Showing papers in "Journal of Analytical Toxicology in 1993"

Hair analysis for drugs of abuse : evaluation of analytical methods, environmental issues, and development of reference materials

Abstract: Methods for extraction of cocaine, some of its metabolites, morphine, and codeine from hair and methods for analyzing the extracts have been investigated. Results of these studies have shown that extractions with 0.1N HCl are efficient at removing the target compounds from hair and appear to be as effective as enzymatic digestions that dissolve the hair. GC/MS with either electron ionization or chemical ionization was found to provide accurate and unambiguous determinations of the target compounds. Tandem mass spectrometry (MS/MS) also provided accurate results when performed on extracts from hair, but results were ambiguous when MS/MS was performed on hair segments directly. Environmental issues, including the removal of powdered and vapor-deposited cocaine from the hair surface and the effect of various hair treatments on the levels of cocaine entrapped in hair, have also been investigated. Removal of cocaine deposited on hair was incomplete by all approaches tested, making differentiation of hair of cocaine users from hair with environmental exposure of cocaine difficult. Cocaethylene, a cocaine metabolite believed to be formed in the liver, was found in the hair of some cocaine users and may be a good marker for proving drug use. Common hair treatments, such as shampoos, conditioners, and peroxide bleaches, reduced cocaine levels in a fortified hair material by 60 to 80% after 30 treatments. Finally, to assist laboratories in evaluating the accuracy of their methods, two human hair reference materials with recommended concentrations of cocaine, benzoylecgonine, morphine, and codeine determined by GC/MS have been developed.

Cocaine and Metabolite Excretion in Saliva under Stimulated and Nonstimulated Conditions

Abstract: The accessibility of saliva for rapid, nonlnveelve sampling makes It an attractlve blologlcel fluld for detectlng drug use. However, llttle Is known about sallvary excratlon patterns of the major cocalne metabolltes, benzoylecgonlne (BE) and ecgonlne methyl ester (EME). Addltlonally, there Is a general lack of Informstlon on the effects of sallvary collecUon condltlons on cocelne excretion In sallva. Thls study documents the profile of cocaine and metabolltee In human sallva under stlmulated and nonstlmulated sallva flow condltlons. Saliva samples were obtained perlodlcally from six healthy volunteers who were administered three, equally spaced, slngle Intravenous doses of 25 mg of cocelne durlng a 6-h test seeslon. On dlfferent days, whole sallva was obtained either under nonstlmulated or stlmulated (sour candy) conditions. The samples were analyzed for cocalne and metabolites by GCIMS. 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 contalned substantially more drug than stimulated samples. The ratio of the area under the curve (AUC) of cocalne in nonstlmulated saliva to that of stimulated saliva was varlable and ranged from 3.0 to 9.5. The AUC ratlos of BE and EME were similar to those observed for cocalne. The lowering of cocalne concentratlon In saliva In the stimulated flow condltlon was likely due to an increase In sallva pH assoclated with Increased sallva flow rate; it Is known that an increase in saliva pH retards cocalne partitioning Into this blologlcal fluid. Generally, the results of this study indicated that cocaine is the predomlnant analyte In saliva and that concentratlons of cocaine and metabolltes can be influenced substantially by the method of collectlon. These factors should be taken Into account In the deslgn of sallva tests for detection of cocaine exposure.

Pharmacokinetics and Pharmacodynamics of Intranasal “Snorted” Heroin

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.

Performance-based testing for drugs of abuse : dose and time profiles of marijuana, amphetamine, alcohol, and diazepam

Abstract: The time courses of the effects of acute doses of amphetamine (5 and 10 mg/70 kg), alcohol (0.3 and 0.6 g/kg), dlszepam (5 and 10 mg/70 kg), and marijuana (2.0% and 3.5% Ag-THC) on performance engendered by each of four computerized behavioral tasks were evaluated in six human subjects. These performance-based tasks have potential commercial utility for drug-use detection in the workplace. Alcohol and marijuana effects were reliably detected for up to three hours following dose administration with most procedures. Amphetamine and diazepam effects were also detected, but the dose effects and time courses were variable. The profile of behavioral effects varied across drugs, suggesting that performance-based testing procedures might be useful in discriminating which drug was administered and the time course of the drug's effects. Results indicate that repeated measurement with performance-based drug detection procedures can provide immediate indications of performance impairment in a cost-effective and

A class-independent drug screen in forensic toxicology using a photodiode array detector.

