The importance of updatable analytical screening methods for new psychoactive substances in clinical and forensic toxicology.

Abstract: The continuous emergence of New Psychoactive Substances enter-ing the illegal markets poses always new challenges to forensic and clinical toxicology laboratories facing the lack of analytical methods capable of determining the newer parent compounds and eventual metabolites. More than 50 new molecules have been reported by the European Monitoring Centre for Drugs and Drug Addiction only in 2021, further expanding the panel of target analytes. However, the implementation of new detection methodologies may not be viable due to time constraints and inadequate resources. To this concern, the updatable analytical screening methods constitute a powerful tool for pharmacotoxicological laboratories investigating both non biological seizure materials and conventional and non conventional matrices coming from intoxications and fatalities of drugs consumers.

The targeted analysis of New Psychoactive Substances in oral fluid through chromatographic-spectrometric methods: review of recent findings.

OBJECTIVE: The oral fluid was demonstrated as an effective matrix to assess drug consumption in forensic settings. Recently, the increasing number of intoxications related to New Psychoactive Substances raised the attention of the scientific community. To this concern, different analytical methods to detect and quantify NPS in oral fluids were developed and validated, most of them based on hyphenated techniques. MATERIALS AND METHODS: A broad-ranging search was conducted on multidisciplinary research databases using "New Psychoactive Substances", "oral fluid", "toxicological analysis", "analytical method", "targeted method", "HPLC-MS/MS", "GC-MS", "GC-MS/MS" alone or in combination as search strings. All research articles published between 2017 and 2021 were considered. RESULTS: Different chromatographic-spectrometric methods to detect and quantify the NPS in oral fluid were reported in the literature. The classes of NPS explored were synthetic cannabinoids, synthetic cathinones, new designer benzodiazepines, synthetic opioids, fentanyl analogues, tryptamines, and phenethylamines. The most used technique was HPLC-MS/MS due to the sensitivity and high throughput. The GC-MS technique was preferred for synthetic cannabinoids, anyway different HPLC-MS/MS methods were developed. Moreover, the LC-HRMS technique was applied for the development of an analytical assay to detect new synthetic opioids and fentanyl analogues. CONCLUSIONS: The analytical interest on oral fluid as an effective matrix to assess drug exposure is increasing. The hyphenated techniques were demonstrated effective in the detection of NPS in oral fluids. The most suitable techniques are HPLC-MS/MS due to the sensitivity and the possibility to include different classes of substances in a single analytical run.

Adulterants in drugs of abuse: a recent focus of a changing phenomenon.

ABSTRACT: The adulteration of drugs of abuse is a well-known practice with alarming public health implications. In fact, the presence of other substances than those declared by the drug dealers may expose the consumers to unexpected adverse effects, even potentially fatal ones depending on the adulterant. Various adulterants are often added during the manufacturing process as cutting agent for deliberate underdosing, such as in case of levamisole for cocaine, or to strengthen the drug ef-fect, as in case of fentanyl for heroin. The authors aimed to elaborate on such unfolding dynamics and concluded that continuous development of effective analytical tools, along with a closely focused evaluation of each case by the medical doctor is essential to clarify the origin of the intoxications, so as to undertake the best therapeutic pathway.

New Psychoactive Substances consumption and their monito- ring during Covid-19 pandemic.

ABSTRACT: Currently, the world is facing an unprecedent change of everyday life, due to the Covid-19 pandemic that has been affecting all the nations for more than one year. The public health systems were restructured in all the countries as a response to the constant emergency status, ne-glecting some services like toxicological analyses. In this scenario, the current spread of the New Psychoactive Substances is less controlled than before and the data on its expected mutation come from seizures analyses. Where the global distribution of drugs of abuse was affected by the restriction, fentanyl seizures did not drop during the pandemic. Moreover, new synthesis of fentanyl analogues resulted in new toxic adulterants as by products. Furthermore, diversion of benzodiazepines and new designer benzodiazepines were reported during the pandemic period. In this scenario, the scientific community and the international agencies should tighten their collaboration in order to monitor the emerging of new unknown substances.

Management of GHB acute intoxications.

