La dimensión espiritual en el anciano desde el modelo de la gerotrascendencia y su abordaje desde la terapia ocupacional: revisión bibliográfica / A dimensão espiritual nos idosos a partir do modelo de gerotranscendência e sua abordagem a partir da terapia ocupacional: uma revisão da literatura / The spiritual dimension in the elderly from the model of gerotranscendence and its approach from Occupational Therapy: a literature review

Resumen Introducción El campo de estudio del envejecimiento conoce un marco teórico conocido como gerotrascendencia. La gerotrascendencia entiende la vejez como la etapa final de una progresión natural que lleva a alcanzar la madurez y la sabiduría. La ancianidad es, así, una etapa de introspección entendida como espiritualidad que se explica también en un contexto religioso. Objetivo Comprobar desde la literatura científica la implementación de la gerotrascendencia y sus aspectos fundamentales en el marco de la terapia ocupacional. Metodología Estudio bibliográfico ejecutado por un estudio previo de Google Trends para conocer el tráfico de datos. Su usan bases de datos especializadas en ciencias de la salud (Pubmed, Medline, EMBASE, CINAHL complete, LILACS, Medes, CUIDENplus, Cuidatge, Cochrane Library, Trip medical database, Epistemonikos e IBECS), en el área de psicología (Psicodoc, PsycArticles, PsycBooks, PsycInfo y Psychology and Behavioral Sciences Collection), y multidisciplinares (Google Scholar, Web of Science y Scopus) a partir de una exploración booleana/frase (operadores: AND, OR, NOT). Se utilizan los tesauros especializados (MeSH, DeCS y HONselect) usando palabras clave en español e inglés: gerotrascendencia, enfermería, terapia ocupacional. El uso de bases de datos y un sistema de cribado basado en objetivos da como resultado el estudio especifico de seis artículos. Resultados La gerotrascendencia conoce una bibliografía consolidada en inglés, pero que es casi inexistente en español. De entre los estudios en español existen estudios sólidos con resultados que plantean una alternativa a los modelos materialistas que plantean un enfoque que se reducen al paradigma biomédico. Estos estudios plantean una ética de las necesidades y un enfoque centrado en la atención al usuario, donde la espiritualidad (religión interna) y su vivencia religiosa externa tienen una gran importancia. Conclusión La terapia ocupacional no puede olvidar los modelos teóricos centrados en la persona, de modo que puedan salir del corsé biomédico, al que pertenecen, pero al que no se reducen. Parece necesario realizar más abordajes teóricos y estudios cuantitativos y cualitativos que revelen la operatividad de la implementación de modelos que entiendan la fenomenología del envejecimiento de forma holística, como es el caso de la gerotrascendencia.

Resumo Introdução O campo de estudo do envelhecimento está familiarizado com uma estrutura teórica conhecida como gerotranscendência. A gerotranscendência entende a velhice como o estágio final de uma progressão natural em direção à maturidade e à sabedoria. A velhice é, portanto, um estágio de introspecção entendido como espiritualidade que também é explicada em um contexto religioso. Objetivo Verificar na literatura científica a implementação da gerotranscendência e seus aspectos fundamentais no âmbito da terapia ocupacional. Metodologia Estudo bibliográfico realizado por um estudo prévio do Google Trends para conhecer o tráfego de dados. Utiliza bases de dados especializadas em ciências da saúde (Pubmed, Medline, EMBASE, CINAHL complete, LILACS, Medes, CUIDENplus, Cuidatge, Cochrane Library, Trip medical database, Epistemonikos e IBECS), na área de psicologia (Psicodoc, PsycArticles, PsycBooks, PsycInfo e Psychology and Behavioral Sciences Collection) e multidisciplinar (Google Scholar, Web of Science e Scopus) com base em uma exploração booleana/frase (operadores: AND, OR, NOT). São usados tesauros especializados (MeSH, DeCS e HONselect) usando palavras-chave em espanhol e inglês: gerotranscendência, enfermagem, terapia ocupacional. O uso de bancos de dados e um sistema de triagem baseado em metas resulta no estudo específico de seis artigos. Resultados A gerotranscendência tem uma bibliografia consolidada em inglês, mas é quase inexistente em espanhol. Entre os estudos em espanhol, há estudos sólidos com resultados que propõem uma alternativa aos modelos materialistas que propõem uma abordagem que se reduz ao paradigma biomédico. Esses estudos propõem uma ética das necessidades e uma abordagem centrada no usuário, em que a espiritualidade (religião interna) e a experiência religiosa externa são de grande importância. Conclusão A terapia ocupacional não pode se esquecer dos modelos teóricos centrados na pessoa, para que ela possa sair do espartilho biomédico, ao qual pertence, mas ao qual não se reduz. Parece necessário realizar mais abordagens teóricas e estudos quantitativos e qualitativos que revelem a operacionalidade da implementação de modelos que compreendam a fenomenologia do envelhecimento de forma holística, como é o caso da gerotranscendência.

