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We developed a highly efficient and low-cost organic solvents-resistant microfluidic paper-based analytical device (μPAD) coupled with paper spray mass spectrometry (PS-MS) for quantitative determination of C 18 -ceramide as a prognostic biomarker for several diseases. Several models of μPAD patterns have been examined to select the most resistant and efficient microchannel barriers, which can provide continuous spray at ionization zone and prevent "coffee ring" effect. Moreover, the developed μPAD has enabled the analysis of low concentration of C 18 -ceramide because of the maximum supply of deposited analyte through microchannel. The MS results confirmed the formation of doubly and singly charged metal ion complexes between ceramide and different metal ions. Notably, the complexation that occurs between lithium ions and C 18 -ceramide showed a high relative abundance compared with other formed complexes. Taking into account the relative abundance of complex [Cer + Li] + at 572.8 m/z, it can be considered as a stable ion and therefore be used for the analysis of C 18 -ceramide at low concentrations. Complexation of C 18 -ceramide and lithium confirmed with quantum chemical calculations. The proposed method represents good linearity with a regression coefficient of 0.9956 for the analysis of C 18 -ceramide and reaches a limit of detection to 0.84 nM. It has been adapted successfully for practical application in human serum samples with high recovery values in range of 92%-105%. The developed μPAD-MS technique provides clear advantages by reducing the experimental steps and simplifying the operation process and enables to identify subnanomolar concentration of C 18 -ceramide in human serum samples., (© 2020 John Wiley & Sons, Ltd.)

We developed a highly efficient and low‐cost organic solvents‐resistant microfluidic paper‐based analytical device (μPAD) coupled with paper spray mass spectrometry (PS‐MS) for quantitative determination of C18‐ceramide as a prognostic biomarker for several diseases. Several models of μPAD patterns have been examined to select the most resistant and efficient microchannel barriers, which can provide continuous spray at ionization zone and prevent "coffee ring" effect. Moreover, the developed μPAD has enabled the analysis of low concentration of C18‐ceramide because of the maximum supply of deposited analyte through microchannel. The MS results confirmed the formation of doubly and singly charged metal ion complexes between ceramide and different metal ions. Notably, the complexation that occurs between lithium ions and C18‐ceramide showed a high relative abundance compared with other formed complexes. Taking into account the relative abundance of complex [Cer + Li]+ at 572.8 m/z, it can be considered as a stable ion and therefore be used for the analysis of C18‐ceramide at low concentrations. Complexation of C18‐ceramide and lithium confirmed with quantum chemical calculations. The proposed method represents good linearity with a regression coefficient of 0.9956 for the analysis of C18‐ceramide and reaches a limit of detection to 0.84 nM. It has been adapted successfully for practical application in human serum samples with high recovery values in range of 92%–105%. The developed μPAD‐MS technique provides clear advantages by reducing the experimental steps and simplifying the operation process and enables to identify subnanomolar concentration of C18‐ceramide in human serum samples. [ABSTRACT FROM AUTHOR]

A novel CuS grafted PEDOT:PSS conducting paper-based electrochemical immunosensor has been successfully fabricated for the detection of EpCAM antigen. Electrical conductivity is further improved by doping the CuS@PEDOT:PSS/WP electrode with different organic solvents such as N,N-dimethyl formamide (DMF), N,N-dimethyl acetamide (DMA), ethylene glycol (EG) and dimetyl sulfoxide (DMSO). It is observed that DMSO doped conducting paper shows the highest conductivity (6.1 × 10−2 S/cm) among all these solvents. This low cost, biodegradable electrochemical paper-based immunosensor (BSA/anti-EpCAM/EP) possesses a broad linear range from 0.01 pg/mL to 1000 ng/mL for EpCAM antigen detection. The immunosensor is found to have high sensitivity of 104.31 μA pg−1 mL. This conducting paper based immunosensor is further used to detect EpCAM antigen in human serum samples, thereby confirming a cost effective clinical tool over other costly conventional techniques. • An electrochemical paper-based immunosensor has been designed using CuS and PEDOT:PSS. • This low cost and flexible platform may be a promising candidate over other costly conventional electrodes. • It exhibits high sensitivity of 104.31 μA pg−1 mL in broad range of 0.01 pg/mL to 1000 ng/mL, for EpCAM detection. • The clinical applicability and validity has been checked with human serum sample. [ABSTRACT FROM AUTHOR]

