Solfrid Hegstad - Academia.edu (original) (raw)
Papers by Solfrid Hegstad
Mutation Research/Environmental Mutagenesis and Related Subjects
Journal of analytical toxicology, Jan 3, 2015
Over the past years, use of synthetic cannabinoids has become increasingly popular. To draw the r... more Over the past years, use of synthetic cannabinoids has become increasingly popular. To draw the right conclusions regarding new intake of these substances in situations of repeated urinary drug testing, knowledge of their elimination rate in urine is essential. We report data from consecutive urine specimens from five subjects after ingestion of synthetic cannabinoids. Urinary concentrations of the carboxylic acid metabolites JWH-018-COOH and JWH-073-COOH were measured by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) with a limit of quantification of 0.1 ng/mL. In these subjects, specimens remained positive over a period of 20-43 (mean 27) days for JWH-018-COOH and over a period of 11-25 (mean 19) days for JWH-073-COOH. Detection times were shorter for subjects that appeared to have ingested only one, or a few, doses prior to urine collection in the study. Creatinine-normalized concentrations (CN-concentrations) slowly declined throughout the follow-u...
Tidsskrift for den Norske lægeforening : tidsskrift for praktisk medicin, ny række, Jan 17, 2008
Buprenorphine is one of the drugs used for treatment of opioid-dependent patients enrolled in reh... more Buprenorphine is one of the drugs used for treatment of opioid-dependent patients enrolled in rehabilitation programs in Norway. Buprenorphine is metabolized in the liver by cytochrome P450 to the active metabolite norbuprenorphine, and further to buprenorphine-glucuronide and norbuprenorphine-glucuronide. The Division of Forensic Toxicology and Drug Abuse at the Norwegian Institute of Public Health has during the past 5 years received an increasing number of urine samples for buprenorphine analysis. All urine samples with question of buprenorphine have since August 2005 been analysed with a new method, which analyses the glucuronides of buprenorphine and norbuprenorphine in urine. This method is fast and simple and saves time and resources in our routine laboratory.
Tidsskrift for Den norske legeforening, 2009
Journal of analytical toxicology, Jan 3, 2015
The toxicodynamics and, to a lesser degree, toxicokinetics of the widely used opiate codeine rema... more The toxicodynamics and, to a lesser degree, toxicokinetics of the widely used opiate codeine remain a matter of controversy. To address this issue, analytical methods capable of providing reliable quantification of codeine metabolites alongside codeine concentrations are required. This article presents a validated method for simultaneous determination of codeine, codeine metabolites codeine-6-glucuronide (C6G), norcodeine and morphine, and morphine metabolites morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in post-mortem whole blood, vitreous fluid, muscle, fat and brain tissue by high-performance liquid chromatography mass spectrometry. Samples were prepared by solid-phase extraction. The validated ranges were 1.5-300 ng/mL for codeine, norcodeine and morphine, and 23-4,600 ng/mL for C6G, M3G and M6G, with exceptions for norcodeine in muscle (3-300 ng/mL), morphine in muscle, fat and brain (3-300 ng/mL) and M6G in fat (46-4,600 ng/mL). Within-run and between-run accu...
Xenobiotica, 1999
1. The excretion of benz[j]aceanthrylene (B[j]A) and the biotransformation products found in faec... more 1. The excretion of benz[j]aceanthrylene (B[j]A) and the biotransformation products found in faeces, urine and bile of rat exposed to [3H]-labelled B[j]A have been studied. 2. About 95% of the administered radioactivity was excreted within 7 days, 79% via faeces and 16% via urine, and most of the radioactivity in urine and faeces was excreted within 2 days. 3. The B[j]A metabolites excreted between days 1 and 2, including those excreted in bile during the first 5.5 h in a separate experiment, were further characterized by HPLC, UV and electrospray/atmospheric pressure chemical ionization mass spectrometry. 4. In faeces, bile and urine, hydroxylated B[j]A metabolites predominated. The major metabolites in faeces were B[j]A-1,2-dihydrodiol-8-hydroxy and B[j]A-1,2-dihydrodiol-10-hydroxy. These metabolites were found as conjugated metabolites in the bile. The glucuronide conjugate of B[j]A-1,2-dihydrodiol-10-hydroxy was also a major metabolite in urine. Two sulphate conjugates of oxidized B[j]A were detected in bile, a sulphate conjugate of a B[j]A-dihydrodiol-phenol and B[j]A-1,2-dihydrodiol-10-sulphate. Trans-B[j]A-1,2-dihydrodiol was detected in urine, faeces and bile. 5. These findings support the hypothesis that epoxidation at the cyclopenta ring is an important biotransformation pathway for B[j]A in vivo. In addition to the characterized metabolites, a large fraction of polar compounds, possibly glutathione conjugates, was also excreted in urine and bile.