Abstract: A class-independent drug screen, that is suitable for use in clinical and forensic toxicology and uses gradient-elution high-performance liquid chromatography and photodiode array detection, is presented. The assay is capable of detecting and identifying therapeutic and toxic amounts of barbiturates, anti-convulsants, diuretics, non-steroidal anti-inflammatory drugs, sulfonylurea anti-diabetic drugs, theophylline, and analgesic drugs. The assay has been successfully used to screen over 1000 postmortem blood specimens.

Determination of Amphetamine and Methamphetamine in Blood by Derivatization with Perfluorooctanoyl Chloride and Gas Chromatography/Mass Spectrometry

Abstract: A relatively simple method for quantitation of amphetamine and methamphetamine in blood was developed. Blood samples were extracted with cyclohexane, and the extracts were derivatized with perfluorooctanoyl chloride prior to gas chromatography/mass spectrometry. Selected ions were monitored at m/z 118 and 440 for amphetamine, m/z 118 and 454 for methamphetamine, and m/z 121 and 443 for the internal standard amphetamine-d3. The imprecision was less than 5% for amphetamine and less than 9% for methamphetamine. Limits of detection were 11 micrograms/L for amphetamine and 13 micrograms/L for methamphetamine, while limits of quantitation were 22 and 34 micrograms/L, respectively. Calibration curves were linear in the ranges 14-2700 micrograms/L and 15-3000 micrograms/L, respectively. Some drugs that were known or suspected to interfere with immunological or chromatographic methods for amphetamine and methamphetamine were tested for interference. None of the drugs tested interfered, except for a substance in putrefied blood samples, which was found to interfere with one ion (m/z 118) for amphetamine but not with the second ion.

Simultaneous Determination of Eight Anticoagulant Rodenticides in Blood Serum and Liver

Abstract: A liquid chromatographic method was developed for the analysis of indandione and 4-hydroxycoumarin anticoagulant rodenticides in blood serum and liver. The method enabled the measurement of serum and liver concentrations of eight anticoagulant rodenticides: brodifacoum, bromadiolone, chlorophacinone, coumafuryl, coumatetralyl, diphacinone, difenacoum, and warfarin. Anticoagulants were extracted from serum and liver with acetonitrile. Extracts were applied to solid-phase extraction columns, which contained mixed packings. Column eluates were evaporated to dryness, reconstituted, and subjected to reversed-phase liquid chromatography. Hydroxycoumarins were detected by fluorescence at an excitation wavelength of 318 nm and an emission wavelength of 390 nm. Indandiones were detected by UV absorption at 285 nm. Extraction efficiencies of greater than 75% for serum and greater than 69% for liver were obtained. The within-run precision (CV) ranged from 2.4 to 8.6% for serum and 2.6 to 8.7% for liver. The between-run precision (CV) ranged from 1.5 to 12.2% for serum and from 2.1 to 11.8% for liver. Hydroxycoumarin rodenticides were detected at 1 ng/mL of serum and 1 ng/g of liver. Indandiones were detected at 10 ng/mL of serum and 10 ng/g of liver.

Solid-Phase Extraction and GC/MS Quantitation of Cocaine, Ecgonine Methyl Ester, Benzoylecgonine, and Cocaethylene from Meconium, Whole Blood, and Plasma

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.

