Detection of drugs in human hair using Abbott ADx, with confirmation by gas chromatography/mass spectrometry (GC/MS).

The authors suggest use of the fluorescence polarization immunoassay (FPIA) technique in evaluation of chronic drug abuse using human hair. Hair was decontaminated in 5 mL of ethanol for 15 min at 37 degrees C and then incubated in 3 mL of 1M sodium hydroxide (NaOH) for 1 h at 100 degrees C. Afterwards, the aliquots were neutralized and analyzed using Abbott ADx for a negative or positive response for the following drugs: benzodiazepines, barbiturates, antidepressants, opiates, cocaine, amphetamine, and cannabis. All the positive samples were confirmed by gas chromatography/mass spectrometry (GC/MS). Only one false positive was detected (caused by interference of a phenothiazine with the antidepressants kit), clearly demonstrating the capability of ADx for toxicological screening of human hair.     

GC/MS、GC/NPD法检测血液中氯胺酮

目的利用GC/MS、GC/NPD与固相萃取（SPE）技术相结合,建立血液中氯胺酮的定性定量分析方法。方法选择4-苯基丁胺为内标,采用Bond-Elut Certify固相柱萃取、二氯甲烷：异丙醇：氨水（78∶20∶2,v/v/v）洗脱的固相萃取分离技术,比较不同pH体系、洗脱溶剂对回收率的影响,建立血液中氯胺酮的GC/MS、GC/NPD定性定量分析方法。结果以GC/NPD分析氯胺酮在6.0～5000ng/mL范围内线性关系良好,GC/MS-Scan定性检测限为20.0ng/mL。方法平均回收率达96.9%,标准偏差小于5%。结论此方法可满足氯胺酮毒品滥用者血液定性定量分析。     

Benzoylecgonine (cocaine metabolite) detection in hair samples of jail detainees using radioimmunoassay (RIA) and gas chromatography/mass spectrometry (GC/MS)

Benzoylecgonine (BE) was detected in hair samples using nonproprietary extraction methodology and modifications of well-established radioimmunoassay (RIA) screening/quantitative gas chromatography/mass spectrometry (GC/MS) confirmation procedures. Samples collected anonymously from a population of 48 jail detainees weighed between 5.3 and 61.2 mg. All of the 22 hair samples which had RIA results indicating the presence of BE or immunologically similar substances above a cutoff amount of 1.25 ng/sample (50 ng/mL) were confirmed by GC/MS. Several varieties of hair color and texture were tested, although in each general category there were samples which contained BE as well as other samples which did not reveal detectable amounts of BE. The range of concentrations in 22 hair extracts that screened positive were 0.26 to 18 ng/mg hair as determined by GC/MS. In comparison with other reports of cocaine-related substances in hair, these data show consistent concentrations.     

The analysis of forensic samples using laser micro-pyrolysis gas chromatography mass spectrometry

Laser micropyrolysis gas chromatography-mass spectrometry is used for the analysis of paint, photocopier toner, and synthetic fiber materials to test the forensic potential of this emerging technology. It uses a laser microprobe to selectively target very small parts of the materials for GC-MS analysis. Whereas the paint and the toner samples were amenable to direct laser pyrolysis, the synthetic fibers proved transparent to the 1064 nm laser radiation. The difficulty with the fibers demonstrates that a specific laser wavelength may not be appropriate for all types of materials. Nevertheless, the fibers were able to be indirectly pyrolyzed by impregnation in a strongly absorbing graphite matrix. A vast array of hydrocarbon pyrolysates was detected from the different materials studied. Unique product distributions were detected from each sample and in sufficient detail to facilitate individual molecular characterization (i.e., molecular fingerprinting). The integrity of the laser data were confirmed by comparison to data obtained from the same samples by the more conventional pyroprobe pyrolysis GC-MS method. The high spatial resolution and selectivity of the laser method may be advantageous for specific forensic applications, however, further work may be required to improve the reproducibility of the data.     

Determination of cocaine in human hair by gas chromatography/mass spectrometry

A qualitative method for the determination of cocaine alone without its metabolites in human hair by gas chromatography/mass spectrometry (GC/MS) was developed. The assay used helium as carrier gas, a 30-m bonded phase fused silica OV-1 capillary column, and solid injection at 290 degrees C evaporator temperature. The cocaine concentrations in hair were determined also by radioimmunoassay (RIA). The values obtained are the sum of cocaine and its metabolites. Both GC/MS and RIA meet the requirements for the determination of drug abuse by two different methods in forensic science.     

