Correlation of the incidence of cocaine and cocaethylene in hair and postmortem biologic samples

Hair samples are useful as a matrix for drug testing because drugs can be detected in hair for longer periods than in blood or urine. The authors report a prospective comparison of the detection of cocaine and cocaethylene in routine postmortem biologic specimens to the detection of cocaine and cocaethylene in hair. The authors collected hair samples from various areas of the head in 53 autopsy cases, prepared them, and analyzed them by gas chromatography/mass spectrometry (GC/MS) for cocaine and cocaethylene. The authors compared the results of hair analysis with the results of toxicologic analysis performed on routine postmortem samples by enzyme multiplied immunoassay technique and GC/MS. Cocaine was found in either biologic fluids or in hair in 16 of 53 samples tested. Nine samples were positive for cocaine in both biologic fluids and hair. Five samples contained cocaine only in biologic fluids, and two contained cocaine only in hair. Cocaethylene was present in two cases. Drug screening of hair provides additional information in some autopsy cases, but the authors have not made hair analysis a routine practice. It may prove useful to save hair samples in all cases for later analysis if warranted by additional history or autopsy findings.     

Coca paste: chemical analysis and smoking experiments

Several samples of Colombian and a sample of Peruvian coca paste were subjected to chemical analysis to ascertain the complexity of these products. A neutral and acid fraction and a basic fraction were analyzed by gas chromatography/flame ionization detection (GC/FID) and gas chromatography/mass spectrometry (GC/MS). The basic fraction was also analyzed as its trimethylsilyl (TMS) derivative. Several gasoline residue components were identified in the neutral fraction. In addition to cocaine (greater than 60% in all cases), other alkaloids were identified. Lead and manganese analyses were carried out on these samples. While all the samples contained no lead (less than 45 ppm), most of the Colombian samples contained significant amounts of manganese (greater than 5%). Preliminary smoking experiments with a Colombian coca paste sample indicated that it behaves more like free cocaine than like a cocaine sulfate salt.     

The detection of cocaine in hair specimens using micro-plate enzyme immunoassay.

The analysis of hair for drugs of abuse is becoming increasingly popular and is under consideration by the Division of Health and Human Services as a possible alternative or adjunct to urinalysis in workplace programs. The detection of cocaine in human hair using a commercially available micro-plate enzyme immunoassay is described for the first time. Sample size and incubation time were the major variables in the optimization of the method. In order to validate the procedure, the method was applied to 105 consecutive hair samples routinely received into our laboratory. The samples were simultaneously analyzed by the Micro-Plate immunoassay (EIA), as well as our current fluorescence polarization immunoassay (FPIA) procedure and gas chromatography-mass spectrometry (GC/MS). The sensitivity of the EIA and FPIA assays were 75% and 67.8% respectively; specificity 97.4% and 80.5% respectively; and efficiency 91.4 and 77.1% respectively. The Micro-Plate EIA was shown to be a valid alternative to other immunoassay screening methods for the detection of cocaine in hair by demonstrating increased sensitivity, specificity and efficiency over our current technique.     

Analytical Fingerprint of Wolframite Ore Concentrates

Ongoing violent conflicts in Central Africa are fueled by illegal mining and trading of tantalum, tin, and tungsten ores. The credibility of document‐based traceability systems can be improved by an analytical fingerprint applied as an independent method to confirm or doubt the documented origin of ore minerals. Wolframite (Fe,Mn)WO₄ is the most important ore mineral for tungsten and is subject to artisanal mining in Central Africa. Element concentrations of wolframite grains analyzed by laser ablation‐inductively coupled plasma‐mass spectrometry are used to establish the analytical fingerprint. The data from ore concentrate samples are multivariate, not normal or log‐normal distributed. The samples cannot be regarded as representative aliquots of a population. Based on the Kolmogorov–Smirnov distance, a measure of similarity between a sample in question and reference samples from a database is determined. A decision criterion is deduced to recognize samples which do not originate from the declared mine site.     

