A hydrophilic interaction liquid chromatography - Tandem mass spectrometry method for the determination of phenylephrine in dried blood spots from preterm infants.

A sensitive and accurate hydrophilic interaction liquid chromatography - tandem mass spectrometry method (HILIC-MS/MS) was developed and validated for the determination of phenylephrine concentration in Dried Blood Spot (DBS) samples from preterm infants, after ocular administration of an ophthalmic solution with phenylephrine. Sample preparation involved the extraction of the analyte from an 85 µL DBS sample with methanol - acetonitrile (50:50, v/v). Chromatographic separation was achieved on an ACQUITY UPLC BEH AMIDE column, under isocratic conditions within a 5 min run. Detection was achieved with a triple quadrupole MS applying electrospray ionization in positive mode. The method was fully validated and proved precise and accurate with in a linear range of 0.59-3.53 ng/ml in blood. The method was developed to provide insights on the level of exposure of infant population to phenylephrine after ocular administration.

Involuntary psychiatric hospitalization of children and adolescents in Northern Greece: Retrospective epidemiological study and related ethical issues.

The aim of the present study was to investigate epidemiological data on involuntary hospitalization of underage patients in psychiatric settings and illustrate the related ethical issues. The medical records of 131 involuntary psychiatric admissions of children and adolescents ordered by public prosecutor between 2005 and 2014 were examined carefully. The examined variables involved the place of origin, the place of residence of minors after discharge, the length of stay in hospitals, the discharge diagnosis, the rate at which the minors were introduced to police and other authorities before their hospitalization, and the results of the neuropsychological assessment (WISC II). Data were analyzed by SPSS (Statistical Package for the Social Sciences). The mean age of the minors was 14.19 years (Male: Female ratio; 1.6:1). First, a high rate of incidences of compulsory admissions was found [5-year period (2005-2009):(2010-2014) ratio; 1:1.85] most likely due to organizational factors, which, however, could have been avoided in a more patient-oriented healthcare system. It is most likely that the criteria used for making decisions in favor of compulsory admissions were disproportionately (unduly) broad. In parallel, it was observed that, during 2010-2014, despite the increase in the rate of the prosecutor's orders, there was a decrease in the duration of coercive hospitalization of minors in psychiatric departments of hospitals in comparison to the period 2005-2009 [5-year period duration of hospitalization (2005-2009):(2010-2014) ratio; 2.33:1]. Furthermore, family was found likely to wield considerable influence on the decision-making for compulsory admissions. In addition, the effectiveness of a compulsory hospitalization of minors in a child and adolescent psychiatry department was found largely dependent on the type of the underlying mental health problem. In that respect, low rates of recidivism (7.6%) indicated that the measure of involuntary hospitalization was necessary and effective. It was also observed that the short-term removal of the minor from the family environment was a potentially relieving strategy for both the child and the family apart from the need for therapeutic intervention. The paper concludes by highlighting the role of a multi-stakeholder decision-making process (which entails shared decision-making as an integral component of providing mental healthcare to minors) in facilitating a decision about involuntary psychiatric hospitalization that is proportional and respectful to patient autonomy.

Conium maculatum intoxication: Literature review and case report on hemlock poisoning.

The aim of this paper is to highlight the symptomatology in three Conium maculatum intoxication incidents, one of which was fatal. A number of studies were reviewed in order to update and summarize the relevant literature on the incidence, sociodemographic variables, method of poisoning, pathophysiology, diagnosis, variables associated with survival and fatality, management, and treatment of C. maculatum intoxication as well as the biosynthesis and biological effects of poison hemlock alkaloids. Results show that hemlock poisoning is relatively rare, although incidence varies in different regions, despite its worldwide distribution. Hemlock poisoning is more common in European and especially Mediterranean countries. The majority of the patients are adult males over 38 years of age. The clinical course of hemlock poisoning includes neurotoxicosis, tremor, vomiting, muscle paralysis, respiratory paralysis/failure, rhabdomyolysis, and acute renal failure. The therapeutic management focuses on absorption reduction, close observation for complications, and supportive therapy (especially for respiration). Acute occurrence is severe and life-threatening, but the survival rate is high if treatment is provided promptly. Recovery is rapid, generally taking only a few days.

Quantification of 15 Psychotropic Drugs in Serum and Postmortem Blood Samples after a Modified Mini-QuEChERS by UHPLC-MS-MS.