Detection of methadone in human hair by gas chromatography/mass spectrometry

Determination of methadone in human hair by gas chromatography/mass spectrometry was described. Helium as carrier gas, a 30-m bonded phase-fused silica DB-1 capillary column and splitless injection at 230 degree C temperature were used. The concentrations of methadone and its metabolites were measured in addition by radioimmunoassay (RIA). Both methods GC/MS and RIA showed the presence of methadone in human hair.     

Simultaneous determination of opiates,cocaine and major metabolites of cocaine in human hair by gas chromotography/mass spectrometry (GC/MS)

A procedure is presented for the simultaneous identification and quantification of morphine (MOR), codeine (COD), ethylmorphine (EM), 6-monoacetylmorphine (6-MAM), cocaine (COC), benzoylecgonine (BZE), ecgonine methylester (EME) and cocaethylene (CE), contained in the hair of opiates and cocaine addicts. The method involves decontamination in dichloromethane, pulverization in a ball mill, heat-acid hydrolysis, addition of deuterated internal standards, liquid-liquid extraction and gas chromatography/mass spectrometry (GC/MS) after silylation. The limit of detection (LOD) was ~0.1–0.8 ng/mg for each drug, using a 30-mg hair sample. The method is reproductible, with a coefficient of variation (CV) of ~8–17%. Cocaine and 6-monoacetylmorphine were the major compounds detected in cases of cocaine (14 cases) and heroin (68 cases) intake. Concentrations were in the range 0.4–78.4 ng/mg (COC), 0.0–36.3 ng/mg (BZE), 0.0–1.6 ng/mg (EME), 0.0–2.1 ng/mg (CE), 0.0–84.3 ng/mg (6-MAM), 0.2–27.1 ng/mg (MOR) and 0.1–19.6 ng/mg (COD). An application in forensic sciences, involving multi-sectional analysis, is given.     

Differentiation of side chain isomers of ring-substituted amphetamines using gas chromatography/infrared/mass spectrometry (GC/IR/MS).

Common analytical methods used for identifying samples obtained from clandestine laboratories were evaluated for their ability to differentiate between possible amphetamine isomers and homologs. A series of ring-substituted (4-methyl, 4-methoxy, and 3,4-methylenedioxy) amphetamine and N-methylphenethylamine isomers was analyzed using color tests, thin-layer chromatography, gas chromatography/mass spectrometry (GC/MS) and GC/infrared (GC/IR). The N-acetyl derivatives of the isomers were analyzed using GC/IR/MS. GC/IR/MS readily differentiated the 4-methylphenylalkylamine isomers. MS and IR spectra were also obtained for each pair of the 4-methoxyphenylalkylamine isomers and the 3,4-methylenedioxyphenylalkylamine isomers, but differentiation via GC/IR/MS was difficult. The N-acetyl derivatives of each pair of isomers could be readily differentiated using GC/IR/MS. Good library researchable spectra for N-acetylamphetamine could be obtained for IR identification with 10 ng (on-column) and MS identification with 2 ng. The spectrometrically independent IR and MS data obtained for the N-acetyl derivatives indicated that the combination of GC/IR/MS can add a significant level of confidence in the analysis of ring-substituted arylalkylamines.     

Evidence of mephedrone chronic abuse through hair analysis using GC/MS

Mephedrone is a synthetic derivative of cathinone which is becoming more common on the recreational drug market. Several intoxications following mephedrone abuse have been reported though published papers have focused essentially on analytical approaches for biological fluids and only one has involved a hair sample. After the development and validation of a new method, the first series of positive results for mephedrone in hair specimens is reported here. After decontamination of the hair strand in methylene chloride, hair segments were cut into small pieces with scissors, weighed and incubated overnight in Soerensen buffer pH 7.0 in the presence of deuterated 3,4-methylenedioxymethamphetamine (MDMA) at 40°C. The incubation medium was extracted using ethyl acetate after alkalinisation with 1N sodium hydroxide (NaOH). Before injection, the dry extract was derivatized using a mixture of heptafluorobutyric anhydride/ethyl acetate (100:50, v/v), evaporated and dissolved in ethyl acetate (25μl). After introduction of 1μl of the extract onto a splitless injector, chromatographic separation was achieved on a HP 6890 gas chromatograph equipped with a 5-MS capillary column. Detection was achieved in single ion monitoring mode (m/z 254-119-210 for mephedrone, m/z 258-213 for MDMA-d5) using a 5973 MSD operating in electron impact mode. Sixty-seven hair specimens were tested for mephedrone. Thirteen of them were found positive for mephedrone with concentrations ranging from 0.2 to 313.2ng/mg with a mean concentration of 26.8ng/mg. It was difficult to compare our findings due to a lack of reference data, nevertheless mephedrone seems well incorporated into hair (concentrations in the ng/mg range) like other stimulant drugs such as amphetamines or cocaine. The aim of this work was to develop a specific and accurate method for mephedrone analysis in hair specimens and its application to a large number of samples (n=67). The developed analytical method appears sensitive enough to reveal occasional to regular use of mephedrone.     