Journal of Chromatography B, 2014
The purpose of this work was to develop and evaluate a fast, robust and specific UPLC-MS/MS scree... more The purpose of this work was to develop and evaluate a fast, robust and specific UPLC-MS/MS screening platform for the determination and quantification of a variety of commonly used drugs of abuse in urine, i.e. a high-throughput quantitative analysis. Substances in the drug classes opioids, central nervous system stimulants and benzodiazepines and related agents were included in addition to cannabis and pregabalin, a total of 35 different analytes. Based on the concentrations and the physico-chemical properties of the substances, three UPLC-MS/MS methods were developed in parallel. Prior to analysis, sample preparation consisted of two different simple dilutions with 60 and 100 L urine, respectively, using a Tecan Freedom Evo pipetting robot platform. A Waters Xevo TQ-S tandem quadrupole mass spectrometer coupled to a Waters I-class UPLC was used for quantitative analysis of one quantitative and one qualifying MRM transition for each analyte, except for tramadol for which the metabolite O-desmethyl-tramadol was included in the MRM method to confirm tramadol identity. Deuterated analogs were included as internal standards. The between-assay relative standard deviations varied from 2% to 11% and the limits of quantification were in the range 1-200 ng/mL for the various analytes. After development and initial testing, the method has been successfully implemented and routinely used at our hospital for quantitative screening of drugs of abuse in more than 35,000 urinary samples.
Journal of Analytical Toxicology, 2009
Cotinine is the main metabolite of nicotine and is used as an indicator of exposure to tobacco sm... more Cotinine is the main metabolite of nicotine and is used as an indicator of exposure to tobacco smoke. A method has been developed for quantification of cotinine in pericardial fluid and whole blood collected from autopsy casework involving cases of infant death. Sample clean-up was achieved by solid-phase extraction with a mixed-mode column. Cotinine was quantified by liquid chromatography-tandem mass spectrometry. Positive ionization was performed in the multiple reaction monitoring mode. Two transitions were monitored for the analyte and one for the internal standard, cotinine-d 3 . The calibration range was 0.9-176 ng/mL for cotinine in both matrixes. The recovery of the analyte ranged from 86 to 92%, and the between-assay precisions ranged from 4 to 6% relative standard deviation. Whole blood and pericardial fluid samples from 95 infant deaths obtained during autopsy were analyzed. A strong correlation (R 2 = 0.97) was found between the cotinine concentrations in pericardial fluid and blood. The correlation was not affected by the postmortem time interval. This study demonstrates that pericardial fluid may be an alternative specimen to blood for quantification of cotinine in forensic autopsies.
Journal of Analytical Toxicology, 2009
An ultra-performance liquid chromatography-tandem mass spectrometry method has been developed and... more An ultra-performance liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of ethylglucuronide (EtG) in oral fluid. Sample clean-up was achieved by solid-phase extraction with a Hyper-SEP SAX column. Negative ionization was performed in the multiple reaction monitoring mode. Two transitions were monitored for the analyte and one for the internal standard EtG-d 5 . The calibration range was 4.4-222 ng/mL. The recovery of the analyte ranged from 86 to 99%, and the between-assay precisions ranged from 5 to 9% RSD. The limit of quantification was found to be 4.4 ng/mL. The concentration of EtG in oral fluid collected 2-14 h after a moderate alcohol intake varied from 13.3 to 57.7 ng/mL.
Journal of Analytical Toxicology, 2013
In cases of sexual assault, victims often present too late for the detection of ethanol in biolog... more In cases of sexual assault, victims often present too late for the detection of ethanol in biological samples. An ultra-performance liquid chromatography -tandem mass spectrometry (UPLC-MS-MS) method was developed and validated for the determination of ethyl glucuronide (EtG) and ethyl sulfate (EtS) in urine. Sample preparation prior to UPLC -MS-MS analysis was a simple sample dilution. The calibration ranges were 0.2 -20 mg/L, and between-assay relative standard deviations were in the range of 0.7-7.0% at concentrations of 0.3, 3.0 and 7.0 mg/L. Urine samples were analyzed from 59 female patients presenting to the Sexual Assault Centre at St. Olav University Hospital in Trondheim, Norway between November 2010 and October 2011. EtG and EtS results were fully concordant, and positive in 45 of the 48 cases with self-reported alcohol intake. In contrast, ethanol was detectable in only 20 of these cases, corresponding to sensitivities of 94 and 42%, respectively. Of the patients reporting no alcohol intake, none had positive EtG/EtS findings. These data show that analysis of EtG and EtS greatly increases the detection window of alcohol ingestion in cases of sexual assault, and may shed additional light on the involvement of ethanol in such cases. The victims' self-reported intake of alcohol seems to be reliable in this study, according to the EtG/EtS findings.
