Research Interests: Chemistry, Analytical Chemistry, Adolescent, Ireland, Humans, and 15 moreAnalytical Toxicology, Blood Pressure, Female, Clinical Forensic Medicine, Jejunum, Clinical Sciences, High Pressure Liquid Chromatography, Cardiopulmonary Resuscitation, Autopsy, Amlodipine, Felodipine, Calcium Channel Blocker, Calcium Channel Blockers, Heart arrest, and Fatal outcome
Introduction A case report involving repeated intoxications investigated by segmented hair analysis is presented. A 41-year-old man intoxicated his 46-year-old wife several times over half a year. During this period of time the husband... more
Introduction A case report involving repeated intoxications investigated by segmented hair analysis is presented. A 41-year-old man intoxicated his 46-year-old wife several times over half a year. During this period of time the husband went away repeatedly for some days while the wife got sick. Often when he was away, she felt dizzy and nauseated and vomited. One night she was attacked by a man, who was later identified as the husband. The Police suspected the husband for several intoxication attempts on his wife. Methods Hair samples were collected from the top (5 cm long light brown hair with bleached stripes) and from the back of the head (2 cm long light brown unbleached hair) of the wife. Hair was sampled the 15-07-2013, so two of the suspected intoxication attempts (4-4 & 13-5-2013) are represented by segment 2 (1.5–4 cm). The segmented hair samples were washed, dissolved and analyzed for drugs and drugs of abuse by UPLC/TOF and quantified by UPLC/MS/MS (C. Montesano, SS. Johansen et al., JPBA 88 (2014) 295–306) Results Drug findings in head hair from the scalp and back (ng/mg) are shown in the Table. (0–1,5 cm) Scalp hair (4–5 cm) Back hair LOQ (1.5–4 cm) (0–2 cm) Caffeine 1.1 0.42 Trace 1.5 0.10 Codeine 0.009 0.031 nd 0.012 0.005 Lidocaine 0.009 0.024 0.0240 0.014 0.005 Morphine nd 0.006 nd nd 0.005 Tramadol 0.013 0.045 nd 0.015 0.010 O-desmethyltramadol Trace 0.008 nd Trace 0.005 Zolpidem 0.004 0.050 nd Trace 0.002 Diazepam nd Nd nd nd 0.002 Desmethyldiazepam 0.002 Nd nd nd 0.002 Segment 2 has highest drug concentrations of opioids and zolpidem. O-desmethyltramadol is a metabolite from tramadol, while morphine due to the low morphine:codeine ratio probably derives from codeine metabolism. After the violent attack the wife took diazepam a few times and the trace of the metabolite desmethyl-diazepam in the inner segment from scalp hair confirmed the ingestion. In general the drug levels in hair were low indicating few intake(s) and not sustained therapeutic use. The drug findings in the inner hair segment (0–1,5 cm) were com-parable with the drug levels found in the back hair, although the back hair was not bleached. Bleaching is reported to reduce the opioid content in hair from 57 to 94%, however the scalp hair was only partly bleached suggesting a lower loss and high ability to retain the drugs. Conclusion The case illustrates that hair is a valuable forensic specimen when natural processes have eliminated the drug from typical biological specimens such as blood and urine. It was possible to verify suspected intoxications by segmented hair analysis. The husband admitted the intoxications by mixing opioids and zolpidem in the wife’ coffee. He was sentenced to prison for 6 years.
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Abstract Three known HPLC methods for the analysis of 1,2-dibromo-2,4-dicyanobutane (BCB) were compared to evaluate the strength of the methods for the routine analysis of BCB in cosmetic products. Reversed- and normal-phase HPLC with UV... more
Abstract Three known HPLC methods for the analysis of 1,2-dibromo-2,4-dicyanobutane (BCB) were compared to evaluate the strength of the methods for the routine analysis of BCB in cosmetic products. Reversed- and normal-phase HPLC with UV detection were not suitable for routine analysis of BCB in cosmetics. Reversed-phase HPLC with electrochemical detection fulfilled the requirements of a method to be used on a routine basis.