The aim of the study was to develop a LC-MS-MS method able to detect and quantify a number of frequently prescribed antipsychotic and antidepressant drugs for toxicological purposes. Separation of compounds was performed on a C-18 RP column by Ultra High-Pressure Chromatography over a 11 min run. A modified single step QuEChERS protocol consisted essentially by the addition of acetonitrile, potassium carbonate and magnesium sulfate in 100 µL of sample, vortexing, centrifugation and evaporation has been selected. The method achieves satisfactory recoveries for 15 psychotropic drugs with a mean R% of 85% and provides efficient purification of the sample from endogenous interferences, simplicity and short sample handling times. The method was validated and provided satisfactory accuracy with recoveries ranging from 85 to 113% and precision with CV ranging from 1.2 to 13.2%. LODs were determined to be from 0.0003 to 0.017 µg/mL while LOQs were from 0.001 to 0.05 µg/mL for the 15 drugs. Matrix effect was below 20% and the analytes were stable in the matrix for 3 weeks. The method proved to be suitable for both analysis of clinical samples for Therapeutic Drug Monitoring and antemortem or postmortem whole blood samples of forensic cases. A number of samples with clinical and forensic interest were successfully analyzed demonstrating the effectiveness of QuEChERS in this field.

An ultra-high pressure liquid chromatography-tandem mass spectrometry method for the quantification of teicoplanin in plasma of neonates.

The development and validation of an ultra-high pressure liquid chromatography (UHPLC) tandem mass spectrometry (MS/MS) method was performed with the aim to be applied for the quantification of plasma teicoplanin concentrations in neonates. Pharmacokinetic data of teicoplanin in the neonatal population is very limited, therefore, a sensitive and reliable method for the determination of all isoforms of teicoplanin applied in a low volume of sample is of real importance. Teicoplanin main components were extracted by a simple acetonitrile precipitation step and analysed on a C18 chromatographic column by a triple quadrupole MS with electrospray ionization. The method provides quantitative data over a linear range of 25-6400ng/mL with LOD 8.5ng/mL and LOQ 25ng/mL for total teicoplanin. The method was applied in plasma samples from neonates to support pharmacokinetic data and proved to be a reliable and fast method for the quantification of teicoplanin concentration levels in plasma of infants during therapy in Intensive Care Unit.

Unusual mercury poisoning from tattoo dye.

BACKGROUND: Tattoos have become very popular in modern societies due to the gradual change in aesthetics and psychosocial acceptability. Consequently, tattoo reactions are seen more commonly than in the past. Cutaneous lesions associated with tattoos can be divided into three major groups: allergic/granulomatous/lichenoid, infectious, and coincidental lesions. Early identification and proper treatment of these skin lesions is challenging and necessitates close coοperation of different medical specialties. CASE DESCRIPTION: We report an unusual case of mercury poisoning in a young person manifested with local skin reactions following amateur tattooing. The tattoo induced inflammatory foreign body reactions and required multiple surgical excisions to be removed. The unique feature of this case is the use of the elemental form of mercury in the tattoo dye and the resulting mercury poisoning. The poisoning was confirmed by detection of mercury in blood, urine, and hair samples. CONCLUSION: This is a rare case of tattoo-associated skin reaction and mercury poisoning by the elemental form of mercury contained in the tattoo dye. In the literature, many conditions have been documented in association with tattoos and the process of tattoo application, especially when red dyes are used, but no similar cases of elemental mercury poisoning from the tattoo dye exist. HIPPOKRATIA 2017, 21(4): 197-200.

Metabolic profile of human coelomic fluid.

AIM: Till now there is very limited knowledge on the molecular content of coelomic fluid and cells. This study presents the first attempt to elucidate the metabolic profile of such samples. METHODOLOGY: Samples were collected via coelocentesis from 41 women during the first trimester of gestation. Metabolic content was assessed using four different analytical platforms. For targeted analysis a hydrophilic interaction chromatography ultra high performance LC-MS/MS method was applied. Holistic analysis performed by GC-MS, NMR spectroscopy and ion cyclotron ultra-high resolution MS (FT-ICR-MS) instrumentation. RESULTS & CONCLUSIONS: Our observations suggest coelomic fluid and cells as promising biosamples, rich in metabolites with potential use in mammalian system biology studies.

A GC-MS method for the detection and quantitation of ten major drugs of abuse in human hair samples.

A sensitive analytical method has been developed in order to identify and quantify major drugs of abuse (DOA), namely morphine, codeine, 6-monoacetylmorphine, cocaine, ecgonine methyl ester, benzoylecgonine, amphetamine, methamphetamine, methylenedioxymethamphetamine and methylenedioxyamphetamine in human hair. Samples of hair were extracted with methanol under ultrasonication at 50°C after a three step rinsing process to remove external contamination and dirt hair. Derivatization with BSTFA was selected in order to increase detection sensitivity of GC/MS analysis. Optimization of derivatization parameters was based on experiments for the selection of derivatization time, temperature and volume of derivatising agent. Validation of the method included evaluation of linearity which ranged from 2 to 350ng/mg of hair mean concentration for all DOA, evaluation of sensitivity, accuracy, precision and repeatability. Limits of detection ranged from 0.05 to 0.46ng/mg of hair. The developed method was applied for the analysis of hair samples obtained from three human subjects and were found positive in cocaine, and opiates.

