title: Simultaneous determination of delta9-tetrahydrocannabinol and 11-nor-9-carboxy-delta9-tetrahydrocannabinol in human plasma by solid-phase extraction and gas chromatography-negative ion chemical ionization-mass spectrometry.
abstract: Delta9-tetrahydrocannabinol (THC) and 11-nor-9-carboxy-delta9-tetrahydrocannabinol (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 0.1 M 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-d0 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 degrees C, 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 degrees C and the presence of 1% sodium fluoride or the cannabinoid receptor antagonist, SR141716, at 1 microg/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.
authors.affiliation: Center for Human Toxicology, University of Utah, Salt Lake City 84112, USA.
authors.forename: W
authors.initials: W
authors.lastname: Huang
authors.forename: D E
authors.initials: DE
authors.lastname: Moody
authors.forename: D M
authors.initials: DM
authors.lastname: Andrenyak
authors.forename: E K
authors.initials: EK
authors.lastname: Smith
authors.forename: R L
authors.initials: RL
authors.lastname: Foltz
authors.forename: M A
authors.initials: MA
authors.lastname: Huestis
authors.forename: J F
authors.initials: JF
authors.lastname: Newton
mesh_terms: D000328:Adult; D002623:Chemistry Techniques, Analytical / Q000379:methods; D013759:Dronabinol / Q000031:analogs & derivatives* / Q000097:blood*; D008401:Gas Chromatography-Mass Spectrometry; D006213:Hallucinogens / Q000097:blood*; D006801:Humans; D008297:Male; D008385:Marijuana Smoking; D008875:Middle Aged; D012680:Sensitivity and Specificity; D013048:Specimen Handling; D015813:Substance Abuse Detection / Q000379:methods
chemical_list: D006213:Hallucinogens; C016780:11-nor-delta(9)-tetrahydrocannabinol-9-carboxylic acid; D013759:Dronabinol