SPE—GC／MS、GC／NPD法检测血液中苯丙胺类毒品

目的利用GC／MS、GC／NPD与固相萃取（SPE）技术相结合,建立血液中苯丙胺类毒品的定性定量分析方法。方法采用Bond—ElutCerti{y固相柱、甲醇淋洗、二氯甲烷／异丙醇／氨水（78／20／2）洗脱固相萃取分离提取,比较了不同PH体系、稀释状态、洗脱溶剂对提取回收率的影响,建立血液中苯丙胺类毒品的GC／MS、GC／NPD定性定量分析方法。结果以GC／NPD分析AM、MA、MDA和MDMA浓度在15ng／mL-2000ng／mL、10ng／mL～1600ng／mL、20ng／mL-3000ng／ml、20ng／mL-3000ng／mL范围内线性关系良好,AM、MA、MDA和MDMA的检测限分别为10ng／mL、8ng／mL、15ng／mL、15ng／mL,方法平均回收率大于85％,标准偏差小于5％,GC／MS-Scan检测限分别为40ng／mL、32．0ng／mL、60．0ng／mL、60．0ng／mL。结论此方法可满足苯丙胺类毒品滥用者的血液定性定量分析。     

Intratracheal gas analysis for volatile substances by gas chromatography/mass spectrometry--application to forensic autopsies

Intratracheal gas analysis was carried out by gas chromatography/mass spectrometry (GC/MS) in 20 burned body cases (13 males and 7 females). Volatile aromatic and aliphatic hydrocarbons were detected by GC/MS using a GS-Q column with the intratracheal gas as well as the blood in 19 cases. The characteristic patterns of mass chromatograms for gasoline, kerosene (gas oil), and liquid petroleum gas could be differentiated from each other using the intratracheal gas. The burned body in one case showed no presence of volatile substances in the intratracheal gas, nor intratracheal soot, although high concentrations (1 microg/g and more) of volatile substances were detected on the clothes. The victim also had normal CO-Hb concentrations (0.1 to 0.2%) in the heart blood. The results of intratracheal gas analysis were consistent with signs of the vital reaction. In conclusion, intratracheal gas analysis provides a supportive method for diagnosing the cause of death in burned bodies, and yields for at least 48 hours valuable information on volatile hydrocarbons (being detected in deliberate or accidental fire cases) to which the body had been exposed just before death.     

Practical GC/MS analysis of oxidation dye components in hair fiber as a forensic investigative procedure

The purpose of this study was to improve the reliability of discrimination (or identification) of dyed hair by analyzing chemical substances present in the hair, as an addition to the conventional macroscopical and microscopical examinations and ABO blood group examination. Oxidation hair-dye components such as p-phenylenediamine (PPDA), toluylene-2,5-diamine (T-2,5-DA), o-aminophenol (OAP), m-aminophenol (MAP), p-aminophenol (PAP) and p-amino-o-cresol (PAOC) were selected as the object of study. After alkaline-digestion, hair samples were adjusted to a pH of 12.6 to 12.8, and applied onto an Extrelut column. After 15 min, the components were simultaneously extracted and derivatized with n-hexane including 1% heptafluoro-n-butyryl (HFB) chloride. Their HFB derivatives within a condensed sample were diluted in ethyl acetate, and analyzed by gas chromatography-mass spectrometry (GC-MS) with full mass scanning or selected ion monitoring. For estimating their levels, 2,4,6-trimethylaniline was used as the internal standard. Standard curves obtained by preparing a 5 cm segment of control hair spiked with authentic substances were linear, ranging from 0.1 to 4.0 micrograms for PPDA and T-2,5-DA, and from 0.01 to 0.4 microgram for OAP, MAP, PAP and PAOC. The coefficient of variation of inter-day precisions (each n = 4) was below 16% for PPDA, below 20% for OAP and PAP and below 24% for T-2,5-DA, MAP and PAOC. These components were detectable even at 6 ng for PPDA, T-2,5-DA, MAP and PAP, 8 ng for OAP, and 4 ng for PAOC. Recovery percents using this procedure ranged from 54 to 86%. By using actual dyed hair samples this method was shown to provide high sensitivity for accurate detection.     

Comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (GC x GC-TOFMS) for drug screening and confirmation

Comprehensive two-dimensional gas chromatography (GC x GC) is applied to analysis of drug standard mixtures containing 78 drugs of interest in forensic samples. For this study, underivatised drugs were employed. While several of the drugs were not detected at the low concentrations employed in the samples, most could be satisfactorily assigned their first and second dimension retentions in the GC x GC retention plane. For this study, time-of-flight mass spectrometry (TOFMS) detection was used. The enhanced separation possible in GC x GC is demonstrated, and typical linearity and apparatus precision are shown for tramadol, diazepam, olanzapine and desipramine using selected qualifier ions. Mass spectral library search quality for the detection of drugs in a selection of authentic forensic cases, along with retention position in the 2D retention plane, is used to support positive identification of the presence of the drugs. The analysis of 'difficult' drugs paracetamol and phenytoin is shown to produce anomalous chromatographic peak shape in the 2D plane, whereas most drugs gave acceptable peak shapes. The GC x GC technique was applied to screening drugs in forensic samples, with either flame ionisation (FID) or TOFMS detection, and compared favourably with conventional single column GC-MS analysis when tested for diazepam in an authentic forensic study.     

Quantitative determination of amphetamines, cocaine, and opiates in human hair by gas chromatography/mass spectrometry

Hair of young subjects (N = 36) suspected for drug abuse was analysed for morphine, codeine, heroin, 6-acetylmorphine, cocaine, methadone, amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA). The analysis of morphine, codeine, heroin, 6-acetylmorphine, cocaine, and methadone in hair included incubation in methanol, solid-phase extraction, derivatisation by the mixture of propionic acid anhydride and pyridine, and gas chromatography/mass spectrometry (GC/MS). For amphetamine, methamphetamine, MDA, MDMA, and MDEA analysis, hair samples were incubated in 1M sodium hydroxide, extracted with ethyl acetate, derivatised with heptafluorobutyric acid anhydride (HFBA), and assayed by GC/MS. The methods were reproducible (R.S.D. = 5.0-16.1%), accurate (85.1-100.6%), and sensitive (LoD = 0.05-0.30ng/mg). The applied methods confirmed consumption of heroin in 18 subjects based on positive 6-acetylmorphine. Among these 18 heroin consumers, methadone was found in four, MDMA in two, and cocaine in two subjects. Cocaine only was present in two, methadone only in two, methamphetamine only in two, and MDMA only in seven of the 36 subjects. In two out of nine coloured and bleached hair samples, no drug was found. Despite the small number of subjects, this study has been able to indicate the trend in drug abuse among young people in Croatia.     

Detection of methamphetamine and amphetamine in a single human hair by gas chromatography/chemical ionization mass spectrometry

A detailed procedure of an extremely sensitive method for quantitation of methamphetamine and amphetamine in human hair by gas chromatography (GC)/chemical ionization (CI) mass spectrometry (MS) is presented. N-methylbenzylamine was used as an internal standard. The samples, after extraction with an organic solvent, were derivatized with trifluoroacetic anhydride before the GC/MS analysis. Quantitation was made with quasi-molecular ions of the derivatives by selected ion monitoring in the CI mode. The detection limit was about 10 pg in an injected volume. The high sensitivity enabled us to measure both stimulants in a single human hair in actual cases.     

A method for forensic identification of vegetable oil stains--rapid analysis of carboxylic acids with methyl esterification using purge-and-trap gas chromatography/mass spectrometry.

A simple method using purge-and-trap gas chromatography/mass spectrometry (P&T-GC/MS) for forensic examination of oil stains was studied. Carboxylic acids, chosen as target components for discrimination of oil samples, were extracted from stains with ether, methyl esterified by tetramethylammonium hydroxide, and analyzed by P&T-GC/MS. Vegetable oils were discriminated according to their carboxylic acid compositions. Carboxylic acid composition was independent of the substrate material of the stain. Although the carboxylic acid composition of the oil changed on exposure to sunlight, identification of oil was possible for oil stains that had been in the shade, if analysis was made within 20 days.     

