Analysis of fentanyl and sufentanil in hair by GC/MS/MS

Fentanyl and sufentanil are potent narcotic analgesics used only in hospitals as anaesthetic agents. The dependence potential of fentanyl is known. As they are given in doses at the microgram level and their elimination half-life is in the order of a few hours, detection in body fluids is possible only for a short time after administration. Radioimmunological methods are the only ones capable of detecting fentanyl in hair, as normal GC/MS methods for hair analysis are not sensitive enough to detect the drugs after doses in the order of micrograms. We therefore chose GC/MS/MS to analyse fentanyl and sufentanil in two cases where the drugs were given under controlled conditions over several days. The concentration was in the order of less than 100 pg/mg hair.     

Detection and quantification of psychotropic drug etaqualone in human hair using GC–MS/MS

In this study, sensitive analytical procedure for detection and quantification of etaqualone in human hair samples using gas chromatography tandem mass spectrometry (GC–MS/MS) was newly established, and applied it to authentic human samples obtained from an abuser. In this method, the hair samples were treated with hydrochloric acid and then extracted with ethyl ether. The ether layer was dried in a warm water bath, and the residue was reconstituted in ethyl acetate, followed by GC–MS/MS analysis. Multiple reaction monitoring (MRM) mode was used for data collection, and quantitative analysis was performed using internal standard method. Good linear relationship within the concentration range of 1–100 pg/mg were obtained in calibrators for the hair samples showing its correlation coefficient value was 0.9993. The lower limit of quantitation in this study was 1 pg/mg and the recovery rate examined ranged from 100.4% to 108.5%. The intra-day precision and accuracy were less than 5.0% and 5.8%, respectively. The inter-day precision and accuracy were lower than 6.4% and 4.6%, respectively. Using this established method, etaqualone could be detected in the hair sample obtained from a suspected user to be level of 65.2 pg/mg. It should be expected that the method established in this study would contribute to rapid detection and identification of psychotropic drug etaqualone among multiple fields including forensic investigation, clinical application and of course public health matters.     

Specific interpretation of hair concentrations in 2 fatal metformin intoxication cases

Hair analysis is very useful for toxicological investigations since, by providing a wider detection window, it gives the possibility to perform a retrospective study on the historical consumption of a substance. Unfortunately, there are no data available for hair concentrations in metformin-related deaths. In this study, the authors present 2 cases of fatal metformin intoxication in which, for the first time, hair analysis was performed using a specific GC–MS/MS method. Metformin was tested positive in femoral blood (112.3 mg/L and 64.7 mg/L respectively) and cardiac blood (226.9 and 203.2 mg/L) of the two subjects. For case 1, other samples were also tested positive, including vitreous humor (31.1 mg/L) and gastric contents (773.5 mg/L). In case 2, metformin was measured at 844.9 mg/L in urine. Metformin hair concentrations were 28.3–44.8 and 22.5 ng/mg for both cases, respectively. The concentrations found in the 2 fatal cases are clearly higher than those obtained in a previous study with subjects under treatment (0.3–3.8 ng/mg) or those found in 3 post-mortem cases where metformin death was excluded (0.6–1.4 ng/mg). Excessive sweating during the agonal phase due to fatal hypoglycemia could explain these elevated concentrations as sweat can have contaminated the hair.     

Review: Drug concentrations in hair and their relevance in drug facilitated crimes

Segmental hair analysis can provide valuable retrospective information on the history of drug exposure in victims of drug facilitated crimes (DFC). This is now possible with availability of sensitive tandem MS techniques such as GC-MS/MS and LC-MS/MS allowing drugs to be detected at pg/mg concentrations after a single dose. In this review hair concentrations of 35 psychoactive drugs given in 20 controlled dose studies are reviewed and compared to the 25 different drugs detected in reported case work. The most common drugs were the benzodiazepines and related hypnotics, gamma-hydroxybutyrate (GHB), ketamine and methamphetamine. Those concentrations reported in DFC were mostly similar or higher than that seen in controlled dose studies. The factors that affecting interpretation of segmental hair results including hair color, growth rates, sample preparation and surface contamination are discussed.     

