Recent Advances in Analytical Techniques for Antidepressants Determination in Complex Biological Matrices: A Review.

Depression is one of the most prevalent but severe of mental disorders, affecting thousands of individuals across the globe. Depression, in its most extreme form, may result in self-harm and an increased likelihood of suicide. Antidepressant drugs are first-line medications to treat mental disorders. Unfortunately, these medications are also prescribed for other in- and off-label conditions, such as deficit/hyperactivity disorders, attention disorders, migraine, smoking cessation, eating disorders, fibromyalgia, pain, and insomnia. This results in an increase in the use of antidepressant medications, leading to clinical and forensic overdose cases that could be either accidental or deliberate. The findings revealed that people who used antidepressants had a 33% greater chance of dying sooner than expected, compared to those who did not take the medications. Analytical techniques for precisely identifying and detecting antidepressants and their metabolic products in a variety of biological matrices are greatly needed to be developed and made available. Hence, this study attempts to discuss various analytical techniques used to identify and determine antidepressants in various biological matrices, which include urine, blood, oral fluid (saliva), and tissues, which are commonly encountered in clinical and forensic science laboratories.

Identifying the effectual-combination ingredients of Zhi-zi-Hou-po decoction based on metabolic difference-oriented network regulation strategy.

Zhi-zi-Hou-po decoction (ZZHPD) has been used to treat depression in the clinic for thousand years in China. However, the pharmacodynamic substance of ZZHPD is still not totally clear due to its complex components. The objective of this study was to identify the effectual combination ingredients (ECIs) of ZZHPD, which could represent the antidepressant effect of the original formula. Firstly, differential plasma absorbed components with different variable importance in projection (VIP) value in chronic unpredictable mild stress (CUMS)-induced depression and control rat were revealed by untargeted metabolomics-driven strategy based on HPLC-ESI-TOF/MS, XCMS online and SIMCA-p software. Secondly, network topology scores (NTS) of plasma absorbed components were exposed by protein-protein interaction (PPI) network analysis based on components-related genes and depression-related genes, which were performed by network pharmacology tools. Finally, the ECIs were screened by considered of VIP value and NTS. Then the bioactivity was evaluated by cell viability and expression of glial fibrillary acid protein (GFAP), tumor necrosis factor α (TNFα) and interleukin 1ß (IL-1ß) of a lipopolysaccharide-induced astrocyte depression model. An effective combination composed with 12 components was filtrated as ECIs of ZZHPD, exposed to which the cell viability effect, the expression of GFAP and IL-1ß in astrocytes were essentially equivalent with original ZZHPD (p > 0.05), and that both characteristic constituents and trace compounds of ZZHPD might exert synergistic actions through multi-targets. The result of this study provided useful information for the clinical application and modern development of ZZHPD, and the proposed strategy could be regard as an alternative solution for effective combination screening of herbal medicines.

Combination of gel-electromembrane extraction with switchable hydrophilicity solvent-based homogeneous liquid-liquid microextraction followed by gas chromatography for the extraction and determination of antidepressants in human serum, breast milk and wastewater.

The current study presents for the first time a combination of the gel electromembrane extraction (GEL-EME) and switchable hydrophilicity solvent-based homogeneous liquid-liquid microextraction (SHS-HLLME) methods which can be used as an efficient hyphenated extraction procedure. This coupled method, which was followed by GC-FID, was applied for quantification of antidepressants (desipramine, clozapine, and citalopram) in biological and wastewater samples. The effective parameters of both GEL-EME and SHS-HLLME procedures were optimized. Using an agarose gel membrane, analytes were extracted from 7.0 mL of the sample solution to 500 µL of the aqueous acceptor solution. The maximum extraction of analytes of interest was obtained under the optimized conditions (pH of acceptor solution, 5.0; pH of gel membrane, 5.0; pH of sample solution, 7.0, voltage value, 30 V; and extraction time, 30 min). Then, the acceptor solution was transferred to the extraction cell and the SHS-HLLME procedure was conducted again under the optimized conditions. Dipropylamine (50 µL) was selected as an extraction solvent. The introduced technique exhibited good linearities with coefficients of determinatin (R2) higher than 0.983 and an acceptable linear range of 5.0-1000 ng/mL. Accordingly, the limit of detection was ≤ 1.0 ng/mL (S/N = 3) for all analytes, and the high enrichment factors were obtained in the range of 178.7-194.8. Moreover, the corresponding repeatability was from 4.0 to 8.7% (n = 3). The proposed method was successfully utilized to determine trace levels of the drugs in human serum, wastewater, and breast milk samples.

