Triangulating abuse liability assessment for flavoured cigar products using physiological, behavioural economic and subjective assessments: a within-subjects clinical laboratory protocol.

INTRODUCTION: In the USA, Food and Drug Administration regulations prohibit the sale of flavoured cigarettes, with menthol being the exception. However, the manufacture, advertisement and sale of flavoured cigar products are permitted. Such flavourings influence positive perceptions of tobacco products and are linked to increased use. Flavourings may mask the taste of tobacco and enhance smoke inhalation, influencing toxicant exposure and abuse liability among novice tobacco users. Using clinical laboratory methods, this study investigates how flavour availability affects measures of abuse liability in young adult cigarette smokers. The specific aims are to evaluate the effect of cigar flavours on nicotine exposure, and behavioural and subjective measures of abuse liability. METHODS AND ANALYSES: Participants (projected n=25) are healthy smokers of five or more cigarettes per day over the past 3 months, 18-25 years old, naive to cigar use (lifetime use of 50 or fewer cigar products and no more than 10 cigars smoked in the past 30 days) and without a desire to quit cigarette smoking in the next 30 days. Participants complete five laboratory sessions in a Latin square design with either their own brand cigarette or a session-specific Black & Mild cigar differing in flavour (apple, cream, original and wine). Participants are single-blinded to cigar flavours. Each session consists of two 10-puff smoking bouts (30 s interpuff interval) separated by 1 hour. Primary outcomes include saliva nicotine concentration, behavioural economic task performance and response to various questionnaire items assessing subjective effects predictive of abuse liability. Differences in outcomes across own brand cigarette and flavoured cigar conditions will be tested using linear mixed models. ETHICS AND DISSEMINATION: The Virginia Commonwealth University Institutional Review Board approved the study (VCU IRB: HM20007848). Dissemination channels for study findings include scientific journals, scientific meetings, and policy briefs. TRIAL REGISTRATION NUMBER: NCT02937051.

A comparison study on a sulfonated graphene-polyaniline nanocomposite coated fiber for analysis of nicotine in solid samples through the traditional and vacuum-assisted HS-SPME.

A simple, rapid, and reliable headspace solid-phase microextraction (HS-SPME) procedure, reinforced by applying vacuum in the extraction vial, was developed. It was applied for the extraction of nicotine in solid samples prior to determination by gas chromatography-flame ionization detection (GC-FID). First, the surface of a narrow stainless steel wire was made porous and adhesive by platinization to obtain a durable, higher surface area, and resistant fiber. Then, a thin film of sulfonated graphene/polyaniline (Sulf-G/PANI) nanocomposite was synthesized and simultaneously coated on the platinized fiber using the electrophoretic deposition (EPD) method. It was demonstrated that the extraction efficiency remarkably increased by applying the reduced-pressure condition in the extraction vial. To evaluate the conventional HS-SPME and vacuum-assisted HS-SPME (VA-HS-SPME) platforms, all experimental parameters affecting the extraction efficiency including desorption time and temperature, extraction time and temperature and moisture content of sample matrix were optimized. The highest extraction efficiency was obtained at 60°C, 10min (extraction temperature and time) and 280°C, 2min (desorption condition), for VA-HS-SPME strategy, while for conventional HS-SPME the extraction and desorption conditions found to be 100°C, 30min and 280°C, 2min, respectively. The Sulf-G/PANI coated fiber showed high thermal stability, good chemical/mechanical resistance, and long lifetime. For analysis of nicotine in solid samples using VA-HS-SPME-GC-FID, linear dynamic range (LDR) was 0.01-30µgg-1 (R2=0.996), the relative standard deviation (RSD%, n=6), for analyses of 1µgg-1 nicotine was calculated 3.4% and limit of detection (LOD) found to be 0.002µgg-1. The VA-HS-SPME-GC-FID strategy was successfully carried out for quantitation of nicotine in hair and tobacco real samples.