Development of a preparation method to produce a single sample that can be applied to both LC–MS/MS and GC–MS for the screening of postmortem specimens

Simple and efficient extraction methods have been developed for the screening of a wide array of drugs in postmortem autopsy specimens. Acidic and basic compounds were targeted with two extraction methods that can be applied to both GC–MS and LC–MS/MS instrumentation. The extractions were achieved by utilizing lipid-removal and solid-phase extraction cartridges while carefully monitoring the pH of the samples to ensure the adequate removal of interfering substances like lipids and amino acid derivatives. These methods were applied to actual autopsy cases, with 94 and 124 compounds detected by GC–MS and LC–MS/MS, respectively. The developed methods could easily be incorporated into a forensic laboratory’s daily routine for screening many different compounds from postmortem samples.     

Determination of dihydrocodeine in hair of opiate addicts by GC/MS

Summary After the examination of more than 300 hair samples of suspected heroin abusers, a large number of which proved positive, we can say that high concentrations of dihydrocodeine can be determined either in addition to, or in the place of, morphine and also frequently in combination with codeine. The opiates were extracted after dissolving the hair samples in NaOH and hydrolysis with HCI. The quantitative determination of dihydrocodeine was achieved by derivatisation with HFBA using GC/MS at m/u = 497. Dihydrocodeine is used in antitussive drugs. After the examination of individual hair samples, it was obvious that some heroin consumers had switched to dihydrocodeine. This may lead to the conclusion that dihydrocodeine itself is used either as an intoxicating drug or to reduce withdrawal symptoms. The increasing number of positive samples should be noted by the legal authorities.     

Identification and quantification of diphenhydramine,haloperidol, and its metabolite,reduced haloperidol in a saponified brain specimen that was immersed in the sea water for more than 10 years

In forensic toxicology, blood and urine specimens are commonly used for detecting and quantifying drugs and their metabolites. When the cadaver is so damaged or decomposed such that the specimens mentioned above cannot be collected, it is necessary to perform drug analysis using alternative specimens such as hair, nails, oral fluids and meconium. Adipocere is resistant to further degradation; it is thus possible to be used as an alternative specimen to analyze drugs and their metabolites. Some researchers indeed have reported drug concentrations in saponified samples that were collected years after decedents’ deaths.In this study, we subjected saponified brain, which remained under sea for over 10 years after death, to forensic toxicological analysis using liquid chromatography/tandem mass spectrometry (LC/MS/MS). Using product ion scan analysis, we confirmed the presence of diphenhydramine, haloperidol, and reduced haloperidol, a metabolite of haloperidol. In addition, drugs and metabolite quantification were performed using the standard addition method. Correlation coefficients of the calibration curves were over 0.98. Analyte concentrations in the saponified brain were as follows: diphenhydramine was 1.84 ng/g, haloperidol was 1.30 ng/g, and reduced haloperidol was 3.02 ng/g. Our results suggest that it can be possible to quantify not only parent drugs but also their metabolites in saponified brain. These findings indicate that saponified tissues could be applied as alternative specimens for forensic toxicology, and could be useful as supporting information for victim identification.     

On-site testing of saliva and sweat with Drugwipe and determination of concentrations of drugs of abuse in saliva, plasma and urine of suspected users