Tandem mass spectrometry: a helpful tool in hair analysis for the forensic expert

The Bavarian State Bureau of Investigation in Munich has the exclusive responsibility for investigation of criminal acts. One considerable expertise is that of hair analysis. According to the legal system in Germany, there is a special interest when some clients' hair tested positive for illicit drugs. An accused with a lot of drugs in his hair will be treated as a supposed addict and will be guaranteed extenuating circumstances. The instrumentation used for hair analysis is a powerful analytical tool: a Varian 3400 gas chromatograph linked to a Finnigan Tandem-MS (TSQ 700). The methanol extraction method is used for the detection of illegal drugs and metabolites: amphetamine, methamphetamine, MDA, MDMA (ecstasy), MDE, MBDB, methadone, THC, EDDP (metabolite of methadone), cocaine, benzoylecgonine, cocaethylene, opiates (dihydrocodeine, codeine, heroin, 6-monoacetylmorphine, morphine, acetylcodeine). For the detection of 9-carboxy-THC by negative chemical ionization the hair sample is hydrolyzed under alkaline conditions. Solid-phase extraction is used for clean-up. The LOQ for the determination of 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic-acid is 0.16 pg/mg hair. An unsurpassed combination for rendering an expert opinion based on hair analysis may be: a forensic expert using diligence and experience, coupled with the performance of a sophisticated analytical instrument.     

Screening and determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry

Benzodiazepines are one of the most widely prescribed drugs for the treatment of a wide spectrum of clinical disorders. They are used as anticonvulsants, anxiolytics, hypnotics or muscle relaxants with different duration of action. In this paper, a simple and sensitive method for the determination of benzodiazepines in whole blood using solid-phase extraction and gas chromatography/mass spectrometry (GC/MS) is described. The drugs spiked in whole blood were extracted with an Oasis HLB solid-phase extraction cartridge (Waters), which contains a copolymer designed to have a hydrophilic-lipophilic balance. GC/MS analysis was performed using a Shimadzu QP-5000 equipped with a BPX5 capillary column (15 mx0.32 mm I.D., film thickness 0.25 microm, SGE). Nineteen benzodiazepines and two thienodiazepines were well separated from each other on their SIM chromatograms and also on the TIC with the exception of oxazolam to cloxazolam separation. The blank extract from whole blood gave no peaks that interfered with all benzodiazepines and thienodiazepines on the chromatogram. The calibration curves for selected benzodiazepines with fludiazepam as an internal standard showed excellent linearity over the concentration range 5-500 ng/ml blood with a correlation coefficients of >0.995. The detection limits ranged from 0.2 to 20 ng/ml blood. The method is simple and sensitive for the determination of benzodiazepines in whole blood and seems to be useful in the practice of forensic science.     

Analysis of 11-nor-9-carboxy-delta(9)-tetrahydrocannabinol in biological samples by gas chromatography tandem mass spectrometry (GC/MS-MS)

Gas chromatography tandem mass spectrometry (GC/MS-MS) analysis of 11-nor-carboxy-delta(9)-tetrahydrocannabinol (delta(9)-THC-COOH), the major metabolite of delta(9)-tetrahydrocannabinol, in biological samples is reported. The proposed method, using deuterated delta(9)-THC-COOH as an internal standard, is able to detect the major metabolite of cannabis derivatives at very low levels (picograms/millilitre) with high specificity. These characteristics render the proposed analytical procedure suitable for confirmatory analysis in drug testing for cannabis use.     

Determination of dopamine and dopamine-derived (R)-/(S)-salsolinol and norsalsolinol in various human brain areas using solid-phase extraction and gas chromatography/mass spectrometry

Using a solid-phase extraction procedure and a gas chromatographic-mass spectrometric (GC/MS) method the levels of dopamine and the levels of dopamine-derived salsolinol (SAL) and norsalsolinol (NorSAL) were determined in human brain areas involved in the etiology of alcoholism, parkinsonism and other diseases. The possibility that biosynthesis of salsolinol occurs through a stereospecific enzymatic reaction was considered. Using a two-step derivatization with N-methyl-N-trimethylsilyltrifluoracetamide (MSTFA) and the chiral reagent (R)-(-)-2-phenylbutyryl chloride, baseline separated peaks of (R)- and (S)-SAL were obtained. Both enantiomers were found in human brain samples with no correlations between levels of salsolinol and dopamine. These findings do not support the hypothesis that only an enantio-selective synthesis of (R)-SAL by a putative salsolinol synthase is responsible for the in vivo formation. In our opinion, non-enzymatic formation of salsolinol via the Pictet-Spengler reaction reveals both salsolinol enantiomers and an additional enzymatic synthesis of only (R)-SAL explains the enantiomer ratio (R)-/(S)-SAL of approximately 2.