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

Forensic Toxicology Use Only

Analysis of Expanded NIDA 5 panel

Using Exactive System

Kevin HeThermo Fisher Scientific

2Forensic Toxicology Use Only

New Guidelines from SAMHSA for Confirmation

New cutoffs, added analytes

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

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

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)

LC Conditions

Hypersil Gold PFP 150 × 2.1 mm, 5 µm

Lockmass Setup

Amphetamine: Calibration Curve

Amphetamine: QC

Methamphetamine: Calibration Curve

Methamphetamine: QC

MDA: Calibration Curve

MDA: QC

MDMA: Calibration Curve

MDMA: QC

MDEA: Calibration Curve

MDEA: QC

BE: Calibration Curve

BE: QC

THCA: Calibration Curve

THCA: QC

PCP: Calibration Curve

PCP: QC

Morphine: Calibration Curve

Morphine: QC

Codeine: Calibration Curve

Codeine: QC

6-MAM: Calibration Curve

6-MAM: QC

Conclusion

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

NIDA-5 drugs

• Excellent quantitation using 0.5 mL processed sample.

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

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