Potential drug users participated voluntarily in a Belgian study on the usefulness of the non-instrumental immunoassay Drugwipe (Securetec, Germany) for the screening of cocaine, opiates, amphetamine and cannabinoids in saliva and sweat. If one of the screening assays (urine, oral fluid, sweat) showed a positive result, blood and saliva were collected. The on-site Drugwipe results were correlated with the Drugwipe results for saliva in the laboratory and with the GC/MS results of the corresponding saliva, plasma and urine samples and pharmacological effects at the time of sampling. The Drugwipe assay proved to be sufficiently sensitive for the detection of recent cocaine (n = 6) and amphetamine (n = 15) abuse, whether the device was wiped on the tongue or on the surface of the body, or when a saliva sample was applied to the wiping part. In five of the six potential cocaine users, the saliva concentrations of cocaine exceeded 1000 ng/ml. In the amphetamine group, the saliva concentrations of amphetamine, MDMA or both were high (> 1000 ng/ml) in 13 subjects. For cocaine and amphetamine, the positive scores for Drugwipe matched the GC/MS results for the three body fluids. Recent heroin abuse (n = 5) could be demonstrated to some extent with Drugwipe on samples from the tongue but only the two subjects with the highest saliva concentrations of MAM (> 500 ng/ml) and morphine (> 500 ng/ml) were positive. If the legal cut-off value for driving under the influence of opiates in Belgium (20 ng/ml of free morphine in plasma) was taken into account, only three subjects would have been legally positive. For cannabinoids (n = 15), false negatives and even some false positives were observed. Saliva can be considered as a useful analytical matrix for the detection of drugs of abuse after recent abuse when analysed with GC/MS. Received: 26 January 1999 / Received in revised form: 19 May 1999 / Accepted: 17 June 1999     

Characterization of dextromoramide (Palfium) abuse by hair analysis in a denied case

By providing information on exposure to drugs over time, hair analysis is useful in verifying the history of drug use. In a clinical case, where drug abuse was denied, it was possible to identify dextromoramide in the hair of the subject. After acid hydrolysis of the hair with 0.1 M HCl, in the presence of SKF 525A as an internal standard, the drug was extracted at pH8.4 with chloroform-isopropanol-n-heptane (50:17:33 v/v) and quantified by gas chromatography/mass spectrometry. The hair strands were cut into 3 sections of 2.5 cm, corresponding to a growth period of 2 months. Concentrations were 1.09, 1.93 and 1.48 ng/mg from the root to the end, respectively. This is the first report on dextromoramide testing in human hair.     

Automated solid-phase extraction and two-step derivatisation for simultaneous analysis of basic illicit drugs in serum by GC/MS

A combination of automated solid-phase extraction (SPE) and subsequent two-step derivatisation has been developed for the simultaneous analysis of basic drugs of abuse and cocaine metabolites in serum samples. Substances included in this procedure are morphine, codeine, methadone, cocaine, benzoylecgonine, methylecgonine, amphetamine, methamphetamine, MDMA, MDEA and MDA. SPE with mixed-mode cartridges (RP-C8 and cation-exchange) was fully automated with a Zymark RapidTrace SPE robot. GC/MS analysis was performed after derivatisation with a new two-step reaction by trifluoroacetic anhydride and 2,2,3,3,3-pentafluoropropanol. High recoveries (> 85%) with high reproducibility (CV 1.1-3.8%) were found for all drugs. High correlation coefficients (r > 0.998) were obtained due to the addition of deuterated standards prior to extraction. Experience obtained over 2 years of applying this method to drug analysis in serum is discussed.     

Preventing misidentification of 25I-NBOH as 2C-I on routine GC–MS analyses

25I-NBOH is a novel psychoactive substance (NPS) recently reported to have been found on blotter paper samples seized on the streets of Brazil, and used as a replacement for the NBOMes now scheduled in many countries. The presence of this NPS on the street market may go undetected, because the most widely and routinely utilised analytical technique for drug sample analyses is gas chromatography–mass spectrometry (GC–MS), which can misidentify 25I-NBOH (and indeed the other members of the NBOH series), because of its degradation into 2C-I (and corresponding 2C for the other members of the series) within the injector, unless a derivatization procedure is employed, which is often non-standard. While direct detection of 25I-NBOH under routine GC–MS conditions is still achieved, a slight adjustment in the standard GC–MS method, including shortening of the solvent delay window, was found to enable the detection of an additional peak due to 25I-NBOH degradation. Consequently, the presence of this secondary early chromatographic peak allowed for the distinction between 25I-NBOH and 2C-I using routine GC–MS without resorting to derivatization (or other analytical processes), thus preventing misidentification of 25I-NBOH as 2C-I.     

