High Sample Throughput Research Articles

Constructing an In Vitro and In Vivo Flow Cytometry by Fast Line Scanning of Confocal Microscopy.

Composed of a fluidic and an optical system, flow cytometry has been widely used for biosensing. The fluidic flow enables its automatic high-throughput sample loading and sorting while the optical system works for molecular detection by fluorescence for micron-level cells and particles. This technology is quite powerful and highly developed; however, it requires a sample in the form of a suspension and thus only works in vitro. In this study, we report a simple scheme to construct a flow cytometry based on a confocal microscope without any modifications. We demonstrate that line scanning of microscopy can effectively excite fluorescence of flowing microbeads or cells in a capillary tube in vitro and in blood vessels of live mice in vivo. This method can resolve microbeads at several microns and the results are comparable to a classic flow cytometer. The absolute diameter of flowing samples can be indicated directly. The sampling limitations and variations of this method is carefully analyzed. This scheme can be easily accomplished by any commercial confocal microscope systems, expands the function of them, and is of promising potential for simultaneous confocal microscopy and in vivo detection of cells in blood vessels of live animals by a single system.

Study on Separation of Rhenium, a Surrogate Element of Fissiogenic Technetium, from Aqueous Matrices Using Ion-Selective Extraction Chromatographic Resins

Long-lived fissiogenic technetium, 99Tc (half-life, 2.11 × 105 yrs), is a byproduct of the 235U and 239Pu fission in nuclear reactors and is a major component in nuclear waste. Hence, the monitoring of 99Tc activity in the environmental samples is essential. Techniques used to measure 99Tc in environmental samples include radiometric and non-radiometric approaches. Inductively coupled plasma mass spectrometry (ICP-MS) has some advantages among the methods due to its high sample throughput, better selectivity, and commercial availability. Rhenium (Re) is often used as the non-isotopic tracer during 99Tc measurement by ICP-MS. From this perspective, studying the separation behavior of Re from the aqueous matrix can provide helpful insight regarding the ecological monitoring of 99Tc. In the current work, three extraction chromatographic resins (ECRs), MetaSEP AnaLig Tc-01, MetaSEP AnaLig Tc-02, and Eichrom TRU, have been used to separate Re from the aqueous matrix. Operating variables, such as solution pH, choice of eluent and eluent concentration, matrix cation effect, and retention capacity, have been studied to optimize the separation protocol. The extraction and recovery behavior of Re was used to interpret the selectivity behavior of the ECRs. The Re separation factor at optimized operating conditions comparing with Mo and Ru (interfering elements during the measurement by ICP-MS) for Meta SEP AnaLig Tc-01, Meta SEP AnaLig Tc-02, and Eichrom TRU resin, respectively, are SFRe/Mo~32, 16.51, and ∞ (metal not retained at all), while SFRe/Ru~∞, 3.25, 5437. In terms of selectivity and retention capacity, MetaSEP AnaLig Tc-01 is the better choice for selective separation of Re, and, assumably so, also Tc, from the aqueous matrices.

High-Throughput Preparation and High-Throughput Detection of Polymer-Dispersed Liquid Crystals Based on Ink-Jet Printing and Grayscale Value Analysis.

In this paper, based on high-throughput technology, polymer dispersed liquid crystals (PDLC) composed of pentaerythritol tetra (2-mercaptoacetic acid) (PETMP), trimethylolpropane triacrylate (TMPTA), and polyethylene glycol diacrylate (PEGD 600) were investigated in detail. A total of 125 PDLC samples with different ratios were quickly prepared using ink-jet printing. Based on the method of machine vision to identify the grayscale level of samples, as far as we know, it is the first time to realize high-throughput detection of the electro-optical performance of PDLC samples, which can quickly screen out the lowest saturation voltage of batch samples. Additionally, we compared the electro-optical test results of manual and high-throughput preparation PDLC samples and discovered that they had very similar electro-optical characteristics and morphologies. This demonstrated the viability of PDLC sample high-throughput preparation and detection, as well as promising application prospects, and significantly increased the efficiency of PDLC sample preparation and detection. The results of this study will contribute to the research and application of PDLC composites in the future.