Fast and efficient analyses of the post-mortem human blood and bone marrow using DI-SPME/LC-TOFMS method for forensic medicine purposes.

For the first time the method DI-SPME/LC-TOFMS was used and developed in order to determine the large antidepressant drugs in real forensic cases. The aim of the study was to optimize the new DI-SPME/LC-TOFMS method for the quantification of the large group of psychotropic drugs such as benzodiazepines, selective serotonin reuptake inhibitors, selective serotonin and noradrenaline reuptake inhibitors, tricyclic antidepressants and sleeping pills "Z". The volume of the sample, adsorption time, post-adsorption purification and desorption time were precisely optimized. The validation parameters such as limit of detection and quantification, linearity, precision during and between days and the matrix effect were determined. All obtained values are within the acceptable range for toxicological analyses. The usefulness of the method was confirmed by analyzing the post-mortem samples. Drug concentrations were determined in real samples with high precision, which gives perspectives for the DI-SPME/LC-TOFMS routine application in toxicological and forensic analyses in the future.

Emerging intervention of antidepressant with DMARD in non-cancerous nociceptive persistent pain associated depression in FCA induced rheumatoid arthritic rats

Rheumatoid arthritis (RA) is a chronic inflammatory disorder that causes pain, systemic complications and premature mortality. Depression has also been identified as a problem for persons with RA. This association remaining significant even after the degree of disease activity is controlled. In the present study, the efficacy of combination therapeutic effect of antidepressant (amitriptyline) with Disease Modifying Anti rheumatoid drug (leflunomide) was determined in rheumatoid arthritis pain associated depression in Freund's complete adjuvant (FCA) induced arthritic rats. Drug treatment was started 9 days after induction of FCA induced arthritis in rats. The antiarthritic activity was assessed by measuring paw volume, weight-bearing, hematological, biochemical, serological parameters, Radiographic analysis and Histopathology of tibiotarsal joints. The antidepressant activity was assessed by Forced swimming test, Rota-rod test and confirmed by estimation of brain neuro transmitters (serotonin and norepinephrine) level. Results of this study revealed that leflunomide and amitriptyline combination showed more significant (p<0.001) antiarthritic and antidepressant action and leflunomide alone treatment showed significant (p<0.001) antiarthritic activity only as compared to arthritic control. The leflunomide and low dose amitriptyline combination found to be more effective in pain associated depression in rheumatoid arthritic rats

Postmortem analysis of human bone marrow aspirate - Quantitative determination of SSRI and SNRI drugs.

For the first time the MAE/UHPLC-TOFMS method was developed and used in order to determine the antidepressant drugs within the human bone marrow aspirate in real forensic cases. The following drugs, belonging to the group of selective serotonin (or serotonin-norepinephrine) reuptake inhibitors, were tested in this study: venlafaxine, citalopram, fluoxetine, sertraline and paroxetine. The sample preparation proposed in the present article included several steps: homogenization in ultrasound bath, liquid-liquid extraction, fat removal and evaporation under nitrogen. The extraction involved microwave-assisted extraction, performed for 15 min at 55 °C, with hexane-isoamyl alcohol (99:1, v/v) mixture, as an extraction solvent. Time and temperature of extraction were optimized using the simplex method. The fat removal step was introduced because of the fatty nature of the bone marrow that resulted in insufficiently purified samples, impossible to analyze using HPLC. It was achieved by adding a mixture of ethanol, water and formic acid to samples consisting of hexane and isoamyl alcohol, so that the analytes could diffuse to polar phase and fat could stay in non-polar phase. The method was validated and parameters such as: LOD, LOQ, linearity, matrix effect, recovery were determined. The validation parameters obtained allowed to recognize the method as quantitative. Due to lack of the data on therapeutic and toxic levels of considered drugs in bone marrow, data regarding serum has been used for reference. Under this assumption, the developed method allows for quantification of all mentioned drugs at therapeutic levels. Moreover this method has been used in real cases. Finally four analytes: venlafaxine (104 ng/mL), fluoxetine (84 ng/mL), paroxetine (about 3.6 µg/mL) and citalopram (68 ng/mL) were found in three case samples.