Estimations and predictors of non-compliance in switchers to reduced nicotine content cigarettes.

BACKGROUND AND AIMS: Clinical trials on the impact and safety of reduced nicotine content cigarettes (RNCs) are ongoing, and an important methodological concern is participant compliance with smoking only RNCs. Our aims were to measure non-compliance biochemically with urine cotinine (COT) and total nicotine equivalents (TNEs), compare with self-reported non-compliance and identify associated covariates. DESIGN: Secondary analysis of a double-blind, parallel, randomized clinical trial. SETTING: Research centers from the United States, enrolling participants from June 2013 to July 2014. PARTICIPANTS: Volunteer sample of 242 participants (55% Caucasian), average age of 41.2 years, smoking at least five cigarettes per day (CPD). INTERVENTION: Smoking very low nicotine cigarettes (VLNCs; 0.4 mg nicotine/g tobacco) for 6 weeks. MEASUREMENTS: The primary outcome was biochemically verified non-compliance, measured as thresholds of COT/CPD and TNE/CPD ratios, considering changes in nicotine content from conventional levels to VLNCs, and as an absolute threshold of week 6 TNEs. Self-reported non-compliance was measured via daily phone calls. Key predictors included age, sex, race, menthol preference, nicotine metabolite ratio, time to first cigarette, dependence, CPD, TNEs, tar level and cigarette evaluation. FINDINGS: Estimates of non-compliance with smoking the VLNCs exclusively include: the biochemical ratios (both 78%), the week 6 TNE threshold (76%) and self-report (39%). Of the key covariates, age, dependence and cigarette evaluations of satisfaction were significant; for age, younger participants more likely to be non-compliant [P = 0.01; odds ratio (OR) = 0.98, 95% confidence interval (CI) = 0.96-0.99]. Dependence was associated significantly with self-reported non-compliance (P = 0.01; OR = 1.28, 95% CI = 1.06-1.55). Cigarette evaluations of satisfaction were associated significantly with non-compliance (P = 0.001; OR = 0.71, 95% CI = 0.61-0.82). CONCLUSIONS: Among smokers volunteering to smoke only very low nicotine cigarettes for 6 weeks, non-compliance was common and biochemical assessments detected more cases of non-compliance than self-report. Despite high levels of non-compliance, smokers reduced their intake of nicotine by an average of 60%.

Simultaneous quantification of tobacco alkaloids and major phase I metabolites by LC-MS/MS in human tissue.

INTRODUCTION: Insurance agencies might request laboratories to differentiate whether a deceased has been a smoker or not to decide about refunding of his nonsmoker rate. In this context, the question on a solid proof of tobacco alkaloids and major metabolites in tissues came up. Currently, an appropriate assay is still lacking to analyze tissue distribution in smokers or nonsmokers. Nicotine (NIC), nornicotine (NNIC), anatabine (ATB), anabasine (ABS), and myosmine (MYO) are naturally occurring alkaloids of the tobacco plant; most important phase I metabolites of NIC are cotinine (COT), norcotinine (NCOT), trans-3'-hydroxycotinine (HCOT), nicotine-N'-oxide (NNO), and cotinine-N-oxide (CNO). An analytical assay for their determination was developed and applied to five randomly selected autopsy cases. METHODS: Homogenates using 500 mg aliquots of tissue samples were analyzed by liquid chromatography/tandem mass spectrometry following solid phase extraction. The method was validated according to current international guidelines. RESULTS: NIC, COT, NCOT, ABS, ATB, and HCOT could be detected in all tissues under investigation. Highest NIC concentrations were observed in the lungs, whereas highest COT concentrations have been found in the liver. MYO was not detectable in any of the tissues under investigation. CONCLUSIONS: The assay is able to adequately separate isobaric analyte pairs such as NIC/ABS/NCOT and HCOT/CNO thus being suitable for the determination of tobacco alkaloids and their phase I metabolites from tissue. More autopsy cases as well as corresponding body fluids and hair samples will be investigated to differentiate smokers from nonsmokers.