Background: Subclinical atherosclerosis may result in fatal cardiovascular (CV) events, but the underlying mechanisms and molecular players leading to disease are not entirely understood. Thus, novel approaches capable of identifying the factors involved in pathological progression and providing a better understanding of the subjacent mechanisms are needed. Extracellular vesicles (EVs) have been shown to have numerous biological functions, and their metabolome has recently generated interest as a source of novel biomarkers. The metabolic content of the exosomes has been so far unexplored in cardiovascular disease (CVD), and here, we developed an analytical strategy aimed at probing urinary exosomal metabolite content and its association to CV risk. Results: Direct analysis of the exosomes without metabolite extraction was evaluated by high-resolution magic angle spinning (1H HR-MAS). Other two methodologies for the analysis of exosomal metabolites by 1H NMR were set up, based on methanol or organic solvents sequential extraction. The three methods were compared in terms of the number of detected signals and signal to noise ratio (S/N). The methanol method was applied to identify altered metabolites in the urinary exosomes of subjects with programmed coronary artery by-pass grafting (CABG) versus a control group. Target mass spectrometry (MS) was also performed for differential analysis. The clinical performance of exosomal metabolites of interest in CVD was investigated, and the added value of the exosomes compared to urine analysis was evaluated. Based on S/N ratio, simplicity, reproducibility, and quality of the spectrum, the methanol method was chosen for the study in CVD. A cardiometabolic signature composed by 4-aminohippuric acid, N-1-methylnicotinamide, and citric acid was identified in urinary exosomes. Directly in urine, 4-aminohippuric acid and citric acid do not show variation between groups and changes in N-1-methylnicotinamide are less pronounced, proving the added value of exosomes. Conclusions: We set up a novel methodology to analyze metabolic alterations in urinary exosomes and identified a cardiometabolic signature in these microvesicles. This study constitutes the first evidence of a role for the exosomal metabolism in CVD and demonstrates the possibility to evaluate the urinary exosomal metabolic content by NMR and MS. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the ototoxic effect of the exposure to different organic solvents and noise using distortion product otoacoustic emissions (DPOAEs). Design: The exposure to different solvents was evaluated by measuring, before and at the end of the work-shift, the urinary concentrations of solvent metabolites used as dose biomarkers. The urinary concentrations of DNA and RNA oxidation products were also measured as biomarkers of oxidative damage. The simultaneous exposure to noise was also evaluated. DPOAEs and pure tone audiometry (PTA) were used as outcome variables, and were correlated to the exposure variables using mixed effect linear regression models. Study sample: Seventeen industrial painters exposed to a solvent mixture in a naval industry. A sample size of 15 was estimated from previous studies as sufficient for discriminating small hearing level and DPOAE level differences (5 dB and 2 dB, respectively) at a 95% confidence level. Results: Statistically significant associations were found between the DPOAE level and the urinary dose biomarkers and the oxidative damage biomarkers. DPOAE level and the logarithm of the metabolite concentration showed a significant negative correlation. Conclusions: DPOAE are sensitive biomarkers of exposure to ototoxic substances and can be effectively used for the early detection of hearing dysfunction. [ABSTRACT FROM AUTHOR]

Organic solvents widely used in paint manufacturing, painting, and shoemaking industries may be associated with hepatotoxicity. We present a case report of a patient with prolonged occupational exposure to organic solvents who developed transient hepatitis. Monitoring contact to these chemicals and early identification of biological markers of occupational exposure should be done. More epidemiological studies on the effects of solvents on the liver should be performed so as to help the policy makers to formulate appropriate measures to prevent detrimental effects of exposure of such chemicals. Additionally, early reporting of such cases of occupational hazards will be helpful in further understanding the incidence and possible mechanisms. [ABSTRACT FROM AUTHOR]

The use of organic solvents such as benzene and toluene, causes in the people with high exposure a series of serious health effects, among which stand out neurotoxicity and cancer. The objective of this study was to determine the levels of phenol and hippuric acid, in the urine, as biological indicators of exposure to benzene and toluene. A descriptive, observational, cross-correlation and field study was carried out in which 20 workers with occupational exposure to benzene and 20 without occupational exposure participated, with prior informed consent. The Theis-Benedict method was used for the determination of phenol, NIOSH 8300 for hippuric acid and Jaffé modified for the determination of creatinine, in urine samples taken at the end of the working day. Phenol/crea, levels in the exposure group ranged from 98.23 to 102.34 mg/g-crea (Mean = 97.89; SD = 3.69) and 8.21 to 33.21 mg/g-creatinine in the group without exposure (Mean = 15.16; SD = 6.23). In addition, a statistically significant difference (p < 0.05) was observed for phenol / crea levels between both groups but not for hippuric acid (p > 0.05). Workers did not wear personal protective equipment (PPE) and work under poor hygiene conditions, which increases exposure to benzene. The low concentration of urinary hippuric acid implies a low toluene exposure. [ABSTRACT FROM AUTHOR]