Abstract Introduction The field of the study of ageing is familiar with a theoretical framework known as gerotranscendence. Gerotranscendence understands old age as the final stage of a natural progression towards maturity and wisdom. Old age is thus a stage of introspection understood as spirituality explained in a religious context. Objective To verify from the scientific literature the implementation of gerotranscendence and its fundamental aspects within the occupational therapy framework. Methodology Bibliographic study carried out by a previous study of Google Trends to know the data traffic. It uses databases specialised in health sciences (Pubmed, Medline, EMBASE, CINAHL complete, LILACS, Medes, CUIDENplus, Cuidatge, Cochrane Library, Trip medical database, Epistemonikos and IBECS) in the area of psychology (Psicodoc, PsycArticles, PsycBooks, PsycInfo and Psychology and Behavioral Sciences Collection), and multidisciplinary (Google Scholar, Web of Science and Scopus) based on a Boolean/phrase exploration (operators: AND, OR, NOT). Specialised thesauri (MeSH, DeCS and HONselect) are used using keywords in Spanish and English: gerotranscendence, nursing, and occupational therapy. The use of databases and a target-based screening system results in the specific study of six articles. Results Gerotranscendence has a consolidated bibliography in English but is almost nonexistent in Spanish. Among the studies in Spanish, there are solid studies with results that propose an alternative to the materialistic models that propose an approach that is reduced to the biomedical paradigm. These studies propose an ethics of needs and a user-centred approach, where spirituality (internal religion) and external religious experience are essential. Conclusion: Occupational therapy must remember the theoretical models centred on the person so that they can get out of the biomedical corset to which they belong but to which they are not reduced. It seems necessary to carry out more theoretical approaches and quantitative and qualitative studies that reveal the operability of implementing models that holistically understand the phenomenology of ageing, as is the case of gerotranscendence.

mtDNA variability determines spontaneous joint aging damage in a conplastic mouse model.

Mitochondria and mtDNA variations contribute to specific aspects of the aging process. Here, we aimed to investigate the influence of mtDNA variation on joint damage in a model of aging using conplastic mice. A conplastic (BL/6NZB) mouse strain was developed with the C57BL/6JOlaHsd nuclear genome and NZB/OlaHsd mtDNA, for comparison with the original C57BL/6JOlaHsd strain (BL/6C57). Conplastic (BL/6NZB) and BL/6C57 mice were sacrificed at 25, 75, and 90 weeks of age. Hind knee joints were processed for histological analysis and joint pathology graded using the Mankin scoring system. By immunohistochemistry, cartilage expression of markers of autophagy (LC3, Beclin-1, and P62) and markers of senescence (MMP13, beta-Galactosidase, and p16) and proliferation (Ki67) were analyzed. We also measured the expression of 8-oxo-dG and cleaved caspase-3. Conplastic (BL/6NZB) mice presented lower Mankin scores at 25, 75, and 90 weeks of age, higher expression of LC3 and Beclin-1 and lower of P62 in cartilage than the original strain. Moreover, the downregulation of MMP13, beta-Galactosidase, and p16 was detected in cartilage from conplastic (BL/6NZB) mice, whereas higher Ki67 levels were detected in these mice. Finally, control BL/6C57 mice showed higher cartilage expression of 8-oxo-dG and cleaved caspase-3 than conplastic (BL/6NZB) mice. This study demonstrates that mtDNA genetic manipulation ameliorates joint aging damage in a conplastic mouse model, suggesting that mtDNA variability is a prognostic factor for aging-related osteoarthritis (OA) and that modulation of mitochondrial oxidative phosphorylation (OXPHOS) could be a novel therapeutic target for treating OA associated with aging.