The Effects of Adulterating Agents on FPIA Analysis of Urine for Drugs of Abuse

Abstract: A variety of chemical agents were evaluated to determine their effects on fluorescence polarization immunoassays for drugs of abuse. Sixteen different agents, at concentrations up to 10%, were tested against urine assays for cannablnolds, cocaine (metabolite), amphetamines, opiates, phencyclidlne (PCP), and barbiturates. The potential to cause both false positive and false negative results was evaluated, and assays were performed one and seven days after sample adulteration to simulate different collection/testing formats. All six drug assays were susceptible to one or more adulterating agents, but the degree varied considerably between assays. The cannabinoid assay was most susceptible to adulterantinduced false negative results, and the barbiturate assay was most susceptible to false positive results. The remaining assays demonstrated relatively few, but characteristic effects, some of which were attributable to drug degradation and others to assay interference. Although the results of pH measurement on adulterated samples verified its utility in identifying some samples adulterated with interfering agents, other adulterants that cause substantial effects would not be identified by pH measurements alone.

Simultaneous HPLC Gradient Analysis of 15 Benzodiazepines and Selected Metabolites in Postmortem Blood

Abstract: The simultaneous identification and quantitation of 15 benzodiazepines and selected metabolites in postmortem blood, serum, or liver homogenate is described. The assay involves extraction with diethylether, followed by an acid clean-up step of the ether. Chromatographic separation was achieved on a Nova-Pak phenyl 18 column using ultraviolet detection at 240 nm. A gradient HPLC system was developed to improve separation of nitro-reduction metabolites from the solvent front and endogenous peaks. The mobile phases consisted of a gradient from 15 to 28% acetonitrile in 40 mM potassium phosphate buffer. Within-run and day-to-day precision were generally 10-15%. The method described is sensitive and reproducible for the analysis of benzodiazepine concentrations in postmortem tissues.

Evaluation of the triage system for emergency drugs-of-abuse testing in urine.

Abstract: The Triage Panel for Drugs of Abuse (DOA) was evaluated for detection of phencyclidine (PCP), amphetamines (AMPH), opiates (OPI), tetrahydrocannabinol (THC), the cocaine metabolite benzoylecgonine (BE), and barbiturates (BARB) in urine. This assay was compared with Syva EMIT. The comparisons were conducted on 606 positive and 325 negative samples. When there was a sufficient volume of sample available for retesting, positive and negative samples with discordant results between these two screening assays were confirmed by quantitative gas chromatography/mass spectrometry (GC/MS). The percent agreement between the two assays for positive samples ranged from 93 to 100%. For negative samples, the agreement ranged from 95 to 100%. For AMPH, 19 out of 27 discordant samples of urine that were positive for EMIT and negative for Triage DOA contained total amphetamine and methamphetamine concentrations of less than 1000 ng/mL by GC/MS. For THC, seven urine samples that were negative for Triage DOA and positive for EMIT contained 9-carboxy THC concentrations at greater than 15 ng/mL by GC/MS. For BARB, three samples that were negative for Triage DOA and positive for EMIT contained barbiturates levels greater than 300 ng/mL. Two of these three samples contained phenobarbital below the concentration that produces a positive result for Triage DOA. For the majority of the urine samples studied, however, the Triage DOA produced identical results to a commercial-instrument-based immunoassay. Language: en

Interpretation of Methamphetamine and Amphetamine Enantiomer Data

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 methamphetamlne. While methamphetamine is a schedule II controlled substance, the l..enantiomer of methamphetamine Is found in the Vlck's Inhaler, which is a product exempted from control. For this reason, while Identification of methamphetamlne and amphetamine In the urine of an Individual can clearly establish the use of methamphetamlne, It does not prove the use of s controlled substance. Use of racemic methamphetamlne can make the Interpretation even more difficult because of the different metabolism and excretion of l- and d-methamphetamlne. Enantiomeric characterization of methamphetamlne may not give unequivocal results. Evaluation of experimentally derived and published data from urine samples containing l- and d,lmethamphetamlne Indicates that use of the enantlomerlc distribution of amphetamine affords unambiguous interpretation. Because the !-enantlomer Is the only possible finding in an individual who Is using the Vlck's Inhaler, detection of the d-enantlomer or �9 mixture of the d-and lenantlomers 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.

