Analytical Procedure for the Determination of the Marijuana Metabolite 11-nor- 9-Tetrahydrocannabinol-9-carboxylic Acid in Oral Fluid Specimens (original) (raw)
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Journal of Analytical Toxicology, 2006
The detection of the marijuana metabolite 11-nor-Agtetrahydrocannabinol-9-carboxyllc acid (THC-COOH) in oral fluid specimens is described, and its contribution to an immunoassay for the detection of cannabinoids is investigated. Oral fluid specimens, screened using an enzyme-linked immunosorbent immunoassay (ELISA), were carried forward to confirmation for both tetrahydrocannabinol (THC) and THC-COOH using gas chromatography-mass spectrometry (GC-MS). One hundred and fifty-three specimens were analyzed, of which 143 screened positive for cannabinoids. Ninety-five (66.4%) of these specimens were positive for both THC and THC-COOH; 14 (9.7%) were positive for THC-COOH only, and 27 (18.8%) were positive for THC only. The GC-MS assay for the detection of THC-COOH in oral fluid was linear to 160 pg/ml, with a limit of quantitation of 2 pg/mL The detection of the marijuana metabolite, THC-COOH, in 76.2% of oral fluid specimens screening positive for cannabinoids is reported. As a potential defense against passive exposure claims, proposed SAMHSA regulations may require the simultaneous collection of a urine sample when oral fluid samples are used. The detection of the metabolite, THC-COOH, is a significant alternative to this approach because its presence in oral fluid minimizes the argument for passive exposure to marijuana in drug testing cases.
Therapeutic Drug Monitoring, 2014
A sensitive and specific method for the quantification of 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (THCCOOH) in oral fluid collected with the Quantisal and Oral-Eze devices was developed and fully validated. Extracted analytes were derivatized with hexafluoroisopropanol and trifluoroacetic anhydride and quantified by gas chromatography-tandem mass spectrometry with negative chemical ionization. Standard curves, using linear least-squares regression with 1/x 2 weighting were linear from 10 to 1000 ng/L with coefficients of determination .0.998 for both collection devices. Bias was 89.2%-112.6%, total imprecision 4.0%-5.1% coefficient of variation, and extraction efficiency .79.8% across the linear range for Quantisal-collected specimens. Bias was 84.6%-109.3%, total imprecision 3.6%-7.3% coefficient of variation, and extraction efficiency .92.6% for specimens collected with the Oral-Eze device at all 3 quality control concentrations (10, 120, and 750 ng/L). This effective high-throughput method reduces analysis time by 9 minutes per sample compared with our current 2-dimensional gas chromatography-mass spectrometry method and extends the capability of quantifying this important oral fluid analyte to gas chromatographytandem mass spectrometry. This method was applied to the analysis of oral fluid specimens collected from individuals participating in controlled cannabis studies and will be effective for distinguishing passive environmental contamination from active cannabis smoking.
Detection of Conjugated 11-nor- 9-Tetrahydrocannabinol-9-carboxylic Acid in Oral Fluid
Journal of Analytical Toxicology, 2007
The presence of the conjugated marijuana metabolite 11-nor-∆ ∆ 9tetrahydrocannabinol-9-carboxylic acid (THCA) glucuronide in oral fluid specimens is described for the first time. Oral fluid specimens were collected using a Quantisal™ device and analyzed for the presence of THCA using two-dimensional gas chromatography (GC) with mass spectrometric (MS) detection both before and after hydrolysis. The nature of the conjugation was determined by analyzing specimens from a marijuana user without hydrolysis, with base hydrolysis, with β β-glucuronidase treatment, and hydrolysis using sulfatase only. Treatment with sodium hydroxide proved to be the most efficient hydrolytic procedure. Specimens collected over 48 h showed an average conjugation of over 64.5%. The specimens were also analyzed for the active component, tetrahydrocannabinol (THC), which was detected in the oral fluid, in most cases, for up to 24 h. Parent THC was not found to be glucuronide bound. Specimens were then subjected to commercially available immunoassays in order to determine their utility as screening procedures. The metabolite, THCA, was detected in all samples up to and including the specimen 48 h after smoking, using the more sensitive screening assay and two dimensional GC-MS. Moreover, proof that the THCA is conjugated in oral fluid minimizes concerns associated with passive inhalation.
Journal of Analytical Toxicology, 2012
This paper describes the determination of tetrahydrocannabinol (THC) and its metabolite, 11-nor-D 9 -tetrahydrocannabinol-9carboxylic acid (THC-COOH) in oral fluid using solid-phase extraction and liquid chromatography with tandem mass spectral detection (LC-MS-MS) and its application to proficiency specimens. The method employs collection of oral fluid with the Quantisal TM device, base hydrolysis, solid-phase extraction and LC -MS-MS in positive ion electrospray mode. Because the concentration of the metabolite in oral fluid is quite low, extremely sensitive analytical methods are necessary. The requisite sensitivity was achieved by a simple, rapid derivatization of the compound after extraction. The derivatization conditions did not affect parent THC. The method was fully validated using standard parameters including linearity, sensitivity, accuracy, intra-day and inter-day imprecision, drug recovery from the collection pad, limit of quantitation, limit of detection and matrix effects. The procedure was applied to oral fluid proficiency specimens previously analyzed to assess the stability of THC-COOH.