Complex suicide with homemade nicotine patches.

Suicide by self-poisoning is rather common around the world. This paper presents an exceptional complex suicide in which nicotine was applied in the form of self-made patches soaked with an extraction from fine-cut tobacco. In addition, the 51-year-old suicide victim took a lethal dose of diphenhydramine. Toxicological analysis also revealed the presence of tetrazepam in subtherapeutic concentrations. The scene of death suggested an autoerotic accident at first, as the body was tied with tapes, cables and handcuffs. As a result of the entire investigations, the fatality had to be classified as a suicidal intoxication by nicotine and diphenhydramine.

Determination of stimulants using gas chromatography/high-resolution time-of-flight mass spectrometry and a soft ionization source.

RATIONALE: The aim of this study was to investigate the mass spectral fragmentation of a small set of stimulants in a high-resolution time-of-flight mass spectrometer equipped with a soft ionization source using vacuum ultraviolet (VUV) photons emitted from different plasma gases. It was postulated that the use of a plasma gas such as Xe, which emits photons at a lower energy than Kr or Ar, would lead to softer ionization of the test compounds, and thus to less fragmentation. METHODS: A set of nine stimulants: cocaine, codeine, nicotine, methadone, phenmetrazine, pentylenetetrazole, niketamide, fencamfamine, and caffeine, was analyzed by gas chromatography/time-of-flight mass spectrometry (GC/TOFMS) in positive ion mode with this soft ionization source, using either Xe, Kr, or Ar as plasma gases. Working solutions of the test compounds at 0.1 to 100 ng/µL were used to establish instrument sensitivity and linearity. RESULTS: All test compounds, except methadone and pentylenetetrazole, exhibited strong molecular ions and no fragmentation with Xe-microplasma photoionization (MPPI). Methadone exhibited significant fragmentation not only with Xe, but also with Kr and Ar, and pentylenetetrazole could not be ionized with Xe, probably because its ionization energy is above 8.44 eV. The Kr- and Ar-MPPI mass spectra of the test compounds showed that the relative intensity of the molecular ion decreased as the photon energy increased. CONCLUSIONS: When coupled to a TOF mass spectrometer this soft ionization source has demonstrated signal-to-noise (S/N) ratios from 7 to 730 at 100 pg per injection (depending on the compound), and a dynamic range of three orders of magnitude (100 pg to 100 ng) for some of the test compounds.

High-throughput simultaneous analysis of buprenorphine, methadone, cocaine, opiates, nicotine, and metabolites in oral fluid by liquid chromatography tandem mass spectrometry.

A method for simultaneous determination of buprenorphine (BUP), norbuprenorphine (NBUP), methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), cocaine, benzoylecgonine (BE), ecgonine methyl ester (EME), anhydroecgonine methyl ester (AEME), morphine, codeine, 6-acetylmorphine (6AM), heroin, 6-acetylcodeine (6AC), nicotine, cotinine, and trans-3'-hydroxycotinine (OH-cotinine) by liquid chromatography tandem mass spectrometry in oral fluid (OF) was developed and extensively validated. Acetonitrile (800 µL) and OF (250 µL) were added to a 96-well Isolute-PPT+protein precipitation plate. Reverse-phase separation was achieved in 16 min and quantification was performed by multiple reaction monitoring. The assay was linear from 0.5 or 1 to 500 µg/L. Intraday, interday, and total imprecision were less than 13% (n = 20), analytical recovery was 92-114% (n = 20), extraction efficiencies were more than 77% (n = 5), and process efficiencies were more than 45% (n = 5). Although ion suppression was detected for EME, cocaine, morphine, 6AC, and heroin (less than 56%) and enhancement was detected for BE and nicotine (less than 316%), deuterated internal standards compensated for these effects. The method was sensitive (limit of detection 0.2-0.8 µg/L) and specific (no interferences) except that 3-hydroxy-4-methoxyamphetamine interfered with AEME. No carryover was detected, and all analytes were stable for 24 h at 22 °C, for 72 h at 4 °C, and after three freeze-thaw cycles, except cocaine, 6AC, and heroin (22-97% loss). The method was applied to 41 OF specimens collected throughout pregnancy with a Salivette® OF collection device from an opioid-dependent BUP-maintained pregnant woman. BUP ranged from 0 to 7,400 µg/L, NBUP from 0 to 71 µg/L, methadone from 0 to 3 µg/L, nicotine from 32 to 5,020 µg/L, cotinine from 125 to 508 µg/L, OH-cotinine from 11 to 51 µg/L, cocaine from 0 to 419 µg/L, BE from 0 to 351 µg/L, EME from 0 to 286 µg/L, AEME from 0 to 7 µg/L, morphine from 0 to 22 µg/L, codeine from 0 to 1 µg/L, 6AM from 0 to 4 µg/L, and heroin from 0 to 2 µg/L. All specimens tested negative for EDDP and 6AC. This method permits a fast and simultaneous quantification of 16 drugs and metabolites in OF, with good selectivity and sensitivity.

