Journal of Analytical Toxicology

Cody, John, T.;Schwarzhoff,, Robert

doi: 10.1093/jat/17.6.321pmid: 8271777

Abstract Interpretation of drug testing results is a challenging and complex task, particularly when the interpretation can result in establishing legitimate use of a drug or illicit use with all of its attendant complications (i.e., loss of job, criminal prosecution, etc.). One of the more challenging drugs to interpret is methamphetamine. While methamphetamine is a schedule II controlled substance, the l-enantiomer of methamphetamine is found in the Vick's inhaler, which is a product exempted from control. For this reason, while identification of methamphetamine and amphetamine in the urine of an individual can clearly establish the use of methamphetamine, it does not prove the use of a controlled substance. Use of racemic methamphetamine can make the interpretation even more difficult because of the different metabolism and excretion of l- and d-methamphetamine. Enantiomeric characterization of methamphetamine may not give unequivocal results. Evaluation of experimentally derived and published data from urine samples containing l- and d,l-methamphetamine indicates that use of the enantiomeric distribution of amphetamine affords unambiguous interpretation. Because the l-enantiomer is the only possible finding in an individual who is using the Vick's inhaler, detection of the d-enantiomer or a mixture of the d-and l-enantiomers clearly establishes the use of a controlled substance. Without a prescription from appropriate medical personnel, this detection would indicate the illicit use of a controlled substance. * The views expressed in this article are those of the authors and do not reflect the official policy of the Department of Defense or other Departments of the U.S. Government. This content is only available as a PDF.

Cone, Edward, J.;Holicky, Barbara, A.;Grant, Terrance, M.;Darwin, William, D.;Goldberger, Bruce, A.

doi: 10.1093/jat/17.6.327pmid: 8271778

Abstract The purity of illicit heroin in the United States has increased steadily over the last several years, while prices have fallen. Associated with this trend, there has been a recent shift among heroin addicts from Intravenous injection to intranasal use (“snorting”). Because of the lack of information on this route of administration, we evaluated the pharmacokinetic and pharmacodynamic properties of intranasal heroin. Results were compared to the effects of heroin by the intramuscular route. Six healthy, male volunteers were administered single doses of intranasal heroin hydrochloride (6 and 12 mg), intramuscular heroin hydrochloride (6 mg), and placebo. Blood levels of heroin, 6-acetylmorphine, and morphine were measured by gas chromatography/mass spectrometry. Simultaneous physiological, behavioral, and performance measures were obtained. Peak blood levels of heroin were attained within 5 min of heroin administration by the intranasal route, similar to those observed for intramuscular administration. Generally, the pharmacokinetic profile of intranasal heroin was equivalent to that for the intramuscular route. Physiological, behavioral, and performance effects following intranasal administration were similar to the effects following intramuscular administration. The relative potency of intranasal heroin was estimated to be approximately one-half that of intramuscular administration. The efficacy of the intranasal route, combined with decreased heroin cost, reduced fear of infection, and the lack of requirements for additional drug paraphernalia, could make this an attractive route of drug administration to naive or infrequent drug users. This content is only available as a PDF.

Kato,, Kenichi;Hillsgrove,, Mary;Weinhold,, Linda;Gorelick, David, A.;Darwin, William, D.;Cone, Edward, J.

