Replacement of immunoassay by LC tandem mass spectrometry for the routine measurement of drugs of abuse in oral fluid.
BACKGROUND
There is increasing interest in the use of oral fluid as the matrix for the detection of drugs of abuse which requires the use of sensitive immunoassays to achieve the low detection limits required. The use of liquid chromatography linked to tandem mass spectrometry (LC/MS/MS) is explored as a possible replacement for immunoassay in screening for drugs of abuse in oral fluid samples.


METHODS
Oral fluid samples collected from 72 subjects attending an addiction clinic were screened for opiates, cocaine, methadone and benzodiazepines using both enzyme-linked immunosorbent assays (ELISA) and LC/MS/MS. The latter analysis used a short gradient elution with individual drugs detected by multiple reaction monitoring using tandem mass spectrometry. Results between the two methods were compared qualitatively using the cut-off concentrations defined by the ELISA assays.


RESULTS
With regard to the ELISA assays which show group specificity, LC/MS/ MS detected the presence of 6-monoacetylmorphine, morphine or dihydrocodeine in all but two of 49 samples positive for opiates. Of 55 samples positive for benzodiazepines by ELISA, all but two were confirmed by LC/MS/MS. Overall, LC/MS/MS compared favourably with ELISA for detection of specific drugs or their metabolites in the case of morphine, methadone and the cocaine metabolite benzoylecgonine. Many of the discrepant results between the two assays were a result of samples with drug concentrations near to the cut-off concentrations and the imprecision of these assays at very low concentrations.


CONCLUSIONS
LC/MS/MS offers a more flexible, specific and sensitive alternative to the screening of oral fluid samples for drugs of abuse than ELISA. A wide range of drugs and metabolites can be detected from a single sample injection.
Department of Clinical Biochemistry, Leeds Teaching Hospitals, Britannia House, Morley, Leeds LS27 0DQ, UK. keith.allen@leedsth.nhs.uk
K R
KR
Allen
R
R
Azad
H P
HP
Field
D K
DK
Blake
D002853:Chromatography, Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D005122:Exudates and Transudates / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009061:Mouth Mucosa / Q000737:chemistry*; D012463:Saliva / Q000737:chemistry*; D012680:Sensitivity and Specificity; D019966:Substance-Related Disorders / Q000175:diagnosis*