Complete investigations (autopsy, toxicology, and histology) in a death due to apixaban overdose.

The dead body of a 54-year-old man was found at home by his partner. He was off work due to depression. A letter with suicidal intention was present on the scene. He was known to be a heavy drinker, and near the body, an empty bottle of whisky was found. In addition, 2 empty blisters of Eliquis (apixaban) 5 mg, corresponding to 40 tablets, were identified. Apixaban is an oral anticoagulant, acting as a factor Xa inhibitor. Autopsy findings were mostly unremarkable, except numerous bruises and some superficial self-inflected wounds. Histology showed hematomas of calyces and renal pelvis and in the liver, several areas of perivenular haemorrhagic necrosis. Others organs were congestive. Femoral venous blood alcohol was 0.11 g/L. In femoral venous blood, a toxic concentration of apixaban was measured at 1184 ng/mL using LC-MS/MS. Other drugs found at therapeutic concentrations included diazepam (99 ng/mL), nordiazepam (171 ng/mL), flecainide (447 ng/mL), and mianserine (65 ng/mL). Using liquid chromatography coupled to high-resolution mass spectrometry, 2 metabolites were identified, O-desmethyl-apixaban (61.8% of the apixaban response) and hydroxyl-apixaban (4.5% of the apixaban response). Long-term therapy was confirmed by a concentration of 10390 pg/mg in pubic hair.

Hidden administration of 5-APB in a dancing club of New Caledonia documented by urine analysis: about 3 cases.

1-Benzofuran-5-ylpropan-2-amine or 5-APB is a new psychoactive substance (NPS) with empathic effects close to ecstasy (MDMA). Although 5-APB has been observed in fatality cases, the drug has not yet been reported in the context of hidden administration for behaviour impairment, also known as drug-facilitated crime. Such a situation was recently observed on 3 separate occasions in the same dancing club of New Caledonia. It involves 3 women, aged 27, 29, and 33 years who presented, after having drunk a cocktail, anxiety, abnormal movements of the inferior jaw, and aggressiveness. No memory loss was noticed. About 12 h after the event, a urine specimen was collected in the 3 cases. Comprehensive toxicology was requested and only 5-APB was identified, at 6, 8, and 14 ng/mL. Urine ethanol tested negative, which is consistent with the limited intake before the event occurred. These results have demonstrated that NPS are circulating in New Caledonia, which was not previously reported, and that 5-APB, like ecstasy, can be used to modify the behaviour of a subject, as it can be done by a chemical weapon.

The use of multiple keratinous matrices (head hair, axillary hair, and toenail clippings) can help narrowing a period of drug exposure: experience with a criminal case involving 25I-NBOMe and 4-MMC.

The objective of this publication is to present the interest of collecting several keratinous specimens in order to document possible drug impairment at the time of the assault, when knowledge solely occurred 7 months after. A subject committed a murder and within minutes after the crime self-inflicted serious wounds. He was charged to the hospital where he slowly recovered. After several weeks, he was sent to prison. During this period, intelligence indicated possible drug impairment at the time of the assault after using 25I-NBOMe and 4-MMC. Head hair (4 cm), axillary hair, and toenails were collected 7 months after the crime. New psychoactive substances were tested in each specimen using LC-MS/MS, which revealed the presence of 25I-NBOMe and 4-MMC in axillary hair (2 and 6 pg/mg) and toenails (1 and 5 pg/mg). However, the perpetrator claimed that the positive findings were due to contamination in prison. Therefore, the head hair was also tested and results returned negative (LOQ at 1 pg/mg), demonstrating absence of contamination during the last 4 months before collection. Combining the window of drug detection in axillary hair (about 4 to 8 months) and the one of toenail clippings (up to 8 months), and excluding drug exposure during the previous 4 months as well as external contamination as the head hair results were negative, allowed us to conclude that the positive findings in axillary hair and toenails are more likely than not consistent with consumption of both 25I-NBOMe and 4-MMC at the time of the crime.

Anabolic steroids and extreme violence: a case of murder after chronic intake and under acute influence of metandienone and trenbolone.