Cortisol Immunosensors: A Literature Review.

Cortisol is a steroid hormone that is involved in a broad range of physiological processes in human/animal organisms. Cortisol levels in biological samples are a valuable biomarker, e.g., of stress and stress-related diseases; thus, cortisol determination in biological fluids, such as serum, saliva and urine, is of great clinical value. Although cortisol analysis can be performed with chromatography-based analytical techniques, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS), conventional immunoassays (radioimmunoassays (RIAs), enzyme-linked immunosorbent assays (ELISAs), etc.) are considered the "gold standard" analytical methodology for cortisol, due to their high sensitivity along with a series of practical advantages, such as low-cost instrumentation, an assay protocol that is fast and easy to perform, and high sample throughput. Especially in recent decades, research efforts have focused on the replacement of conventional immunoassays by cortisol immunosensors, which may offer further improvements in the field, such as real-time analysis at the point of care (e.g., continuous cortisol monitoring in sweat through wearable electrochemical sensors). In this review, most of the reported cortisol immunosensors, mainly electrochemical and also optical ones, are presented, focusing on their immunosensing/detection principles. Future prospects are also briefly discussed.

Sequential Injection Analysis for Rapid Determination of Mercury in Skincare Products Based on Fluorescence Quenching of Eco-Friendly Synthesized Carbon Dots.

This work reports the analysis of mercury using a spectrofluorometric method combined with a sequential injection analysis (SIA) system. This method is based on the measurement of fluorescence intensity of carbon dots (CDs), which is quenched proportionally after adding mercury ions. Herein, the CDs underwent environmentally friendly synthesis using a microwave-assisted approach that provides intensive and efficient energy and shortens reaction time. After irradiation at 750 W for 5 min in a microwave oven, a dark brown CD solution with a concentration of 2.7 mg mL-1 was obtained. The properties of the CDs were characterized by transmission electron microscopy, X-ray diffractometry, X-ray photoelectron spectroscopy, Fourier-transform infrared spectroscopy, and UV-vis spectrometry. We presented for the first time the use of CDs as a specific reagent for the determination of mercury in skincare products with the SIA system to achieve rapid analysis and full automatic control. The as-prepared CD stock solution was diluted 10 times and used as a reagent in the SIA system. Excitation and emission wavelengths at 360 and 452 nm, respectively, were used to construct a calibration curve. Physical parameters affecting the SIA performance were optimized. In addition, the effect of pH and other ions was investigated. Under the optimum conditions, our method showed a linear range from 0.3 to 600 mg L-1 with an R 2 of 0.99. The limit of detection was 0.1 mg L-1. Relative standard deviation was 1.53% (n = 12) with a high sample throughput of 20 samples per hour. Finally, the accuracy of our method was validated by comparison using inductively coupled plasma mass spectrometry. Acceptable recoveries were also presented without a significant matrix effect. This method was also the first time that uses the untreated CDs for the determination of mercury(II) in skincare products. Therefore, this method could be an alternative for mercuric toxic control in other sample applications.

An easy and green assay to determine albendazole and ivermectin in veterinary preparations by micellar liquid chromatography.

A procedure to determine albendazole and ivermectin in veterinary formulations, like tablet, bolus, oral suspensions, and injections by micellar liquid chromatography, has been developed. Sample preparation was a batch solid-to-liquid extraction in mobile phase, consisting of a stirring step (15min), followed by ultrasonication (15min) and filtration of the obtained supernatant, to reach a target concentration of 2mg/L for both analytes. Using a mobile phase of 0.15M sodium dodecyl sulfate-6% 1-pentanol buffered at pH 3 with a 0.01M phosphate salt, running at 1mL/min through a C18 column, both drugs were resolved in less than 10min. Absorbance detection wavelength was 292nm. Procedure was validated by the guidelines of the International Council on Harmonization in terms of specificity, calibration range (0.025-5mg/L), trueness (97.8%-102.6%), precision (<2.2%), and system suitability. The method was found easy-to-handle, low cost, safe, green, and with high sample-throughput, thus useful for routine analysis. Therefore, it represents a valuable alternative for quality control of veterinary formulations. It was applied to samples of veterinary formulations purchased from local chemists and veterinarians, and label claims were inside the acceptance criteria (95%-105%).