Determination of synthetic opioids in oral fluid samples using fabric phase sorptive extraction and gas chromatography-mass spectrometry.

New psychoactive substances (NPS) continue to emerge in the drug market every year, becoming a global threat to public health and safety. These compounds are mostly synthetic cannabinoids and designer cathinones. However, synthetic opioids have appeared on the recreational drug markets in recent years, particularly fentanyl and its derivatives ("fentanyls"). Fentanyl and its analogs are related to harmful intoxications and an increase in opioid-related mortality in many countries, such as in the United States and Europe in the last years. Taking the drug related global crisis into consideration, this work developed and validated an effective and sensitive method based on fabric phase sorptive extraction (FPSE) followed by gas chromatography-mass spectrometry (GC-MS) for the simultaneous determination of 11 fentanyl analogs in oral fluid samples. The extraction was carried out using a sol-gel Carbowax 20 M sorbent immobilized on 100% cellulose fabric substrate and using ethyl acetate as the desorption solvent. The limits of detection (LODs) and quantification (LOQs) ranged from 1 to 15 ng mL-1 and 5 to 50 ng mL-1, respectively. Intra-day and inter-day precision were found within 8.2% and 8.6%, respectively, while accuracy ranged from -5.5 to 9.1%, in accordance with the established criteria. The absolute recovery values were in the range of 94.5%-109.1%. The validated method demonstrated its great potential to detect and quantify fentanyl analogs in possible forensic work and off-site analysis in road traffic cases.

Mitochondrial DNA impact on joint damaged process in a conplastic mouse model after being surgically induced with osteoarthritis.

It has been suggested that mitochondrial dysfunction and mtDNA variations may contribute to osteoarthritis (OA) pathogenesis. However, the causative link to support this claim is lacking. Here, we surgically-induced OA in conplastic mice in order to evaluate the functional consequences of mtDNA haplotypes in their joint degeneration. BL/6NZB strain was developed with C57BL/6JOlaHsd nuclear genome and NZB/OlaHsdmtDNA while BL/6C57, which is the original, was developed with C57BL/6JOlaHsd nuclear genome and C57/OlaHsdmtDNA for comparison. The surgical DMM OA model was induced in both strains. Their knees were processed and examined for histopathological changes. Cartilage expression of markers of autophagy, apoptosis, oxidative stress and senescence were also analyzed by immunohistochemistry. The joints of BL/6NZB mice that were operated presented more cellularity together with a reduced OARSI histopathology score, subchondral bone, menisci score and synovitis compared to those of BL/6C57 mice. This was accompanied with higher autophagy and a lower apoptosis in the cartilage of BL/6NZB mice that were operated. Therefore, the study demonstrates the functional impact of non-pathological variants of mtDNA on OA process using a surgically-induced OA model. Conplastic (BL/6NZB ) mice develop less severe OA compared to the BL/6C57original strain. These findings demonstrate that mitochondria and mtDNA are critical targets for potential novel therapeutic approaches to treat osteoarthritis.

Intraarticular Administration Effect of Hydrogen Sulfide on an In Vivo Rat Model of Osteoarthritis.