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A 79-year-old woman, with Parkinson's disease treated with cabergoline, was admitted to a hospital due to jaundice and weakness. She was found confused, absent minded, and died after 2 weeks. Autopsy showed an extrahepatic bile... more
A 79-year-old woman, with Parkinson's disease treated with cabergoline, was admitted to a hospital due to jaundice and weakness. She was found confused, absent minded, and died after 2 weeks. Autopsy showed an extrahepatic bile duct adenocarcinoma with spread to the gall bladder, the liver, and regional lymphnodes. While cleaning the hospital bed after her death, the nurses found several tablets hidden in the bed. Biological samples obtained at the autopsy were screened for common drugs and narcotics. Several drugs such as buprenorphine, codeine, paracetamol, and propranolol were detected in the blood at therapeutic levels. A method to determine cabergoline in whole blood and other forensic matrices was developed, and further investigations determined cabergoline concentrations in whole blood and liver tissue of 94 and 3100 microg/kg, respectively. The blood concentration was 100 times above the therapeutic level reported on cabergoline in plasma and in combination with her symptoms, suggest she took a fatal overdose of cabergoline.
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A liquid chromatography mass spectrometric (LC/MS/MS) method has been developed for the determination of LSD, iso-LSD and the metabolite 2-oxo-3-hydroxy-LSD in forensic applications. The procedure involves liquid-liquid extraction of the... more
A liquid chromatography mass spectrometric (LC/MS/MS) method has been developed for the determination of LSD, iso-LSD and the metabolite 2-oxo-3-hydroxy-LSD in forensic applications. The procedure involves liquid-liquid extraction of the analytes and LSD-D3 (internal standard) from 1.0 g whole blood or 1.0 ml urine with butyl acetate at pH 9.8. Confirmation and quantification were done by positive electrospray ionisation with a triple quadrupole mass spectrometer operating in multiple reaction monitoring (MRM) mode. Two MRM transitions of each compound were established and identification criteria were set up based on the retention time and the ion ratio. The curves of extracted standards were linear over a working range of 0.01-50 microg/kg for all transitions of LSD and iso-LSD. The limit of quantification was 0.01 microg/kg for LSD and iso-LSD. The method was applied to a case investigation involving a 26-year-old male suspected for having attempted homicide, where blood concentrations of LSD and iso-LSD were determined to 0.27 and 0.44 microg/kg, respectively. 2-Oxo-3-hydroxy-LSD was detected in the urine and confirmed the LSD abuse. The case illustrated the importance of analyte separation before MRM detection of a sample due to identical fragmentation ions of the isomers.
Research Interests: Chemistry, Analytical Chemistry, Chromatography, Mass Spectrometry, Forensic Medicine, and 11 moreForensic Toxicology, Humans, Male, High Pressure Liquid Chromatography, Liquid Chromatography / Electrospray Ionization Mass Spectrometry, Adult, Reproducibility of Results, Sensitivity and Specificity, SELECTED REACTION MONITORING, Biochemistry and cell biology, and Pharmacology and pharmaceutical sciences
Recently there has been a strong public interest as well as politically on the abuse of ecstasy in Denmark. The Department of Forensic Chemistry performs the toxicological analysis on the driving and criminal cases in Denmark and the... more
Recently there has been a strong public interest as well as politically on the abuse of ecstasy in Denmark. The Department of Forensic Chemistry performs the toxicological analysis on the driving and criminal cases in Denmark and the post-mortem cases of East Denmark requested by the police. Screening for amphetamines and ecstasy is performed by RIA (amphetamine double antibody DPC) and confirmation and quantification by GC/MS analysis. In recent years there has been a change in the number of samples tested positive for designer amphetamines including MDMA (ecstasy), MDA, MDEA etc. The first two positive cases of designer ampetamines among 213 traffic cases requested for analysis by the police were detected in 1997 (2/213 ???0,9%). In 1999 4 cases were found, in 2000 were 9 cases observed and in 2001 were 6 positive cases among 201 analysed traffic cases. In these cases the dominating compound was MDMA followed by MDA. A few positive cases of MDEA were also observed and always in mi...