Targeting the Endocannabinoid/CB1 Receptor System For Treating Major Depression Through Antidepressant Activities of Curcumin and Dexanabinol-Loaded Solid Lipid Nanoparticles.

BACKGROUND/AIMS: This study investigated the underlying mechanisms of the antidepressant effects of curcumin and dexanabinol-loaded solid lipid nanoparticles in corticosterone-induced cell and mice depression models. METHODS: Curcumin and dexanabinol-loaded solid lipid nanoparticles (Cur/SLNs-HU-211) were synthesized via an emulsifcation and low-temperature solidification method. Antidepressant activities of nanoparticles in a corticosterone-induced major depression model were investigated by MTT assay, cellular uptake by flow cytometry, behaviour by Forced Swimming Test and rotarod test, neurotransmitters by High Performance Liquid Chromatography, Western blotting, qPCR and immunofluorescence. RESULTS: Treatment with Cur/SLNs-HU-211 induced greater dopamine (DA)/5-hydroxytryptamine (5-HT) release with reduced corticosterone-induced apoptotic cell death in PC12 cells. Additionally, in vivo Cur/SLNs-HU-211 significantly induced recovery from depressive behaviour with increased DA/5-HT levels, CB1 mRNA levels and CB1, p-MEK1 and p-ERK1/2 protein expression levels in the hippocampus and striatum. Cur/SLNs-HU-211 improved CB1 expression and inspired the proliferation of astrocytes in the hippocampus and striatum, exerted neuroprotective effects by preventing corticosterone -induced BDNF/NeuN expression reduction. CONCLUSION: Our study implies that Cur/SLNs-HU-211 may be a useful approach for treatment of major depression.

LC/QTOF profile and preliminary stability studies of an enriched flavonoid fraction of Cecropia pachystachya Trécul leaves with potential antidepressant-like activity.

There is increasing interest in natural antioxidants that are candidates for the prevention of brain damage occurring in major depressive disorders. Cecropia pachystachya is a tropical tree species of Central and South America and a rich source of polyphenols, particularly flavonoids. The aim of this study was to characterize the flavonoid profile of an enriched flavonoid fraction of C. pachystachya (EFF-Cp) and evaluate the antidepressant-like effects of its acute administration in behavior, cytokine levels, oxidative stress and energy metabolism parameters. The EFF-Cp chemical characterization was performed by HPLC/DAD and LC/QTOF. The antidepressant-like effects were performed by the forced swimming test, splash test and open field test. EFF-Cp revealed 15 flavonoids, including seven new glycosyl flavonoids for C. pachystachya. Quantitatively, EFF-Cp showed isoorientin (43.46 mg/g), orientin (23.42 mg/g) and isovitexin (17.45 mg/g) as major C-glycosyl flavonoids. In addition, EFF-Cp at doses 50 and 100 mg/kg reduced the immobility time in the forced swimming test, without changing the locomotor activity and grooming time. In addition, EFF-Cp was able to prevent the oxidative damage in some brain areas. In conclusion, the results of this study suggest that EFF-Cp exerts antidepressant-like effects with its antioxidant properties.

Liquid-Phase Microextraction and Gas Chromatographic-Mass Spectrometric Analysis of Antidepressants in Vitreous Humor: Study of Matrix Effect of Human and Bovine Vitreous and Saline Solution.

