title: Direct detection of Δ9-tetrahydrocannabinol in aqueous samples using a homogeneous increasing fluorescence immunoassay (HiFi).
abstract: The detection of the major active component of cannabis, Δ9-tetrahydrocannabinol (THC), becomes increasingly relevant due to its widespread abuse. For control purposes, some easy-to-use, sensitive and inexpensive test methods are needed. We have developed a fluorescence immunoassay utilising THC-fluorescein conjugate as tracer. Fluorescence spectroscopy of the conjugate revealed an unusual property: The relatively weak fluorescence of a dilute tracer solution was increased by a factor of up to 5 after binding of a THC-specific antibody. Fluorescence lifetime measurements in aqueous solutions suggested two different tracer conformations both associated with quenching of fluorescein fluorescence by the intramolecular THC moiety. After antibody binding, the tracer enters a third conformation in which fluorescence quenching of fluorescein is completely suppressed. Utilising this property, we established a homogeneous competitive immunoassay (homogeneous increasing fluorescence immunoassay) with low detection limits. The test requires only two reagents, the new tracer molecule and an anti-THC antibody. A single test takes only 8 min. The dynamic detection range for THC is 0.5 to 20 ng/mL in buffer, with a limit of detection (LOD) of 0.5 ng/mL. The test also works in diluted saliva samples (1:10 dilution with buffer) with an LOD of 2 ng/mL and a dynamic range of 2-50 ng/mL.
authors.affiliation: Group Biosensors, Fraunhofer Institute for Biomedical Engineering, Am Mühlenberg 13, 14476 Potsdam-Golm, Germany.
authors.forename: Chongxiao
authors.initials: C
authors.lastname: Tan
authors.forename: Nenad
authors.initials: N
authors.lastname: Gajovic-Eichelmann
authors.forename: Rainer
authors.initials: R
authors.lastname: Polzius
authors.forename: Niko
authors.initials: N
authors.lastname: Hildebrandt
authors.forename: Frank F
authors.initials: FF
authors.lastname: Bier
mesh_terms: D013759:Dronabinol / Q000032:analysis*; D019793:Fluorescein / Q000737:chemistry; D005453:Fluorescence; D007118:Immunoassay / Q000379:methods*; D057230:Limit of Detection; D013058:Mass Spectrometry; D015394:Molecular Structure; D014867:Water
chemical_list: D014867:Water; D013759:Dronabinol; D019793:Fluorescein