A polydrug intoxication involving methoxetamine in a drugs and driving case.

Methoxetamine ((RS)2-(3-methoxyphenyl)-2-(ethylamino)cyclohexanone)) is becoming a drug of interest among practitioners of forensic toxicology. In this case report, we describe the case background, standard field sobriety tests, sampling, and analysis of this drug in a whole blood sample as well as screening methods and analysis from a driver operating under the influence of intoxicating substances. Methoxetamine was isolated from the blood sample using mixed mode solid phase extraction. After elution and evaporation, the residue was dissolved in mobile phase (consisting of acetonitrile and aqueous formic acid) for analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry (GC-MS). The case sample was found to contain clonazepam, 7-aminoclonazepam, carboxy-THC, Ddphenhydramine, and MDMA. The case sample was found to contain 10 ng/mL of the drug (methoxetamine) in whole blood. The results of this drug analysis and previous analyses are discussed in terms of this driver operating under the influence of drugs.

Analysis of AM-2201 and metabolites in a drugs and driving case.

This case report describes the analysis of AM-2201 in plant material and its metabolites in human urine obtained from an operator of a motor vehicle in the United States. The samples were taken from the driver because of his illegal driving activities and, his subsequent erratic behaviour. The AM-2201 was extracted from a sample of plant material by sonicating it in methanol, after which an aliquot was taken and diluted with aqueous phosphate buffer (pH 6) and extracted by solid-phase extraction using a C8/aminopropyl SPE cartridge. The cartridges were washed, dried, and eluted with ethyl acetate- methanol solvent system. The urine sample was hydrolyzed with ß-glucuronidase at pH 6.8 before being diluted with aqueous phosphate buffer (pH 6) and extracted with the same type of SPE cartridge. After evaporation of the eluates, the samples were dissolved in mobile phase for analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Analysis of the plant material determined the concentration to be 0.05% (AM2201) by mass of dry material. The concentration of AM-2201 (AM2201-N-(4-hydroxypentyl ) metabolite in the urine was found to be 3.1 ng/ml. The urine also contained 109 ng/ml of delta-9-tetrahydrocannabinol carboxylic acid but no other drugs including JWH-018 metabolites.

Anion exchange SPE and liquid chromatography-tandem mass spectrometry in GHB analysis.

In this study, the extraction of γ-hydroxybutyrate (GHB) from urine using solid-phase extraction (SPE) is described. SPE was performed on anion exchange columns after samples of urine had been diluted with de-ionized water. After application of the diluted samples containing GHB-d(6) as an internal standard, the sorbent was washed with deionized water and methanol and dried. The GHB was eluted from the SPE column with a solvent consisting of methanol containing 6% glacial acetic acid. The eluent was collected, evaporated to dryness, and dissolved in mobile phase (100 µL) for analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in negative multiple reaction monitoring (MRM) mode. Liquid chromatography was performed in gradient mode employing a biphenyl column and a mobile phase consisting of acetontitrile (containing 0.1% formic acid) and 0.1% aqueous formic acid. The total run time for each analysis was less than 5 min. The limits of detection/quantification for this method were determined to be 50 and 100 ng/mL, respectively. The method was found to be linear from 500 ng/mL to 10,000 ng/mL (r(2)>0.995). The recovery of GHB was found to be greater than 75%. In this report, results of authentic urine samples analyzed for GHB by this method are presented. GHB concentrations in these samples were found to be range from less than 500 ng/mL to 5110 ng/mL.

Solid-phase extraction and analysis of THC and carboxy-THC from whole blood using a novel fluorinated solid-phase extraction sorbent and fast liquid chromatography-tandem mass spectrometry.

In this study, solid-phase extraction (SPE) is described using a novel fluorinated [heptadecafluorotetrahydrodecyl (C(10)H(4)F(17))] phase to isolate THC and its primary metabolite carboxy-THC from whole blood samples. SPE was performed in hydrophobic mode after samples of whole blood were precipitated with acetonitrile. After applying the sample to the SPE column in aqueous phosphate buffer (pH 7), the sorbent was washed with deionized water and phosphate buffer (pH 7) and dried. The SPE column was eluted with a solvent consisting of ethyl acetate/hexanes (50:50) containing 2% acetic acid. The eluate was collected, evaporated to dryness, and dissolved in mobile phase (50 microL) for analysis by fast liquid chromatography-tandem mass spectrometry in positive/negative multiple reaction monitoring mode. Chromatography was performed in gradient mode employing a C(18) column and a mobile phase consisting of acetontitrile (containing 0.1% formic acid) and 0.1% aqueous formic acid. The total run time for each analysis was less than 5 min. The limits of detection/quantification for this method were determined to be 0.1 and 0.25 ng/mL, respectively. The method was found to be linear from 0.25 to 50 ng/mL (r(2) >or= 0.995). Recoveries of the individual cannabinoids were found to be greater than 85%. In this report, results of authentic samples analyzed for THC and carboxy-THC are reported using this new methodology.