In forensic bioanalytical methods, there is a general agreement that calibrators should be prepared by fortifying analytes in matrix-based blank samples (matrix-based). However, in the case of vitreous humor (VH), the collection of blank samples for the validation and for routine analysis would require the availability of many cadavers. Besides the difficulty of obtaining enough blank VH, this procedure could also represent an ethical issue. Here, a study of matrix effect was performed taking into consideration human and bovine vitreous and saline solution (SS) (NaCl 0.9%). Tricyclic antidepressants [amitriptyline (AMI), nortriptyline (NTR), imipramine (IMI) and desipramine (DES)] were used as model analytes and were extracted from samples by means of liquid-phase microextraction and detected by gas chromatography-mass spectrometry. Samples of human and bovine VH and SS were prepared in six different concentrations of antidepressants (5, 40, 80, 120, 160 and 200 ng/mL) and were analyzed. Relative matrix effect was evaluated by applying a two-tailed homoscedastic Student's t-test, comparing the results obtained with the set of data obtained with human VH and bovine VH and SS. No significant matrix effect was found for AMI and NTR in the three evaluated matrices. However, a great variability was observed for IMI and DES for all matrices. Once compatibilities among the matrices were demonstrated, the method was fully validated for AMI and NTR in SS. The method was applied to six VH samples deriving from real cases whose femoral whole blood (FWB) was analyzed by a previously published method. An average ratio (VH/FWB) of ∼ 0.1 was found for both compounds.

Antidepressant-like effect of essential oil isolated from Toona ciliata Roem. var. yunnanensis.

Depressive order is one of the most common psychiatric diseases, and Toona ciliata Roem. var. yunnanensis has shown many bioactivities in folk medicine. This study was designed to investigate the antidepressant-like effect of essential oil isolated from T. ciliata Roem. var. yunnanensis. Gas chromatography and mass spectrometry (GC-MS) was used to analyze the compositions of essential oil. The immobility time in the forced swimming test (FST), tail suspending test (TST), and open field test (OFT) were used to evaluate the antidepressive effects of essential oil. Furthermore, chronic mild stress (CMS) rats were established, and contents of dopamine (DA), norepinephrine (NE), and 5-hydroxytryptamine (5-HT) in the brain were determined by high-performance liquid chromatography-electron capture detector (HPLC-ECD). Western blotting was performed to investigate the effects of essential oil on the expressions of brain-derived neurotrophic factor (BDNF) protein in rats' brain. The GC-MS analysis showed that the main components of essential oil were estragole (6.16 %), ß-elemene (24.91 %), ß-cubebene (14.29 %), and γ-elemene (8.05 %). The results from the FST and TST demonstrated that the immobility time could be significantly reduced by essential oil (10, 20, 40, and 80 mg/kg), without accompanying changes in ambulation when assessed in the OFT. Additionally, the contents of DA, NE, 5-HT, and BDNF in the hippocampus of CMS rats could be increased by treatment with essential oil at doses of 20, 40, and 80 mg/kg. All these results suggested that essential oil could be considered as a new candidate for curing depressive disorders.

Redistribuição postmortem de antidepressivos e seus produtos de biotransformação em tecidos biológicos humanos / Postmortem redistribution of antidepressants and their metabolites in human biological tissues