On-line carbon dots synthesis using flow injection analysis. Application to aluminium determination in water samples

An on-line synthesis of CDots is proposed for the first time, using the flow injection analysis (FIA) technique, which was coupled, in a single system, to the analytical determination of aluminium in water samples. The nanoparticles were obtained from the carbonization of glucose and iron(III) in an acidic medium, and their photoluminescence increased in the presence of aluminium ions. Under optimal experimental conditions, the proposed method has shown an acceptable linearity range –between 0.04 and 3.0 mg L−1 (R2 = 0.9999) – and a detection limit of 0.007 mg L−1. The analysis of drinking water and groundwater samples showed good accuracy (recoveries ranged between 91 – 113%) and RSD% < 13. The on-line system exhibited a high sample throughput (36 h−1), since no incubation time was required.

The martini 3 ecosystem for coarse-grained simulations.

Coarse-grained molecular dynamics (MD) simulations using the Martini force-field have become a powerful tool in studying biomolecular systems at near to all-atom resolution. About two years ago we put forward a complete reparameterization of the original Martini force-field. The Martini 3 force-field comprises rebalanced non-bonded interaction parameters as well as an improved parametrization strategy for generating molecular parameters. So far Martini 3 has been successfully applied in numerous studies ranging from protein ligand binding for computer-aided drug design to simulations of large-scale systems such as the SARS-Cov2 viral envelope. As exemplified by these studies, the trend goes towards high-throughput sampling studies as well as large-scale biomolecular systems with the set goal of simulating whole cells. To prepare the Martini universe for the next simulation era, advancement in three aspects is of fundamental importance: 1) development of automated workflows that enable fast and efficient setup of complex systems; 2) extension and improvement of available molecule and force-field parameters; 3) management of feedback from the research community. To this end we created the vermouth python library as a unified framework for both program development and parameter management. Based on vermouth, tools such as martinize2, polyply, or fast-forward have been created, which facilitate setting up Martini simulations. In addition, improved parameters for small molecules and carbohydrates have been released with updates for other classes of biomolecules such as lipids underway. Lastly, by openly hosting all programs and parameters on GitHub, we give the research community an opportinuty to provide feedback and contribute to developments. Here we present how these individual developments make up the Martini 3 ecosystem for coarse-grained simulations and our current goals for future advances.

ASAP─Automated Sonication-Free Acid-Assisted Proteomes─from Cells and FFPE Tissues.

Formalin-fixed, paraffin-embedded (FFPE) tissues are an invaluable resource for retrospective studies, but protein extraction and subsequent sample processing steps have been shown to be challenging for mass spectrometry (MS) analysis. Streamlined high-throughput sample preparation workflows are essential for efficient peptide extraction from complex clinical specimens such as fresh frozen tissues or FFPE. Overall, proteome analysis has gained significant improvements in the instrumentation, acquisition methods, sample preparation workflows, and analysis pipelines, yet even the most recent FFPE workflows remain complex and are not readily scalable. Here, we present an optimized workflow for automated sonication-free acid-assisted proteome (ASAP) extraction from FFPE sections. ASAP enables efficient protein extraction from FFPE specimens, achieving similar proteome coverage as established methods using expensive sonicators, resulting in reduced sample processing time. The broad applicability of ASAP on archived pediatric tumor FFPE specimens resulted in high-quality data with increased proteome coverage and quantitative reproducibility. Our study demonstrates the practicality and superiority of the ASAP workflow as a streamlined, time- and cost-effective pipeline for high-throughput FFPE proteomics of clinical specimens.

High-throughput, dynamic, multi-dimensional: an expanding repertoire of plant respiration measurements.