Drug and Alcohol Dependence, 2010
Background: Naltrexone's usefulness in the treatment of opioid dependence stems from its ability ... more Background: Naltrexone's usefulness in the treatment of opioid dependence stems from its ability to block the action of heroin and other opioids. However, many patients are ambivalent towards naltrexone and often drop out of treatment with orally administered naltrexone. Sustained release naltrexone seems promising in reducing opioid use, but the extent to which patients remain in treatment beyond the first dosage of naltrexone is not clear. Methods: Patients (n = 61) receving treatment with sustained release naltrexone implants were offered a second naltrexone implant after 6 months. Patients who remained in treatment were compared to those who did not, on drug use, mental health, and social problems before and during naltrexone implant treatment. Information was obtained on other treatments sought by patients who discontinued naltrexone. Blood samples were used to verify naltrexone release, and hair samples to confirm opioid intake. Results: Of the patients who received the first naltrexone implant, 51% (n = 31) remained in naltrexone implant treatment. Among those who discontinued treatment, 21% expressed a wish to reimplant but failed to attend for reimplantation and 28% declined reimplantation: 6 non-retained patients initiated maintenance or residential treatment. Remaining in naltrexone treatment was related to pre-study length of employment, illicit drug use, and concern for family problems. Higher levels of substance misuse and criminal activity during naltrexone treatment were negatively related to subsequent retention. Conclusion: Rates of retention among opioid-dependent patients receiving naltrexone implant treatment are encouraging and support this as a feasible long-term treatment option.
The British Journal of Psychiatry, 2009
Despite decades of research confirming naltrexone's pharmacological efficacy in blocking the acti... more Despite decades of research confirming naltrexone's pharmacological efficacy in blocking the actions of heroin, 1 its clinical usefulness has proved limited. 2 Many people who are opiatedependent are reluctant or refuse to take naltrexone; 3,4 many others begin but do not continue treatment. 5-7 Sustained-release opioid antagonist treatment removes the requirement for patients to take daily doses of the medication, 8 and one depot naltrexone formulation has been shown to reduce heroin use safely and effectively for the duration of its 4-week release period. 9,10 Despite these advances, longer-lasting products are desirable in order to minimise patients' opportunity for between-dosage withdrawal and relapse. Preliminary studies indicate that one type of implantable naltrexone provides about 6 months' opioid receptor blockade and has a satisfactory safety profile. No randomised clinical trial of these implants has been done and the question of efficacy is still unanswered. This paper reports the findings of a randomised clinical trial of the safety and effectiveness of naltrexone implants in opioid-dependent patients who had completed in-patient treatment. We predicted that patients with a naltrexone implant would use fewer opioids and have fewer overdoses; also, that they would be less depressed, and would have better outcomes for work, education and criminal behaviour at follow-up compared with patients randomised to usual aftercare.
![Research paper thumbnail of Incorporation of the Food Mutagen 2-Amino-1-Methyl-6-Phenylimidazo[4,5-b]Pyridine (PhIP) into Fur and Correlation with Intestinal Tumourigenesis in Min/+ Mice](https://attachments.academia-assets.com/46956914/thumbnails/1.jpg)
](Pharmacology and Toxicology, 2003
The purpose of this work was to correlate the amount of the food mutagen 2-amino-1-methyl-6-pheny... more The purpose of this work was to correlate the amount of the food mutagen 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) determined in mouse fur with the number of intestinal tumours induced by PhIP, to further evaluate incorporation of PhIP into hair as a putative exposure biomarker for food mutagens. We have previously shown that PhIP increases intestinal tumourigenesis in C57BL/6J-Min/π (Multiple Intestinal Neoplasia) mice. Fur was sampled from mice exposed according to various PhIP protocols, and the amount of PhIP in the fur was quantitated by high performance liquid chromatography -mass spectrometry (HPLC-MS). A quantitative incorporation of PhIP in the fur was demonstrated after a single subcutaneous injection of PhIP, and between one and eight PhIP exposures either via direct subcutaneous injections or through breast milk from PhIP-injected dams. However, after higher exposures, the amount of PhIP in the fur appeared to reach saturation. After low exposures to PhIP, the number of intestinal tumours correlated with the amount of PhIP in the fur of individual mice, whereas after higher exposures, the number of tumours was relatively higher than the amounts of incorporated PhIP in the fur. Other factors, e.g. amounts of reactive PhIP metabolites formed, are also determining the number of intestinal tumours. The demonstrated quantitative incorporation of PhIP into mice fur and the correlation with number of intestinal tumours at the lower exposures, indicate that determination of PhIP in human hair may be a suitable biomarker for exposure to dietary PhIP, which is found in human hair in 10 ª3 lower amounts than in these mice.