Differentiation of regioisomeric fluoroamphetamine analogs by gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry

In recent years, a large number of clandestinely produced controlled-substance analogs (designer drugs) of amphetamine with high structural variety have been detected in forensic samples. Analytical differentiation of regioisomers is a significant issue in forensic drug analysis because, in most cases, legal controls are placed only on one or two of the three isomers. In this study, we used gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) for the differentiation of regioisomers of fluoroamphetamine analogs (fluoroamphetamines and fluoromethamphetamines), which were synthesized in our laboratories. Free bases and their acylated and silylated derivatives were subjected to GC–MS analysis using DB-1ms, DB-5ms, and DB-17ms capillary columns. The separation of free bases was incomplete on all columns. Trifluoroacetyl derivatives of 3- and 4-positional isomers showed slight separation on DB-1ms and DB-5ms. On the other hand, trimethylsilyl derivatization enabled baseline separation of six fluoroamphetamine analogs on DB-1ms and DB-5ms columns, which was sufficient for unequivocal identification. For LC–MS/MS, a pentafluorophenyl column was able to separate six regioisomeric fluoroamphetamine analogs but a conventional C18 column could not achieve separation between 3- and 4-positional isomers. These results show that a suitable choice of derivatization and analytical columns allows the differentiation of regioisomeric fluoroamphetamine analogs.     

Quantification of the benzimidazole opioid analog isotonitazene in human hair using liquid chromatography-tandem mass spectrometry

Benzimidazole opioids were originally developed from the late 1950s to 1970s as analgesics for medical use, although a lot of them could not be approved as licit medicines because of their severe side effects and physical dependence. Such benzimidazole opioid analogs as abused drug, however, have recently been found in illicit drug markets throughout the world. Isotonitazene is one such benzimidazole opioids, whose analgesic potency can be as much as 500 times greater than that of morphine, according to previous animal studies. In line with this potency, a couple of hundred fatalities related to it were reported to date. In this study, a well validated method for the quantification of isotonitazene in human hair samples using liquid chromatography (LC)–tandem mass spectrometry (MS/MS) was established, and could be applied to authentic samples which were seized by the police security bureau. Isotonitazene concentrations in the seized hair averaged 6.11 pg/mg. The LLOQ and LOD of this method were 1.25 and 2.5 pg/mg, respectively; the calibration curve of the substance in hair samples showed a good linearity in the concentration range of 2.5–250 pg/mg (r > 0.999); the extraction recovery rates were 87.3–105% in the tested range; the inter- and intra-day precisions and accuracies (%biases) were not greater than 9.09% for each determination. Isotonitazene in human hair showed good stability at room temperature and under dark storage conditions for 30 days. As for matrix effect in hair samples, moderate ion suppression of target substances could be found. This is the first report for the analysis of isotonitazene in human hair samples.     

Quantitation of sibutramine in human hair using gas chromatography–isotope dilution tandem mass spectrometry

Forensic Toxicology - An analytical method for quantitation of sibutramine in human hair using gas chromatography (GC)–isotope dilution tandem mass spectrometry (MS/MS) was newly established....     

液相色谱串接质谱测定马饲料中的三种生物碱

本文建立了采用高效液相色谱串接质谱(LC-MS/MS)同时快速筛查和定量测定马复合饲料中三种天然痕量生物碱(吗啡、可待因、罂粟碱)的方法。样品经充分粉碎后用1 mmol/L HClO4溶液提取,并通过混合型固相萃取进行净化。采用ZORBAX Eclipse Plus-C18(150 mmL×2.1mm ID,3.5μm)色谱柱以含0.4%甲酸的水溶液和乙腈作为流动相进行梯度洗脱,正离子多反应监测(MRM)模式下进行检测。吗啡的检出限为0.50 ng/g,定量限为1.0 ng/g;可待因的检出限为0.75 ng/g,定量限为1.5 ng/g;罂粟碱的检出限为0.20 ng/g,定量限为0.50 ng/g。三种药物空白饲料高、中、低添加浓度水平下的回收率在77.4~97.8%之间,日内、日间精密度分别不高于6.2%和7.1%,实验结果表明本方法有较好的灵敏度和精密度,可以满足马饲料中该类药物的常规检测分析。     

