Development, validation, and application of a quantitative volumetric absorptive microsampling-based method in finger prick blood by means of LC-HRMS/MS applicable for adherence monitoring of antipsychotics.

Volumetric absorptive microsampling (VAMS), an emerging microsampling technique, is expected to overcome some disadvantages of dried blood spots such as volume inaccuracy and influence of hematocrit (HT). This study aimed to develop and evaluate a VAMS-based strategy for quantification of 13 frequently prescribed antipsychotics in finger prick blood within the scope of adherence monitoring to complement already-established qualitative urine analysis. The final workflow consisted of VAMS tip hydration and subsequent precipitation. Samples were analyzed by using reversed-phase ultra-high-performance liquid chromatography and Orbitrap mass spectrometry operated in parallel reaction monitoring mode. The analytical procedure was successfully validated based on international recommendations at three different HT values (20%, 40%, 60%) for most of the analytes. Selectivity and within/between-run accuracy and precision were in accordance with the recommendations in most cases. Internal standard-normalized matrix factor met recommended criteria for all analytes at HT 40%. For the HT values of 20% and 60%, only four substances did not meet the criteria. Dilution integrity was given for all substances, except for olanzapine, allowing a quantification over the whole therapeutic range of selected antipsychotics. Long-term stability in VAMS tips was tested and revealed degradation of five antipsychotic drugs after 1 week of storage at 24 °C. A proof of concept of the applicability of the method was obtained by quantification of a selection of the 13 antipsychotic drugs in VAMS tips and matched plasma samples. Results were coherent between matrices. Thus, VAMS was shown to be a promising alternative for adherence monitoring of at least the investigated antipsychotics.

Concentrations of aripiprazole and dehydroaripiprazole in hair segments from deceased individuals with mental disorders.

Segmental hair analysis provides information regarding previous long-term drug exposure, which is useful in the evaluation of cause of death for individuals with mental disorders. The aim was to analyze postmortem concentrations of the antipsychotic drug aripiprazole and its active metabolite dehydroaripiprazole in hair segments from individuals with known aripiprazole intake. Hair samples were collected during autopsy. Each sample was segmented into one to six 1cm segments, depending on the length of the hair shaft. Pulverized hair was extracted and analyzed using a previously published ultra-high-performance liquid chromatography-tandem mass spectrometric method. The 10th-90th percentile of aripiprazole concentrations in all hair segments (n=78) from 17 individuals were 0.024ng/mg-11ng/mg with a median of 2.3ng/mg, and the 10th-90th percentile concentrations of dehydroaripiprazole were 0.020ng/mg-11ng/mg, with a median of 2.6ng/mg, in all hair segments (n=71). The metabolite-to-parent drug ratios ranged from 0.21 to 1.5, with a median of 0.72. The administered doses were calculated for each individual based on aripiprazole prescription data and pharmacy pickups, giving dose estimates of 1mg-32mg daily. A positive significant correlation was observed between concentrations in hair and blood, whereas no trends were observed between the concentrations in hair and the estimated doses. Besides aripiprazole, other antipsychotic drugs were found in several hair segments, indicating a high degree of polypharmacy among all subjects. The present study establishes concentrations of aripiprazole and dehydroaripiprazole in hair segments from 17 deceased individuals with long-term aripiprazole use. In addition, hair analysis demonstrates the possibility of evaluating polypharmacy.

Post mortem tissue distribution of quetiapine in forensic autopsies.

