Field Test of On-Site
Drug Detection Devices
Final Report -- October 2000
As part of the Field Test of On-Site Drug Detection Devices funded by the National Highway Traffic Safety Administration, the Center for Human Toxicology performed Mass Spectrometry (MS) confirmations all drugs presumptively identified as positive by any of the devices. In addition, 5% of the samples that tested negative (drug free) were randomly selected for confirmation testing. For each confirmation test, Gas Chromatography/Mass Spectrometry (GC/MS) confirmation cutoff sensitivities at lower concentrations than those of the federal standards (DOT, 1992; DHHS, 1993) were established. A summary of GC/MS methods and testing limits used in this study follows.
Multi-point calibration curves, containing certified negative urine and at least 4 calibrators were generated by fortifying drug-free urine with the target analytes. Verified negative and positive controls were included in each testing batch. Deuterium labeled drug analogs were used as internal standards to ensure accurate relative retention time information for qualitative identification and for internal standard quantitation. LC/MS and LC/MS/MS testing was used to supplement the GC/MS specifically for the detection of low concentrations of cannabinoids and for the identification of additional sympathomimetic amines and opiates.
All MS data were reviewed and certified prior to reporting. Each confirmation test was performed on a standard volume of 1.0 mL of urine. Samples testing at concentrations greater than the described calibration curve were reported as positive greater than the highest calibrator concentration (e.g., THC-COOH > 250 ng/mL).
THC-COOH confirmations were performed to a testing limit of 4 ng/mL. Each urine was hydrolyzed at basic pH, extracted, derivatized with pentafluoropropionic anhydride/ hexafluorisoprapanol (PFPA/HFIP), and subjected to GC/MS analysis by negative ion chemical ionization with selected ion monitoring. Samples that proved unsuitable for GC/MS analyses were confirmed by LC/MS/MS. These samples were hydrolyzed at a basic pH, extracted and subjected to LC/MS/MS analysis using positive ion electrospray ionization with selected reaction monitoring.
A cocaine confirmation consisted of testing for parent cocaine and benzoylecgonine (BZE) to a limit of 50 ng/mL. Each urine was extracted using SPE (solid phase extraction), the BZE derivatized with N-methyl-N-(tert-butyldimethylsilyl)-trifluoroacetamide (MTBSTFA) and subjected to GC/MS analysis for parent cocaine, BZE. The analyses were performed by positive ion chemical ionization with selected ion monitoring. Samples that proved unsuitable for GC/MS analyses were confirmed by LC/MS. These samples were extracted by SPE and subjected to LC/MS analysis for parent cocaine and BZE using positive ion electrospray ionization with selected ion monitoring.
An "amphetamines" confirmation consisted of testing for amphetamine, phentermine, and methamphetamine to a limit of 100 ng/mL. In addition, phenylpropanolamine, ephedrine, pseudoephedrine, or methylenedioxymethamphetamine (MDMA) were qualitatively identified. Initially, urine samples were extracted at basic pH, derivatized with trifluoroacetic anhydride (TFAA), and subjected to GC/MS analysis for the listed drugs by positive ion chemical ionization with selected ion monitoring. However, it quickly became apparent that many samples had high concentrations of phenylpropanolamine, ephedrine, pseudoephedrine, and MDMA that adversely affected the chromatography. Therefore, most samples were extracted at basic pH and subjected to LC/MS analysis for using positive ion electrospray ionization with selected ion monitoring.
An opiate confirmation was performed to a testing limit of 50 ng/mL for total morphine and codeine. In addition, hydrocodone and hydromorphone were qualitatively identified. Each urine sample was hydrolyzed enzymatically, extracted at basic pH, derivatized with TFAA, and subjected to positive ion chemical ionization with selected ion monitoring GC/MS analysis. Many samples contained hydromorphone and hydrocodone. Due to the inherently better analysis characteristics for these drugs by LC/MS, these samples were hydrolyzed enzymatically, extracted by SPE, and subjected to LC/MS analysis for the target opiates using positive ion electrospray ionization with selected ion monitoring.
A phencyclidine confirmation consisted of testing for PCP to a limit of 5 ng/mL. Each urine was extracted at basic pH and subjected to GC/MS analysis for PCP. The analysis used positive ion chemical ionization with selected ion monitoring.