Journal of Analytical Toxicology, 2004
Understanding the relationship of A%tetrahydrocannabinol (THC) concentrations in oral fluid and plasma is important in interpretation of oral fluid test results. Current evidence suggests that THC is deposited in the oral cavity during cannabis smoking. This "depot" represents the primary or sole source of THC found when oral fluid is collected and analyzed. In this research, oral fluid and plasma specimens were collected from six subjects following smoking of cannabis cigarettes containing 1.75% and 3.55% THC. There was at least one week between ~ach cannabis administration. Plasma specimens were analyzed by gas chromatography-mass spectrometry (GC-MS) and paired oral fluid specimens were analyzed by radioimmunoassay (RIA). In addition, one individual's oral fluid specimens were also analyzed by GC-MS. These data are unique in that they represent simultaneous or near simultaneous collection of oral fluid and plasma specimens in subjects following controlled cannabis dosing. The first oral fluid specimen, collected from one subject at 0.2 h following initiation of smoking, contained a THC concentration of 5800 ng/mL (GC-MS). By 0.33 h, the THC concentration in oral fluid bad fallen to 81 ng/mt. From approximately 0.3 h through 4.0 h, the mean (_+ SD) THC ratio of oral fluid to plasma THC concentrations was 1.18 (0.62) with a range of 0.5 to 2.2. Within 12 h, both oral fluid and plasma THC concentrations generally declined below 1 ng/mL. RIA analyses of oral fluid specimens for six subjects demonstrated the same pattern of initial high levels of contamination immediately after smoking, followed by rapid clearing, and a slower decline over 12 h. Mean THC oral fluid concentrations by RIA at 0.2 h were 864 ng/mL and 4167 ng/mL compared to plasma concentrations of 52 ng/mL and 230 ng/mL at 0.27 h following the low-and high-dose cannabis cigarettes, respectively. The similarity in oral fluid and plasma THC concentrations following the dissipation of the initial "contamination" indicates the likelihood of a physiological link between these specimens. Recent studies have shown that sublingual or transmucosal administration of pure THC results in direct absorption of intact THC into the bloodstream, thereby bypassing the gastrointestinal tract. The current study * Author to whom correspondence should be addressed.
Cannabinoids and metabolites in expectorated oral fluid following controlled smoked cannabis
Clinica Chimica Acta, 2012
Background-Δ 9-Tetrahydrocannabinol (THC) in oral fluid (OF) implies cannabis intake, but eliminating passive exposure and improving interpretation of test results requires additional research. Methods-Ten adult cannabis users smoked ad libitum one 6.8% THC cigarette. Expectorated OF was collected for up to 22h, and analyzed within 24 h of collection. THC, 11-nor-9-carboxy-THC (THCCOOH), cannabidiol, and cannabinol were quantified by 2-dimensional-GCMS. Results-Eighty specimens were analyzed; 6 could not be collected due to dry mouth. THC was quantifiable in 95.2%, cannabidiol in 69.3%, cannabinol in 62.3%, and THCCOOH in 94.7% of specimens. Highest THC, cannabidiol, and cannabinol concentrations were 22370, 1000, and 1964 μg/l, respectively, 0.25 h after the start of smoking; THCCOOH peaked within 2 h (up to 560 ng/ l). Concentrations 6h after smoking were THC (0.9-90.4 μg/l) and THCCOOH (17.0-151 ng/l) (8 of 9 positive for both); only 4 were positive for cannabidiol (0.5-2.4 μg/l) and cannabinol (1.0-3.0 μg/l). By 22h, there were 4 THC (0.4-10.3 μg/l), 5 THCCOOH (6.0-24.0 ng/l), 1 cannabidiol (0.3 μg/l), and no cannabinol positive specimens. Conclusions-THCCOOH in OF suggests no passive contamination, and CBD and CBN suggest recent cannabis smoking. Seventeen alternative cutoffs were evaluated to meet the needs of different drug testing programs.