The antipsychotic drug quetiapine is widely used, and increasingly prescribed off-label. Furthermore, quetiapine use has been linked to increased mortality rates, most likely due to a range of cardiovascular and metabolic adverse effects. This makes quetiapine a relevant substance in forensic toxicology casework. Quetiapine is believed to undergo significant post mortem redistribution. Herein, we present tissue distribution and concentration levels of quetiapine in post mortem whole blood, brain tissue, skeletal muscle, and liver tissue in a series of 14 quetiapine-implicated forensic autopsy cases along with the quetiapine concentrations determined in femoral whole blood in conjunction with the autopsies. Quantification was performed using liquid-liquid extraction and a validated UPLC-MSMS method. Six deaths were attributed to intoxication with quetiapine in combination with other substances; there were no quetiapine monointoxications. In eight cases, death was attributed to other causes than drug toxicity. In a majority of the cases, liver tissue contained the highest quetiapine concentrations, while whole blood levels were the lowest. Central (heart) blood concentrations were generally higher than peripheral (femoral) blood levels. Quetiapine concentrations in femoral blood correlated most strongly with concentrations in skeletal muscle. Otherwise, there was no consistent hierarchy of quetiapine tissue concentrations, and the tissue distribution showed no clear relationship with the length of the post mortem interval.

Bioassay: A novel approach in antipsychotic pharmacology.

Accurate identification and measurement of antipsychotic drugs in in-vivo and in-vitro environment has a key role in the development and design of drugs, as any slight change in their dosage can affect the treatment process. Determination of antipsychotics in physiological fluids such as blood, plasma and urine are important in drug tablets and medical approaches. Chromatography is the main method used in the determination of antipsychotic drugs but this technique, despite its relative sensitivity, is a costly and complicated method. Biosensors, as simple, low-cost and highly sensitive and specific tools, can be the best alternative to eliminate the drawbacks of routine methods. For this purpose, various biosensors have been extensively developed in the recent years. The main objective of the present study is to introduce and collect recently developed biosensors for the measurement and detection of antipsychotic drugs. Therapeutic drug monitoring (TDM) is a fundamental principle in pharmaceuticals, with the primary goal being to reduce and eliminate the side effects of drugs. This study shows that biosensors can be a modern and novel approach in the field of TDM.

Analysis of clozapine in its tablets using two novel spectrophotometric reactions targeting its tertiary amino group.

Two simple spectrophotometric methodologies have been proposed and validated for the measurement of an atypical antipsychotic drug Clozapine (CLZ). Method A depends on interaction of CLZ with N-bromosuccinimide(NBS) resulting in formation of a yellowish orange colored product, measured at 320 nm. The linearity range was 5.0-70.0 µg/mL. Method B depends on condensation of the same drug with acetic acid mixed anhydride reagent producing a purple colored product, measured at 319 nm. The linearity range was 8.0-24.0 µg/mL. All parameters affecting the reaction condition (volume of both reagent, temperature, time and the different diluting solvents) were optimized. Both methods were successfully applied to assay CLZ in its pure form and tablets giving mean percentage recoveries of (98.87 ± 1.8 and 100 ± 1.7) for method A, and corresponding values of (98.6 ± 0.96 and 99.5 ± 1) for method B. Besides, the study of reactions stoichiometry was performed and the reaction mechanisms were proposed.

A different insight in hair analysis: Simultaneous measurement of antipsychotic drugs and metabolites in the protein and melanin fraction of hair from criminal justice patients.