A 32-year-old male went to the police to claim he just killed his girlfriend by inflicting several stabs with a kitchen knife. He was very nervous and particularly aggressive. About 90 min after the assault, a blood specimen was collected with natrium fluoride as preservative. The blood was free of alcohol, pharmaceuticals and drugs of abuse, but tested positive by LC-MS/MS for metandienone (32 ng/mL) and trenbolone (9 ng/mL). The perpetrator admitted regular consumption of anabolic steroids to enhance his muscular mass, as he was a professional security agent. To document long-term steroid abuse, a hair specimen was collected 3 weeks after the assault, which tested positive for both drugs. Segmental analyses revealed in the proximal 1.5 cm segment, corresponding to the period of the assault, the simultaneous presence of metandienone (11 pg/mg) and trenbolone (14 pg/mg), while only metandienone (3 pg/mg) was identified in the distal 1.5 cm segment. As aggressiveness and violence can be associated with abuse of anabolic steroids, the aetiology of this domestic crime was listed to be due impulsive behaviour in a context of antisocial lifestyle.

Perspectives in Evaluating Selective Androgen Receptor Modulators in Human Hair: A Short Communication.

BACKGROUND: As hair testing increases the window of drug detection and permits the differentiation of long-term use from a single exposure when performing segmental analyses (which also allows establishing the pattern of use), this matrix should be considered as a suitable complement to standard investigations in clinical, forensic, and sport toxicology. The authors were recently involved in 3 cases where hair analysis was used to demonstrate the use of selective androgen receptor modulators (SARMs), including ligandrol (LGD-4033), andarine (S-4), and ostarine (S-22). SARMs are increasingly being abused as "safe" alternatives to steroids. METHODS: After decontamination using dichloromethane, hair specimens were segmented, cut into very small segments (<1 mm), incubated overnight in a buffer, and extracted using a mixture of organic solvents. Drugs were tested using liquid chromatography-tandem mass spectrometry and confirmed using liquid chromatography/HRMS. RESULTS: The determined concentrations were as follows: ligandrol, 14-42 pg/mg; andarine, 0.1-0.7 pg/mg; and ostarine, 3-21 pg/mg. CONCLUSIONS: To enhance performance, SARMs must be used on a long-term basis, which can have serious clinical consequences, including liver damage, myocardial infarction, and blood clots. Hair testing for SARMs has additional benefits versus urine analysis as it can detect the parent compound and numerous metabolites.

The Importance of Autopsy in a Case of Digestive Perforation Undetected by Postmortem Computed Tomography.

ABSTRACT: Cases of foreign body ingestion in the forensic literature are mainly described in children or psychiatric patients. Postmortem imaging can detect most foreign bodies, but its sensitivity depends, among other things, on the type of item and its location. In some cases, the ingestion of foreign bodies can remain unnoticed and have serious consequences for the patient. We describe the case of a patient who died in a psychiatric seclusion room with no obvious cause and for whom a forensic autopsy was requested. Further investigations showed the existence of a subdural hematoma associated with a midline shift, secondary to a skull fracture that was considered to be the cause of death. Toxicological analyses identified in blood several drugs, including diazepam (24 ng/mL) and its major metabolite nordazepam (24 ng/mL), propranolol (57 ng/mL), paliperidone (9 ng/mL), and loxapine (620 ng/mL). The forensic autopsy revealed the existence of a gastrointestinal perforation after the ingestion of a plastic teaspoon, which the postmortem CT scan had failed to detect. Although technological advances continue to assist the forensic pathologist in his diagnosis, autopsy still has a leading role in forensic investigations and does not yet seem to be replaceable by imaging techniques alone.

Negative hair test result after long-term drug use. About a case involving morphine and literature review.