![Research paper thumbnail of Eumelanin is a Major Determinant for2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) Incorporation into Hair of Mice](https://attachments.academia-assets.com/46956910/thumbnails/1.jpg)
](Pharmacology and Toxicology, 2002
Mice with different hair pigmentation were studied to evaluate the role of melanin in the incorpo... more Mice with different hair pigmentation were studied to evaluate the role of melanin in the incorporation of 2amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) into hair. Mice C57BL/6J-c2j/π (white), C57BL/6J-Ay (yellow), C57L/J (grey), C57BR/cdJ (brown) and C57BL/6J (black) were dosed with PhIP: 7-9 days old (total amount: 0.006 or 0.58 mg/kg b.wt., for 4 days) and adults (total amount 50 mg/kg b.wt. during 8 weeks). Hair was collected either 30 days after the last PhIP administration (new-born mice) or 8 weeks after the first administration (adult mice). PhIP was incorporated into black hair to a greater extent than into brown, grey, yellow and non-pigmented hair. The concentration of PhIP in the hair of new-born mice exposed to 0.58 mg PhIP/kg b.wt. were (mean∫S.D.): 328∫135 (black), 134∫41 (brown), 9.1∫1.2 (yellow) and 5.2∫1.4 (white) ng/g hair. The PhIP concentrations in the hair of adult mice exposed to 50 mg/kg b.wt. were: 4750∫1449 (black), 810∫235 (brown), 541∫119 (grey), 35.5∫4.6 (yellow) and 21.6∫8.8 (white) ng/g, and the eumelanin hair concentration in the same animals decreased in a similar pattern. A linear relationship (r 2 Ω 1.00, PϽ0.0001) between the relative PhIP incorporation and the eumelanin concentration in hair was found.
Addiction, 2010
Aims Naltrexone is a competitive opioid antagonist that effectively blocks the action of heroin a... more Aims Naltrexone is a competitive opioid antagonist that effectively blocks the action of heroin and other opioid agonists. Sustained-release naltrexone formulations are now available that provide long-acting opioid blockade. This study investigates the use of heroin and other opioids among opioid-dependent patients receiving treatment with long-acting naltrexone implants, their subjective experience of drug 'high' after opioid use, and factors associated with opioid use. Methods Participants (n = 60) were opioid-dependent patients receiving treatment with naltrexone implants. Outcome data on substance use, drug 'high', depression and criminal activity were collected over a 6-month period. Blood samples were taken to monitor naltrexone plasma levels, and hair samples to verify self-reported opioid use. Findings More than half [n = 34 or 56%; 95% confidence interval (CI) 44-68%)] the patients challenged the blockade with illicit opioids during the 6-month treatment period; 44% (n = 26; 95% CI 32-56%) were abstinent from opioids. Mean opioid use was reduced from 18 [standard deviation (SD)13] days during the month preceding treatment to 6 days (SD 11) after 6 months. Of the respondents questioned on opioid 'high' (n = 31), nine patients (30%; 95% CI 16-47%) reported partial drug 'high' following illicit opioid use, and three (12%; 95% CI 3-26%) reported full 'high'. Opioid use was associated with use of non-opioid drugs and criminal behaviour. Conclusions Challenging naltrexone blockade with heroin on at least one occasion is common among sustained-release naltrexone patients, but only a minority of patients use opioids regularly. Challenges represent a warning sign for poor outcomes and often occur in the context of polydrug use and social adjustment problems.
Food and Chemical Toxicology, 2002
Various methods of exposure assessment, such as questionnaires, sometimes combined with pictures ... more Various methods of exposure assessment, such as questionnaires, sometimes combined with pictures of cooked meat, have been employed in investigations on the relationship between heterocyclic amines (HA) and health effects. However, as the content of heterocyclic amines vary greatly with cooking conditions, it is difficult to obtain an accurate estimate of the exposure. To improve the exposure assessment, the use of biomarkers has been investigated. The metabolism of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is well characterised. In humans, the major part of the dose is excreted in urine within 24-48 h following a meal. A few percent is excreted as parent compounds, whereas the major part is metabolites. Urinary level of parent HA reflects only recent exposure. However, the pattern of excreted metabolites might indicate the capacity to activate or detoxify HAs. The excretion of glucuronide conjugates of N-hydroxy-PhIP and N-hydroxy-MeIQx could be a marker for the N-hydroxylation capacity and the dose of the proximate metabolites. Recently, we proposed 5-OH-PhIP as a marker for the ultimate reactive metabolite of PhIP, since it is formed from this compound as a by-product along with the formation of PhIP-DNA adducts. In a search for biomarkers reflecting exposure over some time, blood protein adducts with a longer lifespan have been investigated, and PhIP adducts of serum albumin and haemoglobin from meat-consuming humans were recently reported. Many compounds, like drugs, nicotine and narcotics, bind to melanin in hair and give information on exposure for longer time periods. In mice, PhIP is irreversibly incorporated in a dose-dependent manner into hair, and in humans exposed to an ordinary diet, it was found to vary from <50 to 5000 pg PhIP/g hair. The incorporation is also dependent on the content of eumelanin. The use of PhIP in hair as a biomarker of exposure is promising, but needs validation, using other methods of exposure assessment. #
Mutation Research/Environmental Mutagenesis and Related Subjects
Journal of analytical toxicology, Jan 3, 2015
Over the past years, use of synthetic cannabinoids has become increasingly popular. To draw the r... more Over the past years, use of synthetic cannabinoids has become increasingly popular. To draw the right conclusions regarding new intake of these substances in situations of repeated urinary drug testing, knowledge of their elimination rate in urine is essential. We report data from consecutive urine specimens from five subjects after ingestion of synthetic cannabinoids. Urinary concentrations of the carboxylic acid metabolites JWH-018-COOH and JWH-073-COOH were measured by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) with a limit of quantification of 0.1 ng/mL. In these subjects, specimens remained positive over a period of 20-43 (mean 27) days for JWH-018-COOH and over a period of 11-25 (mean 19) days for JWH-073-COOH. Detection times were shorter for subjects that appeared to have ingested only one, or a few, doses prior to urine collection in the study. Creatinine-normalized concentrations (CN-concentrations) slowly declined throughout the follow-u...