BACKGROUND: Previous studies have postulated that four structural compartments may be differentiated in hair: surface protein domain, water-accessible protein domain, water-inaccessible protein domain, and melanin. Drugs contained in blood, sweat, sebum, and environment would be deposited in the first two domains, with primarily drugs in blood being incorporated in the latter two domains during hair synthesis. Drugs in the first two domains would be removed by washing procedures. Use of enzymatic extraction procedures and evaluation of hair for damage from harsh cosmetic treatments might help to separately identify and quantify the drugs incorporated in the second two domains. AIMS: a) Development of an UPLC-MS/MS method for the simultaneous quantification of the following 19 antipsychotic drugs and metabolites in hair: amisulpride, aripiprazole, chlorpromazine, clotiapine, clozapine, desmethylclozapine, desmethylolanzapine, haloperidol, norchlorpromazine, 7-OH-quetiapine, 9-OH-risperidone, olanzapine, pimozine, pimpamperone, quetiapine, risperidone, sertindole, sulpride, and tiapride; b) evaluation of measurement of patient adherence to prescribed medication use, c) determination of the influence of biochemical individuality effects on hair drug content, d) evaluation of relative binding of antipsychotic drugs to protein and to melanin hair structures. METHOD: Approximately 10 mg of intact hair were decontaminated with isopropanol and phosphate buffer, and then enzymatically digested overnight with dithiothreitol. After centrifugation, the supernatant digest (protein fraction) was separated from the remaining melanin hair pellet (melanin fraction). Melanin fraction was washed with water, and the drugs were extracted with dimethyl sulfoxide with ball-mill pulverization. Both fractions were purified with solid-phase cation exchange cartridges and injected in the UHPLC-MS/MS system. RESULTS AND DISCUSSION: Validation of the method was carried out on the protein fraction following international guidelines. The limits of quantification ranged from 1.6-40 pg/mg. The method was applied to 59 head hair samples from prisoners from an antipsychotic compliance study in the criminal justice system in US. The patients were under chlorpromazine, haloperidol, risperidone, olanzapine, or quetiapine multiple antipsychotic treatment, during incarceration. The first head hair centimeter, closest to the scalp, was analyzed. The results were evaluated in relation to the type of hair, colour, hair damage, drug melanin affinity, and prescribed dose. In general, no good correlation between the prescribed dose/concentration in hair was obtained. A wide range of antipsychotic concentrations were observed 'dose mg/day (d); pg/mg protein fraction (A)': chlorpromazine (d:50-400;A:1600) and its metabolite norchlorpromazine (A: 1600), haloperidol (d:4-20;A: 1600) and its metabolite 9-OH-quetiapine (A:

Development of a method of hollow fiber-based solid-phase microextraction followed by ultra performance liquid chromatography-tandem mass spectrometry for determination of five antipsychotics in human whole blood and urine.

This work focused on the development and validation of a method based on hollow fiber-based solid-phase microextraction coupled to ultra-performance liquid chromatography tandem mass spectrometry (HF-based-SPME-UPLC-MS/MS) for the determination of five antipsychotics at a pg mL-1 level in human whole blood and urine. Four types of hollow fiber membrane materials, including polyether sulfone, polypropylene, polyvinyl chloride and polyvinylidene fluoride were investigated. Finally, polyether sulfone hollow fiber without any modification was selected as the adsorption medium for solid-phase microextraction (SPME) with the following extraction procedure: the analytes were adsorbed onto the hollow fiber in the sample bottle with application of ultrasonication. Subsequently, the hollow fiber was transferred into a slim glass tube containing an appropriate solvent, and the analytes were desorbed by ultrasound treatment before detection by UPLC-MS/MS. In order to obtain satisfactory extraction efficiency, extraction parameters such as hollow fiber membrane material, pH, hollow fiber length, extraction time, desorption solvent and desorption time were investigated. Under the optimum experimental conditions, this method allowed for determination of five antipsychotics in human whole blood with excellent limits of quantification (LOQs) (25.0, 12.5, 25.0, 25.0 and 12.5 pg mL-1 for perphenazine, chlorpromazine, chlorprothixene, promethazine and trifluoperazine, respectively). The corresponding LOQs in human urine were 25.0, 12.5, 12.5, 12.5 and 12.5 pg mL-1 for the respective antipsychotics. The precision (RSD) was no more than 13.3%. The extraction recoveries for human whole blood and urine were in the range of 46.4-96.6% and 65.2-101.9%, respectively. The proposed method was compared with other methods from the literature and the results demonstrate that it is a simple, sensitive, efficient and green technique. It is suitable for analyzing trace target analytes in complex matrices such as biological samples and can provide a reliable tool for drug monitoring especially in forensic analysis and case of drug abuse.