Environmental nicotine contamination in latent fingermarks from smoker contacts and passive smoking.

Hydrophobic silica nanopowder has been used as an effective latent fingermark development agent and subsequently as an enhancement agent in the surface-assisted laser desorption/ionisation-time of flight (SALDI-TOF) mass spectrometry for analysis of fingermark components. The technique has been used in the detection of nicotine and cotinine in the fingermarks of smokers. In order to have confidence in concluding that the nicotine in such samples is indicative of cigarette usage, it is necessary to establish that contamination by environmental contact or from hand to hand contact with smokers or from passive smoking does not lead to false identification of non-smokers as smokers. To investigate this possibility, the background level of nicotine in fingermark material from a number of commonly used places was determined. In addition, a series of experiments was carried out to assess the extent to which nicotine can be transferred through handshakes and finger transfer as well as touching of door handles. The rate of loss of nicotine from latent fingermarks was also assessed over a 24-h period under ambient laboratory conditions. Finally, a laboratory-based model system was evaluated to ascertain the possible transport of nicotine in cigarette smoke from a source to adjacent areas to simulate cross-contamination of a non-smoker by passive exposure. It was observed that person-to-person transfer from a smoker to a non-smoker can occur following handshakes but at low levels and that passive cross-contamination from contact with surfaces is possible under simulated conditions. However, levels of nicotine in the wider environment were found to be too low for detection using this technique which may reflect the half-life of nicotine in latent fingermarks which was about 11h. Likewise, transfer via smoke is possible to objects within about 0.1m of the cigarette but it is unlikely that significant secondary nicotine contamination will occur on the faces and hands of adjacent non-smokers.

Exposure to secondhand smoke and associated factors among non-smoking pregnant women with smoking husbands in Sichuan province, China.

BACKGROUND: Secondhand smoke (SHS) exposure harms pregnant women and the fetus. China has the world's largest number of smokers and a high male smoking prevalence rate. OBJECTIVE: To compare exposure to SHS among rural and urban Chinese non-smoking pregnant women with smoking husbands, and analyze factors associated with the level of SHS exposure and hair nicotine concentration. SETTING: Sichuan province, China. POPULATION: In all 1,181 non-smoking pregnant women with smoking husbands recruited from eight district/county Women and Children's hospitals. METHODS: The women completed a questionnaire in April and May 2008. Based on systematic sampling, 186 pregnant women were selected for sampling the nicotine concentration in their hair. Ordinal logistic regression analysis was conducted to examine correlates with self-reported SHS exposure (total and at home); linear regression was conducted for the sub-sample of hair nicotine concentrations. MAIN OUTCOME MEASURES: Secondhand smoking exposure rates, hair nicotine levels. RESULTS: About 75.1% of the non-smoking pregnant women with smoking husbands reported regular SHS exposure. The major source of exposure was through their husband. In the multivariate analysis, the risk of greater SHS exposure (total and at home) and hair nicotine concentration was increased for women who were rural, had a husband with greater cigarette consumption, less knowledge about SHS, less negative attitudes about SHS, and no smoke-free home rules. CONCLUSIONS: The high prevalence rate of SHS exposure suggests that it is important for non-smoking pregnant women, especially rural women, to establish smoke-free home rules and increase knowledge and negative attitudes towards SHS.

