Electrochemical flow injection analysis for the rapid determination of reducing sugars in potatoes.

Non-enzymatic electrochemical sensors for the monitoring of reducing sugars in foods has great potential as a rapid in-situ detection method. This development involved the assembly of a nanoporous platinum structure on a screen-printed carbon electrode (SPCE). The modified electrode was then employed as an amperometric sensing element in a flow injection analysis (FIA) manifold. The system was successfully applied to the rapid detection of reducing sugars in potatoes, without the need for sample preparation. Optimal signals were achieved in phosphate buffer (pH 7.4) at a flow rate of 0.5 mL min-1 and an applied potential of 0.6 V. Experimental results demonstrated the sensor's long-term stability and high selectivity for reducing sugars. This method provides high sample throughput due to a rapid response time of less than five seconds. Reducing sugar values determined were in good agreement with those recorded using a commercially available enzymatic assay kit.

Determination of Nalbuphine Hydrochloride in Pharmaceutical Formulations Using Diperiodatoargentate(III)-Rhodamine-B Chemiluminescence System by Flow Injection Analysis.

A novel method for the analysis of nalbuphine hydrochloride (NAL) is reported based on its enhancement effect on a diperiodatoargentate(III)-rhodamine-B (Ag(III) complex-Rh-B) chemiluminescence (CL) system in an aqueous sulfuric acid medium using flow-injection analysis (FIA). The optimal conditions of the CL reaction were: sulfuric acid, 10-2 M; Ag(III) complex, 2.0 × 10-4 M; Rh-B, 2.0 × 10-5 M; Brij-35, 0.01%; sample loop volume, 300 µL; and flow rate, 3.0 mL/min/stream. The limit of detection (LOD) and limit of quantification (LOQ) were 0.001 and 0.003 mg/L (S/N = 3 and 10); linear calibration range, 5 × 10-3 - 5.0 mg/L (R2 = 0.9999) and injection throughput, 150/h. The relative standard deviation (RSD) was from 0.8 - 3.2% over the range studied. The suggested technique was applied for the determination of NAL in pharmaceutical injections, compared with a reported spectrophotometric method, and obtained results were found to be satisfactory. Based on spectrophotometric studies, the most probable mechanism of the CL reaction has been briefly described and drawn schematically.

A preliminary study of rapid-fire high-throughput metabolite analysis using nano-flow injection/Q-TOFMS.

In this study, we demonstrated nano-flow injection analysis (nano-FIA) with quadrupole time-of-flight mass spectrometry (Q-TOFMS) for 17 highly polar intermediates produced during glycolysis, the tricarboxylic acid (TCA) cycle, and the pentose phosphate pathway (PPP). We optimized the analytical conditions for nano-flow injection/Q-TOFMS, and set the flow rate and ion source temperature to 1000 nL/min and 150 °C, respectively. Under optimal conditions, a single run was finished within 3 min, and the RSD value of 50 sequential injections was 4.2%. The method also showed quantitativity of four stable-isotope-labeled compounds (r2 > 0.99), demonstrating its robustness, high repeatability, and specificity. In addition, we compared three sample-preparation methods for rodent blood samples and found that protein precipitation with threefold methanol was the most effective. Finally, we applied the method to plasma samples from the serotonin syndrome (SS) model and control rats, the results of which were evaluated by principal component analysis (PCA). The two groups showed clearly separated PCA score plots, suggesting that the method could successfully catch the differences in metabolic profiles between SS and control rats. The results obtained from our new method were further validated by using the established gas chromatography/tandem mass spectrometry method, which demonstrated that there were good correlations between the two methods (R = 0.902 and 0.958 for lactic acid and malic acid, respectively, each at p < 0.001), thus proving the validity of our method. The method described here enables high-throughput analysis of metabolites and will be of use for the rapid analysis of metabolic profiles. Graphical abstract.

Automatic On-Line Purge-and-Trap Sequential Injection Analysis for Trace Ammonium Determination in Untreated Estuarine and Seawater Samples.