Hair analysis of antidepressants and antipsychotics-Overview of quantitative data.

Antidepressant and antipsychotic drugs are regularly encountered in different aspects of forensic toxicology, and some cases require the examination of hair samples. In this study, common antidepressant and antipsychotic drugs regarding hair concentrations over the past decades were reviewed. Although numerous publications around method validations, case reports, or controlled dose studies were found, apparently there is a lack of comprehensive data for many substances. Information on the hair length and dosage across the publications varied largely, and case numbers were generally low except for several retrospective controlled dose studies. Many substances were described only in method validations or case reports, and data were obtained from small case numbers. On the contrary, clozapine, haloperidol, amitriptyline, nortriptyline, risperidone and its metabolite, methylphenidate, citalopram, chlorpromazine, chlorprothixene, and quetiapine had a well-founded database as these substances were investigated in controlled dose studies with higher case numbers. Given the advancements made in analytical techniques over the past years, gas chromatography-mass spectrometry and liquid chromatography with tandem mass spectrometry techniques were the methods of choice and allowed the detection of chemical compounds at low concentrations. The controversy around a potential use of hair analysis to estimate the dosage remains as dose-concentration studies provided divergent results. A harmonization on the investigated hair length as well as on the extraction protocol would be of favor to achieve better comparability. Although hair analysis research focused mainly on drug abuse, availability of more data on antidepressants and antipsychotics would help to gain better knowledge and assist other forensic investigators.

Electrochemical determination of atypical antipsychotic drug quetiapine using nano-molecularly imprinted polymer modified carbon paste electrode.

In this research, the advantages of molecularly imprinted polymer (MIP) materials have been used to develop a new electrochemical sensor for determination of quetiapine (QTP) drug. MIP nanoparticles were synthesized by precipitation polymerization method and used as QTP recognition elements in the composition of modified carbon paste electrode (CPE) for selective and sensitive assay of this drug. Cyclic voltammetry (CV) and square wave voltammetry (SWV) techniques were used for electrochemical analysis. Some parameters affecting the sensor performance were optimized and under optimal conditions, the proposed sensor showed linear responses with QTP concentration in the range of 1.6 × 10-8 to 2.5 × 10-6 M (R2 = 0.9964). The limits of detection (LOD) and quantification (LOQ) were calculated 5.04 × 10-9 M and 1.68 × 10-8 M respectively. Also, the amounts of %RSD for evaluation of repeatability and reproducibility of the proposed sensors were respectively obtained 2.19 and 3.02%. The method was successfully applied to determination of QTP in its pharmaceutical formulation and human urine samples.

Application of Ionic Liquids for the Determination of Lipophilicity Parameters Using TLC Method, and QSRR Analysis for the Antipsychotic Drugs.

BACKGROUND: Reversed-phase liquid chromatography may cause difficulties, especially in the case of basic drugs due to the strong silanophilic interactions in the partition mechanism. Recently, imidazolium-based ionic liquids additives appeared interesting and a convenient solution for suppressing the harmful effect of free residuals of silanol groups, allowing remodeling of the stationary/mobile-phase system, and thus improving the lipophilicity assessment process. OBJECTIVE: The aim of the study was to evaluate the retention behavior of basic antipsychotics using various RP-LC systems, and compare them with data obtained from the modified ionic-liquids RP-TLC systems, and perform the QSRR analysis. METHODS: Retention and lipophilicity parameters of diverse antipsychotics have been examined in various RP-LC systems. Lipophilicity indices were compared with miscellaneous computed logP values. Furthermore, a large number of molecular descriptors have been computed and compared using various medicinal chemistry software, in order to contribute to the analysis of QSRR. RESULTS: Designated correlation coefficients showed that lipophilicity parameters from TLC systems without [EMIM][BF4] additive correlates very poor with the calculated logPs indices, whereas the indices from the traditional HPLC and TLC systems (with [EMIM][BF4]) were clearly better. Furthermore, QSRR analysis performed for these experimentally obtained lipophilicity parameters showed significant relationships between the retention constants (ROM, logkw) and the in silico calculated physicochemical molecular descriptors. CONCLUSION: ILs additive may be a significant factor affecting the lipophilicity of basic compounds, thus their use may be favorable in lipophilicity assessment studies. QSRR models with ILs showed that they may be useful in searching/or predicting HPLC/TLC retention parameters for the new/other antipsychotic drugs.

