Journal of Analytical Toxicology

Jones, A. Wayne; Holmgren, Anita; Kugelberg, Fredrik C.

doi: 10.1093/jat/32.4.265pmid: 18430293

Morphine and codeine are frequently identified in blood samples from impaired drivers. But whether these opiates reflect the use of prescription analgesics or abuse of the illicit drug heroin (diacetyl morphine) is not always obvious. Opiates, either alone or together with other drugs, were determined in 2573 blood specimens from impaired drivers by sensitive and specific methods of analysis. The specific metabolite of heroin 6-acetyl morphine (6-AM) was quantifiable in only 52 cases (2%) at mean, median, and highest concentrations of 0.015, 0.010, and 0.10 mg/L, respectively. The mean, median, and highest concentrations of morphine were 0.046, 0.03, and 1.13 mg/L, respectively (N = 2029). The corresponding concentrations of codeine (N = 1391) were 0.047, 0.01, and 2.40 mg/L. Ethyl morphine was identified in 63 cases at a mean concentration of 0.055 mg/L (median 0.03 mg/L). When 6-AM was present in urine (N = 324), the mean morphine/codeine ratio in blood was 7.5 (median 6.7), and this important ratio was less than unity in only two cases. This study finds compelling evidence that ∼90% of apprehended drivers in Sweden with morphine and codeine in their blood had used heroin.

Silva, Lalith K.; Wilburn, Clayton R.; Bonin, Michael A.; Smith, Mitchell M.; Reese, Katherine A.; Ashley, David L.; Blount, Benjamin C.

doi: 10.1093/jat/32.4.273pmid: 18430294

Widespread use of fuel oxygenates, coupled with their high water solubility and slow degradation rate, have led to an increase in the potential for human exposure. We developed an accurate, precise, sensitive, and high-throughput analytical method to simultaneously quantify trace levels (low parts-per-trillion) of four fuel oxygenates in human blood: methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), di-isopropyl ether (DIPE), and tert-amyl methyl ether (TAME). The analytes were extracted from the head space above human blood samples, using solid-phase microextraction, desorbed into the heated injector, and chromatographically resolved by capillary gas chromatography. Analytes were detected by high-resolution mass spectrometry with multiple ion monitoring, and quantified against known standard levels by use of stable isotope-labeled internal standards for recovery correction. The low limits of detection (0.6 ng/L) allowed for measurement of MTBE, ETBE, DIPE, and TAME in parts-per-trillion levels with excellent precision (coefficient of variation ranging from 1.7 to 5.4%) and accuracy (96–100%). This method provides a means to assess fuel oxygenate exposure and study the potential relationship between exposure and adverse health outcomes.

Pappas, R.S.; Stanfill, S.B.; Watson, C.H.; Ashley, D.L.

doi: 10.1093/jat/32.4.281pmid: 18430295

The extent to which smokeless tobacco endangers human health is an ongoing subject of debate. Studies have shown that smokeless tobacco products contain high levels of biologically available nicotine and tobacco-specific nitrosamines. Toxic metals in smokeless tobacco products have been less extensively studied. In this study, concentrations of arsenic, barium, beryllium, cadmium, chromium, cobalt, lead, and nickel were measured in snuff products and iqmik tobacco, a product popular among some Alaska Natives. The average arsenic, cadmium, lead, and nickel concentrations in 17 commercially available brands were 0.23 ± 0.06 µg/g, 1.40 ± 0.31 µg/g, 0.45 ± 0.13 µg/g, and 2.28 ± 0.36 µg/g, respectively. In 17 iqmik tobacco samples, the average arsenic, cadmium, lead, and nickel concentrations were 0.19 ± 0.06 µg/g, 1.41 ± 0.56 µg/g, 0.55 ± 0.19 µg/g, and 2.32 ± 1.63 µg/g, respectively. Using artificial saliva, the extractable levels of beryllium and lead were relatively low and consistent, whereas barium extracted from tobacco samples ranged from 2 to 21%. The group 1 and 2B carcinogens cadmium, cobalt, and nickel were more efficiently extracted by artificial saliva (30–65% of the cobalt, 20–46% of the nickel, and 21–47% of the cadmium).