Os antidepressivos pertencem a uma importante classe de medicamentos investigados na toxicologia forense. Em casos de amostras provenientes de cadáveres, o intervalo entre o óbito e a obtenção da espécie biológica pode proporcionar a redistribuição postmortem destes fármacos. Com o objetivo de elucidar esse fenômeno, métodos analíticos foram desenvolvidos e aplicados utilizando sangue total (ST), humor vítreo (HV) e fígado. Para as amostras de ST e HV, o método de extração escolhido e validado foi a microextração em fase líquida (LPME) trifásica. Fibras ocas constituídas de polipropileno, com a extensão de 8 cm cada, foram tratadas com o solvente orgânico dodecano (fase orgânica), resultando em um membrana com permeabilidade seletiva. No lúmen destas fibras, adicionou-se ácido fórmico 0,1 mol/L (fase aceptora). Em frasco de fundo chato com 5 mL de capacidade, pipetou-se 3,5 mL de NaOH 0,1 mol/L (fase doadora) e 0,5 mL de ST ou HV. Ao término da extração, as amostras foram introduzidas no GC-MS, sem a necessidade de reações de derivatização. O estudo com ST contemplou os antidepressivos amitriptilina (AMI), nortriptilina (NTR), imipramina (IMI), desipramine (DES), clomipramina (CLO), desmetilclomipramina (DMC), fluoxetina (FLU) e norfluoxetina (NFL). Os limites de quantificação para estas substâncias ficaram inferiores aos níveis terapêuticos (20 ng/mL). As médias dos coeficientes de variação intradia e interdia foram, respectivamente, de 9,7 e 9,8%. As curvas de calibração apresentaram linearidade entre as concentrações de 20 até 1200 ng/mL. A validação do parâmetro integridade da diluição assegurou a mensuração de quantidades superiores ao limite apresentado na curva de calibração. O método foi aplicado em sete amostras reais postmortem e em apenas um caso foi observada uma diferença significativa (300%) entre os valores quantificados no ST periférico e central. Os antidepressivos tricíclicos AMI, NTR, IMI e DES foram avaliados no HV e o efeito matriz foi detectado para os dois últimos analitos. O método foi otimizado e validado utilizando solução salina adicionada de AMI e NTR. O limite de detecção igual a 5 ng/mL, foi obtido com a redução da voltagem da fonte de íons do espectrômetro de massa para 50 eV. Coeficientes de variação foram inferiores a 15%. Os procedimentos validados foram aplicados em seis amostras reais de HV. A relação encontrada entre os valores obtidos no ST periférico e HV foi de aproximadamente 0,1. A extração acelerada por solvente (ASE) e, posteriormente, a extração em fase sólida (SPE) foram as técnicas de separação dos analitos da matriz fígado. Ao término das citadas extrações, os antidepressivos foram analisados no GC-MS. Para esta matriz sólida, são necessários mais estudos, pois os valores encontrados nos ensaios analíticos estão em desacordo com as diretrizes utilizadas na validação dos métodos

Antidepressants belong to an important class of drugs investigated in forensic toxicology. In cases of samples from corpses, the interval between death and obtaining the biological specimens can provide the postmortem redistribution of these drugs. Aiming to elucidate this phenomenon, analytical methods were developed and applied using whole blood (WB), vitreous humor (VH) and liver. For samples of WB and HV, the extraction method chosen and validated was the three-phase liquid phase microextraction (LPME). Hollow fibers consist of polypropylene, with a length of 8 cm each were treated with dodecane organic solvent (organic phase) resulting in a membrane with selective permeability. Into the lumen of these fibers was added formic acid 0.1 mol/ L (acceptor phase). In the vial containing 3.5 mL of NaOH 0.1 mol / L (donor phase) was spiked 0.5 ml of biological fluids (WB or VH). Subsequently, the samples were injected in GC-MS without derivatization reactions. The study of the ST included antidepressants amitriptyline (AMI), nortriptyline (NTR), imipramine (IMI), desipramine (DES), clomipramine (CLO), desmethylclomipramine (DMC), fluoxetine (FLU) and norfluoxetine (NFL). The quantification limits for these substances were below the therapeutic levels (20 ng / ml). The mean coefficients of variation and separate intradays were respectively 9.7 and 9.8%. The calibration curves showed linearity between concentrations of 20 to 1200 ng / mL. The validation of the integrity of the dilution parameter assured measurement higher than the limit shown in the calibration curve quantities. The method was applied to seven real postmortem samples and in one case a significant difference (300%) between the measured values in the peripheral and central ST was observed. The tricyclic antidepressants AMI, NTR, IMI and DES were evaluated in VH and the matrix effect was detected in the last two analytes. The method was optimized and validated using saline spiked AMI and NTR. The limit of detection (5 ng/ml) was obtained by reducing the voltage of the ion source of the mass spectrometer 50 eV. Coefficients of variation were below 15%. The procedures were validated in six real samples of HV. The relationship found between the values obtained in the peripheral ST and HV was approximately 0.1. Accelerated solvent extraction (ASE) and subsequently the solid phase extraction (SPE) were the techniques of separation of analytes liver matrix. At the end of the cited extractions, antidepressants were analyzed in GC-MS. To this solid tissue, further studies are needed, because the values found in the analytical tests were not in accordance with the guidelines used in the validation of the methods

Application of response surface methodology to optimise supercritical carbon dioxide extraction of volatile compounds from Crocus sativus.