Tidsskrift for den Norske lægeforening : tidsskrift for praktisk medicin, ny række, Jan 17, 2008
Buprenorphine is one of the drugs used for treatment of opioid-dependent patients enrolled in reh... more Buprenorphine is one of the drugs used for treatment of opioid-dependent patients enrolled in rehabilitation programs in Norway. Buprenorphine is metabolized in the liver by cytochrome P450 to the active metabolite norbuprenorphine, and further to buprenorphine-glucuronide and norbuprenorphine-glucuronide. The Division of Forensic Toxicology and Drug Abuse at the Norwegian Institute of Public Health has during the past 5 years received an increasing number of urine samples for buprenorphine analysis. All urine samples with question of buprenorphine have since August 2005 been analysed with a new method, which analyses the glucuronides of buprenorphine and norbuprenorphine in urine. This method is fast and simple and saves time and resources in our routine laboratory.
Tidsskrift for Den norske legeforening, 2009
Journal of analytical toxicology, Jan 3, 2015
The toxicodynamics and, to a lesser degree, toxicokinetics of the widely used opiate codeine rema... more The toxicodynamics and, to a lesser degree, toxicokinetics of the widely used opiate codeine remain a matter of controversy. To address this issue, analytical methods capable of providing reliable quantification of codeine metabolites alongside codeine concentrations are required. This article presents a validated method for simultaneous determination of codeine, codeine metabolites codeine-6-glucuronide (C6G), norcodeine and morphine, and morphine metabolites morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in post-mortem whole blood, vitreous fluid, muscle, fat and brain tissue by high-performance liquid chromatography mass spectrometry. Samples were prepared by solid-phase extraction. The validated ranges were 1.5-300 ng/mL for codeine, norcodeine and morphine, and 23-4,600 ng/mL for C6G, M3G and M6G, with exceptions for norcodeine in muscle (3-300 ng/mL), morphine in muscle, fat and brain (3-300 ng/mL) and M6G in fat (46-4,600 ng/mL). Within-run and between-run accu...
Xenobiotica, 1999
1. The excretion of benz[j]aceanthrylene (B[j]A) and the biotransformation products found in faec... more 1. The excretion of benz[j]aceanthrylene (B[j]A) and the biotransformation products found in faeces, urine and bile of rat exposed to [3H]-labelled B[j]A have been studied. 2. About 95% of the administered radioactivity was excreted within 7 days, 79% via faeces and 16% via urine, and most of the radioactivity in urine and faeces was excreted within 2 days. 3. The B[j]A metabolites excreted between days 1 and 2, including those excreted in bile during the first 5.5 h in a separate experiment, were further characterized by HPLC, UV and electrospray/atmospheric pressure chemical ionization mass spectrometry. 4. In faeces, bile and urine, hydroxylated B[j]A metabolites predominated. The major metabolites in faeces were B[j]A-1,2-dihydrodiol-8-hydroxy and B[j]A-1,2-dihydrodiol-10-hydroxy. These metabolites were found as conjugated metabolites in the bile. The glucuronide conjugate of B[j]A-1,2-dihydrodiol-10-hydroxy was also a major metabolite in urine. Two sulphate conjugates of oxidized B[j]A were detected in bile, a sulphate conjugate of a B[j]A-dihydrodiol-phenol and B[j]A-1,2-dihydrodiol-10-sulphate. Trans-B[j]A-1,2-dihydrodiol was detected in urine, faeces and bile. 5. These findings support the hypothesis that epoxidation at the cyclopenta ring is an important biotransformation pathway for B[j]A in vivo. In addition to the characterized metabolites, a large fraction of polar compounds, possibly glutathione conjugates, was also excreted in urine and bile.
