NIDA 5 panel Exactive...6 Forensic Toxicology Use Only Sample Preparation Procedure: Modified Add IS...

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Forensic Toxicology Use Only Analysis of Expanded NIDA 5 panel Using Exactive System Kevin He Thermo Fisher Scientific

Transcript of NIDA 5 panel Exactive...6 Forensic Toxicology Use Only Sample Preparation Procedure: Modified Add IS...

Page 1: NIDA 5 panel Exactive...6 Forensic Toxicology Use Only Sample Preparation Procedure: Modified Add IS (4 µg/mL, 10 µL) Add 180 µL MeOH Transfer ~ 400 µL to glass insert, 10, 25

Forensic Toxicology Use Only

Analysis of Expanded NIDA 5 panel

Using Exactive System

Kevin HeThermo Fisher Scientific

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New Guidelines from SAMHSA for Confirmation

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New cutoffs, added analytes

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List of compounds, their m/zs and cutoff vales

Number Name Formula [M+H]+

IS M+H Cutoff (ng/mL) QC1 (-60%) QC2 (+25%)

1 Amphetamine C9H13N 137.11936 141.14298 250 100 312.5

2 Methamphetamine C10H15N 151.13496 155.15858 250 100 312.5

3 MDA C10H13NO2 181.10916 185.13278 250 100 312.5

4 MDMA C11H15NO2 195.12486 199.14848 250 100 312.5

5 MDEA C12H17NO2 209.14046 214.17026 250 100 312.5

6 Benzoylecgonine (BE) C16H19NO4 291.14596 293.15722 100 40 125

7 THCA (cTHC) C21H28O4 346.21336 348.22462 15 6 18.75

8 Phencyclidine (PCP) C17H25N 245.21326 249.23688 25 10 31.25

9 Morphine C17H19NO3 287.15106 289.16232 2000 800 2500

10 Codeine C18H21NO3 301.16666 303.17792 2000 800 250011 6-Acetylmorphine (6-MAM) C19H21NO4 329.16166 331.17292 10 4 12.5

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Calibrators and Controls

Drug 1 2 3 4 5 6 7 8 9 IS

Amphetamine 2 5 10 20 50 100 200 1000 200

Methamphetamine 2 5 10 20 50 100 200 1000 200

MDA 2 5 10 20 50 100 200 1000 200

MDMA 2 5 10 20 50 100 200 1000 200

MDEA 2 5 10 20 50 100 200 1000 200

BE 2 5 10 20 50 100 200 1000 200

THCA 2 5 10 20 50 100 200 200

PCP 2 5 10 20 50 100 200 1000 200

Morphine 2 5 10 20 50 100 200 1000 4000 200

Codeine 2 5 10 20 50 100 200 1000 4000 200

6MAM 2 5 10 20 50 100 200 1000 200

Concentration of calibrators (ng/mL)

Drug Cutoff(ng/mL) QC1 (40%)(ng/mL) QC2 (125%)(ng/mL) IS (ng/mL)

Amphetamine 250 100 312.5 200

Methamphetamine 250 100 312.5 200

MDA 250 100 312.5 200

MDMA 250 100 312.5 200

MDEA 250 100 312.5 200

BE 100 40 125 200

THCA 15 6 18.75 200

PCP 25 10 31.25 200

Morphine 2000 800 2500 200

Codeine 2000 800 2500 200

6MAM 10 4 12.5 200

Page 6: NIDA 5 panel Exactive...6 Forensic Toxicology Use Only Sample Preparation Procedure: Modified Add IS (4 µg/mL, 10 µL) Add 180 µL MeOH Transfer ~ 400 µL to glass insert, 10, 25

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Sample Preparation Procedure: Modified

Add IS (4 µg/mL, 10 µL)

Add 180 µL MeOHAdd 180 µL MeOH

Transfer ~ 400 µL to glass insert,

10, 25 or 50 µL injection

Transfer ~ 400 µL to glass insert,

10, 25 or 50 µL injection

Urine 200 µL in LR plastic tube

Add Spike (10 µL, S1-9, QC1, Cutoff, QC2)

Mix, add glucuronidase (0.1 mL), 60 °C, 30min or longer

Vortex, stand for 20 min,

centrifuge for 5 min at 15K RCF

Note: 50 mL injection (10 mL raw urine equivalent)

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LC Conditions

Hypersil Gold PFP 150 × 2.1 mm, 5 µm

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Lockmass Setup

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Amphetamine: Calibration Curve

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Amphetamine: QC

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Methamphetamine: Calibration Curve

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Methamphetamine: QC

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MDA: Calibration Curve

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MDA: QC

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MDMA: Calibration Curve

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MDMA: QC

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MDEA: Calibration Curve

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MDEA: QC

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BE: Calibration Curve

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BE: QC

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THCA: Calibration Curve

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THCA: QC

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PCP: Calibration Curve

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PCP: QC

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Morphine: Calibration Curve

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Morphine: QC

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Codeine: Calibration Curve

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Codeine: QC

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6-MAM: Calibration Curve

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6-MAM: QC

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Conclusion

• One sample preparation and one injection for confirmatory testing of all

NIDA-5 drugs

• Excellent quantitation using 0.5 mL processed sample.


Combination of epT.I.P.S.® with Eppendorf Pipettes · pearl white 0.5 – 100 µL light yellow 2 – 20 µL yellow 2 – 100 µL yellow 2 – 200 µL yellow 20 – 300 µL orange

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Urea UV SL - POCT ServicesUREA UV SL y Sample Standard Reagent R2 60 µL 60 µL 60 µL Sample

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Immunosuppressant drugs Exactive - Orbitrapplanetorbitrap.com/download.php?filename=5006ffb1e8c26.pdfQuantitative Analysis of 4 Immunosuppressant Drugs in Blood using Exactive System

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10 µl 10 µl 10 µl 20 µl 100 µl 200 µl 300 µl 1.000 µl 1.250 µl … · 2017. 3. 9. · w w .nerbe-lus conformity tested (ISO 8655) Certified RNase/DNase Pyrogen, DNA, ATP

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Primerdesign Ltd TM Mycoplasma · 2019-01-07 · 2 x 105 per µl 2 x 104 per µl 2 x 103 per µl 2 x 102 per µl 20 per µl 2 per µl Standard Curve Copy Number Tube 1 Positive control

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Supporting Information Boosting CRISPR/Cas9 …...Bsa I (10 U/µl, NEB) 0.5 µl T7 DNA Ligase (3000 U/µl, NEB) 0.5 µl Total Volume 20 µl For GG assembly reactions to construct PTG1-PTG8,

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EASY AND EFFICIENTLSD - PerkinElmer · 4 5 EASY ASSAY WORKFLOW 4 To deep 96-well plate (DWP). Add 400 µL EA & 200 µL H2O Mix with pipette. Transfer Liquid & Extract OR DWP centrifuge,

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AUTOMATED LIQUID HANDLING · MICROLAB M Volume 10 µL 25 µL 50 µL 100 µL 250 µL 500 µL 1 mL 2.5 mL 5 mL 10 mL 25 mL Model 1701 1702 1705 1710 1725 1750 1001 1002 1005 1010 1025

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1.Digest remaining DNA with DNAse I 1.7 µl 10x RDD buffer 2.1 µl Superasin RNAse inhibitor 3.2.5 µl DNAse I 2.Leave 30’ @ 37˚ C 3.Add 15 µl 10 M ammonium.

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