Although it has been accepted by most scientists that drugs circulating in blood are eligible to hair incorporation, this cannot be considered as a general statement. A 42-year old man was found dead in his swimming pool. He was living alone, and seen alive 2 days before by a neighbour. Femoral blood, cardiac blood and hair were collected during body examination. Free morphine was identified in femoral blood at 28 ng/mL, corresponding to his treatment for chronic pain (3 × 5 mg daily for 4 months). However, with a limit of quantitation (LOQ) at 10 pg/mg, segmental hair testing (3 × 1 cm) for morphine was negative. In this paper, the author has reviewed the different factors which can be responsible of this discrepancy. Several variables can influence the detection of a drug in hair and the author has listed reasons that can account for the absence of analytical response in hair after drug administration. The drug may not be incorporated in hair. That is the case for large bio-molecules, such as hormones, which cannot be transferred from the blood capillaries to growing cells of hair. Cosmetic treatments (perming, colouring, bleaching) or environmental aggressions (ultraviolet radiation, thermal application) will always reduce the concentrations. In this case, the lack of morphine detection was attributed to the effects of chlorinated water from the swimming pool. A negative hair result is also a result. However, this can be interpreted in three different ways: 1. the owner of the hair did not take or was not exposed to the specific drug, 2. the procedure is not sensitive enough to detect the drug, or 3. something happened after drug incorporation (cosmetic treatment, environmental influence).

Murdered while under the influence of 3-MeO-PCP.

The abuse of new psychoactive substances (NPS) has been dramatically increasing all around the world since the late 2000s. The availability of hundreds of NPS in the past decade is challenging for both public health and global drug policies. A 39-year-old woman, known as a multidrug addict, was murdered by her partner by ligature strangulation. A comprehensive toxicological screening by gas chromatography and ultra-high performance liquid chromatography with tandem mass spectrometry revealed the simultaneous presence of ethanol (1.37 g/L), diazepam (157 ng/mL) and nordiazepam (204 ng/mL), cocaine (25 ng/mL) and benzoylecgonine (544 ng/mL), and (3-methoxy-(1-(1-phenylcyclohexyl)piperidine) or 3-MeO-PCP, a dissociative hallucinogen anesthetic drug. Concentrations of 3-MeO-PCP were 63, 64, and 94 ng/mL in femoral blood, bile, and urine, respectively. Hair tested also positive for 3-MeO-PCP on 3 × 2-cm segments at 731, 893, and 846 pg/mg, indicating long-term abuse of the drug. This seems to be the first ever reported hair concentrations. Major impairment of the victim, including visual hallucinations and alteration of behavior, was attributed to the mixture of all the drugs, with a major contribution of 3-MeO-PCP. The toxicological findings were compared to the few reports available in the medical literature.

Hair testing of GHB: an everlasting issue in forensic toxicology.

BACKGROUND: In this paper, the authors present a critical review of different studies regarding hair testing of endogenous γ-hydroxybutyrate (GHB), concentrations in chronic users, and values measured after a single GHB exposure in drug facilitated sexual assault (DFSA) cases together with the role of a recently identified GHB metabolite, GHB-glucuronide. CONTENT: The following databases (up to March 2017) PubMed, Scopus and Web of Science were used, searching the following key words: γ-hydroxybutyrate, GHB, GHB glucuronide, hair. The main key words "GHB" and "γ-hydroxybutyrate" were searched singularly and then associated individually to each of the other keywords. SUMMARY: Of the 2304 sources found, only 20 were considered appropriate for the purpose of this paper. Summing up all the studies investigating endogenous GHB concentration in hair, a very broad concentration range from 0 to 12 ng/mg was found. In order to detect a single GHB dose in hair it is necessary to commonly wait 1 month for collecting hair and a segmental analysis of 3 or 5 mm fragments and the calculation of a ratio between the targeted segment and the others represent a reliable method to detect a single GHB intake considering that the ratios presently proposed vary from 3 and 10. The only two studies so far performed, investigating GHB-Glucuronide in hair, show that the latter does not seem to provide any diagnostic information regarding GHB exposure. OUTLOOK: A practical operative protocol is proposed to be applied in all suspected cases of GHB-facilitated sexual assault (GHB-FSA).

Assessment of Pregabalin Use by Hair Testing.

