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Rodrigues W.C.,Immunalysis Corporation | Catbagan P.,Immunalysis Corporation | Rana S.,Redwood Toxicology Laboratory | Wang G.,Immunalysis Corporation | Moore C.,Immunalysis Corporation
Journal of Analytical Toxicology | Year: 2013

Synthetic cannabinoids are often referred to as 'Spice' or K2 compounds. Detection of these compounds in oral fluid has, to date, been limited to chromatographic procedures such as liquid chromatography with tandem mass spectrometry detection. We report the first analytical immunoassay for the screening of some synthetic cannabinoids in oral fluid specimens collected with the Quantisal™ device. JWH-200 was chosen as the calibration standard, because parent compounds, not metabolites, are predominantly detected in oral fluid. The immunoassay is capable of detecting JWH-200, JWH-018, JWH-073, JWH-022, AM-2201, AM-2232 and AM-1220. The assay was validated according to accepted laboratory protocols and applied to 32 authentic oral fluid specimens previously analyzed using LC-MS-MS at an accredited laboratory. The assay is sensitive, with a cutoff concentration of 0.25 ng/mL, and has a wide working range from 0.1 to 5 ng/mL. Intra- and interday precision were determined to be <10%. The screening method was completely validated and characterized; critical aspects of the screening included the incorporation of a preincubation step that improves the sensitivity of the assay to allow relevant concentrations of synthetic compounds in oral fluid to be detected. © The Author (2013). Published by Oxford University Press. All rights reserved.

Rana S.,Redwood Toxicology Laboratory | Garg R.K.,Punjabi University | Singla A.,Bundelkhand University
Egyptian Journal of Forensic Sciences | Year: 2014

A sensitive and specific fast gas chromatography-mass spectrometry (FGC-MS) analytical method using hydrogen as a carrier gas is developed for the rapid simultaneous determination of morphine, codeine, hydrocodone and hydromorphone in human urine. Urine samples were spiked with deuterated internal standards, morphine-d3, codeine-d3, hydrocodone-d3 and hydro-morphone-d3, subjected to acid hydrolysis, treated with hydroxylamine to convert the keto-opiates to oximes and then extracted using a positive pressure manifold and silica based solid phase extraction columns. The extracts were derivatized using BSTFA with 1% TMCS.Gas chromatographic-mass spectrometric analysis was performed in electron ionization mode by selective ion monitoring, using hydrogen as a carrier gas, a short narrow bore GC capillary column, and fast temperature program, allowing for a rapid analytical cycle to maximize the instrument time for high throughput laboratories. While maintaining specificity for these drugs, concentrations in human urine ranging from 50 to 5,000. ng/mL can be measured with intraday and interday imprecision, expressed as variation coefficients, of less than 2.3% for all analytes within a run time of less than 3.5. minutes. © 2014 Hosting by Elsevier B.V.

Uralets V.,Redwood Toxicology Laboratory | Rana S.,Redwood Toxicology Laboratory | Morgan S.,Redwood Toxicology Laboratory | Ross W.,Redwood Toxicology Laboratory
Journal of Analytical Toxicology | Year: 2014

The study of 34,561 urine specimens, submitted for designer stimulant testing between February 2011 and January 2013, provided an opportunity: to estimate the range of synthetic cathinones (SC) abused in the USA, to observe multiple examples of metabolic profiles for each drug in various stages of excretion in human urine, to evaluate the extent of metabolism of specific SC and to select metabolites or parent drugs for routine testing. Sixteen SC were found in random patient samples: buphedrone; butylone; 3,4-dimethylmethcathinone; ethcathinone; N-ethylbuphedrone; ethylone; flephedrone; mephedrone; 4-methylbuphedrone; 3,4-methylenedioxypyrovalerone (MDPV); 4-methyl-N-ethylcathinone; methylone; pentedrone; pentylone; α-pyrrolidinobutiophenone (PBP) and α-pyrrolidinopentiophenone (PVP). After liquid/liquid extraction and trifluoroacetylation, specimens were screened by gas chromatography-mass spectrometry (GC-MS) for drugs and metabolites excreted free in urine. Each SC exhibited a characteristic metabolic profile, as shown by multiple examples. Metabolites' structures were postulated on the basis of their mass spectra. A large group of SC appears to metabolize extensively by carbonyl reduction into respective substituted ephedrines and further by N-dealkylation into norephedrines. Abundant metabolites in this group are essential markers of the parent drug use. Unchanged drugs are far less abundant or not found at all. SC with methylenedioxy attachment to the aromatic ring, metabolize by carbonyl reduction to a much lesser extent and are best detected as such in free urine fraction. PBP and PVP can be detected either unchanged or as metabolites, resulting from pyrrolidine ring degradation into primary amine followed by carbonyl reduction. MDPV appears in urine as such with no apparent free metabolites.

Uralets V.,Redwood Toxicology Laboratory | App M.,Redwood Toxicology Laboratory | Rana S.,Redwood Toxicology Laboratory | Morgan S.,Redwood Toxicology Laboratory | Ross W.,Redwood Toxicology Laboratory
Journal of Analytical Toxicology | Year: 2014

2-Ethylamino-1-phenylbutane (EAPB) and 2-amino-1-phenylbutane (APB) were identified by gas chromatography-mass spectrometry in multiple urine samples submitted for stimulant drug testing and screened positive for amphetamines by enzyme immunoassay. Fortytwo samples from all over the USA were found, containing both analytes during a 3-month period May-July 2013. A sports dietary supplement 'CRAZE' has been determined to be one of the sources of EAPB supply. EAPB along with its suggested metabolite APB were detected in a urine sample, obtained from a person known to use 'CRAZE'. © The Author [2014]. Published by Oxford University Press. All rights reserved.

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