Two UK suicides using nicotine extracted from tobacco employing instructions available on the Internet.

Tobacco is one of the most easily accessible and commonly abused drugs world-wide. Nicotine, one of its principal constituents, can cause serious or fatal overdoses. Whilst the deliberate ingestion of this substance appears to be relatively rare, often the important signs of its consumption are not recognised, sometimes with fatal results. Here we describe two cases of intentional fatal ingestion of nicotine. The nicotine was extracted from tobacco using instructions available on the Internet. The first case involved a male aged 19 who died in 2008. The post-mortem blood and urine levels of nicotine were 5.5mg/l and >80 mg/l respectively; the blood level is in line with the generally recognised fatal level of >5mg/l. The levels of nicotine's main metabolite, cotinine, for this case were 2.5mg/l and 7.9 mg/l for blood and urine respectively. A comparative case in 1999 involved a 32 year-old male. The level of post-mortem nicotine in his blood was 1.0mg/l. These are believed to be the first UK suicides by nicotine using instructions from the Internet reported in the literature. Information that nicotine was the agent responsible only became apparent some time after death. There may be more deaths due to this cause that go unrecognised because quantification of nicotine and cotinine levels is not often conducted, due to the wide prevalence of smoking. It is important that all evidence at the scene of a sudden and unexplained death is carefully evaluated, including potential clues on PCs and lap-tops.

Gas chromatography-mass spectrometry assay for the simultaneous quantification of drugs of abuse in human placenta at 12th week of gestation.

We describe the development and validation of a method for the quantification of drugs of abuse, using gas chromatography-mass spectrometry (GC/MS), in human placenta. Concentration ranges covered were 5-500 ng/g for amphetamine, methamphetamine, MDMA, methadone, cocaine, benzoylecgonine, cocaethylene, morphine, 11-nor-9-carboxy-delta-9-tetrahydrocannabinol, nicotine, and cotinine. Intra-assay and inter-assay imprecisions were less than 15.7% for lower quality control samples and less than 14.9% for medium and high quality control samples. Recovery range was 36.2-83.7%. Placenta samples were kept at -80 degrees C until analysis; analytes were stable after three freeze-thaw cycles (samples stored at -20 degrees C). This accurate and precise assay has sufficient sensitivity and specificity for the analysis of specimens collected from women who voluntarily terminated their pregnancy at 12th week of gestation. The method has proven to be robust and accurate for the quantification of the principal recreational drugs of abuse in this period of the prenatal life. This is the first report that highlights the presence of drugs of abuse during the first trimester of gestation.

Assessment of exposure to environmental tobacco smoke in young adolescents following implementation of smoke-free policy in Italy.

We investigated acute and chronic exposure to environmental tobacco smoke (ETS) in a cohort of young adolescents using urinary cotinine and hair nicotine testing after recent implementation of Italian smoke free legislation. Study subjects were 372 Italian young adolescents, between 10 and 16 years of age from the principal city of Sicily, Palermo. Urine and hair samples were collected between November 2005 and May 2006, when the legislation to ban smoking in all the enclosed places of employment (including bars, restaurants, pubs) was completely enforced. An exhaustive questionnaire including sociodemographic characteristics and active and passive exposure to cigarette smoking was completed. Urinary cotinine was analyzed by radioimmunoassay and hair nicotine by a validated GC/MS method. Based on urinary cotinine results, 2.1% and 89% of the study participants, respectively, showed non-exposure and low acute exposure to ETS, whereas only 1.6% presented very high exposure or a hidden active smoking habit in the recent past. Hair nicotine disclosed non-exposure and low exposure to ETS in 11.8% and 65.6% of the young adolescents, respectively, taking into consideration a larger time-window. High repeated exposure, suggesting active smoking in some cases was observed in 8.6% of the study subjects. Hair nicotine was inversely related to educational level of the adolescents' parents. Overall, due to the implementation of smoke-free legislation and information campaign against smoking, a significant trend toward low exposure to ETS was observed in this study cohort with no association between exposure to ETS and respiratory illnesses.