One of the major components of the organic solvent extractable fraction of Cretaceous ambers is 1,6-dimethyl-5-isopentyltetralin, a compound that has been incorrectly or not identified in previous reports on the molecular composition of Cretaceous ambers. Here, the purification and structural identification of 1,6-dimethyl-5-isopentyltetralin is reported from amber samples of the Cretaceous Peñacerrada II deposit at Peñacerrada, Álava (Basque-Cantabrian Basin, Spain). The structural identification of 1,6-dimethyl-5-isopentyltetralin allowed the complete identification of related compounds, comprising the family of 1,6-dimethyl-5-alkyltetralins. This suggests that those compounds, with concentrations up to 27% of the total organic extract of amber from Peñacerrada, are characteristic for labdane-type biomarkers in Cretaceous fossil resins and their distributions indicate different compositional families, related to their botanical and topographic origin. Also, the name amberene is proposed for the main molecule of this group. The relative proportion of the amberene-type compounds suggests three compositional families for the Cretaceous Spanish ambers. [ABSTRACT FROM AUTHOR]

N -alkyl-2-pyrrolidones are important organic solvents for varnishes in industry. This study investigates exposure to N -ethyl-2-pyrrolidone (NEP) in varnishing of hard plastic components in an automobile plant. Two specific biomarkers of exposure, 5-hydroxy- N -ethyl-2-pyrrolidone (5-HNEP) and 2-hydroxy- N -ethylsuccinimide (2-HESI), were analyzed in urine samples of 14 workers. For this purpose, pre-shift, post-shift and next day pre-shift urine samples were collected midweek. Twelve workers performed regular work tasks (loading, wiping and packing), whereas two workers performed special work tasks including cleaning the sprayer system with organic solvents containing N -alkyl-2-pyrrolidones. Spot urine samples of nine non-exposed persons of the same plant served as controls. Median post-shift urinary levels of workers with regular work tasks (5-HNEP: 0.15 mg/L; 2-HESI: 0.19 mg/L) were ∼5-fold higher compared to the controls (0.03 mg/L each). Continuously increasing metabolite levels, from pre-shift via post-shift to pre-shift samples of the following day, were observed in particular for the two workers with the special working tasks. Maximum levels were 31.01 mg/L (5-HNEP) and 8.45 mg/L (2-HESI). No clear trend was evident for workers with regular working tasks. In summary, we were able to show that workers can be exposed to NEP during varnishing tasks in the automobile industry. [ABSTRACT FROM AUTHOR]

The first step in many life detection protocols on Mars involves attempts to extract or isolate organic matter from its mineral matrix. A number of extraction options are available and include heat and solvent assisted methods. Recent operations on Mars indicate that heating samples can cause the loss or obfuscation of organic signals from target materials, raising the importance of solvent-based systems for future missions. Several solvent types are available (e.g. organic solvents, surfactant based solvents and subcritical water extraction) but a comparison of their efficiencies in Mars relevant materials is missing. We have spiked the well characterised Mars analogue material JSC Mars-1 with a number of representative organic standards. Extraction of the spiked JSC Mars-1 with the three solvent methods provides insights into the relative efficiency of these methods and indicates how they may be used on future Mars missions. [ABSTRACT FROM AUTHOR]

Metabolic alteration plays a functional role in kidney allograft complications. Metabolomics is a promising high-throughput approach in nephrology but is still limited by the lack of overlap in metabolite coverage. We performed an untargeted fecal metabolomic analysis of forty stable kidney allograft recipients and twenty non-transplant controls. First, we applied the ultra-high performance liquid chromatography (UHPLC) analysis coupled with the Diod Array detector. The potential biomarkers were then collected and identified by gas chromatography-mass spectrometry (GCMS). In order to allow for complete coverage of the fecal polar and non-polar metabolites, the performance of five organic solvents with increasing polarity was investigated successively. UHPLC analysis revealed that the fecal metabolite profiles following the five extractions were significantly different between controls and kidney allografts. GC-MS analysis showed that the best predictors' metabolites belonged mainly to long-chain fatty acids, phenolic compounds, and amino acids. Collectively, our results showed the efficiency of our pioneer method to successfully discriminate stable kidney-transplant recipients from controls. These findings suggest that distinct metabolic profiles mainly affect fatty acid biosynthesis and amino acid metabolism. In such a context, the novel insights into metabolomic investigation may be a valuable tool that could provide useful new relevant biomarkers for preventing kidney transplant complications. [ABSTRACT FROM AUTHOR]