Journal of Chromatography B, 2014
The purpose of this work was to develop and evaluate a fast, robust and specific UPLC-MS/MS scree... more The purpose of this work was to develop and evaluate a fast, robust and specific UPLC-MS/MS screening platform for the determination and quantification of a variety of commonly used drugs of abuse in urine, i.e. a high-throughput quantitative analysis. Substances in the drug classes opioids, central nervous system stimulants and benzodiazepines and related agents were included in addition to cannabis and pregabalin, a total of 35 different analytes. Based on the concentrations and the physico-chemical properties of the substances, three UPLC-MS/MS methods were developed in parallel. Prior to analysis, sample preparation consisted of two different simple dilutions with 60 and 100 L urine, respectively, using a Tecan Freedom Evo pipetting robot platform. A Waters Xevo TQ-S tandem quadrupole mass spectrometer coupled to a Waters I-class UPLC was used for quantitative analysis of one quantitative and one qualifying MRM transition for each analyte, except for tramadol for which the metabolite O-desmethyl-tramadol was included in the MRM method to confirm tramadol identity. Deuterated analogs were included as internal standards. The between-assay relative standard deviations varied from 2% to 11% and the limits of quantification were in the range 1-200 ng/mL for the various analytes. After development and initial testing, the method has been successfully implemented and routinely used at our hospital for quantitative screening of drugs of abuse in more than 35,000 urinary samples.
Journal of Analytical Toxicology, 2009
Cotinine is the main metabolite of nicotine and is used as an indicator of exposure to tobacco sm... more Cotinine is the main metabolite of nicotine and is used as an indicator of exposure to tobacco smoke. A method has been developed for quantification of cotinine in pericardial fluid and whole blood collected from autopsy casework involving cases of infant death. Sample clean-up was achieved by solid-phase extraction with a mixed-mode column. Cotinine was quantified by liquid chromatography-tandem mass spectrometry. Positive ionization was performed in the multiple reaction monitoring mode. Two transitions were monitored for the analyte and one for the internal standard, cotinine-d 3 . The calibration range was 0.9-176 ng/mL for cotinine in both matrixes. The recovery of the analyte ranged from 86 to 92%, and the between-assay precisions ranged from 4 to 6% relative standard deviation. Whole blood and pericardial fluid samples from 95 infant deaths obtained during autopsy were analyzed. A strong correlation (R 2 = 0.97) was found between the cotinine concentrations in pericardial fluid and blood. The correlation was not affected by the postmortem time interval. This study demonstrates that pericardial fluid may be an alternative specimen to blood for quantification of cotinine in forensic autopsies.
Journal of Analytical Toxicology, 2009
An ultra-performance liquid chromatography-tandem mass spectrometry method has been developed and... more An ultra-performance liquid chromatography-tandem mass spectrometry method has been developed and validated for the determination of ethylglucuronide (EtG) in oral fluid. Sample clean-up was achieved by solid-phase extraction with a Hyper-SEP SAX column. Negative ionization was performed in the multiple reaction monitoring mode. Two transitions were monitored for the analyte and one for the internal standard EtG-d 5 . The calibration range was 4.4-222 ng/mL. The recovery of the analyte ranged from 86 to 99%, and the between-assay precisions ranged from 5 to 9% RSD. The limit of quantification was found to be 4.4 ng/mL. The concentration of EtG in oral fluid collected 2-14 h after a moderate alcohol intake varied from 13.3 to 57.7 ng/mL.
Journal of Analytical Toxicology, 2013
In cases of sexual assault, victims often present too late for the detection of ethanol in biolog... more In cases of sexual assault, victims often present too late for the detection of ethanol in biological samples. An ultra-performance liquid chromatography -tandem mass spectrometry (UPLC-MS-MS) method was developed and validated for the determination of ethyl glucuronide (EtG) and ethyl sulfate (EtS) in urine. Sample preparation prior to UPLC -MS-MS analysis was a simple sample dilution. The calibration ranges were 0.2 -20 mg/L, and between-assay relative standard deviations were in the range of 0.7-7.0% at concentrations of 0.3, 3.0 and 7.0 mg/L. Urine samples were analyzed from 59 female patients presenting to the Sexual Assault Centre at St. Olav University Hospital in Trondheim, Norway between November 2010 and October 2011. EtG and EtS results were fully concordant, and positive in 45 of the 48 cases with self-reported alcohol intake. In contrast, ethanol was detectable in only 20 of these cases, corresponding to sensitivities of 94 and 42%, respectively. Of the patients reporting no alcohol intake, none had positive EtG/EtS findings. These data show that analysis of EtG and EtS greatly increases the detection window of alcohol ingestion in cases of sexual assault, and may shed additional light on the involvement of ethanol in such cases. The victims' self-reported intake of alcohol seems to be reliable in this study, according to the EtG/EtS findings.
