Determination of 9-THC in Whole Blood using Gas Chromatography-Mass Spectrometry (original) (raw)
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Determination of Δ9-THC in whole blood using gas chromatography-mass spectrometry
Journal of analytical toxicology, 2002
A simple and reliable liquid-liquid extraction method for the determination of A%tetrahydrocannabinol (THC) in whole blood utilizing gas chromatography-mass spectrometry in electron impact mode is described. The substance is derivatized with pentafluoropropionic anhydride in pentafluropropanol. The limit of detection is 0.5 ng/mL for a 1-mL specimen, with recovery greater than 70%. The intra-assay coefficient of variation (CV) is 3.1% to 5.2%, and the interassay CV is 6.4% to 9.5%, calculated at THC concentrations of 1, 5, and 25 ng/mL. The accuracy is between 95 and 97%. The optimization of extraction and derivatization conditions is detailed.
Journal of Chromatography B, 2003
A fully validated, highly sensitive and specific method for the extraction and quantification of 9 -tetrahydrocannabinol (THC), 11-hydroxy-9 -tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-9 -tetrahydrocannabinol (THCCOOH) in plasma is presented. This method incorporates Escherichia coli -glucuronidase hydrolysis to cleave glucuronic acid moieties to capture total analyte concentrations, and simultaneous solid phase extraction (SPE) of the three analytes in a single eluant with separation and quantification on a bench-top positive chemical ionization (PCI) gas chromatography-mass spectrometry (GC-MS) in the selected ion monitoring (SIM) mode. Quantitation was achieved by the addition of deuterated analogues for each analyte as internal standards (IS). Limits of quantitation (LOQ) were 0.5, 0.5 and 1.0 for THC, 11-OH-THC and THCCOOH, respectively, with linearity ranging up to 50 ng/ml for THC and 11-OH-THC, and 100 ng/ml for THCCOOH. Absolute recoveries ranged from 67.3 to 83.5% for all three analytes. Intra-assay accuracy and precision ranged from 1.2 to 12.2 and 1.4 to 4.7%, respectively. Inter-assay accuracy and precision ranged from 1.4 to 12.2 and 3.1 to 7.3%, respectively. This method was used to analyze plasma samples collected from individuals participating in a controlled oral THC administration study. Statistically significant (P ≤ 0.05) increases of 40% for 11-OH-THC and 42% for THCCOOH concentrations were found between hydrolyzed and non-hydrolyzed results. This method will be utilized in ongoing controlled cannabinoid administration studies and may be a useful analytical procedure for the fields of forensic toxicology and cannabinoid pharmacology.
Journal of Chromatography A, 2007
A two-dimensional (2D) gas chromatography/electron impact-mass spectrometry (GC/EI-MS) method for simultaneous quantification of 9 -tetrahydrocannabinol (THC), 11-hydroxy-9 -tetrahydrocannabinol (11-OH-THC), and 11-nor-9 -tetrahydrocannabinol-9-carboxylic acid (THCCOOH) in human plasma was developed and validated. The method employs 2D capillary GC and cryofocusing for enhanced resolution and sensitivity. THC, 11-OH-THC, and THCCOOH were extracted by precipitation with acetonitrile followed by solid-phase extraction. GC separation of trimethylsilyl derivatives of analytes was accomplished with two capillary columns in series coupled via a pneumatic Deans switch system. Detection and quantification were accomplished with a bench-top single quadrupole mass spectrometer operated in electron impact-selected ion monitoring mode. Limits of quantification (LOQ) were 0.125, 0.25 and 0.125 ng/mL for THC, 11-OH-THC, and THCCOOH, respectively. Accuracy ranged from 86.0 to 113.0% for all analytes. Intra-and inter-assay precision, as percent relative standard deviation, was less than 14.1% for THC, 11-OH-THC, and THCCOOH. The method was successfully applied to quantification of THC and its 11-OH-THC and THCCOOH metabolites in plasma specimens following controlled administration of THC.
