • Urine based testing

  • Blood/tissue testing

  • Summary of experience with the instrument

  • Future work


Rti set up l.jpg
RTI set up

  • JEOL TOF

  • DART

  • LEAP Technologies CombiPal autosampler

  • Mass Center software

  • More recently trying the Schraeder software


Accutof dart l.jpg

Snorkel

LEAP Autosampler

Ion Source DART

AccuTOF DART




Accutof dart strengths l.jpg
AccuTOF- DART Strengths

  • Minimal sample preparation

  • Broad range of sensitivity

  • Rapid analysis

  • Simultaneous determination of multi-drug analytes

  • Sufficient mass accuracy for formula determination


Reference materials l.jpg
Reference materials

  • Examined 112 compounds

    • drugs and metabolites

  • Methanolic standard materials

  • Directly introduced




Urine method l.jpg
Urine Method

  • Drug standards diluted in blank human urine

  • Glass probe dipped in sample

  • Analyzed with DART in positive mode

  • DART temperature @ 300°C

  • Mass calibrated using polyethylene glycol

  • Orifice 1 voltage set at 20V

  • Archived previously confirmed postmortem samples analyzed


Urine components l.jpg
Urine Components

Urea+H

Creatinine

Dimer +H

Creatinine+H


Cocaine 100 g ml m h 304 1548 l.jpg
Cocaine 100 µg/mL M + H = 304.1548



Triazolam 100 g ml m h 343 0508 l.jpg
Triazolam 100 µg/mL M + H = 343.0508



Creatinine issues l.jpg
Creatinine Issues

Oxazepam (10 µg/mL) in DI water

(M+H 287.0578)

Oxazepam (10 µg/mL) with 10 µg/mL creatinine


Creatinine issues20 l.jpg
Creatinine Issues

Oxazepam (10 µg/mL) with 100 µg/mL creatinine

Oxazepam (10 µg/mL) with 200 µg/mL creatinine


Archived case example l.jpg
Archived Case Example

EDDP=278.190

278.20765

+17mmu

310.21327

Methadone=310.216

-3mmu



Minimal blood tissue method l.jpg
“Minimal” Blood/Tissue Method

  • 2 mL of ACN added to 1 mL of sample

  • Mixture was shaken and vortexed

  • Spun in centrifuge at 2500 rpm for 5 minutes

  • ~100uL of ACN saved for DART analyses

  • Remainder poured off and dried down at 45°C

  • Reconstituted in 100 uL of ACN and analyzed



Blood method l.jpg
Blood method

  • Acetonitrile added to blood


Blood method26 l.jpg
Blood Method

  • Vortexed


Blood method27 l.jpg
Blood method

  • Centrifuged


Blood method28 l.jpg
Blood method

  • Decanted off supernatant


Blood method29 l.jpg
Blood method

  • Dry down under N2


Blood method30 l.jpg
Blood method

  • Reconstitute



Human blood l.jpg
Human Blood

Cholestadiene

Monosaccharide

Urea

Monosaccharide

Plasticizer


Methadone l.jpg

310.21838

Methadone

100 ug/mL Methadone in Blank Blood Unextracted

M + H =310.216

100 ug/mL Methadone in Blank Blood Extracted in 1:3 ACN


Methadone34 l.jpg
Methadone

100 ug/mL Methadone in Blank Blood Extracted in 1:3 ACN/dried/reconstituted in 100 ug/mL

1 ug/mL Methadone in Blank Blood Extracted in 1:3 ACN/dried/reconstituted in 100 ug/mL ACN

310. 215


Postmortem aorta blood specimen l.jpg
Postmortem Aorta Blood Specimen

Un-extracted

Previously reported at 2.8 mg/mL

Nondetected

Extracted in 1:3 ACN


Postmortem aorta blood specimen36 l.jpg
Postmortem Aorta Blood Specimen

Extracted in 1:3 ACN/dried/reconstituted in 100 ug/mL ACN


Postmortem liver specimen l.jpg
Postmortem Liver Specimen

Un-extracted

Previously reported at 6.6 ug/mL

Extracted in 1:3 ACN


Postmortem liver specimen38 l.jpg
Postmortem Liver Specimen

Extracted in 1:3 ACN/dried/reconstituted in 100 ug/mL


Cocaine l.jpg

304.15962

Cocaine

1 ug/mL Cocaine in blank blood extracted in 1:3 ACN

M+H=304.154

1 ug/mL Cocaine in Blank Blood Extracted in 1:3 ACN/dried/reconstituted in 100 ug/mL ACN


Cocaine40 l.jpg
Cocaine

0.1 ug/mL Cocaine in Blank Blood Extracted in 1:3 ACN/dried/reconstituted in 100 ug/mL ACN


Postmortem aorta blood specimen41 l.jpg

304.16913

Postmortem Aorta Blood Specimen

Extracted in 1:3 ACN

Previously reported at 0.52 mg/mL

Extracted in 1:3 ACN/dried/reconstituted in 100 ug/mL ACN


Postmortem cocaine case l.jpg

? BE=290.138

Norpropoxyphene=326.211

Propoxyphene=340.226

Postmortem Cocaine Case

  • All drugs not found

  • Could not see without concentrating

SPE Extraction


Postmortem case by lc ms and spe l.jpg
Postmortem case by LC/MS and SPE

BE

BE-d3

cocaine

cocaine-d3

cocaethylene

cocaethylene-d3

BE 1253 ng/mL, Cocaine 8.8 ng/mL, CE 2.7 ng/mL




Comparison to traditional ms platforms l.jpg
Comparison to “Traditional” MS platforms

* No quantitation attempted on the trap

** Source was saturated, resulting in poor quantitation


Table of drugs analyzed in pm specimens l.jpg
Table of Drugs Analyzed in PM Specimens

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a

a

a

a

a

a


Conclusions l.jpg
Conclusions

  • Reasonable sensitivity for some drugs

  • Greater sensitivity for some drugs over others

  • Creatinine appears to interfere with ionization of some drugs

  • Requirement for some sample preparation to mitigate interference from matrix

  • Auto sampler helps with more consistent sample introduction


Conclusions49 l.jpg
Conclusions

  • At least minimal drug extraction appears necessary to achieve best sensitivity

  • Extensive (solid phase extraction), does not necessarily improve things


Conclusions50 l.jpg
Conclusions

  • Drug detected in spiked blood have better sensitivity than in postmortem specimens at the same or greater concentrations

  • Not ideal for detecting therapeutic drug levels

    • Caveat the post mortem samples used are old and compound degradationmay have occurred


Conclusions51 l.jpg
Conclusions

  • Strong potential for the instrument

  • Issues for implementation:

    • Sensitivity

    • Interferences

    • Software (Mass Center)

      • Mass calibration

      • Stability

      • Retrieval of spectra


Conclusions52 l.jpg
Conclusions

  • Sensitivity

    • GIST/Vapur addition may improve sensitivity

  • Software

    • Schraeder software addresses many issues

      • Mass calibration

      • Retrieval of spectra

    • Still acquire using Mass Center

      • Potential stability issues

      • Be proactive about disk management


Acknowledgement l.jpg
Acknowledgement

  • Jeri Ropero-Miller

  • Nichole Bynum

  • National Institutes of Justice

    • Award #2006-DN-BX-K014

  • LEAP Technologies

  • Maricopa County, Washington State Police and OCME-NC

  • SPEWare


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