doi: 10.1093/jat/17.6.338pmid: 8271779

Abstract The accessibility of saliva for rapid, noninvasive sampling makes it an attractive biological fluid for detecting drug use. However, little is known about salivary excretion patterns of the major cocaine metabolites, benzoylecgonine (BE) and ecgonine methyl ester (EME). Additionally, there is a general lack of information on the effects of salivary collection conditions on cocaine excretion in saliva. This study documents the profile of cocaine and metabolites in human saliva under stimulated and nonstimulated saliva flow conditions. Saliva samples were obtained periodically from six healthy volunteers who were administered three, equally spaced, single intravenous doses of 25 mg of cocaine during a 6-h test session. On different days, whole saliva was obtained either under nonstimulated or stimulated (sour candy) conditions. The samples were analyzed for cocaine and metabolites by GC/MS. Cocaine, BE, and EME were detected and quantitated in the saliva of all subjects. Cocaine was the predominant analyte identified in all samples. Nonstimulated saliva contained substantially more drug than stimulated samples. The ratio of the area under the curve (AUC) of cocaine in nonstimulated saliva to that of stimulated saliva was variable and ranged from 3.0 to 9.5. The AUC ratios of BE and EME were similar to those observed for cocaine. The lowering of cocaine concentration in saliva in the stimulated flow condition was likely due to an increase in saliva pH associated with increased saliva flow rate; it is known that an increase in saliva pH retards cocaine partitioning into this biological fluid. Generally, the results of this study indicated that cocaine is the predominant analyte in saliva and that concentrations of cocaine and metabolites can be influenced substantially by the method of collection. These factors should be taken into account in the design of saliva tests for detection of cocaine exposure. This content is only available as a PDF.

Fitzgerald, Robert, L.;Rexin, Douglas, A.;Herold, David, A.

doi: 10.1093/jat/17.6.342pmid: 8271780

Abstract The application of gas chromatography/mass spectrometry (GC/MS) for the simultaneous quantitation of seven commonly encountered urinary benzodiazepine metabolites is described. After comparison of the signal-to-noise ratios of high mass ions of benzodiazepines using electron impact (EI), positive chemical ionization (PCI), and negative chemical ionization (NCI), NCI was chosen because of its increased sensitivity, which ranged from four to several thousand times that of either PCI or EI. This method is novel because NCI spectra for many of these compounds have not been described. For quantitation of benzodiazepines in urine, sample preparation consisted of enzymatic hydrolysis, liquid-liquid extraction, and reaction with a silylating reagent to form trimethylsilyl derivatives. The extraction efficiency of the method was greater than 70% (range, 73–89%) for nordiazepam, oxazepam, temazepam, iorazepam, N-1-hydroxyethylflurazepam, α-hydroxyal-prazolam, and (α-hydroxytriazolam; the linear range for these compounds was from 50 to 2000 ng/mL. Within-run precision was less than 6% for all analytes in the range 50–2000 ng/mL; however, run-to-run precision ranged from 3 to 21%, depending on the analyte and concentration. Quantitation was based on area ratio of high mass ions relative to deuterated internal standards, acquired by scanning the mass range from m/z 250 to 450. Because these studies were performed in the scan mode, if desired, the sensitivity could be increased by using selected ion monitoring. This content is only available as a PDF.

Steele, Bernard, W.;Bandstra, Emmalee, S.;Wu,, Niou-Ching;Hime, George, W.;Hearn, W., Lee

doi: 10.1093/jat/17.6.348pmid: 8271781

Abstract Meconium has been reported to be a more suitable specimen than maternal or neonatal urine for detecting fetal exposure to cocaine. In a study comparing various immunoassays with gas chromatography/mass spectrometry (GC/MS), several unexplained discrepancies among the assays were noted. Using methanol extracts of meconium samples, an immunoreactive spot that was more polar than benzoylecgonine was detected by thin-layer chromatography (TLC). An extract of this spot analyzed by GC/MS yielded a fragmentation pattern indicative of an aryl hydroxylated benzoylecgonine. Standards of m-hydroxybenzoylecgonine, o-hydroxybenzoylecgonine, and p-hydroxybenzoylecgonine were synthesized; it was determined that m-hydroxybenzoylecgonine had the same retention time and ion ratios as the TLC immunoreactive spot. Furthermore, m-hydroxybenzoylecgonine proved to be immunoreactive. Ten meconium samples immunoreactive for benzoylecgonine were analyzed by GC/MS. Results before and after hydrolysis with β-glucuronidase (type IX) showed free m-hydroxybenzoylecgonine comprising 59 to 94% of the total m-hydroxybenzoylecgonine and showed total m-hydroxybenzoylecgonine values ranging from 0.2 to 6.3 times as high as benzoylecgonine. Therefore, m-hydroxybenzoylecgonine appears to be a quantitatively important cocaine metabolite in meconium, which is responsible for a significant portion of the discrepancy between benzoylecgonine concentrations in meconium extracts as measured by immunoassay and GC/MS. This content is only available as a PDF.

