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Rapid Screening of Amphetamine Drugs in Urine by Positive Ion Electrospray LC/MS/MS

Z. Yang and S. Sadjadi, Varian, Inc.

Introduction

Amphetamine drugs are often abused and misused in sports, in the work place, and by recreational users. Forensic, clinical, and doping laboratories are frequently asked to analyze for the presence of amphetamines in urine. Urine samples are most common because large sample volumes can be collected non-invasively. These drugs generally remain detectable in urine for two to three days longer than in blood. For most clinical and forensic applications, initial screening is done by immunoassay with presumptive positive samples confirmed by a second, more specific method such as gas chromatography/ mass spectrometry (GC/MS).

A simple and sensitive LC/MS/MS method is described below for high throughput identification and quantitation amphetamine drugs in urine. A rapid and effective solid-phase extraction (SPE) procedure using FocusTM was used to extract amphetamines from urine samples.

Instrumentation

Varian ProStar 410 AutoSampler

Varian ProStar 210 Isocratic Solvent Delivery Modules

Varian 1200L LC/MS equipped with ESI source

Materials and Reagents

Standard solutions: 1.0 mg/mL (()-Amphetamine, ()-Methamphetamine, 1S,2R(+)-Ephedrine, ()-MDMA, ()-MDA and ()-MDEA), from Cerilliant Corp., Texas, USA.

Internal standard (IS) solutions: 0.1 mg/mL (()-Amphetamine-D5, ()-Methamphetamine-D5, 1S,2R(+)-Ephedrine-D3 HCl, ()-MDA-D5, ()-MDMA-D5 and ()-MDEA-D5), from Cerilliant Corp., Texas, USA.

All other chemicals are reagent grade or HPLC grade.

FocusTM Solid Phase Extraction Cartridges (Varian Part No. A5306021).

In-house vacuum or vacuum pump (Varian Part No. WL2012B01).

Vac Elut 20 Manifold with the standard Glass Basin (Varian Part No. 122345
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