An innovative automatic purge-and-trap (P&T) system coupled with fluorimetric sequential injection (SI), for the on-line separation and preconcentration of volatile compounds, is presented. The truth of concept is demonstrated for the ammonium fluorimetric determination in environmental water samples with complex matrices without any pretreatment. The P&T flow system comprises a thermostated purge-vessel where ammonium is converted into gaseous ammonia and a trap-vessel for ammonia collection. This configuration results in matrix removal as well as analyte preconcentration, avoiding membrane-associated problems. All the main parameters affecting the efficiency of a P&T system were studied and optimized. The proposed method is characterized by a working range of 2.7-150.0 µg L-1 of NH4+, with a detection and quantification limit of 0.80 and 2.66 µg L-1, respectively, for a 10-mL sample consumption. The accuracy of the method was assessed by recovery assays in seawater, estuarine, and lake water samples as well as by the analysis of standard reference material.

Novel Approach to Sample Preconcentration by Solvent Evaporation in Flow Analysis.

A preconcentration module operated in flow mode and integrated with a sequential injection system with spectrophotometric detection was developed. Using the system, preconcentration was performed in continuous mode and was based on a membraneless evaporation process under diminished pressure. The parameters of the proposed system were optimized and the system was tested on the example of the spectrophotometric determination of Cr(III). The preconcentration effectiveness was determined using the signal enhancement factor. In the optimized conditions for Cr(III), it was possible to obtain the signal enhancement factors of around 10 (SD: 0.9, n = 4) and determine Cr(III) with precision and intermediate precision of 8.4 and 5.1% (CV), respectively. Depending on the initial sample volume, signal enhancement factor values of about 20 were achieved. Applicability of the developed preconcentration system was verified in combination with the capillary electrophoresis method with spectrophotometric detection on the example of determination of Zn in certified reference materials of drinking water and wastewater. Taking into account the enhancement factor of 10, a detection limit of 0.025 mg L-1 was obtained for Zn determination. Zn was determined with precision less than 6% (CV) and the results were consistent with the certified values.

Performing flow injection chromatography using a narrow open tubular column.

Flow injection chromatography (FIC) or sequential injection chromatography (SIC) is a low-pressure liquid chromatography technique that uses flow injection or sequential injection hardware. Due to the constraints of this hardware, the separation resolution is low; often no more than 3-5 components are resolved. We have recently demonstrated the excellent resolving power of narrow open tubular (OT) columns for various biomolecules, and only moderate elution pressures are needed to carry out these separations. In this paper, we incorporate a narrow OT column with FIC and construct an FIC system using a pressure chamber and two injection valves to implement gradient elution. The resultant system not only improves the resolution but also reduces the system cost. When we use the system to separate peptides from trypsin-digested cytochrome C, we can resolve dozens of peptides (with resolutions of 0.5 or greater) at a speed of 12 samples per hour. When we use this system to separate a mixture containing 3 amino acids, we can base-line resolve these compounds at a speed of 1800 sample per hour.

Fast determination of peroxymonosulfate by flow injection chemiluminescence using the Tb(III) ligand in micelle medium.

Based on the chemiluminescence (CL) phenomenon of peroxymonosulfate (PMS) and Tb(III) enhanced by its ligand in a micelle microenvironment, a fast and sensitive flow injection CL method for PMS detection was proposed and applied to the analysis of different samples and PMS decomposition. Under the optimized conditions, a linear range was obtained from 4.0 × 10-6  mol L-1 to 2.0 × 10-4  mol L-1 with a high correlation coefficient (r = 0.9997), detection limit of 5.0 × 10-7  mol L-1 (S/N = 3) and relative standard deviation of 2.4% for 1.0 × 10-5  mol L-1 PMS (n = 9). This was successfully applied to the determination of PMS in Virkon powder, tap water, and swimming pool water samples with satisfactory recoveries from 94.8% to 104.8%. In particular, the analytical frequency could be as fast as five samples per minute because there was no reaction step before analysis and the CL phenomenon was instantaneous. Therefore, this CL method has also been successfully applied to investigate the PMS decomposition profiles in carbon material (carbon nanotubes, carbon nanofibres, activated carbon and graphene oxide) catalysis systems, which followed pseudo-first-order kinetics with good correlation coefficients (r > 0.9305). Quenching experiments and electron spin resonance spectra verified that the CL phenomenon was due to the formation of singlet oxygen, and that hydroxyl and sulfate radicals might be important in the generation of singlet oxygen. Tb(III) is the luminescent emitter according to the characteristics emission bands of the fluorescence and CL spectra in different media.