Abusada, Gabi, M.;Abukhalaf, Imad, K.;Alford, Dempsey, D.;Vinzon-Bautista,, Imelda;Pramanik, Arun, K.;Ansari, Naser, A.;Manno, Joseph, E.;Manno, Barbara, R.

doi: 10.1093/jat/17.6.353pmid: 8271782

Abstract A selective solid-phase extraction technique has been applied to the analysis of cocaine and selected cocaine metabolites in meconium, whole blood, and plasma. This technique uses a mixed-mode Bond Elut Certify® column that utilizes the characteristics of hydrophobic and polar interactions and ion exchange chromatography. Following extraction, cocaine, ecgonine methyl ester, benzoylecgonine, and cocaethylene were identified and quantitated using GC/MS. Linear quantitative response curves have been generated for the metabolites over a concentration range of 0–1000 ng/g for meconium and 0–1000 ng/mL for whole blood and plasma. The overall extraction efficiencies, depending on the metabolite, were between 58.1 and 99.7% for meconium, 95.6 and 124.0% for blood, and 86.9 and 128.9% for plasma. Linear regression analyses of the standard curve for the four analytes exhibited correlation coefficients ranging from 0.850 to 0.946 for meconium, 0.939 to 0.993 for whole blood, and 0.981 to 0.996 for plasma. Because of its capability to detect cocaethylene in meconium, blood, and plasma, the procedure can be used to determine if drug exposure occurred during the latter stages of gestation and if it involved only cocaine or a combination of cocaine and ethanol. This content is only available as a PDF. Author notes * G.M. Abusada and I.K. Abukhalaf were equal contributors. † Present address: Med Tox Laboratories, 402 West County Road D, Saint Paul, MN 55112.

Kalasinsky, Kathryn, S.;Levine,, Barry;Smith, Michael, L.;Magluilo,, Joseph;Schaefer,, Teresa

doi: 10.1093/jat/17.6.359pmid: 8271783

Abstract New methods for identification of amphetamines have been employed using gas chromatography/Fourier transform infrared spectroscopy (GC/FTIR). These methods have provided identification of the drug and its metabolites, with detection at the low picogram levels or less than 10 ng/mL. Developments in cryogenic sample deposition for GC/FTIR spectroscopy have increased the sensitivity of GC/FTIR to levels that match or surpass that of gas chromatography/mass spectrometry (GC/MS). This advancement in technology has allowed the highly selective ability of infrared spectroscopy to be used for identification and quantitation in studies where the analytes are in low concentrations. The limits of detection (LOD), quantitation (LOQ), and linearity (LOL), and precision have been determined in this study, and these instrumental parameters have been compared with those of established techniques. * The opinions or assertions contained herein are the private views of the authors and are not to be construed as being official or as reflecting the views of the Department of Defense. This content is only available as a PDF.

Huang,, Wei;Moody, David, E.;Andrenyak, David, M.;Rollins, Douglas, E.