可卡因及其代谢物苯甲酰爱康宁胶体金检测试剂盒的研制

目的根据胶体金免疫层析的原理,建立一种可以在现场快速检测可卡因及其代谢物苯甲酰爱康宁的方法。方法将胶体金标记的抗可卡因单克隆抗体均匀浸涂在玻璃纤维膜上,用点膜仪将可卡因的完全抗原以及羊抗鼠多抗在硝酸纤维素膜上划线,分别作为可卡因的检测区（T）和质控区（C）。并且对样品垫用0.2%Tween 20和0.5%PVA封闭处理。样本（尿样、唾液、血样等）中游离的可卡因及其代谢物同包被的完全抗原分别免疫竞争结合胶体金标记的抗可卡因单抗。肉眼观察试剂盒的质控区和检测区上有无紫红色带来判读结果。结果可卡因胶体金检测试剂盒对65种毒品、药物的特异性测试中仅识别可卡因及其代谢物。对人体尿样中的可卡因和苯甲酰爱康宁检测阈值均为300ng/mL。与GC/MS对照试验,其准确性分别为98.9%和98.6%。在常温下能稳定保存2年。结论该可卡因检测试剂盒在5min内可以检测出样本中的可卡因及其代谢物成分。     

Hair analysis by using radioimmunoassay, high-performance liquid chromatography and capillary electrophoresis to investigate chronic exposure to heroin, cocaine and/or ecstasy in applicants for driving licences

The present paper describes an integrated diagnostic strategy to check the physical fitness of subjects, formerly users of illicit drugs, to obtain a driving license, after having quit their addiction. According to the Italian law, applicants for a driving license with a history of drug abuse must give evidence to have quit this behaviour and to show no risk of relapse in the future. To prove this, at our institute, they undergo medical examination, hair analysis and a urinalysis program on eight seriate samples, collected over about 40 days. About 700 subjects per year are investigated with this strategy. The hair samples are screened for opiates (morphine), cocaine and ecstasy, the most abused illicit substances in our region, by using commercial radioimmunoassays adopting cut-off levels of 0.1 ng/mg. All positive samples and about 10% of negatives are confirmed by high-performance liquid chromatography. Further confirmation of results can be carried out by capillary electrophoresis (and/or GC/MS or MS/MS). In 1998, the prevalence of positives for morphine, cocaine and ecstasy was 4.8, 11.3 and 2.6%, respectively. In this year, for the first time, the percentage of hair samples positive for cocaine was greater than that for opiates. The results of this integrated diagnostic strategy are presented and discussed, with particular emphasis on the comparison between hair analysis on a single sample and seriate urinalyses (on eight samples).     

Levels of cocaine and its metabolites in washed hair of demonstrated cocaine users and workplace subjects

In a study of subjects in drug rehabilitation programs, cocaine and cocaine metabolite levels were determined in the hair of 75 subjects who had produced cocaine-positive urine results. The hair was analyzed after being washed with the 3.75 h wash procedure developed by this laboratory. In addition, results of testing 73 non-users are presented, as well as levels of cocaine, benzoylecgonine (BE), cocaethylene, and norcocaine from workplace population samples. The data support a recommendation of reporting as positive a sample with cocaine of 500 pg/mg hair and either a 5% ratio of benzoylecgonine (BE) to cocaine in samples, or the presence of cocaethylene at 50 pg/mg hair, or norcocaine at 50 pg/mg hair for samples < or =2000 pg cocaine/mg hair. For samples with cocaine present at >2000 pg/mg hair, the data indicate that a ratio of 5% BE may be an overly conservative approach. In appropriately washed hair samples, cocaine users can produce hair levels of <5% BE and thus a minimum BE cutoff in lieu of a ratio could be considered.     

The rapid determination of cocaine and other local anesthetics using field tests and chromatography

The application of field tests and chromatography to the detection of cocaine and some other local anesthetics that have been used to adulterate cocaine is described. Initial screening of samples by field tests, followed by concurrent TLC and GC, enables rapid identification of these compounds to be achieved. In particular, the use of flow-programmed GC shortens the time for analysis compared with conventional GC and requires negligible equilibration time between consecutive runs [12]. The method gives reliable quantitative data.     