Fluorescence Polarization Immunoassay Detection of Amphetamine, Methamphetamine, and Illicit Amphetamine Analogues*

Abstract: The Abbott Diagnostics Amphetamine/Methamphetamine II and Amphetamine Class reagents were evaluated on the Abbott TDx for cross-reactivity to amphetamine and methamphetamine stereoisomers, several of their metabolites, and various illicit analogues, including 2-methoxyamphetamine, 4-hydroxymethamphetamine, 2,5-dimethoxy-amphetamine (DMA), 4-bromo-2,5-dimethoxyamphetamine (DOB), 4-bromo-2,5-dimethoxy-beta-phenethylamine (BDMPEA), 3,4,5-trimethoxyamphetamine (TMA), 3,4-methylenedioxy-amphetamine (MDA) N,N-dimethyl-3,4-methylenedioxy-amphetamine, N-hydroxy-3,4-methylenedioxyamphetamine (N-OH MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), 2,5-dimethoxy-4-ethylamphetamine (DOE), 2,5-dimethoxy-4-methylamphetamine (DOM), and mescaline in concentrations ranging from 100 to 100,000 ng/mL. Results demonstrate the utility of this assay for detection of several of the above compounds; unfortunately many are still not detectable. Significant differences were observed between the Amphetamine/Methamphetamine II and Amphetamine Class reagents, particularly regarding their cross-reactivity to over-the-counter medications. Detection of the drugs amphetamine, methamphetamine, and the illicit analogues is not enhanced with the Amphetamine Class reagents, and unless detection of the over-the-counter compounds is of interest, these reagents are a poor choice compared to the Amphetamine/Methamphetamine II reagents. Cross-reactivity of some of the illicit analogues is such that the assay can reliably be used for the routine screening of these compounds.

Serial Plasma Concentrations of Cocaethylene, Cocaine, and Ethanol in Trauma Victims

Abstract: Cocaethylene (EC), cocaine (COC), and ethanol (ETOH) concentrations were measured in 46 serial plasma samples from 15 male trauma victims. In all but two patients EC detected on admission to the hospital. In the remaining two, EC was not detected throughout hospitalization. However, these two patients had the lowest ETOH concentrations In the series (200 and 300 mg/L). Plasma concentrations ranged up to 128 lag/L for EC, up to 421 ~g/L for COC, and up to 5,100 mg/L for ETOH. Ratios of EC to COC concentration (EC/COC) were as high as 4.1. Concentrations of EC showed statistically significant correlation with those of COC (p < 0.01). The plasma half-life (Ttt=) of EC was found to be longer than that reported for COC (1 h) and was estimated to be 3.5, 4.5, and 5.5 h In three patients. fects (5). At the same time, the LDs0 of EC is markedly less than that of COC in mice (6). Taken together, these facts suggest that there may be a substantially increased risk of morbidity and mortality as users attempt to titrate their COC intake based on the weaker stimulant effect of EC, during which time additional COC is converted to the more toxic EC (5). Finally, EC demonstrates essentially the same hepatotoxicity as COC in the mouse model (7). The structures of EC, COC, and BE are shown in Figure 1. Published reports containing plasma concentrations of EC have been mostly confined to postmortem material (1,2,8). To date there have been few reports of EC plasma concentrations in living patients, and those reports have been limited to single concentrations per patient (9,10). This study was designed to examine serial plasma concentrations in actual patients in order to estimate the plasma time-concentration profile and half-life of EC.