Journal of Analytical Toxicology, 2001
Ag.Tetrahydrocannabinol (THC) and 11-nor.9-carboxy-A9tetrahydrocannabinol (THCA) in human plasma can be simultaneously detected using solid.phase extraction with gas chromatography and negative ion chemical ionization mass spectrometry. THC-d3 and THCA-d3 are added as internal standards; protein is precipitated with acetonitrile and the resulting supernatants diluted with 0.1M sodium acetate (pH 7.0) prior to application to the solid-phase extraction columns. THC and THCA were eluted separately and then pooled, dried under air, and derivatized with trifluoroacetic anhydride and hexafluoroisopropanol. The derivatized THC-d0 gives abundant molecular anions (m/z 410), and the derivatized THCA-do gives abundant fragment ions (m/z 422) formed by loss of (CF3)2CHOH from its molecular anion. The recoveries of THC and THCA were 74% and 17%, respectively. The lower and upper limits of quantitation were 0.5 and 100 ng/mL for THC and 2.5 ng/mL and 100 ng/mL for THCA. The within-run accuracy and precision for THC (measured at 0.5, 1, 10 and 75 ng/mL) ranged from 98 to 106% (% target) and 4.1 to 9.5 (%CV), respectively. For THCA, the within-run accuracy and precision (measured at 2.5, 5, 10, and 75 ng/mL) ranged from 89 to 101% and 4.3 to 7.5%, respectively. The between-run accuracy and precision for THC ranged from 92 to 110% and 0.4 to 12.4%, respectively. The between-run accuracy and precision for THCA ranged from 97 to 103% and 6.5 to 12.3%, respectively. In processed samples stored in reconstituted form at -20~ THC and THCA were stable for at least three days. THC and THCA stored in plasma were stable following three freeze/thaw cycles. THC and THCA in whole blood at room temperature for 6 h, or in plasma stored at room temperature for 24 h, did not show significant change. Storage in polypropylene containers for 7 days at -20~ and the presence of 1% sodium fluoride or the cannabinoid receptor antagonist, SR141716, at I pg/mL did not interfere with the quantitation of THC and THCA. In three individuals who smoked marijuana under controlled dosing conditions, peak THC concentrations of 151, 266, and 99 ng/mL were seen in the first plasma samples drawn immediately after the end of smoking, and corresponding peak THCA concentrations of 41, 52, and 17 ng/mL occurred at 0.33 to 1 h after cessation of smoking.
Journal of Chromatography B, 2008
Cannabis is considered to be the most widely abused illicit drug in Europe. Consequently, sensitive and specific analytical methods are needed for forensic purposes and for cannabinoid pharmacokinetic and pharmacodynamic studies. A simple, rapid and highly sensitive and specific method for the extraction and quantification of 9 -tetrahydrocannabinol (THC), 11-hydroxy-9 -tetrahydrocannabinol (11-OH-THC) and 11-nor-9-carboxy-9 -tetrahydrocannabinol (THC-COOH) in blood is presented. The method was fully validated according to international guidelines and comprises simultaneous liquid-liquid extraction (LLE) of the three analytes with hexane:ethyl acetate (90:10, v/v) into a single eluant followed by separation and quantification using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Chromatographic separation was achieved using a XBridge C 18 column eluted isocratically with methanol:0.1% formic acid (80:20, v/v). Selectivity of the method was achieved by a combination of retention time, and two precursor-product ion transitions. The use of the LLE 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 250 L of blood. The method was linear over the range investigated (0.5-40 g/L for THC, 1-40 g/L for 11-OH-THC, and 2-160 g/L for THC-COOH) with excellent intra-assay and inter-assay precision; relative standard deviations (RSDs) were <12% for THC and 11-OH-THC and <8% for THC-COOH for certified quality control samples. The lower limit of quantification was fixed at the lowest calibrator in the linearity experiments. No instability was observed after repeated freezing and thawing or in processed samples. The method was subsequently applied to 63 authentic blood samples obtained from toxicology cases. The validation and actual sample analysis results show that this method is rugged, precise, accurate, and well suited for routine analysis.