BACKGROUND: The detection and analysis of drugs in hair has progressively emerged as a consequence of the enhanced sensitivity of analytical techniques used in forensic toxicology; a greater advantage in using this matrix respect to classical ones (i.e., urine and blood) is an easier and noninvasive sample collection, even when the careful supervision of law-enforcement officers is required to avoid the risk that the sample may be adulterated or replaced. Moreover, according to the length of the hair, the history of drug exposure can be retrospectively monitored from few weeks up to months or years since sample collection. OBJECTIVE: Given the potential negative effects of pregabalin, an antiepileptic and analgesic drug with a high risk of misuse and abuse, the laboratory was asked to test for the drug in hair. METHOD: A new ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry was developed. The method involves incubation of 25 mg of cut hair in acetonitrile for 2 h in an ultrasonic bath and separation on an Acquity HSS C18 column (150 × 2.1 mm × 1.8 µm) maintained at 50°C in a thermostatically controlled oven. A gradient elution was performed. RESULTS: The method was fully validated according to international standards. The limit of quantitation of the test was 10 pg/mg. Five authentic cases of pregabalin in hair segments were tested using the method and the results were in the range 17-1487 pg/mg. CONCLUSION: This new method was found suitable to monitor both patients under pregabalin therapy and dependent subjects.

Testing for Drugs in Exhaled Breath Collected With ExaBreath in a Drug Dependence Population: Comparison With Data Obtained in Urine After Liquid Chromatographic-Tandem Mass Spectrometric Analyses.

BACKGROUND: Exhaled breath is commonly used in alcohol testing but has been recently demonstrated by scientists from Sweden, Switzerland, and the United States to contain a large number of both volatile and nonvolatile substances that can be measured using dedicated devices. ExaBreath is a sampling device that collects the bio-aerosols particles from the donor. Approximately 1-2 minutes exhaled breath is enough for the test. The device collects the very small bio-aerosols on a filter, which is consecutively incubated into methanol to release the drugs at the laboratory. METHODS: Eighteen drug addicts from a methadone substitution program were recruited for this study. There were 5 women and 13 men, aged 25-50 years. The daily methadone dosage ranged from 10 to 120 mg, mostly as syrup. Urine (in plastic tubes with no preservative) and exhaled breath were simultaneously collected. In both fluids, methadone and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) were tested using a specific liquid chromatographic-tandem mass spectrometric method, whereas all other compounds were screened by liquid chromatographic-tandem mass spectrometric method using a home made library of more than 800 compounds. Linearity, imprecision, and limit of quantitation were established. In each analytical batch, low and high controls were included. RESULTS: All 18 urine specimens tested positive for methadone and EDDP. Several other compounds were also identified, including morphine, THC-COOH, benzoylecgonine, nicotine, some antidepressants, and neuroleptics. Methadone and EDDP were identified in exhaled breath from all 18 patients, with concentrations in the range 11-1470 and 29-818 pg per filter, respectively. In 13 cases, the ratio methadone/EDDP in exhaled breath was >1 (range 0.4-2.8). Except nicotine (n = 7), no other substance was detectable in exhaled breath. CONCLUSION: This study gives further support to the possibility of using exhaled breath as a new matrix to document exposure to drugs.

Drug transfer during intimate moments: A key issue in doping control that can be documented by hair tests of the athlete and the partner.

The presence of a prohibited substance or its metabolites or its markers in an athlete's sample constitutes the more frequent anti-doping rules violation. In the world anti-doping code, it is indicated (point 10.5) that if someone establishes in an individual case that the athlete bears no fault or negligence, then the otherwise applicable period of ineligibility shall be eliminated. The conditions that have to be met to fix the no fault or negligence evidence are described in several other points of the code. The following two points are of paramount importance: 1. the athlete or his/her legal representative must present verified circumstances of contamination and the source of contamination must be identified; and 2. there must be verified claims by the athlete about the fact that he/she did not knowingly take the prohibited substance, i.e., that the violation was not intentional.In recent years, several cases of contamination involving drug transfer during intimate moments have been reported. This later situation was first reported in 2009 with the Richard Gasquet case. Since that time, several athletes have been allowed to return to competition with no charge based on strong evidence that the source of contamination was drug transfer during intimate moments. As some of these cases are public and because the author performed hair tests for the majority of the international athletes involved in such procedures, the strategy of the defence and the scientific bases of discussion are reviewed in this article.