Osteoarthritis (OA) is the most common articular chronic disease. However, its current treatment is limited and mostly symptomatic. Hydrogen sulfide (H2S) is an endogenous gas with recognized physiological activities. The purpose here was to evaluate the effects of the intraarticular administration of a slow-releasing H2S compound (GYY-4137) on an OA experimental model. OA was induced in Wistar rats by the transection of medial collateral ligament and the removal of the medial meniscus of the left joint. The animals were randomized into three groups: non-treated and intraarticularly injected with saline or GYY-4137. Joint destabilization induced articular thickening (≈5% increment), the loss of joint mobility and flexion (≈12-degree angle), and increased levels of pain (≈1.5 points on a scale of 0 to 3). Animals treated with GYY-4137 presented improved motor function of the joint, as well as lower pain levels (≈75% recovery). We also observed that cartilage deterioration was attenuated in the GYY-4137 group (≈30% compared with the saline group). Likewise, these animals showed a reduced presence of pro-inflammatory mediators (cyclooxygenase-2, inducible nitric oxide synthase, and metalloproteinase-13) and lower oxidative damage in the cartilage. The increment of the nuclear factor-erythroid 2-related factor 2 (Nrf-2) levels and Nrf-2-regulated gene expression (≈30%) in the GYY-4137 group seem to be underlying its chondroprotective effects. Our results suggest the beneficial impact of the intraarticular administration of H2S on experimental OA, showing a reduced cartilage destruction and oxidative damage, and supporting the use of slow H2S-producing molecules as a complementary treatment in OA.

Effect of balneotherapy in sulfurous water on an in vivo murine model of osteoarthritis.

Osteoarthritis (OA) is a chronic joint disease that results in progressive cartilage destruction and subsequently joint dysfunction. Growing evidence indicates beneficial impact of balneological interventions in OA; however, their mechanisms of action are still unclear. Here, we evaluate the effect of balneotherapy in sulfurous water in an OA experimental model. Experimental OA was induced in Wistar rats by transection of the medial collateral ligament and removal of the medial meniscus of the left knee. Animals were randomized into three groups: non-treated (control) and balneotherapy using sulfurous water (SW) or tap water (TW). Macroscopic evaluation was performed, as well as evaluation of pain levels and analysis of motor function by rotarod test. Histopathological changes in articular cartilage and synovium were also evaluated. The presence of matrix metalloproteinase-13 (MMP-13) and oxidative damage markers was assessed by immunohistochemistry. Joint destabilization induced joint thickening, loss of joint flexion, and increased levels of pain. At day 40, animals from SW group presented lower pain levels than those from control group. Experimental OA also affected motor function. Balneotherapy in sulfur-rich water significantly improved joint mobility in relation to that in tap water. Besides, we observed that cartilage deterioration was lower in SW group than in the other two groups. Likewise, SW group showed reduced levels of MMP-13 in the cartilage. Conversely, we failed to observe any modulation on synovial inflammation. Finally, balneotherapy in sulfurous water diminished the presence of oxidative damage markers. Our results suggest the beneficial effect of balneotherapy in sulfur-rich water on an experimental model of OA, showing a reduced cartilage destruction and oxidative damage. Thus, these findings support the use of balneotherapy as a non-pharmacological treatment in OA.

Microextraction by packed sorbent followed by ultra high performance liquid chromatography for the fast extraction and determination of six antidepressants in urine.

The growing use of antidepressants in recent years has led to their increasing presence in forensic analyses. In this work, microextraction by packed sorbent followed by ultra-performance liquid chromatography with photodiode array detection provided a fast method for determining the antidepressants mirtazapine, venlafaxine, escitalopram, fluoxetine, fluvoxamine, and sertraline in human urine. The microextraction conditions (viz., type of sorbent, number of draw-eject extraction cycles or strokes, sample volume and pH, and type and volume of washing solution and eluent) were optimized by using an experimental design. The ensuing analytical method was validated in terms of linearity (25-1000 ng/mL urine), limit of detection (lower than 7.1 ng/mL), limit of quantification (25 ng/mL), precision (4.7-15.1% as relative standard deviation), and accuracy (80.4-126.1% as mean recovery for four replicate determinations). The proposed method allowed the six target antidepressants to be determined at concentrations from therapeutic to toxic levels. The application to small volumes (300 µL) of urine afforded fast extraction of the analytes and provided results on a par with those of existing clinical and forensic alternatives.