Segmental analysis of antidepressant and antipsychotic drugs in the hair of schizophrenic patients.

Hair analysis is useful for documenting long-term exposure to drugs. The potential of hair analysis for therapeutic drug monitoring within the forensic field has been studied, but reference values for some antidepressants and antipsychotics in the hair of individuals undergoing chronic therapy are still lacking. In the present study, a method was developed and validated for the determination of 23 analytes, including antidepressants, antipsychotics, and related metabolites, in human hair by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Hair samples (10 mg) were extracted with a 25:25:50 (v/v/v) mixture of methanol/acetonitrile/2 mM ammonium formate (8% acetonitrile, pH 5.3) utilizing cryogenic grinding. The present method demonstrated sufficient selectivity, robustness, and accuracy. Sixteen analytes in hair were reported in 46 psychiatric patients receiving fixed drug dosages. To the best of our knowledge, the hair concentrations of perphenazine and norolanzapine, as well as the concentrations of amisulpride, aripiprazole and its metabolite dehydroaripiprazole, olanzapine, and sulpiride, in hair from individuals receiving fixed dosages is reported for the first time. A significant relationship between the administered dose and the concentration in the proximal hair segment was found only for clozapine, norclozapine, and chlorpromazine. The results confirmed that the idea of using hair concentrations to monitor a daily dose is inapplicable.

Determination of Antidepressants and Antipsychotics in Dried Blood Spots (DBSs) Collected from Post-Mortem Samples and Evaluation of the Stability over a Three-Month Period.

An LC-MS/MS method for the identification and quantification of antidepressants and antipsychotics was developed on dried blood spots (DBSs). Moreover, analyte stability on DBSs within a 3-month period was monitored. Aliquots of 85 µL of blood from autopsy cases were pipetted onto DBS cards, which were dried and stored at room temperature. DBSs were analyzed in triplicate immediately, within the following 3 weeks, and after 3 months. For each analysis, a whole blood stain was extracted in phosphate buffer and purified using Solid Phase Extraction (SPE) cartridges in order to avoid matrix effects and injected in the LC-MS/MS system. Thirty-nine molecules were screened. Limits of detection (LODs) ranged between 0.1 and 3.2 ng/mL (g) and 0.1 and 5.2 ng/mL (g) for antidepressants and antipsychotics, respectively. Limits of quantification (LOQs) varied from 5 to 10.0 ng/mL for both. Sixteen cases among the 60 analyzed resulted positive for 17 different analytes; for 14 of these the method was fully validated. A general good agreement between the concentrations on DBSs and those measured in conventional blood samples (collected concurrently and stored at -20 °C) was observed. The degradation/enhancement percentage for most of the substances was lower than 20% within the 3-month period. Our results, obtained from real post-mortem cases, suggest that DBSs can be used for routine sample storage.

Sonochemical synthesis of iron-graphene oxide/honeycomb-like ZnO ternary nanohybrids for sensitive electrochemical detection of antipsychotic drug chlorpromazine.