Optimum methamphetamine profiling with sample preparation by solid-phase microextraction

Solid-phase microextraction (SPME) is a relatively new technique in which a small, polymer-coated fiber is employed to extract volatile and semivolatile organic compounds from the sealed headspace above a questioned sample. SPME, coupled with gas chromatography/mass spectrometry (GC/MS), was used to characterize impurities in illicit methamphetamine samples. Trace impurities present in a specimen were tentatively identified using mass-spectral databases and included 1,2-dimethyl-3-phenyl-aziridine (indicating synthesis via a halogenated ephedrine intermediate), ethyl vanillin (a flavoring compound), and caffeine (a stimulant used as cutting agent). The types and numbers of organic compounds sampled by SPME were compared with those collected by various solvent extraction protocols. In addition to unambiguously confirming the presence of methamphetamine, SPME-GC/MS analyses detected approximately 30 more organic analytes than were found by GC/MS following the ethyl acetate extraction method adopted by the United Nations International Drug Control Programme. SPME-GC/MS is a superior method for generating material "fingerprint" profiles in methamphetamine samples. The detection and characterization of increased points of comparison in drug samples provide more detailed chemical signatures for both intelligence and operational information.     

A quantitative densitometric determination of heroin and cocaine samples by high-performance thin-layer chromatography

The traditional thin-layer chromatography (TLC) is a sensitive technique frequently used in chemical analysis for semiquantitative determinations. In the last years a high-performance thin-layer chromatography system (HPTLC) has been developed. It assures quantitative determinations with direct densitometric measurements of chromatographic spots in situ on the plate. Here, we wish to report a method for the drug determination in street samples, that employs a CAMAG TLC/HPTLC Scanner Photodensitometer and a CAMAG LINOMAT III for sample application. Samples of illicit drug traffic (50 samples of street heroin and 30 samples of street cocaine) are tested and the gas-liquid chromatography (GLC) and HPTLC determinations are compared. The HPTLC method has proven to be sensitive, precise and accurate for quantitative determinations of drugs in street samples. Furthermore this method may be useful in forensic chemistry to show a "fingerprint" of each sample.     

Heroin abuse and a gas chromatographic method for determining illicit heroin samples in Singapore.

The abuse of heroin (diacetylmorphine) in Singapore escalated sharply in 1975 and 1976, as indicated by the 35-fold increase in the number of heroin seizures and the 20-fold increase in the urine samples containing morphine since 1974. A rapid and simple GC method has been described to estimate diacetylmorphine (and caffeine). Monoacetylmorphine and acetylcodeine may be ascertained by an additional step involving acetylation. All gas chromatograms of a large number of samples analyzed consistently had the same pattern, indicating that they possibly had a common origin. This GC "fingerprint," together with the quantitative data, appears to be characteristic of the illicit Asian or Chinese type of heroin found in Singapore. The proportions of the four major ingredients in some twelve typical samples have been tabulated. Statistical data confirming the accuracy and reproducibility of the analytical method have also been presented.     

Validity testing of commercial urine cocaine metabolite assays: IV. Evaluation of the EMIT d.a.u. cocaine metabolite assay in a quantitative mode for detection of cocaine metabolite

The EMIT d.a.u. cocaine metabolite assay (EMIT dau) was evaluated in a quantitative mode for analysis of clinical specimens obtained after controlled cocaine administration to human subjects. The quantitative results showed high concordance with those of gas chromatography/mass spectrometry (GC/MS) assays of the same specimens for benzoylecgonine, and no false positive or false negative results were obtained. The evaluation also included analysis of standardized solutions containing benzoylecgonine, cocaine, and other cocaine metabolites and isomers. The EMIT dau antibody demonstrated high selectivity for benzoylecgonine. The precision was somewhat less than that reported earlier for other commercial cocaine metabolite immunoassays. Quantitation of initial screening results from EMIT dau testing can serve as a useful guide for confirmation by GC/MS in forensic science urine testing.     