Response surface methodology for the optimization of dispersive liquid-liquid microextraction of chloropropanols in human plasma.

Chloropropanols are processing toxicants with a potential risk to human health due to the increased intake of processed foods. A rapid and efficient method for the determination of three chloropropanols in human plasma was developed using ultrasound-assisted dispersive liquid-liquid microextraction. The method involved derivatization and extraction in one step followed by gas chromatography with tandem mass spectrometry analysis. Parameters affecting extraction, such as sample pH, ionic strength, type and volume of dispersive and extraction solvents were optimized by response surface methodology using a pentagonal design. The linear range of the method was 5-200 ng/mL for 1,3-dichloro-2-propanol, 10-200 ng/mL for 2,3-dichloro-2-propanol and 10-400 ng/mL for 3-chloropropane-1,2-diol with the determination coefficients between 0.9989 and 0.9997. The limits of detection were in the range of 0.3-3.2 ng/mL. The precision varied from 1.9 to 10% relative standard deviation (n = 9). The recovery of the method was between 91 and 101%. Advantages such as low consumption of organic solvents and short time of analysis make the method suitable for the biomonitoring of chloropropanols.

Articular chondrocyte network mediated by gap junctions: role in metabolic cartilage homeostasis.

OBJECTIVE: This study investigated whether chondrocytes within the cartilage matrix have the capacity to communicate through intercellular connections mediated by voltage-gated gap junction (GJ) channels. METHODS: Frozen cartilage samples were used for immunofluorescence and immunohistochemistry assays. Samples were embedded in cacodylate buffer before dehydration for scanning electron microscopy. Co-immunoprecipitation experiments and mass spectrometry (MS) were performed to identify proteins that interact with the C-terminal end of Cx43. GJ communication was studied through in situ electroporation, electrophysiology and dye injection experiments. A transwell layered culture system and MS were used to identify and quantify transferred amino acids. RESULTS: Microscopic images revealed the presence of multiple cellular projections connecting chondrocytes within the matrix. These projections were between 5 and 150 µm in length. MS data analysis indicated that the C-terminus of Cx43 interacts with several cytoskeletal proteins implicated in Cx trafficking and GJ assembly, including α-tubulin and ß-tubulin, actin, and vinculin. Electrophysiology experiments demonstrated that 12-mer oligonucleotides could be transferred between chondrocytes within 12 min after injection. Glucose was homogeneously distributed within 22 and 35 min. No transfer was detected when glucose was electroporated into A549 cells, which have no GJs. Transwell layered culture systems coupled with MS analysis revealed connexins can mediate the transfer of L-lysine and L-arginine between chondrocytes. CONCLUSIONS: This study reveals that intercellular connections between chondrocytes contain GJs that play a key role in cell-cell communication and a metabolic function by exchange of nutrients including glucose and essential amino acids. A three-dimensional cellular network mediated through GJs might mediate metabolic and physiological homeostasis to maintain cartilage tissue.

Application of dispersive liquid-liquid microextraction for the determination of phosphatidylethanol in blood by liquid chromatography tandem mass spectrometry.

Phosphatidylethanol (PEth) is a phospholipid which requires for its metabolic formation the presence of relatively high ethanol levels. PEth is thus a promising marker to quentify ethanol abuse. Dispersive liquid-liquid microextraction has become a popular technique because it is fast, inexpensive, easy to operate and consumes low volume of organic solvent. In this method, the appropriate mixture of extraction solvent (230 µL dichloromethane) and disperser solvent (630 µL acetone) are injected into the sample by syringe, rapidly. The liquid chromatography method using a reversed phase-C8 column and a negative ion mode electrospray ionization tandem mass spectrometry detection instrument was developed for the determination of small amounts of PEth that might be present in blood samples, using phosphatidylbutanol (PBut) as an internal standard. The sensitivity of detection obtained with tandem MS was better than that of previous methods. Good linearity was obtained for a range of LOQ-10 µg/mL for PEth, whereas all of the deviations in precision and accuracy were less than 15% except for the LLOQ, where it should not exceed 20%. A set of 50 blood samples were analyzed by such method and whole blood concentrations of PEth 16:0/18:1 ranged from LLOQ to 1.71 µg/mL.