Detection of a GC/MS Artifact Peak as Methamphetamine

Abstract: Recently, certain laboratories reported the presence of methamphetamine in several negative urine specimens. We have demonstrated an artifact peak, by SIM GC/MS, in negative urine specimens, that frequently matches the retention time and identity ion ratio criteria for methamphetamine. This peak resulted from the presence of high levels of pseudoephedrine (PS) or ephedrine (EP) and was produced after derivatization with 4-carbethoxyhexafluorobutyryl chloride (CB), heptafluorobutyric anhydride (HFB), and N-trifluoroacetyl-l-prolyl chloride (TFAP). Use of N-methyl trideuterated PS and two deuterated EP compounds in spiked samples provided additional proof that PS and EP can produce the artifact peak. The above results were achieved using a GC injection port temperature of 300 degrees C and low split and isothermal conditions. Thermal transformation in the injector is involved because lowering and raising the injector temperature results in the disappearance and subsequent reappearance of the artifact peak. Proof that the artifact peak is actually methamphetamine was provided by full-scan GC/MS data of the artifact peak produced after use of all three derivatizing reagents. GC/MS analysis has demonstrated that urine (65 specimens) with concentrations of PS or EP ranging from 100,000 to 1,100,000 ng/mL will test negative for methamphetamine, while the same specimens will trigger positive results by radioimmunoassay and the Emit monoclonal assay. Full-scan GC/MS of the CB derivative for PS and EP reveals what appears to be both mono- and diderivatized products. SIM analysis can differentiate between PS and EP in urine specimens because the relative amounts of mono- and diderivatized products are very different for PS than for EP.(ABSTRACT TRUNCATED AT 250 WORDS)

Concentrations of acetone in venous blood samples from drunk drivers, type-I diabetic outpatients, and healthy blood donors

Abstract: Headspace gas chromatography (HSGC) was used to measure the concentrations of acetone in samples of venous whole blood from drunk drivers (n = 500), hospital outpatients with type-I diabetes mellitus (n = 250), and healthy blood donors (n = 288). The standard deviation (SD) of blood-acetone determination by HSGC was 0.048 mg/L at a mean concentration of 2.34 mg/L (2.1%). The concentration of acetone in blood did not change significantly when the samples were stored at 4 degrees C for eight days. The ratio of the concentrations of acetone in plasma and whole blood was 1.23:1 (SD 0.229, n = 22). The frequency distributions of blood-acetone concentrations were markedly skewed to the right. The median concentration of acetone in blood from drunk drivers was 2.03 mg/L and the 2.5 and 97.5 percentiles were 0.80 and 12.8 mg/L, respectively. In patients with type-I diabetes mellitus, the median blood-acetone concentration was 1.90 mg/L and the 2.5 and 97.5 percentiles were 0.40 and 11.1 mg/L, respectively. In healthy blood donors, the median blood-acetone level was 1.26 mg/L and the 2.5 and 97.5 percentiles were 0.37 and 4.69 mg/L, respectively. The concentrations of acetone in blood did not differ appreciably among these three groups of subjects.

Benzodiazepine analysis by negative chemical ionization gas chromatography/mass spectrometry

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, lorazepam, N-1-hydroxyethylflurazepam, alpha-hydroxyal-prazolam, and alpha-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.

Determination of 1-Hydroxypyrene in Human Urine by High-Performance Liquid Chromatography

Abstract: A high-performance liquid chromatography (HPLC)/fluorescence method for quantitative analysis of 1-hydroxypyrene in urine was developed. The method validation analysis showed the method to be in analytical control. No significant systematical errors could be demonstrated. The entire run time of chromatography was 10 min using isocratic elution (acetonitrile-water, 70:30), and the retention time for 1-hydroxypyrene was 3.5 min. The short run time in combination with the low limit detection (1.37 nmol/L) makes the method potentially applicable for surveillance of pyrene exposure in work environments. The developed method is presently used for measurement of 1-hydroxypyrene in urine samples from workers exposed to a low airborne level of polycyclic aromatic hydrocarbons, generally less than 25 micrograms/m3. The urine samples of exposed workers (n = 122) showed a range of 1-hydroxypyrene from the limit of detection (LD) up to 39 nmol/L, whereas unexposed control individuals (n = 108) showed a range from LD to 3.3 nmol/L.

Three fatal drug overdoses involving bupropion

Abstract: Bupropion is a "second generation" antidepressant agent structurally related to the phenethylamines. Postmortem toxicology data are presented from three suicidal drug overdoses in which bupropion was detected. In two cases in which bupropion was the major toxicology finding, peripheral blood levels of the parent drug were 4.0 and 4.2 mg/L and total metabolite levels were 15 and 16.6 mg/L. Lethal doses in both cases were estimated by history to be less than 10 g.