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In this investigation about 900 saliva samples, collected from drivers not suspected to be under the influence of drugs or alcohol were screened by Cozarts "Drugs of Abuse Microplate EIA" for benzodiazepines, amphetamine,... more
In this investigation about 900 saliva samples, collected from drivers not suspected to be under the influence of drugs or alcohol were screened by Cozarts "Drugs of Abuse Microplate EIA" for benzodiazepines, amphetamine, cannabis, cocaine/benzoylecgonine and opiates. The screening only includes few of the benzodiazepines on the market in Denmark, which means, that only a part of the saliva samples positive for benzodiazepines will be detected by the screening, and that the number of positive benzodiazepine samples will be underestimated. All the saliva samples positive for drugs by the screening were analysed by specific methods, mostly gaschromatography-mass spectrometry (GC-MS). Totally 2% of the investigated saliva samples were positive for drugs. 0.7% (6) of the investigated saliva samples were positive for benzodiazepines, 0.8% (7) for cannabis (tetrahydrocannabinol-THC), 0.3% (3) for opiates, 0.1 % (1) for amphetamine and 0.1% (1) for cocaine/benzoylecgonine. An evaluation of the importance of the saliva concentration will be described. (A) For the covering abstract of the conference, see ITRD Abstract No. E201067.
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A 59-year-old woman who intentionally ingested 100-200 ml Basagran was taken to the hospital with a cardiac arrest 2 days after she had consumed the herbicide. During this period she suffered vomiting, urination and diarrhoea and she was... more
A 59-year-old woman who intentionally ingested 100-200 ml Basagran was taken to the hospital with a cardiac arrest 2 days after she had consumed the herbicide. During this period she suffered vomiting, urination and diarrhoea and she was drowsy with a muddled speech. Biological samples obtained at the autopsy were analysed and presence of bentazone, alcohol and an active metabolite of citalopram were detected. Blood concentrations of bentazone, alcohol and desmethyl-citalopram were 625 mg/kg, 0.62 g/l and 0.03 mg/kg, respectively.
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AbstractWe present a series of forensic cases measuring concentrations in hair of γ-hydroxybutyric acid (GHB) and its glucuronide. The compounds were extracted from hair by incubation for 1.5 h in a 25:25:50 (v/v/v) mixture of... more
AbstractWe present a series of forensic cases measuring concentrations in hair of γ-hydroxybutyric acid (GHB) and its glucuronide. The compounds were extracted from hair by incubation for 1.5 h in a 25:25:50 (v/v/v) mixture of methanol/acetonitrile/2 mM ammonium formate (8 % acetonitrile, pH 5.3). The compounds were determined by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). The limits of quantitation (LOQ) were 0.32 and 0.48 ng/mg hair for GHB and its glucuronide, respectively, and linearity up to 50 ng/mg. Extraction recoveries were 62–92 % for the analytes, and the accuracy was 90–108 %. Precision values expressed as relative standard deviations (%) obtained from daily controls were 9.1–11.3 % for GHB and 12.2–17.9 % for GHB glucuronide. Endogenous levels were up to 1.0 ng/mg for GHB and 1.2 ng/mg for GHB glucuronide in control subjects. In two GHB abusers, GHB was highly elevated (461–591 ng/mg), whereas GHB glucuronide was only slightly elevated in one of the subjects (1.7–3.1 ng/mg) and below the LOQ in the other. Thus, GHB glucuronide is not incorporated in hair in any appreciable amounts in GHB abusers and does not contribute diagnostically with regard to detection of exogenous exposure. To our knowledge, this is the first report to present GHB glucuronide in human hair.
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Differentiation between consumption of illegal and prescription drugs remains an important aspect in forensic toxicology. While illicit amphetamine is most often racemic, the medicinal drugs marketed in Denmark for the treatment of... more
Differentiation between consumption of illegal and prescription drugs remains an important aspect in forensic toxicology. While illicit amphetamine is most often racemic, the medicinal drugs marketed in Denmark for the treatment of attention-deficit hyperactivity disorder contain the pure (S)-enantiomer or a prodrug thereof. In this study, we present a simple and efficient analytical workflow to provide information about the origin of amphetamine consumed in forensic cases concerning driving under the influence of drugs (DUID). Following quantification of amphetamine and methamphetamine using our conventional multi-target ultra-high performance liquid chromatography–tandem mass spectrometry method, determination of (R)- and (S)-amphetamine was performed by reinjecting the sample extract on a Phenomenex LUX® AMP chiral column using the same analytical instrument and mobile phases. Chiral separation was performed isocratic within a run time of 6 min. The analytical workflow was applie...