Optimization of microwave-assisted extraction of analgesic and anti-inflammatory drugs from human plasma and urine using response surface experimental designs.

The performance of microwave-assisted extraction and HPLC with photodiode array detection method for determination of six analgesic and anti-inflammatory drugs from plasma and urine, is described, optimized, and validated. Several parameters affecting the extraction technique were optimized using experimental designs. A four-factor (temperature, phosphate buffer pH 4.0 volume, extraction solvent volume, and time) hybrid experimental design was used for extraction optimization in plasma, and three-factor (temperature, extraction solvent volume, and time) Doehlert design was chosen to extraction optimization in urine. The use of desirability functions revealed the optimal extraction conditions as follows: 67°C, 4 mL phosphate buffer pH 4.0, 12 mL of ethyl acetate and 9 min, for plasma and the same volume of buffer and ethyl acetate, 115°C and 4 min for urine. Limits of detection ranged from 4 to 45 ng/mL in plasma and from 8 to 85 ng/mL in urine. The reproducibility evaluated at two concentration levels was less than 6.5% for both specimens. The recoveries were from 89 to 99% for plasma and from 83 to 99% for urine. The proposed method was successfully applied in plasma and urine samples obtained from analgesic users.

A rapid ultrasound-assisted dispersive liquid-liquid microextraction followed by ultra-performance liquid chromatography for the simultaneous determination of seven benzodiazepines in human plasma samples.

A simple and efficient ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) method has been developed for the determination of seven benzodiazepines (alprazolam, bromazepam, clonazepam, diazepam, lorazepam, lormetazepam and tetrazepam) in human plasma samples. Chloroform and methanol were used as extractant and disperser solvents, respectively. The influence of several variables (e.g., type and volume of dispersant and extraction solvents, pH, ultrasonic time and ionic strength) was carefully evaluated and optimized, using an asymmetric screening design 3(2)4(2)//16. Analysis of extracts was performed by ultra-performance liquid chromatography coupled with photodiode array detection (UPLC-PDA). Under the optimum conditions, two reversed-phases, Shield RP18 and C18 columns were successfully tested, obtaining good linearity in a range of 0.01-5µgmL(-1), with correlation coefficients r>0.996. Quantification limits ranged between 4.3-13.2ngmL(-1) and 4.0-14.8ngmL(-1), were obtained for C18 and Shield RP18 columns, respectively. The optimized method exhibited a good precision level, with relative standard deviation values lower than 8%. The recoveries studied at two spiked levels, ranged from 71 to 102% for all considered compounds. The proposed method was successfully applied to the analysis of seven benzodiazepines in real human plasma samples.

Chromatographic determination of drugs of abuse in vitreous humor using solid-phase extraction.

A simple method is presented for the simultaneous determination of morphine, 6-acetylmorphine, codeine, cocaine, benzoylecgonine, cocaethylene, methadone and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in vitreous humor by high-performance liquid chromatography with photodiode array detector after solid-phase extraction with Oasis® HLB cartridges and dichloromethane as eluent. The chromatographic process was carried out using an XTerra® RP8 column (250 × 4.6 mm i.d., 5 µm particle size) and a mobile phase composed of acetonitrile and pH 6.5 phosphate buffer in gradient mode. A linear response from the detector was obtained within the concentration range of 0.1-4 µg ml(-1) , with correlation coefficients higher than 0.99. The limits of detection were lower than 30 ng ml(-1) for all the drugs studied, the coefficients of variation fluctuated between 0.1 and 12.4%, and the average recoveries were higher than 78% for all the drugs except for EDDP, with a value of 66.4%. Finally, the proposed method was applied to 15 vitreous humor samples coming from individuals who had died from opiate and/or cocaine overdose, showing consumption of cocaine in 14 cases, methadone in five cases and heroin in three cases. Average concentrations of 0.30 µg ml(-1) for morphine, 0.24 µg ml(-1) for 6-acetylmorphine, 0.10 µg ml(-1) for codeine, 0.81 µg ml(-1) for cocaine, 1.26 µg ml(-1) for benzoylecgonine, 0.15 µg ml(-1) for cocaethylene, 0.11 µg ml(-1) for methadone and 0.68 µg ml(-1) for EDDP were obtained.