Effect of Freezing on the Concentration of Drugs of Abuse in Urine

Abstract: Benzoylecgonine, 11-nor-delta 9-tetrahydrocannabinol-9-carboxylic acid (THC-acid), phencyclidine, codeine, morphine, amphetamine, methamphetamine, and lysergic acid diethylamide were dissolved in urine, stored frozen in plastic tubes at -16 to -18 degrees C, defrosted, transferred to other tubes, and analyzed by gas chromatography/mass spectrometry (GC/MS); no significant loss of compound was observed, except for THC-acid, which showed an average loss of 11%, ranging from 0 to 34% of the total concentration. The loss is attributed to the decrease in solubility of the compound and to adherence to the side of the container during freezing.

A Rapid Gas Chromatographic Method for Determination of Ethylene Glycol in Serum and Urine

Abstract: Intoxications with the antifreeze constituent ethylene glycol (EG) occur infrequently, but may be fatal If not recognized and treated promptly. The aim of the present work was to develop an analytical method for rapid diagnosis of EG poisoning and for monitoring EG removal by hemodlalysls. EG was measured by gas chromatography upon direct Injection of serum or urine samples (50 ILL diluted In 200 ptL of disUlled water containing 2,3-butanediol as Internal standard). A 2-m x 2-mm glass column with Chromosorb 101 (801100 mesh) separates these glycols within four minutes at 200~ using nitrogen as the carrier gas. The glycols 1,2- and 1,3-propanediol were separated from EG and the internal standard. Acetone, methanol, and isopropanol did not interfere with the analysis. The limit of quantitatlon of EG was close to 0.5mM. Because no derivatizatlon, extraction, or concentration procedures were necessary, EG may be determined quantitatively within 30 min, allowing for monitoring of hemodialysis, which should be performed for 15 h in severe cases. The diagnosis of ethylene glycol intoxication in a late stage may be secured by analysis of urine collected on admission.

Pitfalls and Solutions in the Development of a Fully Automated Solid-Phase Extraction Method for Drug Screening Purposes in Plasma and Whole Blood

Abstract: A fully automated solid-phase extraction (SPE) method for drug screening is described. The extraction of 19 toxicologically relevant drugs from pretreated plasma and pretreated whole blood was accomplished automatically by s Gilson ASPEC system equipped with disposable 2.8-mL Bond Elut Certify columns. The automated extraction procedure includes 11 fundamental steps: column preconditioning; sample application; column washing; pH adjustment; elutlon of drugs by two eluents, which were collected into two separated tubes; addition of the chromatographic standard solution; and several SPE column rack movement steps. After evaporation, the drugs were quantitated by gas chromatography. Water was chosen as the transfer liquid in the ASPEC system because it was cheap and, more importantly, because it caused no protein precipitation problem. The effects of the sample and eluent flow rates were investigated, and it was found that low flow rates were necessary to recover the drugs maximally. In the study, the optimal flow rates of sample application, acetone-chloroform elutlon, and ammoniated ethyl acetate elution were 1.5, 0.72, and 0.33 mL/min, respectively. The absolute recoveries of 19 drugs from whole blood exceeded 82%, with relative standard deviations less than 5% at 2 pg/mL.

Forensic Drug Testing For Opiates. V. Urine Testing for Heroin, Morphine, and Codeine With Commercial Opiate Immunoassays

Abstract: Urine specimens collected after heroin, morphine, and codeine administration were tested by four commercial opiate immunoassays (TDx, CAC, ABUS, and EMIT) and GC/MS. Quantitative immunoassay results (morphine equivalents) were compared with results by GC/MS for total morphine, free morphine, or total codeine. Mean detection times for the broadly cross-reacting immunoassays (TDx, ABUS, and EMIT, 300 ng/mL cutoff) ranged from 15-44 hours following heroin and morphine administration and 33-54 hours following codeine administration. Detection times obtained with CAC (25 ng/mL cutoff) tended to be somewhat shorter as a result of the high selectivity of the antibody for free morphine. High correlations over a wide concentration range were obtained for TDx, CAC, and ABUS versus GC/MS, with specimens collected after heroin and morphine administration. EMIT showed a high correlation over a narrow concentration range (0-1000 ng/mL) with heroin and morphine specimens, but responses plateaued at higher concentrations. There was substantial variability in immunoassay responses with specimens collected after codeine administration. Generally, this study demonstrated that immunoassay responses for opiate urine testing can be used as a semi-quantitative guide for GC/MS confirmation; however, the presence of codeine increased variability and diminished the accuracy of the immunoassay response.