Drug and Alcohol Dependence, 2010
Background: Naltrexone's usefulness in the treatment of opioid dependence stems from its ability ... more Background: Naltrexone's usefulness in the treatment of opioid dependence stems from its ability to block the action of heroin and other opioids. However, many patients are ambivalent towards naltrexone and often drop out of treatment with orally administered naltrexone. Sustained release naltrexone seems promising in reducing opioid use, but the extent to which patients remain in treatment beyond the first dosage of naltrexone is not clear. Methods: Patients (n = 61) receving treatment with sustained release naltrexone implants were offered a second naltrexone implant after 6 months. Patients who remained in treatment were compared to those who did not, on drug use, mental health, and social problems before and during naltrexone implant treatment. Information was obtained on other treatments sought by patients who discontinued naltrexone. Blood samples were used to verify naltrexone release, and hair samples to confirm opioid intake. Results: Of the patients who received the first naltrexone implant, 51% (n = 31) remained in naltrexone implant treatment. Among those who discontinued treatment, 21% expressed a wish to reimplant but failed to attend for reimplantation and 28% declined reimplantation: 6 non-retained patients initiated maintenance or residential treatment. Remaining in naltrexone treatment was related to pre-study length of employment, illicit drug use, and concern for family problems. Higher levels of substance misuse and criminal activity during naltrexone treatment were negatively related to subsequent retention. Conclusion: Rates of retention among opioid-dependent patients receiving naltrexone implant treatment are encouraging and support this as a feasible long-term treatment option.
The British Journal of Psychiatry, 2009
Despite decades of research confirming naltrexone's pharmacological efficacy in blocking the acti... more Despite decades of research confirming naltrexone's pharmacological efficacy in blocking the actions of heroin, 1 its clinical usefulness has proved limited. 2 Many people who are opiatedependent are reluctant or refuse to take naltrexone; 3,4 many others begin but do not continue treatment. 5-7 Sustained-release opioid antagonist treatment removes the requirement for patients to take daily doses of the medication, 8 and one depot naltrexone formulation has been shown to reduce heroin use safely and effectively for the duration of its 4-week release period. 9,10 Despite these advances, longer-lasting products are desirable in order to minimise patients' opportunity for between-dosage withdrawal and relapse. Preliminary studies indicate that one type of implantable naltrexone provides about 6 months' opioid receptor blockade and has a satisfactory safety profile. No randomised clinical trial of these implants has been done and the question of efficacy is still unanswered. This paper reports the findings of a randomised clinical trial of the safety and effectiveness of naltrexone implants in opioid-dependent patients who had completed in-patient treatment. We predicted that patients with a naltrexone implant would use fewer opioids and have fewer overdoses; also, that they would be less depressed, and would have better outcomes for work, education and criminal behaviour at follow-up compared with patients randomised to usual aftercare.
Pharmacology and Toxicology, 2003
The purpose of this work was to correlate the amount of the food mutagen 2-amino-1-methyl-6-pheny... more The purpose of this work was to correlate the amount of the food mutagen 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) determined in mouse fur with the number of intestinal tumours induced by PhIP, to further evaluate incorporation of PhIP into hair as a putative exposure biomarker for food mutagens. We have previously shown that PhIP increases intestinal tumourigenesis in C57BL/6J-Min/π (Multiple Intestinal Neoplasia) mice. Fur was sampled from mice exposed according to various PhIP protocols, and the amount of PhIP in the fur was quantitated by high performance liquid chromatography -mass spectrometry (HPLC-MS). A quantitative incorporation of PhIP in the fur was demonstrated after a single subcutaneous injection of PhIP, and between one and eight PhIP exposures either via direct subcutaneous injections or through breast milk from PhIP-injected dams. However, after higher exposures, the amount of PhIP in the fur appeared to reach saturation. After low exposures to PhIP, the number of intestinal tumours correlated with the amount of PhIP in the fur of individual mice, whereas after higher exposures, the number of tumours was relatively higher than the amounts of incorporated PhIP in the fur. Other factors, e.g. amounts of reactive PhIP metabolites formed, are also determining the number of intestinal tumours. The demonstrated quantitative incorporation of PhIP into mice fur and the correlation with number of intestinal tumours at the lower exposures, indicate that determination of PhIP in human hair may be a suitable biomarker for exposure to dietary PhIP, which is found in human hair in 10 ª3 lower amounts than in these mice.