BACKGROUND: Crocus sativus has been used as a traditional Chinese medicine for a long time. The volatile compounds of C. sativus appear biologically active and may act as antioxidants as well as anticonvulsants, antidepressants and antitumour agents. In order to obtain the highest possible yield of essential oils from C. sativus, response surface methodology was employed to optimise the conditions of supercritical fluid carbon dioxide extraction of the volatile compounds from C. sativus. Four factorswere investigated: temperature, pressure, extraction time and carbon dioxide flow rate. Furthermore, the chemical compositions of the volatile compounds extracted by supercritical fluid extraction were compared with those obtained by hydro-distillation and Soxhlet extraction. RESULTS: The optimum extraction conditions were found to be: optimised temperature 44.9°C, pressure 34.9 MPa, extraction time 150.2 min and CO2 flow rate 10.1 L h⁻¹. Under these conditions, the mean extraction yield was 10.94 g kg⁻¹. The volatile compounds extracted by supercritical fluid extraction and Soxhlet extraction contained a large amount of unsaturated fatty acids. CONCLUSION: Response surface methodology was successfully applied for supercritical fluid CO2 extraction optimisation of the volatile compounds from C. sativus. The study showed that pressure and CO2 flow rate had significant effect on volatile compounds yield produced by supercritical fluid extraction. This study is beneficial for the further research operating on a large scale.

Nickel (II) incorporated AlPO-5 modified carbon paste electrode for determination of thioridazine in human serum.

In this approach, synthesis of nickel (II) incorporated aluminophosphate (NiAlPO-5) was performed by using hydrothermal method. The diffuse reflectance spectroscopy (DRS), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) techniques were applied in order to characterize synthesized compounds. The NiAlPO-5 was used as a modifier in carbon paste electrode for the selective determination of thioridazine which is an antidepressant drug. This research is the first example of an aluminophosphate being employed in electroanalysis. The effective catalytic role of the modified electrode toward thioridazine oxidation can be attributed to the electrocatalytic activity of nickel (II) in the aluminaphosphate matrix. In addition, NiAlPO-5 has unique properties such as the high specific surface area which increases the electron transfer of thioridazine. The effects of varying the percentage of modifier, pH and potential sweep rate on the electrode response were investigated. Differential pulse voltammetry was used for quantitative determination as a sensitive method. A dynamic linear range was obtained in the range of 1.0×10(-7)-1.0×10(-5)mol L(-1). The determination of thioridazine in real samples such as commercial tablets and human serum was demonstrated.

Simultaneous quantitation of venlafaxine and its main metabolite, O-desmethylvenlafaxine, in human saliva by HPLC.

Venlafaxine is used for the treatment of major depression and generalized anxiety disorders. Because its active metabolite, O-desmethylvenlafaxine, has also a similar activity, the purpose of this work was to develop a simple method for simultaneous quantitation of both drugs using HPLC with UV detection. The saliva was chosen as diagnostic material because of its easy accessibility and possibility of sampling by patients, for example, at home. The sample pretreatment by liquid-liquid extraction allows to separate both compounds from this diagnostic material with a high recovery, varying between 92.65 and 104.78%. The major advantage of the validated method lies in its sensitivity, reproducibility, and specificity for routine quantitation of the venlafaxine and O-desmethylvenlafaxine in the human saliva. The low detection and quantification values (2.8-3.1 and 9.4-10.2 ng/mL, respectively) enable to quantify both species excreted with saliva at the nanogram level. The applicability of the method was verified by analysis of the saliva obtained from depressed women treated with venlafaxine. The results suggest that the method could be used for therapeutic drug monitoring in patients undergoing treatment with venlafaxine, especially when metabolic anomalies or low compliance are suspected, or in the case of polypharmacy.

Antidepressant activity of Litsea glaucescens essential oil: identification of ß-pinene and linalool as active principles.