GC/MS/MS时间编程在火灾现场助燃剂检测中的应用

运用Varian SATURN2000气质联用仪通过GC/MS、GC/MS/MS、时间编程 GC/MS/MS对汽油样品进行分析比对,发现利用时间编程GC/MS/MS,对检测火灾现场残留汽油效果很好,可彻底排除样品中基体的背景干扰,大大提高检测灵敏度;同时对混合助燃剂（汽油+煤油、汽油+柴油）进行了实验探索。     

HS/GC/ECD分析生物检材样品中的氰化物

目的 建立生物样品中氰化物的衍生化定性定量分析方法。方法 用氯胺T衍生化,HS/GC/ECD分析衍生物CICN。结果 在1ml血中,添加0.2μg氰化钾,回收率为84.6％,RSD为6.39％;在1g肝中添加0.5μg氰化钾,回收率为67.3％,RSD为5.05％;血中检出限为5ng/ml。结论 所建方法能定性定量分析生物样品中的氰化物。     

Comparison analysis of illicit cocaine samples.

A rapid method for comparison analysis of illicit cocaine samples has been developed. The raw data are obtained by capillary gas chromatography using a nitrogen-phosphorus detector. The area ratios of four alkaloids (tropacocaine, norcocaine, cis-cinnamoylcocaine, and trans-cinnamoylcocaine) to cocaine are calculated for each sample. These ratios are compiled in a computer database which allows easy comparison of samples and makes possible reliable conclusions regarding their commonality of origin.     

GC/MS和GC法定性定量分析可卡因

目的建立用于可卡因案件检验鉴定的GC和GC/MS定性、定量分析方法。方法通过选择和优化,建立GC、GC/MS法检验可卡因的最佳分析条件;用分别含0.6mg/ml地西泮为内标的0.10、0.20、0.40、0.60、0.80、1.00、1.20mg/ml可卡因标准品乙醇液,考察线性范围和方法检测限。结果分析方法线性方程:GC/FID,Y=1.055X-0.0021,R2=0.9999,GC/NPD,Y=0.556X-0.0016,R2=0.9996;可卡因检测限:GC/FID法10ng,GC/NPD法2ng;分别以所建GC/FID、GC/NPD分析方法和内标法对案件中缴获的可卡因毒品进行定量分析,结果为72%±2.3%,且两方法定量重现性良好。结论本文所建方法可以用于可卡因涉毒案件的检验鉴定。     

Impurities,adulterants and cutting agents in cocaine as potential candidates for retrospective mining of GC-MS data

Cocaine is one of the most widely used illicit drugs worldwide. Cocaine powders seized by the Police may contain numerous other substances besides the drug itself. These can be impurities originating from the coca plant or the production process, or be purposely added to the drug formulation as adulterants and cutting agents. In forensic laboratories, identification of cocaine is routinely done through GC-MS analysis, but other components are often ignored even if the method allows for their detection. Yet, they can provide valuable insight into the history of a seizure and its potential connection to other samples. To explore this idea, an extensive review of common impurities and adulterants encountered in cocaine is presented. Based on their incidence, concentration in the end product and compatibility with GC-MS methods, their overall usefulness as candidates for the statistical investigation of existing forensic data is evaluated. The impurities cis- and trans-cinnamoylcocaine, tropacocaine, norcocaine and N-benzoylnormethylecgonine as well as the adulterants lidocaine, procaine, tetracaine, benzocaine, caffeine, acetylsalicylic acid, phenacetin, ibuprofen, levamisole, hydroxyzine and diltiazem are promising candidates to provide additional forensic intelligence. Future research on optimized routine GC-MS methods, signal reproducibility, comparison, statistics and databases is suggested to facilitate this concept. Ultimately, such an approach may significantly advance the amount of information that is extracted from routine casework data, elucidate developments in the cocaine markets in the past and facilitate Police work in the future. Preliminary assessment of existing data from the forensic laboratory of the Amsterdam Police has been included to show that the detection of the identified target impurities is feasible, and that small adjustments to the analysis method could significantly increase the detectability of these analytes in prospective drug screenings. Forensic intelligence based on retrospective data mining of cocaine containing casework samples may thus be realized with minimal additional laboratory efforts by using already available instrumentation, samples and data.     