We report a novel electrochemical sensor for the sensitive and selective determination of the antipsychotic drug chlorpromazine (CPZ) based on the iron (Fe) nanoparticles-loaded graphene oxide (GO-Fe)/three dimensional (3D) honeycomb-like zinc oxide (ZnO) nanohybrid modified screen printed carbon electrode (SPCE). The 3D hierarchical honeycomb-like ZnO was synthesized using a novel aqueous hydrothermal method and the GO-Fe/ZnO nanohybrid was prepared based on an inexpensive and fast sonochemical method using a high-intensity ultrasonic bath (Delta DC200H, 200 W, 40 KHz). Characterizations including scanning electron microscopy, elemental mapping, transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and Raman spectroscopy were carried out as part of this work. The electrocatalytic oxidation behavior of CPZ at various electrodes was investigated using the cyclic voltammetry technique, through which the GO-Fe/ZnO modified SPCE was identified as the best performing electrode. The quantitative determination of CPZ was then performed using the differential pulse voltammetry technique. The as-prepared GO-Fe/ZnO/SPCE sensor exhibited a quick and sensitive response towards the oxidation of CPZ with linear concentration ranges from 0.02 to 172.74 µM and 222.48 to 1047.74 µM. The modified SPCE sensor displayed a low detection limit (LOD) of 0.02 µM and a high sensitivity of 7.56 µA µM-1 cm-2. The proposed sensor also showed remarkable operational and storage stability, reproducibility, and repeatability. Furthermore, the practicability of the GO-Fe/ZnO/SPCE sensor has been verified with real sample analysis using commercial antipsychotic CPZ tablets and human urine samples, and adequate recovery has been achieved.

[A Forensic Autopsy Case in which Sulpiride and Estazolam Were Detected in an Adipoceratous Cadaver by LC-MS/MS].

We had a forensic autopsy case in which drugs were detected in a cadaver that had been stored in a cold and wet condition for 5 years. The skin of the cadaver was hard, and the color was partly whitish or dark brown. Though the cadaver had transformed into adipocere in the wet and cold condition, QuEChERS extraction and LC-MS/MS revealed the presence of sulpiride and estazolam in the femoral muscle and bone marrow. The concentrations of sulpiride and estazolam in the femoral muscle were 10.6 ng/g and 39.9 ng/g, respectively. The result of a drug screening test led not only to the cause of death but also to the personal identification of the cadaver. The individual had a history of drug taking, which had been stored in his medical records at the hospital for a long time. The fact of taking sulpiride and estazolam at the same time was characteristic, and it was useful in identifying the cadaver in this case. The progress in analytical technology has made possible the detection of particle drugs from old or adipoceratous cadavers, but there have been no reports of particle drugs being detected in a cadaver that had been dead for 5 years and had transformed to adipocere, as in our present case. The analytical results by LC-MS/MS were certainly important for the diagnosis of the cause of death, and, moreover, they were useful for the purpose of personal identification.

Determination of antipsychotic drugs in oral fluid using dried saliva spots by gas chromatography-tandem mass spectrometry.

The present work describes the optimization and validation of an analytical method for the determination of six antipsychotic drugs (chlorpromazine, levomepromazine, cyamemazine, clozapine, haloperidol, and quetiapine) in oral fluid samples after solvent extraction from dried saliva spots, by gas chromatography coupled to tandem mass spectrometry. The method was fully validated, and the included parameters were selectivity, linearity, limits of quantification, precision and accuracy, stability, and recovery. The method was linear for all compounds from 10 to 400 ng/mL, except for haloperidol (5-100 ng/mL), presenting coefficients of determination higher than 0.99. Inter- and intra-day precision and accuracy were in conformity with the criteria usually seen in bioanalytical method validation; i.e., coefficients of variation were lower than 15% and an accuracy of 15% or better for all studied drugs. The recoveries obtained with this miniaturized technique ranged from 63 to 97%. The herein described method is the first to be reported using the dried saliva spots approach for the analysis of these antypshychotic drugs, proving great sensitivity apart from its simple and fast procedure. The method was considered a good alternative to the conventional techniques to be applied in clinical and toxicological analyses, even more taking into account the extremely low sample volume used (50 µL). Graphical abstract.

Intra-Nasally Administered Oligopeptide Lunasin Acts as a Possible Anti-Psychotic Agent in Mice Models.