Pharmacology and Toxicology, 2002
Mice with different hair pigmentation were studied to evaluate the role of melanin in the incorpo... more Mice with different hair pigmentation were studied to evaluate the role of melanin in the incorporation of 2amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) into hair. Mice C57BL/6J-c2j/π (white), C57BL/6J-Ay (yellow), C57L/J (grey), C57BR/cdJ (brown) and C57BL/6J (black) were dosed with PhIP: 7-9 days old (total amount: 0.006 or 0.58 mg/kg b.wt., for 4 days) and adults (total amount 50 mg/kg b.wt. during 8 weeks). Hair was collected either 30 days after the last PhIP administration (new-born mice) or 8 weeks after the first administration (adult mice). PhIP was incorporated into black hair to a greater extent than into brown, grey, yellow and non-pigmented hair. The concentration of PhIP in the hair of new-born mice exposed to 0.58 mg PhIP/kg b.wt. were (mean∫S.D.): 328∫135 (black), 134∫41 (brown), 9.1∫1.2 (yellow) and 5.2∫1.4 (white) ng/g hair. The PhIP concentrations in the hair of adult mice exposed to 50 mg/kg b.wt. were: 4750∫1449 (black), 810∫235 (brown), 541∫119 (grey), 35.5∫4.6 (yellow) and 21.6∫8.8 (white) ng/g, and the eumelanin hair concentration in the same animals decreased in a similar pattern. A linear relationship (r 2 Ω 1.00, PϽ0.0001) between the relative PhIP incorporation and the eumelanin concentration in hair was found.
Addiction, 2010
Aims Naltrexone is a competitive opioid antagonist that effectively blocks the action of heroin a... more Aims Naltrexone is a competitive opioid antagonist that effectively blocks the action of heroin and other opioid agonists. Sustained-release naltrexone formulations are now available that provide long-acting opioid blockade. This study investigates the use of heroin and other opioids among opioid-dependent patients receiving treatment with long-acting naltrexone implants, their subjective experience of drug 'high' after opioid use, and factors associated with opioid use. Methods Participants (n = 60) were opioid-dependent patients receiving treatment with naltrexone implants. Outcome data on substance use, drug 'high', depression and criminal activity were collected over a 6-month period. Blood samples were taken to monitor naltrexone plasma levels, and hair samples to verify self-reported opioid use. Findings More than half [n = 34 or 56%; 95% confidence interval (CI) 44-68%)] the patients challenged the blockade with illicit opioids during the 6-month treatment period; 44% (n = 26; 95% CI 32-56%) were abstinent from opioids. Mean opioid use was reduced from 18 [standard deviation (SD)13] days during the month preceding treatment to 6 days (SD 11) after 6 months. Of the respondents questioned on opioid 'high' (n = 31), nine patients (30%; 95% CI 16-47%) reported partial drug 'high' following illicit opioid use, and three (12%; 95% CI 3-26%) reported full 'high'. Opioid use was associated with use of non-opioid drugs and criminal behaviour. Conclusions Challenging naltrexone blockade with heroin on at least one occasion is common among sustained-release naltrexone patients, but only a minority of patients use opioids regularly. Challenges represent a warning sign for poor outcomes and often occur in the context of polydrug use and social adjustment problems.
Food and Chemical Toxicology, 2002
Various methods of exposure assessment, such as questionnaires, sometimes combined with pictures ... more Various methods of exposure assessment, such as questionnaires, sometimes combined with pictures of cooked meat, have been employed in investigations on the relationship between heterocyclic amines (HA) and health effects. However, as the content of heterocyclic amines vary greatly with cooking conditions, it is difficult to obtain an accurate estimate of the exposure. To improve the exposure assessment, the use of biomarkers has been investigated. The metabolism of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is well characterised. In humans, the major part of the dose is excreted in urine within 24-48 h following a meal. A few percent is excreted as parent compounds, whereas the major part is metabolites. Urinary level of parent HA reflects only recent exposure. However, the pattern of excreted metabolites might indicate the capacity to activate or detoxify HAs. The excretion of glucuronide conjugates of N-hydroxy-PhIP and N-hydroxy-MeIQx could be a marker for the N-hydroxylation capacity and the dose of the proximate metabolites. Recently, we proposed 5-OH-PhIP as a marker for the ultimate reactive metabolite of PhIP, since it is formed from this compound as a by-product along with the formation of PhIP-DNA adducts. In a search for biomarkers reflecting exposure over some time, blood protein adducts with a longer lifespan have been investigated, and PhIP adducts of serum albumin and haemoglobin from meat-consuming humans were recently reported. Many compounds, like drugs, nicotine and narcotics, bind to melanin in hair and give information on exposure for longer time periods. In mice, PhIP is irreversibly incorporated in a dose-dependent manner into hair, and in humans exposed to an ordinary diet, it was found to vary from <50 to 5000 pg PhIP/g hair. The incorporation is also dependent on the content of eumelanin. The use of PhIP in hair as a biomarker of exposure is promising, but needs validation, using other methods of exposure assessment. #