doi: 10.1093/jat/17.6.365pmid: 7903726

Abstract The ability of commercial benzodiazepine immunoassays to detect nordiazepam, lorazepam, alprazolam, and triazolam in blood samples was investigated. Two radioimmunoassays (RIA) (Abuscreen [RIA-a] and the Diagnostic Products Corporation serum kit [RIA-d]), two enzyme immunoassays (EIA) (Emit d.a.u. [EIA-u] and Emit TOX serum assay [EIA-a]), and two fluorescence polarization immunoassays (FPIA) (X-systems urine [FPIA-u] and X-systems serum assay [FPIA-s]) were evaluated for their ability to detect benzodiazepines in fortified drug-free human or bovine blood. Prior extraction of the blood was necessary for analysis on the equipment used for EIA and FPIA. Extraction with an organic solvent, such as butyl chloride used in this study, was preferable to precipitation with methanol or zinc sulfate. For all these assays, extraction eliminated matrix effects and offered the possibility of increased sensitivity by reconstitution of the extract in a smaller volume. Extraction was necessary in bovine blood, as there is some nonextractable substance(s) in this matrix that increases non-specific binding. Using extraction with reconstitution in one-half the original volume, the apparent limit of detection for nordiazepam in blood ranged from 3 ng/mL, with RIA-d, to 30 ng/mL, with EIA-s and FPIA-s. These limits of detection were improved by further reduction of the reconstitution volume. RIA-a, EIA-s, and FPIA-s had cross-reactivity for alprazolam that was equivalent to or slightly better than nordiazepam. RIA-d had enhanced cross-reactivity for alprazolam at 30–300 ng/mL. The crose-reactivity for lorazepam at 70 ng/mL was 24, 7, 38, and 42% for RIA-a, RIA-d, EIA-s, and FPIA-s, respectively. For triazolam at 70 ng/mL, it was 37, 13, 66, and 71% for the same assays. Detection by any of these assays was not affected by aging benzodiazepine-fortified human blood at room temperature for up to five weeks. Commercial immunoassays can be used to detect benzodiazepines in blood. When the blood is extracted with an oganic solvent and reconstituted in a reduced volume, benzodiazepines can be detected at low concentrations. * Portions of this work were presented at the 22nd Annual Meeting of the Society of Forensic Toxicologists, Cromwell, CT, October 1992. This content is only available as a PDF.

Sreenivasam, Renee, C.;Sneath, Thomas, C.;Jain, Naresh, C.

doi: 10.1093/jat/17.6.370pmid: 8271784

Abstract An evaluation study of Syva Emit® II reagents using the Chem 1 was performed for the following drugs: barbiturates, benzodiazepines, cannabinoids, benzoylecgonine, opiates, and phencyclidine. The Emit II reagents (100-mL bottles) were reconstituted to 70 mL and evaluated against the Emit d.a.u. reagents. A minimum of 446 samples were run for each drug. For all drugs tested, there were a total of 11 discordant results between the two reagents. The Emit II reagent was found to be correct on 8 of the 11 discordances after retesting by FPIA or GC/MS. The CV of within-run and day-to-day precision of the Emit II was 1.8% or less and 12.3% or less, respectively. This content is only available as a PDF.

Jortani, Saeed, A.;Poklis,, Alphonse

doi: 10.1093/jat/17.6.374pmid: 8271785

Abstract Quanitation of thioridazine (TRZ) enantiomers, (+)-TRZ and (−)-TRZ, in patient serums was performed by a sequential achiral and chiral HPLC method. The lower limit of quantitation was 50 ng/mL for each enantiomer, and the method was linear up to 1000 ng/mL. The within-run and between-run precision at 125 and 500 ng/mL of each enatiomer yielded coefficients of variation (CV) of less than 10% and less than 12%, respectively. Absolute recovery of 250 ng/mL racemic TRZ added to serum (n = 9) yielded mean recoveries of 77.2% and 77.8% for (+)-TRZ and (−)-TRZ, respectively. Absolute recovery of 1000 ng/mL racemic TRZ added to serum (n = 9) yielded mean recoveries of 74.0% and 74.7% for (+)-TRZ and (−-TRZ, respectively. Significantly different TRZ enantiomer concentrations were present in patient serums. The mean serum concentration was 110 ng/mL for (+)-TRZ (n = 18), and 317 ng/mL for (−)-TRZ (n = 21). The ratios of (−)-TRZ to (+)-TRZ ranged from 2.2 to 5.3, with a mean of 3.3. Variations in the concentration of TRZ enantiomers may contribute to the lack of correlation between serum TRZ values and therapeutic effect. This content is only available as a PDF.