Death due to ingestion of nicotine-containing solution: case report and review of the literature.

Nicotine, a lipid-soluble alkaloid obtained from the dried leaves of Nicotiana, is most frequently encountered in tobacco products for smoking, chewing or sniffing as well as in a limited number of pesticides. Though nicotine is one of the most toxic drugs of abuse, it has rarely led to fatalities. Sudden death can be caused by cardiovascular arrest, respiratory muscle paralysis and/or central respiratory failure. A 42-year-old man was found dead by his wife. He was lying on the floor, next to a box containing many empty bottles of beer and vodka. Some labeled chemical bottles found at the scene contained various substances, including nicotine and brucine. Gross examination of the organs at autopsy revealed no specific findings. The toxicological examination failed to disclose any lethal toxic agents other than a high concentration of nicotine and its primary metabolite cotinine in femoral venous blood (2.2 microg/mL). Blood alcohol was determined to be 2.1 g/L in femoral venous blood. Only a paucity of fatal cases of nicotine poisoning has been reported in the literature so far.

Forensic implication of muscle level of nicotine in an adipocere body found in the sea.

We reported previously that blood levels of nicotine in suicidal smokers tend to be significantly higher than those in non-suicidal smokers, and blood level of nicotine seems to be a useful criterion for discriminating suicide from other types of death. In this paper, we report nicotine and cotinine levels in various tissues of an adipocere body found in the sea. The cause of death was drowning, and the postmortem time interval was approximately 5 months at autopsy. His driver's license was concealed in his sock, which seemed to suggest that he committed to suicide. In toxicological analysis by gas chromatography, nicotine and cotinine in the femoral muscle were detected at concentrations of 213 and 488 ng/g, respectively, and these substances were also detected in the brain, liver and kidney. For evaluating the tissue levels of nicotine and cotinine in the adipocere body, we analyzed these levels in blood and various tissues of 13 autopsy cases of smokers. Nicotine and cotinine levels in blood were the most similar to those in skeletal muscle. Although the postmortem time interval, the formation process of adipocere and the environmental condition in water may affect nicotine and cotinine levels in the femoral muscle, the high muscle level of nicotine in the present case seem to implicate suicidal death.

Surveillance of moist snuff: total nicotine, moisture, pH, un-ionized nicotine, and tobacco-specific nitrosamines.

In 2005, approximately 2.3% of U.S. adults used smokeless tobacco. Moist snuff leads all types of smokeless tobacco in revenues and marketing expenditures. The U.S. Surgeon General has concluded that smokeless tobacco use can lead to nicotine addiction. The National Toxicology Program of the National Institutes of Health has classified smokeless tobacco as a human carcinogen. Tobacco-specific nitrosamines (TSNAs) are potent carcinogens in smokeless tobacco products, and the pH of the product influences the content of un-ionized nicotine which is the form of nicotine most rapidly absorbed in the mouth. The Centers for Disease Control and Prevention analyzed 40 top-selling brands of moist snuff to measure nicotine, moisture, pH, un-ionized nicotine, and TSNAs, including 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL). The study findings indicate that moist snuff brands varied widely in content of rapidly absorbed, addictive un-ionized nicotine (500-fold range) and of carcinogenic TSNAs (18-fold range). Product characteristics such as packaging and moisture content appeared to be correlated with concentrations of un-ionized nicotine, and flavor characteristics of low-priced brands may correlate with TSNA concentrations. These findings warrant further study in light of (a) the marketing of smokeless tobacco for use in places where smoking is prohibited, (b) the promotion of smokeless tobacco as a harm-reduction product, and (c) the ever-expanding number of highly flavored smokeless varieties brought to the market.