Intoxications related to γ-Hydroxybutyric acid (GHB), arising from its success as recreational drug due to its psychotropic properties, are significantly and alarmingly jeopardizing public health, posing major challenges to medical staff. In recent years, GHB's prodrug γ-butyrolactone (GBL) has often supplanted GHB in recreational settings, owing to its lower cost and the ease with which it can be obtained, mainly due to its various legal industrial applications. The Authors intend to stress that symptoms should be assessed and confirmed by timely toxicological analyses by highly-trained, expert professionals. Such tests aimed at analytical confirmation are instrumental in providing physicians valuable indications in terms of the proper pharmacological treatments in order to revert the adverse, or even fatal, side effects, particularly when the overall intoxication picture looks ambiguous. At the time being, little is known about the pharmacological therapies effective in GHB intoxication cases; further comprehensive research is therefore essential, if we are to tackle such a burgeoning public health emergency before it is too late.

Fentanyl analogues potency: what should be known.

Fentanyl is a full synthetic opioid acting as a strong µ-opioids receptor agonist. As other opioids, it exerts effects on central nervous systems like euphoria, sedation, anesthesia and respiratory depression at high dosage. It is the parent compound of the high potent opioids class, characterized by a potency up to 10,000 fold higher than mor- phine, currently prescribed as anesthetic and pain killers. Anyway, the diversion of fentanyl analogues has been reported since their appear- ance on the market, rising until alarming rate. Every year, new synthetic alternatives to the controlled fentanyl are proposed on the black market causing an increasing number of fatalities all over the World. Due to the high potency of this class of substances, it may be difficult to ana- lytically detect the molecules in biological matrices and find the actual cause of the deaths. Moreover, an additional analytical challenge is represented by the emergence of newly synthesized derivatives. In this concern, the harmonization of international guidelines, the adoption of common legal responses and the enforcement of international collaboration is desirable to face this alarming public health threat.

Drug-facilitated sexual assaults (DFSA): a serious underestimated issue.

OBJECTIVE: Drug-facilitated sexual assault (DFSA) is a nonconsensual sexual act in which the victim is incapacitated or unconscious due to the effects of alcohol, a drug and/or other intoxicating substances. Dozens of drugs (including ethanol) can potentially be used to commit sexual assaults, but γ-Hydroxybutyric acid (GHB) and flunitrazepam are the most common "date rape drugs". MATERIALS AND METHODS: Multidisciplinary databases were browsed using the following search terms: "drug-facilitated sexual assault", "chemical submission", "date rape", "rape drugs", and "drink-spiking". Moreover, a search for reports was conducted on Institutional websites to identify documentation published by international agencies or institutions. Articles and reports were independently evaluated by each author. RESULTS: There are no accurate estimates of the number of DFSA occurring each year, although assaults are increasingly reported. Many DFSA, however, are still not reported. Victims are reluctant to report incidents because of embarrassment, guilt or perceived responsibility, or because they do not clearly remember the assault. Moreover, most of the drugs typically used in sexual assaults are rapidly metabolized, making them undetectable in routine drug screenings. CONCLUSIONS: Most of the substances involved in DFSA, with the exception of alcohol, are under international control and scheduled under the United Nations Single Convention on Narcotic Drugs of 1961 and the Convention on Psychotropic Substances of 1971. However, several psychotropic substances and antihistamines used in sexual assaults are still not under international control, allowing for trafficking, often via the Internet and courier. The absence of international control makes it difficult to obtain accurate data on the nature and the extent of the problem.

Comparison of cheeses from goats fed 7 forages based on a new health index.

This study proposed the General Health Index of Cheese (GHIC) as an indicator for the presence of health-promoting compounds in cheese and compared the antioxidant capacity and phenolic and fatty acid contents of cheeses from goats consuming 7 forage species. Ninety-one homogeneous Red Syrian goats were randomly assigned to 1 of 7 feeding treatments (Festuca arundinacea, Hordeum vulgare, Triticosecale, Pisum sativum, Trifolium alexandrinum, Vicia sativa, and Vicia faba minor). The housed goat groups received the scheduled forage ad libitum. Forage species affected the antioxidant capacity, the phenolic and fatty acid contents, the Health Promoting Index, and the GHIC. Trifolium alexandrinum, Triticosecale, and Hordeum vulgare showed a clear advantage in terms of beneficial fatty acids content in goat cheese. Cheese from the Triticosecale group also showed a high antioxidant capacity value even if its polyphenol content was intermediate compared with others. Trifolium alexandrinum and Triticosecale had the highest value of the new index GHIC. This comparison suggests that there are important differences in fatty acid profile and polyphenol content among cheeses from goats fed grasses and legumes commonly used in the Mediterranean area. In this first approach, GHIC index, which combines the positive components found in cheese, seems to be a useful tool to provide an indication concerning the general health value of the product.