Evidence of ostarine cross-contamination via sweat in 2 athletes sharing the same neoprene hamstring sleeves. An original situation of drug transfer where the anti-doping rule violation was suspended by the sport authorities.

The presence of ostarine, a selective androgen receptor modulator (SARM) in an athlete's sample constitutes one of the most frequent anti-doping rules violation. It is possible to challenge this violation but it is the athlete who has to demonstrate he / she is innocent. The conditions to evidence no fault or negligence are mostly based on 2 points: 1. the athlete or his/her legal representative must present verified circumstances of contamination and the source of contamination must be identified; and 2. there must be verified claims by the athlete about the fact that he / she did not knowingly take the prohibited substance, i.e. that the violation was not intentional. During a 2-weeks period, a male athlete tested two times positive for ostarine in urine (<0.1 ng/ml) and he challenged these results. His hair and nail tests returned negative (LOQ at 0.5 pg/mg). He admitted using two neoprene hamstring sleeves of another athlete who confessed abusing ostarine. This was confirmed in his hair (190 pg/mg), his fingernail clippings (780 pg/mg) and his toenail clippings (45 pg/mg). To document the presence of ostarine in the hamstring sleeves and therefore possible drug transfer, the hamstring sleeves were analysed. Ostarine was identified in 12 different selected pieces (about 1 g) of the sleeves at concentrations ranging from 3 to 142 pg/g. Sport authorities (USADA) agreed that the most likely source of contamination was the hamstring sleeves, thus confirming the scenario of drug transfer and gave the athlete a no fault.

Evidence of ostarine excretion in oral fluid after a single controlled oral administration.

The presence of ostarine, a selective androgen receptor modulator (SARM) in an athlete's urine specimen constitutes one of the most frequent anti-doping rules violation as the drug is listed as a member of the S1.2 class "other anabolic agents" of the World Anti-doping Agency Prohibited List, forbidden in- and out-competition. It is possible to challenge this violation but it is at the charge of the athlete to prove innocence. The conditions to evidence no fault or negligence are mostly based on 2 points: 1. the athlete must present verified circumstances of contamination and the source of contamination must be identified; and 2. there must be verified claims by the athlete that the violation was not intentional. Some months before the Olympic games, a female athlete was suspended by a national anti-doping agency because of an adverse analytical finding for ostarine. She claimed that her violation was due to drug transfer when kissing her boyfriend, who did not inform her about his ostarine daily intake. To document this claim (excretion of ostarine in oral fluid in sufficient amounts), a male volunteer ingested 17.3 mg of ostarine (dose verified by 1H NMR). Oral fluid was collected over 8 h using the NeoSal™ collection device and was tested by liquid chromatography coupled to tandem mass spectrometry. Maximal ostarine concentration was 468 ng/mL at T + 15 min, which can also be partially attributed to mouth contamination. Ostarine was detectable during the whole period of test, with concentrations at 1-2 ng/mL after T + 4 h. These results support drug transfer during kissing and subsequent possible contamination of the partner.

Evidence of exposure to flecainide in a newborn by keratinous matrices testing and interpretation of the findings.

Pediatric poisoning represents a serious problem all around the world. Abuse or neglect of children by adults must be highlighted in children exposed to drugs to which they would not normally have access. Usually, segmental hair analysis would allow in these contexts to determine whether the exposure was unique or repetitive. Hair and nail samples from a 9-month-old girl were received in our laboratory for analysis, after the child was hospitalized due to severe dehydration caused by her mother's neglect. At the admission, flecainide, an antiarrhythmic never prescribed to the child, was identified in the daughter urine. Using an LC-MS/MS method, flecainide tested positive in the child's hair at the following concentrations: 66 pg/mg (root to 1 cm), 61 pg/mg (1-2 cm), and 125 pg/mg (2-3 cm). Traces below the limit of quantification (1 pg/mg) were also present in the nail clippings. These concentrations are much lower than those obtained in adults under daily treatment. Given the different pharmacokinetic and dynamic parameters in children, the different rate of hair growth, and the greater porosity of the hair, which makes it more prone to external contamination, the interpretation of hair findings in children remains very complicated. In this case, it can be assumed that the presence of the drug in the urine indicates systemic incorporation and that administration had occurred for some months (three positive segments). The interpretation of hair tests from young children needs a global review of all the findings, as a positive result cannot stand alone to claim repetitive exposures.