A recent burst of technological innovation and adaptation has greatly improved our ability to capture respiration rate data from plant sources. At the tissue level, several independent respiration measurement options are now available, each with distinct advantages and suitability, including high-throughput sampling capacity. These advancements facilitate the inclusion of respiration rate data into large-scale biological studies such as genetic screens, ecological surveys, crop breeding trials, and multi-omics molecular studies. As a result, our understanding of the correlations of respiration with other biological and biochemical measurements is rapidly increasing. Difficult questions persist concerning the interpretation and utilization of respiration data; concepts such as allocation of respiration to growth versus maintenance, the unnecessary or inefficient use of carbon and energy by respiration, and predictions of future respiration rates in response to environmental change are all insufficiently grounded in empirical data. However, we emphasize that new experimental designs involving novel combinations of respiration rate data with other measurements will flesh-out our current theories of respiration. Furthermore, dynamic recordings of respiration rate, which have long been used at the scale of mitochondria, are increasingly being used at larger scales of size and time to reflect processes of cellular signal transduction and physiological response to the environment. We also highlight how respiratory methods are being better adapted to different plant tissues including roots and seeds, which have been somewhat neglected historically.

Systematic analysis of in-source modifications of primary metabolites during flow-injection time-of-flight mass spectrometry

Flow-injection mass spectrometry (FI-MS) enables metabolomics studies with a very high sample-throughput. However, FI-MS is prone to in-source modifications of analytes because samples are directly injected into the electrospray ionization source of a mass spectrometer without prior chromatographic separation. Here, we spiked authentic standards of 160 primary metabolites individually into an Escherichia coli metabolite extract and measured the thus derived 160 spike-in samples by FI-MS. Our results demonstrate that FI-MS can capture a wide range of chemically diverse analytes within 30 s measurement time. However, the data also revealed extensive in-source modifications. Across all 160 spike-in samples, we identified significant increases of 11,013 ion peaks in positive and negative mode combined. To explain these unknown m/z features, we connected them to the m/z feature of the (de-)protonated metabolite using information about mass differences and MS2 spectra. This resulted in networks that explained on average 49 % of all significant features. The networks showed that a single metabolite undergoes compound specific and often sequential in-source modifications like adductions, chemical reactions, and fragmentations. Our results show that FI-MS generates complex MS1 spectra, which leads to an overestimation of significant features, but neutral losses and MS2 spectra explain many of these features.

Rapid Determination of Selected PFAS in Textiles Entering the Waste Stream.

Due to new European legislation, products entering the waste stream containing some perfluoro alkyl substances (PFAS) are subject to "low persistent organic pollutant concentration limits". Concentrations of restricted PFAS must be below this limit for them to be legally recycled or disposed of. A rapid extraction and clean-up method was developed for the determination of 21 PFAS in various polymers used in soft furnishings and upholstery. The optimised method used vortexing and ultrasonication in methanol (0.1% NH4OH), followed by a dilution and syringe filter clean-up step. PFAS were subsequently determined via UPLC-TripleTOF/MS. Good recoveries (80-120%) of target analytes were obtained with tall and narrow chromatogram peaks. The method was validated using control matrix samples spiked with target analytes. Repeated measurements of concentrations of target compounds showed good agreement with the spiked concentrations demonstrating good accuracy and precision. The resultant extracts provided low noise levels resulting in low limits of quantification ranging from 0.1 to 0.4 mg/kg. The developed method was applied successfully to real consumer products and it provided various advantages over traditional methods, including a substantially reduced analysis time, consumables and solvent consumption, and a high sample throughput which is critical to comply with implemented and proposed legislation.

Technical note: Validation of energy dispersive X-ray fluorescence for determination of indigestible markers in ruminant fecal and rumen fluid samples.