Occupational exposure to cocaine involving crime lab personnel.

The possibility of exposure to cocaine as a result of analyzing it or handling material contaminated by it has been a major concern of laboratory personnel. Several different work environments and simulated situations were examined to assess the likelihood of this type of exposure occurring. Urine specimens were collected and evaluated for cocaine and benzoylecgonine using the Syva ETS System (EMIT). Each specimen was analyzed for the two substances using gas chromatography/mass spectrometry (GC/MS). Urine specimens of laboratory-management personnel not working with drug samples showed no trace of cocaine or benzoylecgonine. A urinary benzoylecgonine level of 227 ng/mL was found in the specimen from one narcotics criminalist who was working on a routine case of 2 kilos of cocaine hydrochloride in the Narcotics Laboratory. A maximal urinary benzoylecgonine concentration of 1570 ng/mL was determined in the urine specimen from one narcotics criminalist who was sampling a case containing 50 kilos of cocaine hydrochloride over a period of 3 h. Decreasing the levels of airborne cocaine dust appears to minimize the amount of cocaine absorbed by the criminalists. Gloves, face masks, and goggles prove to be effective in minimizing exposure.     

Simultaneous hair testing for opiates, cocaine, and metabolites by GC-MS: a survey of applicants for driving licenses with a history of drug use

A sensitive GC-MS method for the simultaneous determination of opiates, cocaine, and metabolites in hair at a cut-off level of 0.1 ng/mg was adopted to assess past exposure to these drugs in applicants for driving licenses with a history of drug use. The sampling protocol consisted of collection of one hair (sample A, 5-cm length) and one urine sample. When hair and urine (EMIT Syva, cut-off levels: 0.3 mg/l for opiates, 0.15 mg/l for cocaine, GC-MS confirmation of positives) were both positive or negative the protocol was concluded. In the other cases, the assessment of 'current exposure' to drugs was carried out, in order to avoid seriated random urinalysis, by collecting a second hair sample (sample B) 6 weeks later and analysing the proximal 1-cm segment. Out of the 214 'A' hair samples analyzed, 14 (6.5%) tested positive for morphine and/or 6-acetylmorphine (6AM), and 26 (12%) for cocaine and/or benzoylecgonine (BE), whereas none of the samples tested positive for both drugs. Levels between 0.1 and 1 ng/mg of the single analytes were found in eight out of the 14 morphine-6AM positives (57%) and in 18 out of the 26 cocaine-BE positives (69%). The time course of positive cases showed a progressive decrease of morphine-6AM positives and a corresponding increase of cocaine-BE positives within the study period September 1995-February 1999. No cases with positive urine and negative hair were observed. Among the 40 positive cases, seven (four and three for opiates and cocaine, respectively) were found to be 'currently exposed to drug', four by urinalysis (three and one) and three by analysis of the hair sample B (1 and 2).     

Cocaine profiling for strategic intelligence purposes, a cross-border project between France and Switzerland. Part I. Optimisation and harmonisation of the profiling method

Optimisation and harmonisation of analytical and statistical methodology have been carried out between two forensic laboratories (Lausanne, CH and Lyon, F) in order to provide drug intelligence for cross-border cocaine seizures. The aim was to improve the gas chromatographic analysis of cocaine samples for profiling. Some important validation parameters were tested to verify the developed method and demonstrate its profiling capacity: the selectivity of the method with retention time reproducibility, the choice of a derivatisation agent improving the chromatography (MSTFA, BSA, TMSI and BSTFA+TMCS 1%), the cutting agents influence (matrix effect), the influence of the sample storage conditions and the sample quantity to weigh for analyses. Eight main alkaloids, which represent the sample signature, have been selected: ecgonine methyl ester, ecgonine, tropacocaine, benzoylecgonine, norcocaine, cis- and trans-cinnamoylcocaine and 3,4,5-trimethoxycocaine. Their stability in the solvent used (CHCl(3)/pyridine) was demonstrated. In order to reach the final objective, which is the comparison of samples seized and analyzed in two different laboratories, the harmonisation of the profiling method between the two laboratories had to be ensured and is the subject of ongoing research.