ETHNOPHARMACOLOGICAL RELEVANCE: Litsea glaucescens (Lauraceae) has been used in Mexican Traditional Medicine to relieve illness related to central nervous system, such as epilepsy, fright and sadness. In this study, L. glaucescens essential oil properties on central nervous system were evaluated in mice using different behavioral tests. MATERIALS AND METHODS: The essential oil was obtained by hydrodistillation and analyzed by GC/MS. Identification of major compounds was also carried out by comparison with authentic samples. The psychopharmacological profile of L. glaucescens essential oil, and some its major compounds, were evaluated in mice using several experimental models: forced swimming test (FST: Antidepressant-like activity), open field test (OFT: Spontaneous locomotor activity), elevated plus-maze (EPM: Anxiolytic-like activity), exploratory cylinder (ECT: Sedative-like activity), rotarod (motor coordination) and traction performance (myo-relaxant effect) the essential oil and active principles was administered intraperitoneally. RESULTS: The essential oil showed antidepressant-like activity at doses of 100 and 300 mg/Kg. The monoterpenes ß-pinene and linalool were identified as the two main active principles of the essential oil, and showed antidepressant-like and sedative-like activity. Eucalyptol, limonene and α-pinene they did not show antidepressant-like activity, and were not further tested. CONCLUSIONS: L. glaucescens essential oil showed antidepressant activity, ß-pinene and linalool were identified as its active principles. These results support the use of L. glaucescens in Mexican Traditional Medicine for the treatment of sadness.

Selective uptake and biological consequences of environmentally relevant antidepressant pharmaceutical exposures on male fathead minnows.

Antidepressant pharmaceuticals have been reported in wastewater effluent at the nanogram to low microgram-per-liter range, and include bupropion (BUP), fluoxetine (FLX), sertraline (SER), and venlafaxine (VEN). To assess the effects of antidepressants on reproductive anatomy, physiology, and behavior, adult male fathead minnows (Pimephales promelas) were exposed for 21 days either to a single concentration of the antidepressants FLX, SER, VEN, or BUP, or to an antidepressant mixture. The data demonstrated that exposure to VEN (305 ng/L and 1104 ng/L) and SER (5.2 ng/L) resulted in mortality. Anatomical alterations were noted within the testes of fish exposed to SER and FLX, both modulators of the neurotransmitter serotonin. Additionally, FLX at 28 ng/L induced vitellogenin in male fish--a common endpoint for estrogenic endocrine disruption. Significant alterations in male secondary sex characteristics were noted with single exposures. Effects of single compound exposures neither carried over, nor became additive in the antidepressant mixtures, and reproductive behavior was not affected. Analysis of brain tissues from the exposed fish suggested increased uptake of FLX, SER and BUP and minimal uptake of VEN when compared to exposure water concentrations. Furthermore, the only metabolite detected consistently in the brain tissues was norfluoxetine. Similar trends of uptake by brain tissue were observed when fish were exposed to antidepressant mixtures. The present study demonstrates that anatomy and physiology, but not reproductive behavior, can be disrupted by exposure to environmental concentrations of some antidepressants. The observation that antidepressant uptake into fish tissues is selective may have consequences on assessing the mode-of-action and effects of these compounds in future studies.

A fatal case involving extremely high levels of ethylene glycol without elevation of its metabolites or crystalluria.

A unique case of an intentional overdose of ethylene glycol resulting in a fatality is described. The decedent had a very high concentration of ethylene glycol without elevated concentrations of its metabolites or crystalluria. The ethylene glycol concentrations in blood, urine, and vitreous fluid were 2340, 2261, and 1028 mg/dL, respectively. Osmolality of blood and vitreous fluid was also very high at 1426 and 534 mOsm/kg, respectively. No crystals were found in the urine. Furthermore, on the urine organic acids profile the ethylene glycol metabolites oxalic, glycolic, and glyoxylic acids were within the reference ranges. In addition to ethylene glycol, the decedent had an elevated level of mirtazapine, an antidepressant, and a low level of bupropion. It was estimated that the subject consumed 1034 g of ethylene glycol. To our knowledge, this is the first case of death from severe ethylene glycol poisoning in the absence of ethylene glycol metabolites or crystalluria.

Occurrence and removal of pharmaceuticals in a municipal sewage treatment system in the south of Sweden.