Determination of digestibility and passage rate is important for further understanding of nutrient utilization and thereby aids in improving nutrient utilization efficiency. Titanium dioxide and chromium ethylenediaminetetraacetic acid are commonly used as indigestible markers for determining passage rates of diets to aid in determination of digestibility. Analyzing Ti and Cr involves the use of procedures such as acid digestions, inductively coupled plasma spectroscopy, and atomic absorption. These commonly used methodologies involve hazardous chemicals, destruction of samples, and low sample throughput. The objective of this experiment was to develop and validate an accurate and precise method for measuring both Ti and Cr using energy dispersive X-ray fluorescence (ED-XRF). Energy dispersive X-ray fluorescence is an analytical technique used for analyzing elements in various sample types. The samples were added to the ED-XRF machine and irradiated with X-rays. The intensity of the X-rays emitted (termed fluorescent X-rays) was used for calculation of the concentration of the element. The method for Ti was constructed using fecal samples from cattle consuming three different diet types (finishing diet, dairy lactation diet, and grazing native range pasture). The Cr method was developed for rumen fluid analysis. We compared the machine-calculated concentrations of each element to the concentration calculated by a standard curve. For both the Ti and Cr, the standard curve-calculated value had a lower percent difference overall at 4.56% and 12.59%, respectively, compared to the machine percent difference of 8.35% and 16.38% for Ti and Cr, respectively. To determine accuracy and precision of the method, samples were spiked with various amounts of Ti or Cr and measured for their respective compounds with percent recovery and inter- and intra-assay CV-calculated thereafter. The average recovery for Ti across all diet types was 100.3%, and the recovery for Cr in rumen fluid was 95.7%. The average inter- and intra-assay CV for Ti, across all diet types, were 9.70% and 2.16%, respectively. For Cr, the average inter- and intra-assay CV were 5.42% and 8.45%, respectively. The ED-XRF method requires minimal additional chemicals, is cost-effective, and allows for sample preservation as well as a high throughput of samples. Our results indicate utilization of ED-XRF is an accurate and precise method for determination of Ti in feces and Cr in rumen fluid.

Evaluation of two methods for detection of viable Bacillus anthracis simulant spores in maritime environmental samples.

Analytical methods exist to detect biothreat agents in environmental samples during a response to biological contamination incidents. However, the coastal zone facilities and assets of the US Coast Guard (USCG), including response boats in diverse geographical areas and maritime environmental conditions, can pose complex and unique challenges for adapting existing analytical detection methods. The traditional culture (TC) and the rapid viability polymerase chain reaction (RV-PCR) methods were evaluated for their compatibility for maritime environmental surface and grab sample analysis to detect spores of Bacillus thuringiensis subspecies kurstaki (Btk), a surrogate for Bacillus anthracis. The representative samples collected from a USCG installation included surfaces, such as aluminum on boats, nonskid tread on decks of watercraft, computer touchscreens, and concrete piers, and grab samples of boat washdown water, soil, vegetation, and gravel from surrounding areas. Replicate samples were spiked with Btk spores at two to three tenfold increasing levels and analyzed. Out of a total of 150 samples collected and analyzed, the TC method gave 10 false-positive and 19 false-negative results, while the RV-PCR method-based analysis resulted in 0 false-positive and 26 false-negative results. An abundance of microbial background and particulates in some samples interfered with true results, while both methods gave similar results for samples with low microbial background and particulates. Improved and high-throughput sample processing methods are needed for analysis of complex environmental samples.

Development of High-Throughput Sample Preparation Procedures for the Quantitative Determination of Aflatoxins in Biological Matrices of Chickens and Cattle Using UHPLC-MS/MS.

Aflatoxins (AFs) frequently contaminate food and animal feeds, especially in (sub) tropical countries. If animals consume contaminated feeds, AFs (mainly aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1), G2 (AFG2) and their major metabolites aflatoxin M1 (AFM1) and M2 (AFM2)) can be transferred to edible tissues and products, such as eggs, liver and muscle tissue and milk, which ultimately can reach the human food chain. Currently, the European Union has established a maximum level for AFM1 in milk (0.05 µg kg-1). Dietary adsorbents, such as bentonite clay, have been used to reduce AFs exposure in animal husbandry and carry over to edible tissues and products. To investigate the efficacy of adding bentonite clay to animal diets in reducing the concentration of AFB1, AFB2, AFG1, AFG2, and the metabolites AFM1 and AFM2 in animal-derived foods (chicken muscle and liver, eggs, and cattle milk), chicken and cattle plasma and cattle ruminal fluid, a sensitive and selective ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method has been developed. High-throughput sample preparation procedures were optimized, allowing the analysis of 96 samples per analytical batch and consisted of a liquid extraction using 1% formic acid in acetonitrile, followed by a further clean-up using QuEChERS (muscle tissue), QuEChERS in combination with Oasis® Ostro (liver tissue), Oasis® Ostro (egg, plasma), and Oasis® PRiME HLB (milk, ruminal fluid). The different procedures were validated in accordance with European guidelines. As a proof-of-concept, the final methods were used to successfully determine AFs concentrations in chicken and cattle samples collected during feeding trials for efficacy and safety evaluation of mycotoxin detoxifiers to protect against AFs as well as their carry-over to animal products.