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Comprehensive drug‐screening performed by liquid chromatography−high resolution mass spectrometry (LC−HRMS) enables identification of hundreds to thousands of drug compounds in a single analysis. Forensic drug screening is generally... more
Comprehensive drug‐screening performed by liquid chromatography−high resolution mass spectrometry (LC−HRMS) enables identification of hundreds to thousands of drug compounds in a single analysis. Forensic drug screening is generally performed with positive electrospray ionization (ESI+), targeting basic drugs; however, a few toxicologically important drugs such as barbiturates, may require analysis by negative ESI. In this work, screening targets for barbiturates were determined using our LC−HRMS screening with ESI+. For several years, our forensic whole blood samples have been analyzed using the LC−HRMS−ESI+ screening in parallel with a multi‐target LC–MS/MS−ESI− method. From 2014 to 2018, 23 samples were positive for phenobarbital (0.5−81 mg/kg). Retrospective data analysis of 4816 blood samples (15 positive) revealed several potential screening targets for phenobarbital. The targets were tentatively identified by exact mass and isotopic pattern as uncommon adducts of phenobarbita...
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Research Interests: Forensic Science, Medicine, Multidisciplinary, Forensic Toxicology, Quantitative analysis, and 15 moreHumans, Blood sampling, Variation, Drug Addiction, Tandem Mass Spectrometry, Coefficient of Variation, High Performance Liquid Chromatography, Legal Medicine, Dominance, Methadone, Femoral Vein, Narcotics, Quantitative Analysis, Metabolite, and Measurement Uncertainty
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We investigated toxicological and pharmacogenetic factors that could influence methadone toxicity using postmortem samples. R- and S-methadone were measured in femoral blood from 90 postmortem cases, mainly drug users. The R-enantiomer... more
We investigated toxicological and pharmacogenetic factors that could influence methadone toxicity using postmortem samples. R- and S-methadone were measured in femoral blood from 90 postmortem cases, mainly drug users. The R-enantiomer concentrations significantly exceeded that of the S-enantiomers (Wilcoxon's test, p < 0.001). The samples were divided into four groups according to other drugs detected (methadone only, methadone and strong analgesics, methadone and benzodiazepines, or methadone and other drugs). There was no significant difference in any of the R-methadone/total methadone ratios among the four groups. The median R/S ratio was 1.38, which tends to be higher than that reported for the plasma of living subjects. In addition, we investigated whether small nucleotide polymorphisms in the MDR1 gene that encode the drug transporter P-glycoprotein were associated with the concentrations of R- and S-methadone and its metabolite 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine. No significant association was detected.
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A multi-target method that can detect a broad range of drugs of abuse in human hair, such as hypnotics, anxiolytics, analgesics, benzodiazepines, antihistamines, antidepressants, antipsychotics, and anticonvulsants, was developed based on... more
A multi-target method that can detect a broad range of drugs of abuse in human hair, such as hypnotics, anxiolytics, analgesics, benzodiazepines, antihistamines, antidepressants, antipsychotics, and anticonvulsants, was developed based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The drugs were extracted from 10 mg of washed hair by incubation for 18 h in a 25:25:50 (v/v/v) mixture of methanol/acetonitrile/2 mM ammonium formate (8% acetonitrile, pH 5.3). For 51% of the basic drugs, the lower limits of quantification (LLOQs) were in the range of 0.05-0.5 pg/mg, and the majority (98%) were ≤ 5 pg/mg. Linearity ranged from LLOQs to 100-500 pg/mg for all of the basic drugs. For acid and neutral drugs, the LLOQs ranged from 0.4 to 500 pg/mg, and linearity ranged from LLOQs to 80-40,000 pg/mg. According to published reports on concentrations attained in single dose control studies, the present method is sensitive enough to detect single dose drug exposure for many of the drugs. The accuracy was within 75-125% for the majority of drugs. Good precision was observed (relative standard deviations [RSD%] < 25%) for most of the compounds, including the prepared quality control (QC) hair samples. The method was applied to forensic cases and concentrations of rarely reported drugs in hair in 25 postmortem forensic cases were presented. Hair concentrations of amisulpride, gabapentin, mianserin, mepyramine, orphenadrine, and xylometazoline have not been previously reported.