Fu, Shanlin; Lewis, John

doi: 10.1093/jat/32.4.292pmid: 18430296

An automated extraction method for extracting the major urinary metabolite of cannabis, 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) was developed on the four-probe Gilson ASPEC XL4™ solid-phase extraction (SPE) system. The method works on liquid-liquid extraction principles but does not require the use of SPE cartridges. The limits of detection and quantitation and the upper limit of linearity (ULOL) of the developed method were found to be 1, 2, and 1500 ng/mL, respectively. There was no detectable carry over after 10,000 ng/mL analyte. For a batch of 76 samples, the process uses less than 100 mL methanol, 450 mL extracting solvent hexane/ethyl acetate (5:1, v/v) and 1 L rinsing solvent, 30% methanol in water. The automated extraction process takes 5 h to complete. Precision and accuracy of the method are comparable to both manual liquid-liquid extraction and automated SPE methods. The method has proven to be a simple, speedy, and economical alternative to the currently popular automated SPE method for the quantitative analysis of urinary THC-COOH.

De Paoli, Giorgia; Bell, Suzanne

doi: 10.1093/jat/32.4.298pmid: 18430297

GHB and its related compounds have been known for years in forensic toxicology because of their illicit use in drug-facilitated sexual assault, and to a lesser extent, as party drugs. A sensitive and specific method for the identification and quantification of gamma-hydroxybutyric acid (GHB) in saliva has been developed using gas chromatography-mass spectrometry with selective ion monitoring mode. One microliter of synthetic saliva was spiked with 1.0 µL of GHB-d6 as the internal standard and 1.0 µL of 1,7-heptanediol as a surrogate spike to all samples. After a silyl-derivatization, the sample was injected at a split ratio of 10:1. The following ions were monitoring: 233 and 234 for GHB; 239, 240, and 241 for GHB-d6; and 55, 73, and 97 for 1,7-heptanediol. The limit of quantitation was determined to be 0.5 mg/L with a linear dynamic range of 0.5–50.0 mg/L. Quality control samples (5.0, 20.0, and 30.0 mg/L) were prepared for the evaluation of precision. Analytical precision measured by coefficients of variation ranged from 2.1% to 12.50% in both intraday and day-to-day experiments. Surrogate recovery from saliva samples fell in the range of 94.6% to 100% with an average of 98.37% and a corresponding percent relative standard deviation of 1.2%.

Hamelin, G.; Charest-Tardif, G.; Krishnan, K.; Cyr, D.G.; Charbonneau, M.; Devine, P.J.; Haddad, S.; Cooke, G.M.; Schrader, T.; Tardif, R.

doi: 10.1093/jat/32.4.303pmid: 18430298

A sensitive and reproducible procedure using gas chromatography coupled with mass spectrometry is described for the determination of p-tert-octylphenol (OP), a persistent degradation product of alkylphenol ethoxylates that binds to the estrogen receptor in blood and tissues. The first step involved the extraction of blood (200 µL) or tissue homogenate (400 µL) with methyl tert-butyl ether, including p-tert-butylphenol (BP) as internal standard. After extraction, the sample was evaporated to dryness with a gentle stream of nitrogen at 45°C, and OP and BP were derivatized with an acetylation reaction involving acetic anhydride and catalyzed by pyridine. Samples were then analyzed by a gas chromatograph equipped with a mass spectrometer (single ion monitoring) with a Varian VF-5ms capillary column. The limit of detection and the limit of quantification of the method in blood were 4.6 and 15.5 ng/mL, respectively. The linearity and reproducibility of the method were acceptable, with coefficients of variation of approximately 10% for blood and ranging between 9% and 27% for tissues. This method was applied to the determination of unchanged OP in blood and tissues obtained from Sprague-Dawley rats after oral and IV OP administration.