Hair and dietary supplements testing to identify contamination with roxadustat in an adverse analytical finding.

Roxadustat is an oral inhibitor of hypoxia-inducible factor (HIF) prolyl hydroxylase, which increases endogenous erythropoiesis. WADA has included roxadustat and other HIF stabilizers on its list of prohibited substances. We describe here the case of an elite athlete (female, 31 years old, 168 cm and 53 kg) with an adverse analytical finding (AAF) with concentration of roxadustat in her urine at 0.289 ng/mL in the A sample and 0.529 ng/mL in the B sample (83% higher than A). A stability study was carried out, showing total stability of roxadustat at this concentration in urine exposed to light for 50 h, so photoisomerization degradation cannot explain the difference in concentration. Her urine had been completely negative in a control test carried out three days previously, while roxadustat had been shown to be present in urine for at least 20 days after administration of pharmacologically effective doses to an athlete. Hair concentration was 0.39 and 0.35 pg/mg in the segments corresponding to the presumed period of intake, with few adjacent segments also positive (0.29-0.33 pg/mg), likely explained by cosmetic treatments. Concentrations found in a patient treated with a pharmacologically active dose (between 100 and 120 mg 3 days a week) were more than 100 times higher (between 41 and 57 pg/mg). Numerous supplements and pharmaceuticals taken by the athlete were analyzed. Only collagen powder showed the presence of roxadustat, at a very low but highly variable concentration (100 pg/g-1000 pg/g). A female volunteer (58 years old, 169 cm and 65 kg), taking this powder at the same doses as the athlete (10 g of powder 5 times for 6 days) presented 7 roxadustat-positive urine samples (although lower than those observed in the athlete) out of 34 sampled over 7 days, the difference in powder sampling location, age, weight, height, pharmacokinetic parameters variability and level of sporting activity between the athlete and the volunteer probably explaining the difference in concentrations observed. All these results could be consistent with an AAF due to contamination by dietary supplements, which are becoming increasingly common due to the current exposome of athletes in our society.

Hair analysis in postmortem investigations: Case of a skeletonized body.

Analysis of hair collected from putrefied or skeletal bodies is always complex and must take into account several pitfalls, such as external contamination and contamination by biological fluids. This work presents a case of particular complexity. A skeletonized body was discovered on a country road. A tuft of brown hair, detached from the scalp, irregular in length, non-oriented, in contact with soil and vegetation, was removed. An anthropological examination was carried out and genetic samples were taken from the right femoral shaft. After about 10 washes with warm water and dichloromethane, the tuft of hair was analyzed without segmentation. General unknown screening was performed by liquid chromatography system coupled to a high-resolution mass spectrometry (LC-HRMS) after incubation in pH 9.5 borate buffer and liquid-liquid extraction. Specific Multiple Reaction Monitoring (MRM) methods for date rape drugs were carried out by liquid chromatography system coupled to a tandem mass spectrometry (LC-MS/MS). The anthropological examination allowed to determine that the victim was a female individual, over 60 years old, the death dating from 3 months to 1 year. Comparison of the DNA results with the Missing Persons Index led to the identification, a 60-year-old woman who disappeared 5 months earlier. Hair analysis showed the presence of oxazepam (361 pg/mg), nordiazepam (54 pg/mg), and alimemazine (5 pg/mg). The interpretation of these concentrations is extremely difficult due to the risk of degradation of the hair cuticle during prolonged stay in the soil, as well as of contamination by putrefactive fluids. The authors discuss the value of using multiple biological and non-biological matrices in this context to improve the interpretation of the results.

Testing for clomiphene in keratinous matrices using LC-MS/MS in doping purpose: Is a single intake of clomiphene detectable in hair and nail clippings?