Background and Objectives: Previously we have shown that synthetic lunasin, a 43 amino acid residue-containing peptide, after its central (intracisternal) administration in mice demonstrated antagonism against dopaminergic drug behavioural effects, indicating a putative antipsychotic/anti-schizophrenic profile of lunasin. The aims of the present studies were: to test whether lunasin would show an influence on the dopaminergic system after intranasal administration, and to examine the effect(s) of lunasin on serotonin and glutamatergic systems, which could play an essential role in antipsychotic action. Materials and Methods: Lunasin was administered intra-nasally at doses 0.1 and 1 nmol/mouse in ICR mice (n = 7-8) and tested in an open field on hyperlocomotion caused by amphetamine; serotonin 5-HT 2A/2C receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)- 2-aminopropane (DOI); and glutamate NMDA receptor antagonist phencyclidine. Following behavioural testing, the contents of neurotransmitters and their metabolites in brain hemispheres (n = 6-8) were assessed by ultra-high-performance liquid chromatography-time of flight mas-spectrometry (UHPLC-TOF-MS) method. Also, lunasin binding to serotonin receptors was assessed. Results: Lunasin intra-nasally fully normalized hyper-locomotion and brain monoamine levels in amphetamine- and DOI-treated mice brains. Phencyclidine behavioural effects were not influenced. In vitro receptor binding data demonstrated a low affinity of lunasin (at µM concentrations) compared with DOI (nM concentrations) for the 5-HT2A and 5-HT2C receptors. Conclusions: These results demonstrated, for the first time, that the intranasal administration of oligopeptide lunasin normalized mice behaviour and brain monoamine levels in experimental psychosis mice models. Its neuro-regulatory effects indicated a usefulness of this peptide molecule for the design of novel psychotropic agents.

Rapid degradation of clozapine by heterogeneous photocatalysis. Comparison with direct photolysis, kinetics, identification of transformation products and scavenger study.

In this study TiO2-mediated photocatalytic degradation of the persistent drug clozapine under the simulated solar radiation was studied for the first time. The experiments were conducted both in the ultrapure and river water, which enabled the assessment of the organic matrix impact. The direct and indirect photolysis experiments were conducted for a comparison. Influence of the catalyst loading on the efficiency of the process was also assessed, and the highest catalyst loading (300 mg L-1) was found to be the most effective. The TiO2 photocatalysis was extremely effective for clozapine degradation - the decomposition was almost 300 times faster in comparison to the direct photolysis (t1/2 = 1.7 min, neither clozapine, nor the intermediates were detected after 20 min of irradiation), and presence of the organic matrix did not negatively affect the process. Nevertheless the photocatalytic process turned out to be highly sensitive to act of the ROS scavengers. Thirteen transformation products (TPs) were found and their structures were elucidated by the means of high resolution mass spectrometry. Properties - toxicity, biodegradability, BCF and BAF - of TPs and the parent molecule were estimated with the use of computational methods. Identified TPs were found as generally less toxic and more biodegradable than clozapine.

High-Performance Thin-Layer Chromatography Method for Simultaneous Determination of Antipsychotic and Medicinally Important Five ß-Carboline Alkaloids.