Nicotine and cotinine in infants dying from sudden infant death syndrome.

The aim of this component of the German Study on Sudden Infant Death was to determine (1) nicotine concentrations in hair (NCH), as a marker of long standing exposure to tobacco, (2) cotinine concentrations in pericardial fluid (CCP) and (3) cotinine concentrations in liquor cerebrospinalis (CCL), the latter measures being markers of recent exposure to tobacco in the last few hours of life. The results obtained were compared with data on parental smoking revealed from interviews. In 100 cases of sudden infant death syndrome, material was taken at autopsy to determine NCH. In 41 cases, NCH and CCP, and in 70 cases, NCH and CCL were determined. Infants of mothers who stated having smoked during pregnancy had higher NCH than infants of non-smoking mothers (p = 0.008). Furthermore, there was a weak but statistically significant relationship between NCH's and the daily cigarette consumption of the mother during pregnancy (n = 64, r = 0.24, p = 0.05). In 43% of infants, nicotine could be detected in their hair, although the mothers had said at the interview that they did not smoke during pregnancy. On the other hand, in 33% of infants whose mother stated they had smoked during pregnancy nicotine was not detectable in the infant's hair. CCP's were strongly correlated with CCL's (r = 0.62, p = 0.0027). For this reason, both parameters were treated as equivalent for the detection of tobacco smoke exposure in the last hours before death. The influence of breast-feeding was evaluated by comparison of the nicotine concentrations in breast fed and non-breast-fed infants from smokers and non-smokers. Fivefold higher nicotine concentrations were determined in non-breast-fed infants of parents who smoked as compared to all other groups. It can be concluded that nicotine intake by passive smoking is much more important than by breast-feeding. We conclude that both interview data and biochemical measures should be sought to understand the true exposure to tobacco smoke.

Sweat testing in addicts under methadone treatment: an Italian experience.

In the last years the interest in monitoring drug exposure with human sweat as alternative biological fluid, is increasing. Sweat collection is convenient, less invasive and difficult to adulterate compared to traditional specimens. The objective of this study was to determine the excretion profile of methadone and other drugs into human sweat. Pharmscope sweat patches (Medical Europe Diagnostic, Madrid, Spain) were used on heroin abusers under methadone treatment. Sweat patches were applied to 10 heroin addicts and 3 drug free volunteers admitted into the study. Sweat patches were worn for about 1 week; urine, saliva and hair samples were collected at the time of the removal of patches. After the extraction, sweat eluates were directly analyzed by GC/MS for the presence of nicotine, cotinine, caffeine, methadone, EDDP and cocaine. The extracts were subsequently derivatized to detect benzoylecgonine, ecgonine methyl ester, morphine, codeine and 6-acetylmorphine. No false positive results were obtained on the drug free samples. All the patches showed positive results for methadone. Cocaine was detected in two cases. Mainly the parent drug was identified rather than the metabolites. The results obtained show the usefulness of sweat as complementary specimen to saliva and urine providing a longer detection window. Moreover, sweat testing offers the advantage of being a non-invasive means of obtaining information about drug exposure.

Relationship of human toenail nicotine, cotinine, and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol to levels of these biomarkers in plasma and urine.