GC/MS Analysis of Five Common Benzodiazepine Metabolites in Urine as tert-Butyl-dimethylsilyl Derivatives

Abstract: A method for the GC/MS confirmation of oxazepam, nordlazepam, deselkylflurazepam, temazepam, and (xhydroxyelprazolem in urine Is described. The method Incorporates a solid phase extraction technique developed by Varlan which was found to extract all five metabolltes with recoveries exceeding 73%. The extracted metabolltes were derivatlzed with N-methyl-N-(tert-butyldlmethylsilyl)trlfluoroecetamlde (MTBSTFA), resulting in the formation of stable tert-butyldlmethylsllyl (TBDMS) derivatives. Quantitatlon was performed by GC/MS using oxazepam-Ds, nordlazepamDs, and (x-hydroxyalprezolam-Ds as internal standards. The method was found to be linear over the range of 50-2000 ng/mL. Within-run precision, measured as the coefficient of variation at 100 ng/mL, was less than 3% for all analytes except temezepem which was 6% (N = 20). prior to GC/MS analysis. This method is convenient because the acid hydrolysis serves both to form the benzophenone derivative and cleave the urinary conjugates, all in the same step; however, not all benzodiazepines form benzophenones, and since many benzodiazepines form the same benzophenone, interpretation of test results is difficult. A number of derivatization procedures have been used to test the intact metabolites following treatment with glucuronidase (5,6). The unconjugated metabolites are purified by liquid-liquid extraction and derivatized to form TMS or acyl derivatives. These procedures are more complex and time consuming but lend themselves to a more direct interpretation. The objective of this work was to utilize recent advances in solid phase extraction technology and new deuterium labeled standards to develop a new GC/MS confirmatory assay with performance characteristics consistent with NIDA testing requirements.

Radioimmunoassay Screening of Dried Blood Spot Materials for Benzoylecgonine

Abstract: used as urinary screening tests followed by confirmation by GC/MS. Immunoassay tests have been used primarily on urine specimens because of the ease of specimen collection and the relative abundance of the metabolite in urine following cocaine use. Levels of BE in blood are lower than those found in urine in concurrent samples, but they remain in the detectable range of immunoassays. The predictable half-life of BE in blood for extended periods has not been calculated, and the relationship between levels in blood and urine has not been correlated. Moreover, metabolism of BE in the blood during pregnancy and its half-life in the blood of newborns have not been investigated. Previously, investigators have described several techniques for the extraction of BE from blood using organic solvent extraction and subsequent assay in an aqueous or methanolic medium (5-7). Here, the authors describe the use of a commercial urinary BE immunoassay procedure with standards, calibrators, and quality control materials (QC) made up in aqueous eluates from blood spots or BE spiked into whole blood, spotted onto filter paper, and then eluted in aqueous solution. Postmortem blood samples were quantitated as blood spots for BE, and quantitation was confirmed by GC/MS analysis. The correlation of blood and urine levels of BE was measured on samples from pregnant women. Blood samples were collected from mothers; cord and blood spots from neonates were measured for BE. Finally, the quantitative immunoassay was used to measure BE levels in more than 500 anonymous residual blood spots obtained routinely from infants born in three states, at hospitals where high prevalence of cocaine use among childbearing women is suspected.