Analytical and Bioanalytical Chemistry, 2010
A sensitive analytical method for simultaneous quantification of cannabidiol (CBD), Δ 9tetrahydrocannabinol (THC), 11-hydroxy-THC (11-OH-THC), and 11-nor-9-carboxy-THC (THCCOOH) in plasma is presented for monitoring cannabinoid pharmacotherapy and illicit cannabis use. Analytes were extracted from 1 mL plasma by solid phase extraction, derivatized with N, O,-bis(trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane, and analyzed by two-dimensional gas chromatography mass spectrometry (2D-GCMS) with cryofocusing. The lower calibration curve was linear from 0.25-25 ng/mL for CBD and THC, 0.125-25 ng/mL for 11-OH-THC and 0.25-50 ng/mL for THCCOOH. A second higher linear range from 5-100 ng/ mL, achieved through modification of injection parameters, was validated for THC, 11-OH-THC and THCCOOH and was only implemented if concentrations exceeded the lower curve upper limit of linearity. This procedure prevented laborious re-extraction by allowing the same specimen to be re-injected for quantification on the high calibration curve. Intra-and inter-assay imprecision, determined at four quality control concentrations, were <7.8% CV. Analytical bias was within ±9.2% of target and extraction efficiencies were >72.9% for all analytes. Analytes were stable when stored at 22°C for 16h, 4°C for 48h, after three freeze-thaw cycles at −20°C and when stored on the autosampler for 48h. This sensitive and specific 2D-GCMS assay provides a new means of simultaneously quantifying CBD, THC and metabolite biomarkers in clinical medicine, forensic toxicology, workplace drug testing, and driving under the influence of drugs programs.
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. #
Journal of Analytical Toxicology, 2008
9tetrahydrocannabinol (THC), 11-nor-9-carboxy-∆ ∆ 9tetrahydrocannabinol (THCA), and 11-hydroxy-∆ ∆ 9tetrahydrocannabinol (11-OH-THC) in whole blood has been developed and validated using liquid chromatography with tandem mass spectral detection (MS). Cannabinoids present in the blood samples were quantified using solid-phase extraction followed by MS detection in positive electrospray ionization mode. For confirmation, two transitions were monitored and one ratio determined. Samples being reported as positive were required to have both transitions present, the ratio of quantifying transition to qualifying transition being within 20% of that determined from known calibration standards. The monitoring of the qualifying transition and requirement for its presence within a specific ratio to the primary ion has the potential of limiting the sensitivity of the assay, however, the additional confidence in the final result as well as forensic defensibility were considered to be of greater importance. The limit of quantitation was 0.5 ng/mL for THC, 5 ng/mL for THCA, and 2 ng/mL for 11-OH-THC. The limit of detection was 0.5 ng/mL for THC, 4 ng/mL for THCA, and 1 ng/mL for 11-OH-THC. The percentage recovery of the cannabinoids from whole blood at a concentration of 5 ng/mL was 71.5% for THC, 64.5% for 11-OH-THC, and 61.2% for THCA (n = 3).
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.
Journal of Pharmaceutical and Biomedical Analysis, 2008
Recently, electrospray ionization mass spectroscopy (ESI-MS) has been widely used for the identification of drugs of abuse and their metabolites in biological samples. However, the sensitivity and selectivity of this technique are commonly inadequate for the analysis of tetrahydrocannabinol (THC) and its metabolites at very low levels, such as those sometimes required in forensic and clinical-legal applications. We coupled electrospray ionization and surface-activated chemical ionization (ESI-SACI) to various types of mass analyzers (ion trap, triple quadrupole and orbitrap) (ESI-SACI-MS) to improve the detection of 11-nor-9-carboxy-tetrahydrocannabinol (THC-COOH), the most common marker of THC abuse. The benefits of this approach in terms of sensitivity and selectivity compared with a common ESI-MS approach are clearly demonstrated.