The occurrence and removal rate of seven pharmaceuticals (ibuprofen, naproxen, diclofenac, fluoxetine, ofloxacin, norfloxacin, ciprofloxacin), two metabolites (norfluoxetine, clofibric acid), one degradation product (4-isobutylacetophenone) and 3 estrogens (17alpha-ethinylestradiol, 17beta-estradiol, estrone) were studied in the inlet and outlet of a tertiary sewage treatment plant (STP) in Sweden as well as between different treatment steps in the STP which includes a conventional activated sludge step. Pharmaceuticals in raw household and raw hospital sewage streams leading to the STP were as well investigated. Hydraulic retention times (HRT) of each treatment step was considered for sampling and for the calculation of the removal rates. These rates were above 90%, except for diclofenac, clofibric acid, estrone and ofloxacin. However, only diclofenac and naproxen showed significant effluent loads (>145 mg/d/1000 inh). Diclofenac was not eliminated during the treatment and in fact even higher concentrations were found at the effluent than in the inlet of the STP. 17alpha-Ethinylestradiol was not detected in any of the samples. Results indicate that a STP such as the one in Kristianstad, Sweden, with a tertiary treatment is sufficient to remove significantly most of the investigated pharmaceuticals. The chemical treatment improved the removal of several pharmaceuticals especially the antibiotics, which showed step removal rates between 55 and 70%. The expected concentration levels of the pharmaceuticals in the surface water (dilution 1 to 10) close to the outlet of the STP are below the no-observed effect-concentration (NOEC). However, despite that this would imply no important effects in the aquatic environment one cannot rule out negative consequences nearby the STP because most of the NOEC values are derived from acute toxicity data. This may underestimate the real impact of pharmaceuticals in the aquatic ecosystem.

Determination of antidepressants in human postmortem blood, brain tissue, and hair using gas chromatography-mass spectrometry.

A gas chromatographic-mass spectrometric (GC-MS) method in positive ion chemical ionization mode in combination with a solid phase extraction was optimized for new-generation antidepressants and their metabolites in postmortem blood, brain tissue, and hair. Twelve antidepressants and their active metabolites (i.e., mirtazapine, viloxazine, venlafaxine, citalopram, mianserin, reboxetine, fluoxetine, fluvoxamine, sertraline, maprotiline, melitracen, paroxetine, desmethylfluoxetine, desmethylmianserin, desmethylmirtazapine, desmethylsertraline, desmethylmaprotiline, desmethylcitalopram, and didesmethylcitalopram) could be quantified. In this article, in addition to the validation of the GC-MS method, four postmortem cases are discussed to demonstrate the usefulness of the described method in forensic toxicology. In these cases, sertraline, fluoxetine, citalopram, and trazodone in combination with their active metabolites were quantified. Blood concentrations ranged from subtherapeutic to toxic concentrations, while brain to plasma ratios ranged from 0.8 to 17. Hair concentrations ranged from 0.4 to 2.5 ng/mg depending on the compound and hair segment.

Recent studies of the electrospray ionisation behaviour of selected drugs and their application in capillary electrophoresis-mass spectrometry and liquid chromatography-mass spectrometry.

This review is concerned with recent studies of electrospray ionisation-mass spectrometry (ESI-MS) of selected small molecular mass drugs and their application in qualitative and quantitative analytical methods using the techniques liquid chromatography mass spectrometry (LC-ESI-MS) and capillary electrophoresis mass spectrometry (CE-ESI-MS). The publications reviewed are taken from the Web of Knowledge database for the year 2006. The drugs have molecular mass less than 1000 Da and are chosen according to selected drug classifications in which they give ESI signals primarily as [M+H]+ ions. The drug classifications are antibiotics/antibacterials, steroids, anti-tumour drugs, erectile dysfunction agents, anti-epileptic drugs, antiasthmatic drugs, psychoactive drugs and miscellaneous drugs. Details are given on the fragmentations, where available, that these ionic species exhibit in-source and in ion trap, triple quadrupole and time-of-flight mass spectrometers. Analytical methods for the detection and determination of these small molecular mass drug molecules are also discussed, where appropriate, under the particular drug classifications. Analytical information on, for example, sample concentration techniques, separation conditions, recoveries from biological media and limits of detection/quantitation (LODs and LOQs) are provided.