Recently, we developed sensitive and quantitative methods for analysis of the biomarkers of tobacco smoke exposure nicotine, cotinine, and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in human toenails. In this study, we further evaluated the newly developed toenail biomarkers by investigating their relationship to demographic factors, reported exposure, plasma nicotine, cotinine, and trans-3'-hydroxycotinine, and urinary NNAL. Toenails of 105 smokers, mean age 38.9 years (range, 19-68), were analyzed. Fifty-five (53.4%) were male, with approximately equal numbers of Whites and African-Americans. The average number of cigarettes smoked per day was 18 (range, 5-50). There was no effect of age or gender on the toenail biomarkers. Toenail NNAL was higher in White than in African-American participants (P = 0.019). Toenail nicotine and toenail cotinine correlated significantly with cigarettes smoked per day (r = 0.24; P = 0.015 and r = 0.26; P = 0.009, respectively). Toenail nicotine correlated with plasma nicotine (r = 0.39; P < 0.001); toenail cotinine correlated with plasma cotinine (r = 0.45; P < 0.001) and plasma trans-3'-hydroxycotinine (r = 0.30; P = 0.008); and toenail NNAL correlated with urine NNAL (r = 0.53; P = 0.005). The results of this study provide essential validation data for the use of toenail biomarkers in investigations of the role of chronic tobacco smoke exposure in human cancer.

Light cigarette smoking impairs coronary microvascular functions as severely as smoking regular cigarettes.

BACKGROUND: Smoking is the most prevalent and most preventable risk factor for cardiovascular diseases. Smoking low-tar, low-nicotine cigarettes (light cigarettes) would be expected to be less hazardous than smoking regular cigarettes owing to the lower nicotine and tar yield. OBJECTIVE: To compare the chronic and acute effects of light cigarette and regular cigarette smoking on coronary flow velocity reserve (CFVR). METHODS: 20 regular cigarette smokers (mean (SD) age 24.8 (5.0)), 20 light cigarette smokers (mean age 25.6 (6.4)), and 22 non-smoker healthy volunteers (mean age 25.1 (4.2)) were included. First, each subject underwent echocardiographic examination, including CFVR measurement, after a 12 hour fasting and smokeless period. Two days later, each subject smoked two of their normal cigarettes in a closed room within 15 minutes. Finally, within 20-30 minutes, each subject underwent an echocardiographic examination, including CFVR measurement. RESULTS: Mean (SD) CFVR values were similar in light cigarette and regular cigarette smokers and significantly lower than in the controls (2.68 (0.50), 2.65 (0.61), 3.11 (0.53), p = 0.013). Before and after smoking a paired t test showed that smoking two light cigarettes acutely decreased the CFVR from 2.68 (0.50) to 2.05 (0.43) (p = 0.001), and smoking of two regular cigarettes acutely decreased CFVR from 2.65 (0.61) to 2.18 (0.48) (p = 0.001). CONCLUSION: Smoking low-tar, low-nicotine cigarettes impairs the CFVR as severely as smoking regular cigarettes. CFVR values are similar in light cigarette and regular cigarette smokers and significantly lower than in controls.

[Second-hand smoke exposure in Florence and Belluno before and after the Italian smoke-free legislation]. / Esposizione a fumo o passivo a Firenze e a Belluno prima e dopo l'entrata in vigore del divicto di fumo in Italia.

INTRODUCTION: a law banning smoking in enclosed public places entered into force in Italy on January 10th 2005. OBJECTIVE: to compare the SHS exposure before and after the coming into force of the new anti-smoking law, with direct measurements in public venues and workplaces. METHODS: vapour-phase nicotine was measured using passive samplers, with personal and environmental sampling. SETTING: samples were collected in 10 municipality offices, 10 industry buildings and 11 public venues (4 discos and 7 pubs) in two towns (Florence and Belluno) from november 2004 to march 2005. RESULTS: during the pre-ban period a wide range of nicotine concentrations was observed. Offices and industry sector exhibited very low concentrations, 0,47 e 0,40 microg/m3 in median, respectively. Highest concentrations were measured in pubs and discos (35,59 and 127,16 microg/m3). After the smoking ban, a noteworthy reduction in nicotine concentrations was found with a stronger effect in pubs and discos (95% of decrease). DISCUSSION: the introduction of a national smoking ban led to a clear reduction in SHS exposure, with stronger results in environments less protected by previous regulations.