Effects of Sulla forage (Sulla coronarium L.) on the oxidative status and milk polyphenol content in goats.

Twelve milking Girgentana goats were allocated into 3 groups and fed 3 diets ad libitum in a 3×3 Latin square design with 3 periods. The diets were Sulla fresh forage ad libitum (SUL), Sulla fresh forage ad libitum and 800g/d of barley meal (SULB), and mixed hay ad libitum and 800g/d of barley meal (HB). Changes were observed in total polyphenol intake, plasma oxidative status, plasma polyphenol content, metabolic status, milk polyphenol content, and total antioxidant capacity. The fresh forage diets (SUL and SULB) increased dry matter, total polyphenol, nontannic polyphenol and tannin intakes, and the milk total protein, casein, milk total polyphenols, and milk free polyphenols compared with the HB diet. The intake of condensed tannins was higher in the SUL than in the HB group, and an intermediate value was recorded for the SULB group. Plasma from the SUL- and SULB-fed goats exhibited greater total antioxidant capacity and greater total polyphenol and free polyphenol levels compared with the plasma from the HB group. Positive correlations between plasma total antioxidant capacity and condensed tannins intake (r=0.43), plasma total polyphenol and total polyphenol intake (r=0.61), and plasma free polyphenol and milk total polyphenols (r=0.38) were observed. The correlation between milk free polyphenols and whey total antioxidant capacity (r=0.42) highlights the role of free polyphenols in the determination of the antioxidant activity of milk. These results indicate that Sulla fresh forage exerts an antioxidant activity due to its secondary compounds that provide additional antioxidant value and that Sulla forage appears to be a promising strategy for improving product quality.

Influence of fresh forage-based diets and αs1-casein (CSN1S1) genotype on nutrient intake and productive, metabolic, and hormonal responses in milking goats.

Polymorphism at the αS1-casein locus (CSN1S1) in goats influences several milk production traits. Milk from goats carrying strong alleles, which are associated with high αS1-casein (αS1-CN) synthesis, has higher fat and casein contents, longer coagulation time and higher curd firmness than milk from goats with weak alleles linked to low αS1-CN content. Nutrition also affects these milk properties; therefore, it is important to better understand the interaction between dietary characteristics and the CSN1S1 genotype in goats. This study aimed to investigate the effect of fresh forage based diet or energy supplement on feeding behavior, milk production, and metabolic and hormonal parameters of Girgentana goats with different genotypes at CSN1S1 loci. From a group of goats genotyped by PCR at the DNA level, 12 were selected because they had the same genotype for αS2-CN, ß-CN, and κ-CN but a different genotype for αS1-CN: 6 were homozygous for strong alleles at the CSN1S1 loci (AA) and 6 were heterozygous for a weak allele (AF). Goats of each genotype were allocated to 3 subgroups and fed 3 diets ad libitum in a 3×3 Latin square design. The diets were sulla (Hedysarum coronarium L.) fresh forage, sulla fresh forage plus 800 g/d of barley meal (SFB), and mixed hay plus 800 g/d of barley meal (MHB). Diet had a stronger effect than CSN1S1 genotype. The SFB diet led to the highest energy intake, dry matter (DM) digestibility, and milk yield. The fresh forage diets (SFF and SFB) increased DM and crude protein (CP) intake, CP digestibility, and milk CN compared with the MHB diet. The diets supplemented with energy (SFB, MHB) reduced milk fat and urea, improved CP utilization for casein synthesis, and limited body fat mobilization, in accordance with a lower level of nonesterified fatty acids and higher levels of glucose and IGF-1. With regard to CSN1S1 genotype, AA goats showed higher CP digestibility and lower free thyroxine hormone and cholesterol levels than AF goats. Significant diet × genotype interactions indicated how AA goats, compared with AF goats, showed higher DM digestibility and milk yield when fed the SFB diet, which had more energy. A reduction in free triiodothyronine hormone occurred in AF goats fed the MHB diet, whereas no differences were observed in AA goats. These results demonstrate how goats with a higher capacity for αS1-CN synthesis exhibit more efficient energy and protein utilization, evident at the digestive level, and better productive responses to high-nutrition diets.