A new method for quantifying prenatal exposure to ethanol by microwave-assisted extraction (MAE) of meconium followed by gas chromatography-mass spectrometry (GC-MS).

Ethanol is a legal and widely available substance. There are health and social consequences associated with its abuse. One of the most important problems is related to alcohol consumption during pregnancy. In fact, prenatal ethanol exposure can be associated with fetal alcohol spectrum disorder (FASD), a term used to describe a wide range of potentially lifelong effects that include physical, mental, behavioral, and learning disabilities. Fatty acid ethyl esters (FAEEs), which are non-oxidative metabolites of ethanol, are currently used as biomarkers of direct ethanol consumption in different matrices, including hair, blood, skin surface, and meconium. Analysis of these compounds in meconium reveals exposure to alcohol during the second and third trimesters of pregnancy. An important finding for evaluation of gestational ethanol exposure is the fact that FAEEs do not cross the placenta. Because they accumulate in the fetal gut from approximately the 20th week of gestation until birth, this provides a wide window of detection of chronic exposure to alcohol. The sum of the concentrations of all the FAEEs, with a cutoff of 2 nmol g(-1) or 600 ng g(-1) meconium, has been recommended as evidence of maternal alcohol use. We introduce a novel technique to quantify ethyl myristate, ethyl palmitate, ethyl stearate, and their deuterated analogues (as internal standards, IS) in meconium using microwave-assisted extraction (MAE) coupled with gas chromatography-mass spectrometry (GC-MS). Limits of detection and quantification were 50 and 100 ng g(-1) for all analytes except ethyl stearate (LOD 100 ng g(-1) and LOQ 500 ng g(-1)). Calibration curves were linear from the LOQ to 5000 ng g(-1). The validated method was applied to the analysis of 81 meconium samples.

A rapid analytical method based on microwave-assisted extraction for the determination of drugs of abuse in vitreous humor.

Robust and simple validated analytical methods are required in postmortem toxicology to confirm immunoassay screening analysis of drugs of abuse. In this work, microwave-assisted extraction (MAE) was evaluated as an alternative method for extraction of target compounds such as cocaine, benzoylecgonine, cocaethylene, morphine, codeine, 6-monoacetylmorphine, methadone, and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine from vitreous humor. The MAE procedure parameters, namely, extraction temperature, time, and solvent volume, were optimized using a central composite design and applying desirability functions. The optimal conditions for extraction were 80 °C, 8 min, and 15 mL of dichloromethane solvent. The MAE-high-performance liquid chromatography-diode-array detection method was validated, showing its capability for the detection of concentrations in the range from 33 to 76 ng mL(-1) and recoveries in the range from 87 to 99.3% for all drugs. The MAE-based method was tested for 15 vitreous humor samples from forensic cases and its performance was compared with that of a solid-phase extraction method previously developed by our group. In general, better recovery and precision were achieved with the use of the MAE-based procedure.

Determination of fentanyl, metabolite and analogs in urine by GC/MS.

A rapid and sensitive method for the simultaneous determination of alfentanyl, sufentanyl and fentanyl (and its major metabolite norfentanyl) in urine was developed and validated. The method involved a liquid-liquid extraction in alkaline conditions, derivatization with pentafluoropropionic anhydride to improve the sensitivity for norfentanyl and subsequent analysis in GC/MS. The LODs are 0.08 ng ml(-1) for all substances (0.04 ng ml(-1) for alfentanyl). Intra- and inter-day precision coefficient of variation was always below 15%; mean relative error (accuracy) was always below 15%. The method was linear for all analytes, with quadratic regression of calibration curves always higher than 0.99. The method was applied to real samples of subjects who had received therapeutic doses of fentanyl, showing its suitability for the determination of low levels of these substances. The method was also applied to a subject whose death was attributed to fentanyl overdose.