Heitland, Peter; Köster, Helmut D.

doi: 10.1093/jat/32.4.308pmid: 18430299

A fast and reliable high-pressure liquid chromatography (HPLC)-inductively coupled plasma-mass spectrometry (ICP-MS) routine method was developed for the determination of inorganic arsenic [As(III) and As(V)], organic monomethylarsonate [MMA(V)], dimethylarsinate [DMA(V)], and arsenobetaine (As-B) in human urine. The complete method validation is described, including internal and external quality assurance. Limits of quantification for the As species are 0.1 µg/L, which is sufficient to determine background concentrations of the arsenic species in human urine. Additionally, total As in all urine samples was determined by conventional ICP-MS. Mean concentrations for 82 non-exposed inhabitants from northern Germany are 12.7, 5.9, 4.0, 0.23, 0.52, and 0.17 µg/L for total As, As-B, DMA(V), As(III), MMA(V), and As(V), respectively. Approximately 15% of the total As was not identified by the anion exchange HPLC-ICP-MS method, and could be other As metabolites in urine. Two case studies underline the need for As speciation, especially when total urinary arsenic concentrations are elevated. In the first case, we investigated the effect of seafood consumption on the concentration of different arsenic species in urine for different persons. A maximum enhancement of total As from 1 up to 2200 µg/L (2000 µg/L for As-B) was observed after a normal fish meal. The second case describes the exposure of a 7-year-old child to As(III) by inhalation of calcium arsenite powder. Five hours after exposure, the concentrations in the child's urine for As-B, DMA(V), As(III), MMA(V), and As(V) were < 0.1, 189, 304, 229, and 27 µg/L, respectively, and these concentrations were reduced to normal background values after 4 days.

Reisfield, Gary M.; Haddad, Judella; Wilson, George R.; Johannsen, Laura M.; Voorhees, Kathryn L.; Chronister, Chris W.; Goldberger, Bruce A.; Peele, James D.; Bertholf, Roger L.

doi: 10.1093/jat/32.4.315pmid: 18430300

Cone, Edward J.; Caplan, Yale H.; Moser, Frank; Robert, Tim; Black, David

doi: 10.1093/jat/32.4.319pmid: 18430301

A minor pathway for the biotransformation of morphine to hydromorphone has been identified in humans. Recently, an unsubstantiated claim that morphine is metabolized to hydromorphone and then to oxymorphone was published. The goal of this study was to determine if credible evidence that oxymorphone is a metabolite of either morphine or hydromorphone exists. Urine specimens from pain patients who were treated exclusively with high daily doses of morphine (N = 34) or hydromorphone (N = 26) were analyzed by liquid chromatography-tandem mass spectrometry for oxymorphone, hydromorphone, and morphine (LOD = 25 ng/mL). Specimens were also tested for a variety of other medications. Criteria for inclusion of patients' specimens were as follows: 1. patients were undergoing exclusive dosing with either morphine or hydromorphone; 2. non-prescribed medications were not detected; and 3. urine concentrations of morphine were > 100,000 ng/mL for the high-dose morphine group and > 1000 ng/mL of hydromorphone for the high-dose hydromorphone group. Consistent with earlier reports, hydromorphone was detected in patients treated with high-dose morphine. The ratio of hydromorphone to morphine ranged from 0.2 to 2.2%. Oxymorphone was not detected in any specimen from high-dose morphine or high-dose hydromorphone patients. The authors conclude, based on these data, that oxymorphone is not a metabolite of morphine or hydromorphone.

Garg, Uttam; Frazee, C. Clinton; Kiscoan, Mike; Scott, David; Peterson, Bonita; Cathcart, David

doi: 10.1093/jat/32.4.324pmid: 18430302

The decomposed body of a 45-year-old female was found, face down, in a mobile home, along with a suicide note and two antifreeze containers. Analysis of the body fluid collected from the decedent showed the presence of 58 mg/dL ethanol, but suspected ethylene glycol was not found in the sample. However, an unusually large peak of internal standard, 1,3-propanediol, was found in the sample. Gas chromatography-mass spectrometry analysis confirmed the presence of 1,3-propanediol in the sample. Using gas chromatography-flame-ionization detection, the concentration of 1,3-propanediol was determined to be 445 mg/dL. To our knowledge, this is the first report involving 1,3-propanediol as the cause of death. The study also highlights the importance for the close scrutiny of data, as 1,3-propanediol is a frequently used internal standard for the assay of glycols.