TIAFT Young Scientists "Workshop 2001"

Measurement of Amphetamines in Biological Samples by LC-MS/MS

Michelle Wood*, Michael Morris*, Donald P Cooper*, Gert De Boeck**, Nele Samyn**, Jan Claereboudt***

* Micromass UK Ltd., Floats Rd, Manchester, M23 9LZ, UK.
** National Institute for Criminalistics and Criminology (NICC), Brussels, Belgium.
*** Micromass Belgium, Vilvoorde, Belgium.

The identification and quantification of drugs of abuse in biological specimens is of great importance. Traditionally, this has been a two-step procedure i.e. initial screening for classes of substances e.g. amphetamines (using immunoassay) followed by confirmation of the particular drug (s) using a more specific method. The sensitivity and specificity of gas chromatography/mass spectrometry (GC-MS) has made this the technique of choice for confirmation in most laboratories. However, the procedure is very labour-intensive and time-consuming particularly as sample preparation i.e. solid phase extraction and derivatisation are unavoidable.

Here we present the development of an alternative method. Amphetamines were isolated from plasma using a simple methanol extraction procedure and subsequently analysed using reversed phase HPLC-MS/MS. A Quattro Ultima (Micromass) tandem mass spectrometer fitted with a z-spray ion source was used for the analyses. The instrument was operated in electro spray positive ionisation mode and was coupled to a Waters 2690 Alliance HPLC system. The mobile phase (A: 10mM ammonium acetate / B: 95% acetonitrile, 5% 10mM ammonium acetate) (85/15) was delivered at a flow rate of 0.3 ml/min. A Hypersil BDS C18 column (100 mm x 2.1 mm, 3.5 microm) was used.

Quantification of the drugs and their deuterated analogues was performed using multiple reaction monitoring (MRM). The developed method, which has a chromatographic run time of less than 10 minutes, enables the simultaneous quantitation of MDMA, MDA, MDEA, amphetamine, methamphetamine and ephedrine in a single analysis. Limits of detection of 1 ng/ml plasma or better were achieved.

In addition, to assess the feasibility of using saliva as a convenient, non-invasive marker of drug abuse, the method has been used for the quantitation of the commonly abused amphetamines in saliva samples collected from current users. These levels will be compared to corresponding levels in plasma and urine as determined by GC-MS.

ABSTRACTS

• Pitfalls associated with the use of a simple sample preparation in the analysis of saliva with LC-ESI-MS/MS.
  K.A. Mortier, K.M. Clauwaert, W.E. Lambert, J.F. Van Bocxlaer, E.G. Van den Eeckhout, C.H. Van Peteghem, and A.P. De Leenheer
   
  
• Oral Fluid Testing in Road Traffic - The German Experience within the ROSITA Project
  S. Steinmeyer, H. Ohr, and M. R. Moeller
   
  
• The use of oral fluid and sweat wipes for the detection of drugs of abuse in impaired drivers
  Nele Samyn, Bart Viaene, Bart Laeremans, Gert De Boeck
   
  
• Detection of Fluoxetine and Norfluoxetine in Saliva
  Fiona M. Wylie and John S. Oliver
   
  
• Measurement of Amphetamines in Biological Samples by LC-MS/MS
  Michelle Wood, Michael Morris, Donald P Cooper
   
  
• Immunochromatographic teststrips for saliva analysis
  Jörg Zimmermann1, Nele Samyn, Maria da Costa, Thomas Wuske, Rainer Polzius, Hans-Jürgen Duchstein1
   
  
• Determination of MDMA ('Ecstasy') in saliva: a controlled study in recreational users
  Monica Navarro, Magi Farré, Simona Pichini, Jordi Ortuño, Pere Nolasc Roset, Ester Menoyo, Jordi Segura, Rafael de la Torre
   
  
• Determination of MDMA ('Ecstasy') in saliva with Drugwipe analytical device: a controlled study in recreational users
  Monica Navarro, Magi Farré, Simona Pichini, Piergiorgio Zuccaro, Roberta Pacifici, Jordi Ortuño, Pere Nolasc Roset, Ester Menoyo, Jordi Segura, Rafael de la Torre
   
  
• Elimination kinetics of oral nandrolone in urine and saliva
  V. Cirimele, P. Kintz B. Ludes
   
  
• Determination of Cocaine and its Metabolites in Saliva by GC/MS-EI
  P. Campora O.; A.M. Bermejo B.; M.J. Tabernero and P. Fernandez.
   
  
• The Finnish experience from the ROSITA project
  Marielle Grönholm and Pirjo Lillsunde