Ethico-legal issues related to ovarian tissue transplantation.

Ovarian tissue transplantation (OTT) is a promising experimental method which may soon become well-established. In cases of minor oncology, where patients' fertility is seriously threatened by treatment, it may be applied as a unique fertility preservation option. OTT has a dual nature ('organ' and 'gamete'). Many stakeholders are involved, including donor, recipient, child, health-care providers and society at large. There is considerable uncertainty about the long-term consequences of the application of OTT and OT cryopreservation (OTC). Thus, application of OTT gives rise to a number of very different ethico-legal issues and dilemmas which are hard to solve coherently through a principlism-based bioethical approach. This study focuses on such dilemmas and attempts to review them. The role of virtue ethics, which may be combined with principlism, is essential to solve such dilemmas coherently and reasonably. Dealing with conflicts of ethical principles equivalent between them, or moral dilemmas without available answers and mind-sharing in a difficult interpersonal process of decision making, requires a virtue-based ethical approach. Besides, ethico-legal issues related to OTC/OTT are complex issues requiring a multidisciplinary approach (ethical considerations, medical, psychological and social evaluations etc.). We stress the crucial role of multidisciplinary Ethics Committee which is considered indispensable for each reproductive health-care unit practicing OTC/OTT.

Determination of venlafaxine in post-mortem whole blood by HS-SPME and GC-NPD.

Venlafaxine is a phenethylamine derivative widely prescribed for the treatment of depression which inhibits both serotonin and norepinephrine reuptake (SNRI). In treatment with antidepressants of patient with depression and other psychiatric disorders there is also increased risk of suicidal thought and behaviour. Several lethal intoxications involving venlafaxine usually among psychotic patients have been reported in the literature. Sample preparation is of the greatest significance for a successful toxicological analysis. The development of simple, effective and rapid extraction procedures of drugs from post-mortem biological samples is a challenge. Headspace-solid phase microextraction (HS-SPME) offers significant advantages such as simplicity, low cost, compatibility with analytical systems, automation and solvent-free extraction. The aim of our work was the optimization of a HS-SPME procedure for the determination of venlafaxine in post-mortem biological samples by gas chromatography (GC) with nitrogen-phosphorous detection (NPD). Venlafaxine was extracted on 100 µm Polydimethylsiloxone Coating-Red (PDMS) SPME fiber and determined by GC-NPD. Salt addition, extraction temperature, preheating and extraction time were optimized to enhance the recovery of the extraction from aqueous solution spiked with venlafaxine. Finally the developed procedure was applied to post-mortem biological samples of a fatally poisoned woman by venlafaxine. The drug was quantified in post-mortem blood gastric and oesophagus contents of the deceased woman. A simple and rapid procedure using HS-SPME was developed for sample preparation of venlafaxine in post-mortem biological samples prior to GC-NPD determination. Validation data was satisfactory, thus enabling application in the toxicological analysis of forensic samples.

Carbamate poisoning in a dairy goat herd: clinicopathological findings and therapeutic approach.

CASE HISTORY: Approximately 1 hour after the consumption of carnations from a nearby glasshouse 55 animals from a dairy goat herd exhibited signs of possible poisoning. CLINICAL FINDINGS: Upon clinical examination affected animals exhibited signs of salivation, tympany, tachypnoea, polydipsia, urination, diarrhoea, bradycardia, miosis, tremor and convulsions. As poisoning from an acetylcholinesterase-inhibiting insecticide was suspected, treatment with atropine sulphate was initiated at a dose of 0.3 mg/kg bodyweight. The treatment was repeated for some animals that relapsed, and was effective in all cases, with the exception of one goat kid that died. DIAGNOSIS: Necropsy of the goat kid showed pulmonary oedema and congestion of internal organs. Toxicological analysis of stomach contents and liver of the dead animal, as well as of the carnations, revealed high concentrations of carbamates. CLINICAL RELEVANCE: Carbamate poisoning after consumption of polluted feedstuffs or glasshouse products can be fatal for dairy goats. Atropine sulphate, at 0.3 mg/kg, can be useful in treating this condition.

Solid phase microextraction applied to the analysis of organophosphorus insecticides in fruits.