Triazolam blood concentrations in forensic cases in Canada

Abstract: Triazolam has been a controversial drug since its appearance on world markets as a hypnotic more than ten years ago. Whole blood concentrations of triazolam as found in forensic cases are cited in several categories; that is, impaired driving: 17 cases; sexual assault: four cases; death due to drugs: 45 cases; drug-related death (drugs contributed to the death but were not the ultimate cause): 20 cases; drug-involved death (drugs were present but were not felt to be a contributing factor): six cases; miscellaneous: one case. The data was gleaned from a forensic toxicology database designed and used by the Forensic Toxicology Sections of the Royal Canadian Mounted Police (RCMP) laboratories in Canada. Triazolam concentrations from selected references are included for comparison.

Determination of Cocaine and Its Major Metabolite, Benzoylecgonine, in Amniotic Fluid, Umbilical Cord Blood, Umbilical Cord Tissue, and Neonatal Urine: A Case Study

Abstract: The determination of cocaine abuse among pregnant women has become a topical issue for both legal and emotional reasons. The analysis of urine from both mother and baby using immunoassay procedures is the most common method of determining the presence of cocaine and its metabolites. However, unless exposure to cocaine has occurred recently, false negatives are frequently obtained from the analysis of neonatal urine (1). More recently, meconium has been used to determine cocaine abuse. Because meconium is cumulative throughout the pregnancy after about 16 weeks, it has been suggested that it may be a more accurate sample for determining cocaine use than the more commonly used urine (2,3). Amniotic fluid, umbilical cord blood, and umbilical cord tissue have not been analyzed for cocaine and its metabolites even though they are present for the entire pregnancy. A study of the available maternal and fetal biological fluids following a premature delivery in order to determine cocaine abuse during pregnancy was carded out. The maternal urine tested positive for cocaine metabolite using immunoassay. Ease of sample collection, extraction, and determination were all considered to be important factors in sample analysis. Solid-phase extraction using XTrackT | high-flow sorbents (Worldwide Monitoring) allowed the easy extraction of amniotic fluid, homogenized umbilical cord, and umbilical cord blood. The more common Clean Screen | sorbents were used for urinalysis. The extracts were subsequently analyzed and quantitated using HPLC with multiwavelength UV detection (4,5). Results are shown in Table I. Neonatal urine showed the highest concentration of benzoylecgonine, which is probably indicative of recent Table I. Results of HPLC Analyses Benzoylecg0nine Cocaine Sample

Retrospective Analysis of Some l-Methamphetamine/l-Amphetamine Urine Data

Abstract: A limited analysis is presented of data accumulated over about three years for specimens containing either nonracemic L-methamphetamine/L-amphetamine or racemic mixtures of the D and L stereoisomers. 55 data points for both nonracemic L-methamphetamine and racemic mixtures show that it is possible to report a specimen containing L-methamphetamine positive for illegal D-methamphetamine based on current guidelines unless chiral assays are performed on selected methamphetamine positives. Generation of only 45 racemic specimens from chiral analysis of about 5,000 urine methamphetamine positives demonstrates the overwhelming prevalence of nonracemic illegal D-methamphetamine in Southern California.

m-Hydroxybenzoylecgonine : an important contributor to the immunoreactivity in assays for benzoylecgonine in meconium

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 beta-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.

The Use of Bone Marrow in the Study of Postmortem Redistribution of Nortriptyline

Abstract: Thirty-two New Zealand Albino rabbits (1.5-2.0 kg) were dosed on a daily basis with 20 mg/kg nortriptyline (NT) prior to feeding for a period of five days. On the fifth day of dosing, the animals were sacrificed approximately 1.5 h after the final dose. A comparison was made of nortriptyline concentrations in the blood and bone marrow at the time of sacrifice, and between bone marrow collected at the time of sacrifice and bone marrow collected at 2, 4, 6, 12, 24, 36, and 48 h after sacrifice. The results indicate that a linear relationship exists between blood and bone marrow NT concentrations, with an average marrow-to-blood ratio of 29.98 +/- 3.91 and a correlation coefficient of 0.956. Additionally, there was no significant difference (p > 0.05) observed between NT concentrations in bone marrow at the time of sacrifice and its concentration up to 24 h after sacrifice. The results indicate that bone marrow may be used to predict blood concentrations of NT up to 24 h after death when a suitable blood sample is not available.