Microwave-assisted extraction: a simpler and faster method for the determination of ethyl glucuronide in hair by gas chromatography-mass spectrometry.

Alcohol is the most frequently abused "addictive substance" that causes serious social problems throughout the world; thus, alcoholism is of particular interest in clinical and forensic medicine. Alcohol biomarkers are physiological indicators of alcohol exposure or ingestion and may reflect the presence of an alcohol use disorder. The glucuronide conjugation is a minor pathway of ethanol metabolism. Ethyl glucuronide (EtG) is a marker of recent alcohol consumption that detects alcohol use reliably over a definite time period. The present paper describes a new method for the determination of EtG in hair. It is based both in the microwave-assisted extraction (MAE), to extract the analyte from hair samples, and gas chromatography-mass spectrometry (GC-MS), to identify and quantify the EtG in selected ion monitoring (SIM) mode. The method was applied to 15 hair samples from occasional alcohol users, obtaining positive results in all cases. It was fully validated, including a linear range (0.3-10 ng/mg) and the main precision parameters. In summary, the use of microwave-assisted extraction turned out to be a substantially simpler, faster, and a more sensitive procedure than any other conventional sample preparations.

Microwave-assisted extraction and HPLC-DAD determination of drugs of abuse in human plasma.

A sample preparation procedure using microwave energy is proposed for the determination of morphine, 6-acetylmorphine, codeine, cocaine, cocaethylene, benzoylecgonine, methadone, and 2-ethylene-1,5-dimethyl-3,3-diphenylpyrrolidine in human plasma. A screening asymmetrical factorial design was used to identify the most suitable extraction conditions as regards solvent, temperature, and extraction time. The target drugs were quantified by high-performance liquid chromatography with diode-array detection. The use of microwave energy was found to reduce solvent consumption and extraction time compared with solid-phase extraction. The detector response was linear over the drug concentration range of 0.05-2.0 microg/mL in human plasma. The precision and accuracy were good, with values less than 8% and 7%, respectively. Drug recoveries from spiked samples ranged from 69 to 81%. The proposed method was successfully applied to a number of forensic cases.

Microwave assisted extraction of drugs of abuse from human urine.

Microwave assisted extraction (MAE) was used to extract drugs of abuse from urine samples for the simultaneous determination of morphine, codeine, 6-acetylmorphine (6AM), cocaine, cocaethylene, benzoylecgonine (BEG), methadone and 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) by high performance liquid chromatography with diode array detector (HPLC-DAD). The working wavelengths used were 285 nm for morphine, codeine and 6AM; 233 nm for cocaine, cocaethylene and BEG; and 292 nm for methadone and EDDP. The mobile phase was a gradient of acetonitrile and phosphate buffer at pH 6.5. A full factorial design was used to identify the most suitable extraction conditions as regards pH, solvent and time of extraction. The optimum conditions thus established provided analyte recoveries from 61% to 109%. The detector response was linear over the drug concentration range 0.1-4.0 microg ml(-1) in urine. The within- and between-day precision and accuracy were less than 8% and 6.5%, respectively. The method was applied successfully to samples obtained from Galician hospitals and courts.

Determination of cocaine and cocaethylene in plasma by solid-phase microextraction and gas chromatography-mass spectrometry.

The present paper describes a method for the simultaneous determination of cocaine and cocaethylene in plasma. It was based in the extraction of the analytes by solid-phase microextraction (SPME), and gas chromatography-mass spectrometry (GC-MS) was used to identify and quantify the analytes in selected ion monitoring (SIM) mode. The method showed to be very simple, rapid and sensitive. The method was validated for the two compounds, including linearity (range 25-1000 ng/mL) and the main precision parameters. It was applied to ten plasma samples from cocaine and alcohol users, obtaining positive results in all cases.