Journal of Chromatography A, 2005
A rapid and sensitive method for the analysis of 9 -tetrahydrocannabinol (THC) in preserved oral fluid was developed and fully validated. Oral fluid was collected with the Intercept, a Food and Drug Administration (FDA) approved sampling device that is used on a large scale in the U.S. for workplace drug testing. The method comprised a simple liquid-liquid extraction with hexane, followed by liquid chromatography-tandem mass spectrometry (LC-MS-MS) analysis. Chromatographic separation was achieved using a XTerra MS C 18 column, eluted isocratically with 1 mM ammonium formate-methanol (10:90, v/v). Selectivity of the method was achieved by a combination of retention time, and two precursor-product ion transitions. The use of the liquid-liquid extraction was demonstrated to be highly effective and led to significant decreases in the interferences present in the matrix. Validation of the method was performed using both 100 and 500 L of oral fluid. The method was linear over the range investigated (0.5-100 ng/mL and 0.1-10 ng/mL when 100 and 500 L, respectively, of oral fluid were used) with an excellent intra-assay and inter-assay precision (relative standard deviations, RSD <6%) for quality control samples spiked at a concentration of 2.5 and 25 ng/mL and 0.5 and 2.5 ng/mL, respectively. Limits of quantification were 0.5 and 0.1 ng/mL when using 100 and 500 L, respectively. In contrast to existing GC-MS methods, no extensive sample clean-up and time-consuming derivatisation steps were needed. The method was subsequently applied to Intercept samples collected at the roadside and collected during a controlled study with cannabis.
Biological Mass Spectrometry, 1988
A gas chromatographic/mass spectrometric electron impact method is presented for the detection and quantification of 1 l-nor-9-carboxy-A9-tetrahydrocannabinol (THC-COOH) in urine, for use in the confirmation of presump tive results obtained by other techniques. Four extraction procedures, two solid-liquid and two liquid-liquid, have been compared. A comparison of two trimethylsilylating methods demonstrates that the best results are obtained by the use of a mixture containing N-methyl-N-trimethylsilyl-trifluoroacetamide, trimethyliodosilane and dithioerithritol (100:0.2:1) v/v/w. The use of ketoprofen as a new internal standard for the quantification of THC-COOH has proved to be very effective. Both spiked samples and samples from cannabis users have been successfully analysed. It has also been demonstrated that the presence of other drugs of abuse in urine samples do not interfere with cannabis quantification by the method reported here.
Forensic Science International, 2005
An analytical method using solid-phase extraction (SPE) and high-performance liquid chromatography-mass spectrometry (LC-MS) has been developed and validated for the confirmation of D 9 -tetrahydrocannabinol (THC) in oral fluid samples. Oral fluid was extracted using Bond Elut LRC-Certify solid-phase extraction columns (10 cm 3 , 300 mg) and elution performed with n-hexane/ethyl acetate. Quantitation made use of the selected ion-recording mode (SIR) using the most abundant characteristic ion [THC + H + ], m/z 315.31 and the fragment ion, m/z 193.13 for confirmation, and m/z 318.00 for the protonated internal standard, [d 3 -THC + H + ]. The method proved to be precise for THC, in terms of both intra-day and inter-day analyses, with coefficients of variation less than 10%, and the calculated extraction efficiencies for THC ranged from 76 to 83%. Calibration standards spiked with THC between 2 and 100 ng/mL showed a linear relationship (r 2 = 0.999). The method presented was applied to the oral fluid samples taken from the volunteers during the largest music event in Portugal, named Rock in Rio-Lisboa. Oral fluid was collected from 40 persons by expectoration and with Salivette 1 . In 55% of the samples obtained by expectorating, THC was detected with concentration ranges from 1033 to 6552 ng/mL and in 45% of cases THC was detected at concentrations between 51 and 937 ng/mL. However, using Salivette 1 collection, 26 of the 40 cases had an undetectable THC. #
Medicolegal Aspects of Cannabis and Its Metabolites Detection Window in Oral Fluid And Urine
Mansoura Journal of Forensic Medicine and Clinical Toxicology
Ghanem et al ... is approximately 10% as potent as THC (El-Sohly, 2002). Oral fluid (OF) drug testing in workplace, pain management, drug treatment, and driving under the influence of drugs programs is increasing (Lee et al., 2012). In contrast to urine, the advantages include observable sample collection, difficulty to adulterate, and demonstration of recent drug use. However, many immunoassays for detecting THC in oral fluid do not have high reliability. This is mainly due to THC adsorption to the collection device, INTRODUCTlON Cannabis is one of the oldest and most commonly abused substances in the world and its use is associated with physical and behavioral toxicity. The plant Cannabis Sativa contains more than 400 chemicals but the cannabinoid δ-9-tetrahydrocannabinol (THC) is the major psychoactive constituent (Fitzgerald et al., 2013). THC is metabolized to 11-hydroxy-THC (11-OH-THC) and 11-nor-9-carboxy-THC (THCCOOH) (Huestis, 2005). Cannabinol, a degradation product of THC oxidation, MEDICOLEGAL ASPECTS OF CANNABIS AND ITS METABOLITES DETECTION WINDOW IN ORAL FLUID AND URINE