Molecularly Imprinted Plasmonic-Based Sensors for Environmental Contaminants—Current State and Future Perspectives

The increase of production and consumption persistently introduce different pollutants into the environment. The constant development and improvement of analytical methods for tracking environmental contaminants are essential. The demand for high sample throughput analysis has hit the spotlight for developing selective sensors to avoid time-consuming sample preparation techniques. In addition, the sensor’s sensitivity should satisfy the rigorous demands of harmful compound tracking. Molecularly imprinted plasmonic-based sensors are excellent candidates to overcome selectivity and sensitivity issues. Molecularly imprinted polymers are robust, stable in aqueous and organic solvents, stable at extreme pHs and temperatures, and include a low-cost synthesis procedure. Combined with plasmonic-based techniques, they are the perspective choice for applications in the field of environmental protection. Plasmonic-based sensors offer a lower limit of detection, a broad linearity range, high sensitivity, and high selectivity compared to other detection techniques. This review outlines the optical plasmonic detection of different environmental contaminants with molecularly imprinted polymers as sensing elements. The main focus is on the environmental pollutants affecting human and animal health, such as pesticides, pharmaceuticals, hormones, microorganisms, polycyclic aromatic hydrocarbons, dyes, and metal particles. Although molecularly imprinted plasmonic-based sensors currently have their application mostly in the biomedical field, we are eager to point them out as a highly prospective solution for many environmental problems.

Simultaneous detection of dopamine and ascorbic acid by using a thread-based microfluidic device and multiple pulse amperometry.

This study presents a novel approach for the simultaneous detection of ascorbic acid (AA) and dopamine (DA) using an affordable and user-friendly microfluidic device. Microfluidic devices, when combined with electrochemical detectors like screen-printed electrodes (SPEs), offer numerous advantages such as portability, high sample throughput, and low reagent consumption. In this study, a 3D-printed microfluidic device called a μTED was developed, utilizing textile threads as microfluidic channels and an unmodified SPE as the amperometric detector. The method employed multiple pulse amperometry (MPA) with carefully selected potential values (+0.65 V and -0.10 V). The reduction current signals generated by dopamine o-quinone were used to calculate a correction factor for the oxidation signals of ascorbic acid, enabling simultaneous quantification. The developed microfluidic device ensured a stable flow rate of the carrier solution at 1.19 μL s-1, minimizing the consumption of samples and reagents (injection volume of 2.0 μL). Under the optimized experimental conditions, a linear range from 50 to 900 μmol L-1 was achieved for both DA and AA. The obtained sensitivities were 2.24 μA L mmol-1 for AA and 5.09 μA L mmol-1 for DA, with corresponding limits of detection (LOD) of 2.60 μmol L-1 and 1.54 μmol L-1, respectively. To confirm the effectiveness of the proposed method, it was successfully applied to analyze AA and DA in a commercial blood serum sample spiked at three different concentration levels, with a medium recovery rate of 70%. Furthermore, the MPA technique demonstrated its simplicity by enabling the simultaneous determination of AA and DA without the need for prior separation steps or the use of chemically modified electrodes.

Urine and serum metabolic profiling combined with machine learning for autoimmune disease discrimination and classification.

