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Amphetamines in Urine by LC/MS/MS on Gemini 5µm C18 150 x 3.0

Sample Preparation Details

Part No:8B-S029-TAK
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 30 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load:200µl at different concentrations of amphetamines and metabolites (in water) + 100µl of internal standards + 200µl acid hydrolyzed urine (adjusted pH to neutral) + 100µl of 1M acetic acid + 400µl water
Final Prep and Analysis: to prevent the loss of amphetamine and methamphetamine during solvent evaporation, add two drops of conc. HCl before the evaporation step
Inject: 5 µL on HPLC / Mass Spectrometer (MS) @ amu (450 °C)

Analytes:

Name Spiked
Conc.
Log P pKa % Rec %RSC (n=0)
1 4-Hydroxy-3-methoxymethamphetmine 250 
2 Amphetamine 500 
3 Methamphetamine 250 
4 3,4-Methylenedioxyamphetamine (MDA)-d5 50 
5 3,4-Methylenedioxyamphetamine (MDA) 500 
6 3,4-Methylendioxymethamphetamine-d5 (MDMA)-d5 25 
7 3,4-Methylendioxmethamphetamine (MDMA) 250 

Application Detail (App ID: 17900)

Part No:00F-4435-Y0
Dimensions:150 x 3 mm ID
Elution Type:Isocratic
Elution A:10mM Ammonium formate w/0.1% formic acid
Elution B:Acetonitrile
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 90 10

Flow Rate:0.6 mL/min
Col. Temp: ambient
Detection: Mass Spectrometer (MS) @ amu (450°C)
Detector Info: AB Sciex API 3000™ LC/MS/MS ESI TurboIonSpray®
heater gas flow 8000cc/min
Heater temperature 450 °C; Positive Ion Mode
Note:SecurityGuard™ Guard Cartridge System extends column lifetime.
- SecurityGuard Cartridges, Gemini C18 4 x 2.0mm, 10/Pk Part No.: AJ0-7596
- Holder Part No.: KJ0-4282

Analyte Details

Analyte Detail not available

1
4-Hydroxy-3-methoxymethamphetmine
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2
Amphetamine
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3
Methamphetamine
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4
3,4-Methylenedioxyamphetamine (MDA)-d5

5
3,4-Methylenedioxyamphetamine (MDA)
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6
3,4-Methylendioxymethamphetamine-d5 (MDMA)-d5

7
3,4-Methylendioxmethamphetamine (MDMA)
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