Analysis of total caffeine and other xanthines in specialty coffees using mixed mode solid-phase extraction and liquid chromatography-diode-array detection after microwave digestion.

In this study, solid-phase extraction (SPE) in mixed mode operation was employed to isolate xanthines including caffeine and theobromine from milled caffeinated and decaffeinated coffee samples after microwave digestion. 8-Chlorotheophylline was used as an internal standard. SPE was performed in hydrophobic mode using ethyl acetate/methanol (90:10, 2 mL) as the first elution solvent and in ionic exchange mode using ethyl acetate/ acetonitrile/ammonium hydroxide (78:20:2, 3 mL) as the second elution solvent. The eluates were combined, evaporated to dryness and dissolved in aqueous formic acid for analysis. Liquid chromatography with photodiode-array detection was used in isocratic mode employing a C18 column and a mobile phase consisting of acetonitrile/formic acid (0.1% aqueous). The limits of quantitation and detection for this method were 1 and 0.1 mg/L, respectively. The method was linear from 1 to 200 mg/L (r2 > 0.999) with recoveries of the individual xanthines greater than 95%. The decaffeinated coffees contained caffeine at levels less than 0.5 mg/g (range 0.23 to 0.49 mg/g) and caffeinated samples had wide range of levels of caffeine (5.18 to 12.21 mg/g).

Extraction and analysis of clonazepam and 7-aminoclonazepam in whole blood using a dual internal standard methodology.

In this paper, a simple and robust method for the determination of clonazepam and its primary metabolite (7-aminoclonazepam) in whole blood is described. Clonazepam (klonopin) is a popular prescription drug that has been implicated in the field of drug facilitated sexual assaults (DFSA). Clonazepam, 7-aminoclonazepam and the internal standards (deuterated analogues for GC-MS analysis and nitrazepam for analysis by LC-PDA/GC-MS) were spiked into blood samples. The samples were buffered with a pH 6-phosphate solution (5 mL) and extracted from phenyl spe columns. The columns were washed with 5% acetonitrile in pH 6-phosphate buffer (3 mL) and eluted with ethyl acetate (2 x 3 mL). The eluents were evaporated for further chromatographic analysis. For GC-MS, the samples were derivatized prior to analysis. When performed with LC-PDA the samples were reconstituted in distilled water. From this method LOQs of 5 ng/mL of sample is easily achievable by either chromatographic system. By using GC-MS in EI mode, 1 ng/mL of sample can be detected. Data is presented here to show the simplicity and efficiency of the extraction scheme. By employing the properties of GC-MS and LC-PDA, this extraction and analysis procedure further extends the number of tools open to the forensic toxicologist for the analysis of this drug.

Evaluation of solid-phase sorbents for the analysis of ropinirole in whole blood.

In this paper, the extraction and analysis of ropinirole from whole blood using solid-phase cartridges is presented. Previously published methods for the analysis of this drug have employed plasma samples using C(18) cartridges. Liquid-liquid extraction has been employed for analysis of postmortem samples. In the method, drug free blood was spiked with ropinirole (0 to 10 ng) and an internal standard (quinidine). The samples were buffered with distilled water and centrifuged. The supernatant liquid was applied to previously conditioned endcapped C(6), C(18), and C(8)/SCX solid-phase extraction columns. The columns were washed, dried, and eluted with various solvents systems. The eluants were collected and evaporated. The residue was dissolved in 100 microL of aqueous 0.1% trifluoroacetic acid and analyzed by liquid chromatography using a C(18) (4.6 x 150 mm, 5-microm particle size) column and monitored at 250 nm, using diode-array detection. A mobile phase consisting of methanol/0.1% TFA in distilled water (22:78 v/v) was employed. The data was collected and appraised. It was found that 3-mL 200-mg CEC06 C6 (Hexyl endcapped) solid-phase columns that had been washed with 3 x 3 mL water and 3 x 3 mL acetonitrile and eluted with a solvent system consisting of 95:5 v/v acetonitrile/ammonia performed best. The linear range for this analysis was found to be from 0 to 10 ng/mL. The limit of detection was determined to be 1 ng/mL with a limit of quantification of 2.5 ng/mL.

Extraction and analysis of flunitrazepam/7-aminoflunitrazepam in blood and urine by LC-PDA and GC-MS using butyl SPE columns.

The forensic toxicology community has recognized flunitrazepam and its metabolite (7-aminoflunitrazepam) as compounds of concern for several years. In this procedure, the analytes were extracted from whole blood and urine onto single mode solid phase cartridges (butyl) using nitrazepam as an internal standard. The columns were washed with distilled water and hexane. All three compounds were eluted from the sorbent using an ethyl acetate-methanol solvent mixture. After collection and evaporation of the solvent, the residue was dissolved in A, 0.1% (v/v) aqueous trifluoroacetic acid for HPLC-PDA analysis or B, ethyl acetate for derivatization with pentafluoropropionic anhydride (PFPA) for analysis by gas chromatography-mass spectrometry (selected ion monitoring, SIM). A limit of quantitation for this method using HPLC-PDA was found to be 5 and 1.0 ng mL(-1) by SIM.