Abuse of gamma-hydroxybutyrate (GHB) as a recreational drug is becoming increasingly more prevalent due to the general feeling of euphoria it produces when ingested in moderate levels. At higher levels, unconsciousness and coma can result making this the drug of choice in "date-rape" crimes. Because of this, the United States Drug Enforcement Administration has declared it a Schedule I Controlled Substance. The goal of this proposal is to create a rapid, accurate, and simple-to-use test strip capable of detecting GHB in bodily fluids. Such a test strip does not currently exist and therefore it is difficult for emergency room personnel and emergency service providers to tailor their treatment of intoxicated patients when the nature of the intoxicant is not known. The test strip will be based on an enzyme that is capable of selectively oxidizing GHB, but not ethanol, to produce a colored, easily detectable product. Ethanol is commonly found in bodily fluids of GHB overdose patients and therefore a test such as this should not also react with ethanol as false positives will result. An additional goal of this work will be to develop a test strip capable of detecting not only GHB but also the just as potent analogs gamma-butyrolactone (GBL) and 1,4-butanediol (BD). These GHB analogs are legal in many states and their frequency of abuse rivals that of GHB. They are quickly metabolized to GHB in vivo and therefore are just as dangerous. This test strip will be marketed towards consumers to allow them to quickly, easily and discreetly test their beverage for these compounds before they become a victim of sexual assault. The Phase I goals of this Fast-Track proposal will be to screen our extensive lines of commercial enzymes for activity towards GHB, GBL and BD, and to construct and evaluate a prototype GHB-detection test strip. Phase II will concentrate on mass producing the GHB test strip, and evaluating inter- and intra- batch reproducibility and storage stability. An additional goal of Phase II is to develop a multiple sensing GHBGBL- BD detection strip for individual consumers. The latter test strip will also produced in multiple batches and reproducibility and storage stability will be ascertained