Trichloroethylene (TCE), an important volatile organic solvent, causes a series of toxic damage to human. Conventional genetic mechanisms cannot fully explain its toxicity and carcinogenicity, indicative of the possible involvement of epigenetic mechanisms. Our study was intended to investigate the epigenetic toxicity and underlying mechanisms of TCE. Data showed that 0.3 mM TCE treatment for 24 h increased the growth of L-02 cells transiently. In contrast, subacute exposure to TCE inhibited cell growth and induced the genomic DNA hypomethylation and histone hyperacetylation. Further studies have revealed the TCE-induced DNA hypomethylation in the promoter regions of tumor-related genes, N-Ras , c-Jun , c-Myc , c-Fos and IGF-II , promoting their protein levels in a time-dependent manner. These results reveal there is a negative relationship existing between DNA hypomethylation and protein expression in tumor-related gene after TCE exposure under specific epigenetic microenvironment, serving as early biomarkers for TCE-associated diseases. • Low-dose TCE inhibited the growth of L-02 cells by subacute exposure. • TCE induced global DNA hypomethylation and histone hyperacetylation. • Specific DNA hypomethylation upregulated its expression of tumor-related gene. [ABSTRACT FROM AUTHOR]

Most bioanalytical LC-MS/MS methods are developed for determination of single drugs or classes of drugs, but a multi-compound LC-MS/MS method that can replace several methods could reduce both analysis time and costs. The aim of this study was to develop a high-throughput LC-MS/MS method for determination of the alcohol biomarker phosphatidylethanol 16:0/18:1 (PEth 16:0/18:1) and 33 other compounds from eight different drug classes in whole blood. Whole-blood samples were prepared by 96-well supported liquid extraction (SLE). Chromatographic separations were performed on a biphenyl core shell column with a mobile phase consisting of 10 mM ammonium formate, pH 3.1 and methanol. Each extract was analyzed twice by LC-MS/MS, injecting 0.4 μL and 2 μL, in order to obtain narrow and symmetrical peaks and good sensitivity for all compounds. Stable isotope-labeled internal standards were used for 31 of the 34 compounds. A 96-well SLE reversed phase LC-MS/MS method for determination of PEth 16:0/18:1 and 33 other compounds from eight different drug classes was developed and validated. By using an organic solvent mixture of isopropanol/ methyl tert-butyl ether (1:5, v:v), all compounds, including the polar and ampholytic compounds pregabalin, gabapentin and benzoylecgonine, was extracted by 96-well SLE. For the first time an LC-MS/MS method for the determination of alcohol biomarker PEth 16:0/18:1 and drugs and metabolites from several different drug classes was developed and validated. The developed LC-MS/MS method can be used for high-throughput analyses and sensitive determinations of the 34 compounds in whole blood. • Quantification of alcohol biomarker PEth 16:0/18:1 and 33 other compounds by LC-MS/MS • First method for quantification of PEth and drugs from several drug classes • Acidic, basic and ampholytic compounds determined by LC-MS/MS • First supported liquid extraction of pregabalin and gabapentin from blood • High throughput sample preparation by 96-well supported liquid extraction [ABSTRACT FROM AUTHOR]

• Determination of occupational exposure of VOC in the University community. • Development and validation of bioanalytical method for simultaneous exposure solvents. • Biological evaluation as tool for assessing occupational exposure. Occupational exposure to volatile organic compounds (VOC) might generate serious worker health damages. Therefore, biological monitoring is essential to evaluate exposure biomarkers from highly toxic chemicals, ensuring better attention to the worker health. In this study was developed and validated a bioanalytical method based on high-performance liquid chromatography coupled to photodiode array (HPLC-PDA) for the quantification of VOC biomarkers in urine samples from Federal University of Goias (UFG) workers. Samples were collected from 30 occupationally exposed subjects after application of a questionnaire survey. The following biomarkers hippuric acid, methyl-hippuric acid, mandelic acid, phenylglyoxylic acid and phenol were quantified, representing exposition to toluene, xylene, styrene, ethylbenzene, benzene and phenol solvents, respectively. Hippuric acid levels were found close to or above the reference values, although a subject had levels higher than preconized by Biological Limit Values (BLV) guideline of 4.0 mg/g creatinine. Five subjects had 3 and 4-methylhippuric acid ranging from 0.1 to 1.0 mg/g creatinine. These results indicate a moderate to high VOC exposure from UFG workers. Multivariate analysis generated four clusters and indicated that histotechnicians and graphic workers need especial attention on occupational VOC exposure. The results from this study reinforce the need for reliable methods able to the biological monitoring as an important tool for assessing occupational exposure. [ABSTRACT FROM AUTHOR]