Simultaneous determination of tryptamine analogues in designer drugs using gas chromatography–mass spectrometry and liquid chromatography–tandem mass spectrometry

In recent years, a large number of tryptamine-based designer drugs have been encountered in forensic samples. We have developed simultaneous analytical methods for 14 tryptamine analogues using gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–tandem mass spectrometry (LC–MS–MS). Trimethylsilyl (TMS) derivatives of the analytes were separated on a DB-1ms column within 15 min. The structural isomers could be differentiated by electron ionization GC–MS. LC–MS–MS with a C₁₈ column could separate structural isomers of tryptamines except for a combination of 5-methoxy-N,N-diethyltryptamine and 5-methoxy-N-methyl-N-isopropyltryptamine. Higher collision energy gave different product ion spectra between the structural isomers. The results indicate that GC–MS is the first choice for identification of tryptamines, preferably after TMS derivatization, and LC–MS–MS can be used as a complementary approach for the unequivocal differentiation of tryptamine isomers.     

A gas chromatography–mass spectrometry based study on serum metabolomics in rats chronically poisoned with hydrogen sulfide

Hydrogen sulfide poisoning is a common occupational hazard, whose mortality and incidence rates are first and second, respectively, among occupational poisoning incidents in China. The main target organs of its toxicity are in the central nervous system and respiratory system. However, there are currently no specific direct tests that can be used to diagnose poisoned patients. In this study, we developed a serum metabonomic method using orthogonal partial least squares-discriminate analysis (OPLS-DA), based on gas chromatography–mass spectrometry (GC/MS) to evaluate the effect of chronic poisoning by hydrogen sulfide in rats. The OPLS-DA data demonstrated that the model group (n = 60) differed significantly from the control group (n = 30), suggesting that the metabolic profiles of the two groups are markedly different. Alterations in the levels of some metabolites such as citrate, galactose, lactate, mannose, inositol, urea, phosphate, alanine and valine were detected by OPLS-DA analysis. We observed changes in metabolic pathways including lipid metabolism, energy metabolism and amino metabolism in the model group. Our results indicate that GC/MS-based metabonomic methods may provide novel detection means for chronic hydrogen sulfide poisoning.     

Phencyclidine and cannabinoids in vitreous humor

Vitreous humor (VH) is routinely collected at autopsy for the testing of electrolytes and ethanol. In recent years drugs of abuse have been detected in this specimen. In this study the authors assayed 30 VH samples for phencyclidine (PCP) and 50 specimens for cannabinoids. Specimens were screened by immunoassay and then assayed for PCP by GC-FID and cannabinoids by GC/MS. Eighteen (60%) specimens screened positive for PCP using a cutoff of 25ng/mL. Quantitative analysis showed PCP concentrations in VH that screened positive ranged 30-290ng/mL. Corresponding blood concentrations were 50-600ng/mL. VH PCP concentrations in the 12 cases which screened negative were 40-470ng/mL. False negative results were probably due to matrix effects. All VH specimens screened for cannabinoids were negative. Ten negative screening specimens assayed by GC/MS yielded 1 11-nor-9-carboxy-delta-9-tetrahydrocannabinol positive result at 2ng/mL. These data indicated that VH maybe a useful specimen for the detection of PCP but not for cannabinoids.     

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

Summary 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°C evaporator temperature.The cocaine concentrations in hair were determined also by radio-immunoassay (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.     

禁用药物β_2-激动剂的分析研究

采用灵敏度很高的气相色谱 /负离子化学电离质谱 (GC/NICIMS)对 9种国际奥委会禁用药物 β2 -激动剂进行分析研究 ,比较了不同衍生化方法得到的产物在NICIMS上的响应。确定了五氟丙酸酐衍生化产物有最佳的灵敏度 ,适合此类药物的检测要求。重点对克伦特罗尿样进行了检测 ,其结果符合实际工作的需要