This is the first report on the development and validation of high-performance thin-layer chromatography (HPTLC) method for simultaneous analysis of five antipsychotic and medicinally important ß-carboline alkaloids (ßCAs), namely, harmalol, harmaline, harmine, harmane and norharmane. These ßCAs occurs in both plant and animal system including human being. In the present investigation, their best separation was achieved using an optimized mobile phase, chloroform: methanol: glacial acetic acid (7.8:2.2:0.2, v/v/v) on aluminum TLC plates precoated with silica gel 60 F254. The quantification was performed by densitometric scanning in fluorescence mode at 366 nm. The calibration curves were drawn using linear regression, plotted over the range 25-250 ng band-1 of standard ßCAs with correlation coefficient (R2) between 0.97 and 0.992. Accuracy in terms of recovery (83.95-112.40%), repeatability of application (0.61-2.42%), repeatability of measurement (1.94-3.05%) and intermediate precision (0.62-11.16%) of developed method were simultaneously determined. The limit of detection and limit of quantification were between 4.95-6.59 and 16.50-21.93 ng band-1, respectively. The method was validated according to ICH guidelines and was simple, cost-effective, precise, sensitive and specific for the determination of ßCAs in herbs, Fagonia schweinfurthii, Peganum harmala and Tribulus terrestris. The developed HPTLC method would have importance in forensic and industrial chromatographic analysis and fingerprinting of various herbs and drug formulations containing ßCAs.

Antipsychotic drugs in hospital wastewater and a preliminary risk assessment.

The residues of pharmaceutical and personal care products are the cause of increasing concern around the world. The aim of this study was to carry out the quantification of six antipsychotic drugs in hospital wastewater with the aid of liquid chromatography-mass spectrometry and, subsequently, make a preliminary assessment of the environmental risk posed. Dispersive liquid-liquid microextraction and solid phase extraction were optimized by multivariate design and validated in compliance with international guidelines. The extraction procedures were successfully applied to the quantification of the six selected antipsychotics in samples that were formed each day and collected at two main sampling points of the sewage network over the period of a week, in December 2017. Olanzapine (0.31─0.52 µg L-1), clozapine (0.56─0.97 µg L-1), haloperidol (1.43─2.73 µg L-1), risperidone (0.92─0.98 µg L-1) and chlorpromazine (0.52 µg L-1) were found in at least one sampling point. In the case of most analytes, the highest concentrations were determined at sampling point A, which are derived from the psychiatric wing. The environmental risk quotient for clozapine, chlorpromazine and risperidone was ˃600, a very high-risk index, which signals the need for a better control of the emission of antipsychotics and an improvement of the wastewater treatment, especially, with regard to wastewater discharged from the hospital psychiatric wing.

Toxicological findings in suicides - frequency of antidepressant and antipsychotic substances.

The role of psychoactive substances in the treatment of mental disorders and the risk of suicide are major public health issues. This cross-sectional study examined the prevalence of antidepressants and antipsychotics detected in toxicological screenings in suicides. Cases from the Institute of Legal Medicine of the Charité-University Medicine Berlin were reviewed over a 4-year-period. All cases (n = 477) with positive toxicology for antidepressants and antipsychotics in blood or organ tissue were included. Frequencies of the detected substances in non-suicide cases (n = 212; male n = 177, 55.2%; female n = 95, 52.5%) and suicide cases (n = 235; male n = 149, 63.4%; female n = 86, 36.6%) were examined. Tricyclic antidepressants (48.1%) were found most frequently in suicides, followed by atypical neuroleptics (37.0%), selective serotonin reuptake inhibitors (28.1%), typical neuroleptics (17.4%), tetracyclic antidepressants (16.2%) and other substances (8.9%). Alcohol was detected in 37.2% of suicides. The leading cause of death was drug poisoning (35.6%) followed by polytrauma (26.8%) and death by hanging (18.5%). A mental disorder (depression, schizophrenia, bipolar disorder, suicidality) was known in 22.9% of suicides. The most common location of death was the person's own house (63.8%) followed by public places (28.1%) and hospitals (8.1%) The five most common substances in the suicide group were doxepin (20%) citalopram (15.3%), mirtazapine (14.9%), quetiapine (13.6%) and amitriptyline (12.3%). Toxicological findings from cross-sectional studies provide insight into how often certain types of antidepressants and antipsychotics are associated with suicide. A complementary approach is valuable for assessing the risk of suicide during medical treatment because the various available approaches (analysis of suicidal behavior/ideation, toxicity of drugs) each have strengths and limitations.