Precision diagnosis and classification of autoimmune diseases (ADs) is challenging due to the obscure symptoms and pathological causes. Biofluid metabolic analysis has the potential for disease screening, in which high throughput, rapid analysis and minimum sample consumption must be addressed. Herein, we performed metabolomic profiling by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) in urine and serum samples. Combined with machine learning (ML), metabolomic patterns from urine achieved the discrimination and classification of ADs with high accuracy. Furthermore, metabolic disturbances among different ADs were also investigated, and provided information of etiology. These results demonstrated that urine metabolic patterns based on MALDI-MS and ML manifest substantial potential in precision medicine.

Ribbon-shaped supramolecular solvents: Synthesis, characterization and potential for making greener the microextraction of water organic pollutants

Liquid-liquid microextraction (LLME) techniques have experienced a tremendous growth over the last years but still face major challenges related to the use of more efficient and environmentally friendly solvents. Supramolecular solvents (SUPRASs) have proved outstanding efficiency in LLME, but many of the experimental conditions required for SUPRAS formation and/or application cannot be considered green or experimentally convenient. This paper was intended to make greener both SUPRAS formation and their application to the LLME of low-concentration organic pollutants in environmental waters. For this purpose, a variety of SUPRASs were produced at room temperature by simply mixing alkyl phosphonates (A6-12PO3H− and A6-12PO3−2) and tetrahexylammonium (He4N+) ions in aqueous media. Among them, the SUPRASs produced from decyl hydrogen phosphonate (DePO3H−) and He4N+ allowed, for the first time, the development of SUPRAS-based LLMEs where the SUPRAS previously synthesized was added to the liquid sample, instead of being formed in situ as usual, which was proved particularly advantageous for analyses involving large sample/SUPRAS volume ratios. At near equimolar amounts of DePO3H− and He4N+, the amphiphile arranged in the SUPRAS as planar ribbons consisting of water (21 ± 3%, w/v) and DePO3H− and He4N+ in the concentration range 1.0–1.4 M. The application of these SUPRASs to LLMEs was proved by extracting carcinogenic polycyclic aromatic hydrocarbons (CPAHs) from drinking (tap and bottled) and natural (river, reservoir and underground) water (recoveries between 84 and 117% with standard deviations varying between 1 and 14%). The developed method was simple (it only required the addition of 500 μL of SUPRAS to 75 mL of sample, stirring and centrifugation), sensitive (method quantitation limits were below the maximum allowed limits set by the EU; were 0.6–7.1 ng L−1) and selective (SUPRAS extracts were directly analyzed by liquid chromatography-fluorimetry). This research proves that SUPRASs can be operationally used in LLMEs similarly to conventional solvents, which should favor their routine application in high-sample throughput laboratories.

Simple determination of dichlorvos in cases of fatal intoxication by gas Chromatography-Mass spectrometry

Dichlorvos (DDVP) is an organophosphorous insecticide which is classified as “highly hazardous” Class 1B chemical by World Health Organization (WHO) and largely misused for the purpose of self-poisoning in developing countries. Forensic toxicology laboratories are routinely encountering cases of pesticide poisoning due to their fatal intoxication. Herein; a method is described based on vortex-assisted dispersive liquid–liquid microextraction (VA-DLLME) coupled with Gas Chromatography-Mass Spectrometry (GC–MS) for the determination of an organophosphorous insecticide; dichlorvos (DDVP) in human autopsy samples (blood, stomach content and liver). Under the optimum conditions, the method was found to be linear in the range of 0.5–10 µg mL−1 and 1.5–10 µg g−1 for blood and tissue samples, respectively. Limit of quantification was set at 0.55 µg mL−1 and 1.1 µg g−1 for blood and tissue samples, respectively. Intraday and inter-day precisions were less than 8 and 12 %, respectively. Good recoveries in the range of 86–95 % were obtained for the proposed procedure. The method has been satisfactorily applied for the determination of DDVP in autopsy samples from two different cases received in our laboratory. In comparison to previous methods; the proposed method is relatively short, high sample throughput, inexpensive and adheres to the principles of green analytical chemistry (GAC) for determination of DDVP in human autopsy samples. The method can be adopted in forensic toxicological laboratories for analysis of DDVP in autopsy samples. In addition, the green character of the proposed method was evaluated using ComplexGAPI procedure.