Clomiphene is a selective estrogen receptor modulator. It is indicated for the treatment of female infertility issues but in sport, it can be misused to stimulate endogenous testosterone secretion in men. Therefore, it has been prohibited at all times by the World Anti-doping Agency. The aim of this study was to get data to be able to interpret concentrations in athletes. A healthy volunteer (male, 62 years-old) ingested a single therapeutic dose of clomiphene (Clomid™, 50 mg). Strands of hair (blond, 4 cm) were collected one month after the ingestion. Body hair (beard, axillary, pubic and chest hair), and finger and toenails were collected over 4-5 months. A previous method was modified to identify and quantify clomiphene in keratinous matrices. 30 mg of specimen were sonicated and incubated in 1 mL of methanol, in presence of 200 pg of clomiphene-D5 (internal standard). After centrifugation and evaporation of the organic phase, the samples were analyzed using LC-MS/MS. Linearity was verified in hair and nail clippings between 1 and 500 pg/mg. The limits of detection and quantification were determined at 0.3 and 1 pg/mg respectively. The study demonstrated that clomiphene tested positive in all the analyzed specimens at 9 pg/mg in head hair, from 28 to 486 pg/mg (body hair) and from 4 to 57 pg/mg (nails). Clomiphene was identified for the first time in multiple keratinous matrices. This study demonstrated that a single oral therapeutic dose is detectable in keratinous matrices over a long period of time.

Window of detection of cocaine-related alkaloids in oral fluid collected with the FloqSwab™ after coca tea consumption.

Coca tea is a popular drink in some countries of South America, where it is presented as a safe energy preparation, based on a limited total content of cocaine of ∼3-5 mg. Tea bags can be bought with no legal considerations in these countries both by locals and tourists, but its consumption can have consequences when consumed overseas. Driving under the influence of cocaine is banned in most of the places in the world and can be documented by oral fluid testing. A study was implemented with coca tea bags (Coca & Muna) purchased in Peru, after a French attorney-at-law contacted the laboratory to assess the involvement of coca tea in the positive oral fluid results of a driver. Ten healthy volunteers consumed 250 mL of coca tea containing 4.5 mg of cocaine. No volunteer reported any change in behavioral effects after consumption of the coca tea. Oral fluid was collected with a swab (FloqSwab™, Copan) over 8 h to follow the elimination of cocaine and its major metabolites (benzoylecgonine and ecgonine methylester). This is the procedure used by the French police. All samples were analyzed by UHPLC-MS-MS after Quantisal™ buffer desorption. As the device does not allow measurement of the amount of collected fluid, the results are qualitative. This is in accordance with the French law that requires a yes or no response about the presence of cocaine, with a minimum required performance level of 10 ng/mL of cocaine or benzoylecgonine. Parent cocaine was identified for 30-120 min. Benzoylecgonine and ecgonine methylester were identified between 1 and 8 h, with a large inter-individual variation. Although it is generally accepted that a 4-5 mg cocaine dose has no significant pharmacological effect, the consumption of coca tea can lead to the suspension of a person's driving license due to a positive oral fluid test.

In vitro characterization of protonitazene metabolites, using human liver microsomes, and first application to two urines collected from death cases.

Protonitazene, or N,N-diethyl-5-nitro-2-[(4-propoxyphenyl)methyl]-1H-benzimidazole-1-ethanamine, is a novel synthetic opioid, which belongs to the nitazene family. Over the last four years, nitazenes have re-emerged on the new psychoactive substances market and have been reported in several fatal intoxication cases. The metabolism of several nitazene analogues have already been studied, but to date, no data exists regarding protonitazene. The aim of the study was the detection of protonitazene and its metabolites in authentic human urine collected in two fatal intoxication cases, comparing the data after in vitro incubation with human liver microsomes, and subsequent analysis by ultra-performance liquid chromatography-tandem mass spectrometry and ultra-performance liquid chromatography-high-resolution mass spectrometry. Protonitazene metabolites, including N-desethyl-protonitazene, 5-amino-protonitazene and 4-hydroxy-nitazene, were characterized in vitro and were identified in the urine of both cases. The ratios between metabolites and parent protonitazene, higher than 1, were calculated to estimate the proportionality of metabolites. The results suggest that testing protonitazene metabolites should increase the window detection of exposure to protonitazene.