Trace amounts of organophosphorus pesticides (OPs) were determined in various fruits by headspace solid phase microextraction (HS-SPME) and gas chromatography-nitrogen phosphorous detection (GC-NPD). Sampling from the headspace enhanced method selectivity, whereas at the same time improved fiber life time and method sensitivity. Diazinon, parathion, methyl parathion, malathion and fenithrothion were determined in various fruits: more than 150 samples of 21 types of fruits were studied. SPME-GC-NPD provided a useful and very efficient analytical tool: method linearity ranged from 1.2 to 700 ng/ml. Limits of detection (LODs) and quantitation (LOQs) ranged from 0.03 to 3 ng/ml and 0.12 to 10 ng/ml respectively, values well below the residue limits set by the EU. Less than 2% of the samples were found positive containing amounts higher than the EU limits. The effect of fruit peeling and washing was also investigated.

Headspace solid phase microextraction for the gas chromatographic analysis of methyl-parathion in post-mortem human samples. Application in a suicide case by intravenous injection.

A simple and rapid procedure for the determination of methyl-parathion (m-p) in post-mortem biological samples was developed using headspace solid phase microextraction (SPME) and gas chromatography (GC) with nitrogen-phosphorous detection (NPD). Methyl-parathion was extracted on 85 microm polyacrylate SPME fiber. Salt addition, extraction temperature, and extraction time were optimized to enhance the sensitivity of the method. The linearity (y = 0.0473x - 0.0113, R2 = 0.9992) and the dynamic range (0.1-40 microg/ml) were found very satisfactory. The recoveries of methyl-parathion were found to be 46% in spiked human whole blood, 53% in spiked homogenized liver tissue, and 54% in spiked homogenized kidney tissue compared with samples prepared in water. The coefficients of variations for 2, 4, and 20 microg/ml of methyl-parathion in blood ranged from 0.9 to 5.1%, whereas the detection limit of the method was satisfactory (1 ng/ml in aqueous samples, 50 ng/ml in whole blood). The developed procedure was applied to post-mortem biological samples from a 21-year-old woman fatally poisoned (suicide) by intravenous injection of methyl-parathion. The intact insecticide was found in the post-mortem blood at a concentration of 24 microg/ml. No methyl-parathion was detected in the liver, kidneys, and gastric contents.

Amphetamine derivative related deaths in northern Greece.

Until 1997, only one amphetamine related derivatives (AMPs) fatality had been reported in Greece. Since then, amphetamine (AMP) or AMPs have been found in seven out of 1,500 post-mortem toxicological cases. The cause and manner of death of these seven cases were: 3,4-methylenedioxy-N-methamphetamine (MDMA) and 3,4-methylenedioxy-N-ethylamphetamine (MDEA) poisoning (n = 1), drowning in water (n = 4), cranial injuries caused by a traffic accident (n = 1) and heart failure (n = 1). In the case where the use of AMP or AMPs was considered, the immediate cause of death post-mortem toxicological analysis revealed 2 microg/ml MDMA and 0.7 microg/ml MDEA in blood. MDMA was identified in two cases of drowning (2 microg/ml in blood in the first case and 1.7 microg/g in liver in the second case) and in the traffic accident case (0.4 microg/g in liver). Methamphetamine was detected in two cases of drowning (2.5 microg/ml in blood in the first case and 6 microg/g in liver in the second case). AMP was found in the heart failure case (0.2 microg/g in liver). Alcohol was present, together with AMP or AMPs, in four cases. These findings indicate an increase in the illegal abuse of AMPs in Greece. Because of this, we now routinely screen for AMPs.

Determination of opiates in postmortem bone and bone marrow.

Bone and bone marrow of a fatally poisoned heroin addict were analyzed by FPIA and GC-FID, immediately after death. A piece of the bone from the above case was buried for 1 year and analyzed by the same procedure. Morphine was detected in all specimens at concentrations of 195, 340 and 155 ng/g for bone marrow, bone and buried bone, respectively. A loss of 54.4% of morphine concentration was observed during 1-year burial. Such findings have potential forensic value in cases of skeletonized remains.

Modelling of phosphorus removal from aqueous and wastewater samples using ferric iron.

Batch laboratory scale experiments were conducted to investigate the removal of phosphate from aqueous and municipal wastewater samples by addition of FeCl(3).6H(2)O. The effect of pH, Fe-dose and initial phosphate concentration were assessed. Optimum phosphate removal, 63% for 1:1 molar addition of Fe(III) was observed at pH 4.5. However, a 155% excess of Fe-dose was necessary for complete phosphorus removal. Phosphorus removal from municipal wastewater was slightly higher than that observed for the aqueous solutions. A chemical precipitation mathematical model was developed and tested with the available experimental data. The model included a total of 15 chemical reactions and 4 solid phases with the option of single-phase precipitation or two-phase co-precipitation. The resulting system of non-linear algebraic equations was solved numerically, using the Wijngaarden-Dekker-Brent method.