A Decoupled Automation Platform for Hydrogen/Deuterium Exchange Mass Spectrometry Experiments.

Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a biophysical technique well suited to the characterization of protein dynamics and protein-ligand interactions. In order to accurately define the rate of exchange, HDX experiments require the repeated measure of deuterium incorporation into the target protein across a range of time points. Accordingly, the HDX-MS experiment is well suited to automation, and a number of automated systems for HDX-MS have been developed. The most widely utilized platforms all operate an integrated design, where robotic liquid handling is interfaced directly with a mass spectrometer. With integrated designs, the exchange samples are prepared and injected into the LC-MS following a "real-time" serial workflow. Here we describe a new HDX-MS platform that is comprised of two complementary pieces of automation that disconnect the sample preparation from the LC-MS analysis. For preparation, a plate-based automation system is used to prepare samples in parallel, followed by immediate freezing and storage. A second piece of automation has been constructed to perform the thawing and LC-MS analysis of frozen samples in a serial mode and has been optimized to maximize the duty cycle of the mass spectrometer. The decoupled configuration described here reduces experiment time, significantly improves capacity, and improves the flexibility of the platform when compared with a fully integrated system.

Multiple signal amplification chemiluminescence immunoassay for chloramphenicol using functionalized SiO2 nanoparticles as probes and resin beads as carriers.

A novel, rapid and convenient competitive immunoassay for ultrasensitive detection of chloramphenicol residues in shrimp and honey was established combined with flow injection chemiluminescence. The carboxylic resin beads were used as solid phase carriers to load with more coating antigen due to their larger specific surface area and good biocompatibility. The surface of the silica dioxide nanoparticles was modified with aldehyde group to combine with more horseradish peroxidase and the chloramphenicol antibody. There was a competitive process between the chloramphenicol in solution and the immobilized coating antigen to combine with the limited binding site of antibody to form the immunocomplex. Silica dioxide nanoparticles played an important role in enhancing chemiluminescence signal, because the horseradish peroxidase on SiO2 effectively catalyzed the system of luminol-PIP-H2O2. Under optimal conditions, the chemiluminescence intensity decreased linearly with the logarithm of the chloramphenicol concentration in the range of 0.0001 to 100 ng mL-1 and the detection limit (3σ) was 0.033 pg mL-1. This immunosensor demonstrated acceptable stability, high specificity and reproducibility. The horseradish peroxidase-silica dioxide nanoparticle-chloramphenicol antibody complex successfully prepared in this article was firstly applied to the detection of chloramphenicol, and had extremely important meanings for the application of nanoparticles and enzymatic catalysis in the field of chemiluminescence.

Microfluidic sample delivery for serial crystallography using XFELs.

Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) is an emerging field for structural biology. One of its major impacts lies in the ability to reveal the structure of complex proteins previously inaccessible with synchrotron-based crystallography techniques and allowing time-resolved studies from femtoseconds to seconds. The nature of this serial technique requires new approaches for crystallization, data analysis, and sample delivery. With continued advancements in microfabrication techniques, various developments have been reported in the past decade for innovative and efficient microfluidic sample delivery for crystallography experiments using XFELs. This article summarizes the recent developments in microfluidic sample delivery with liquid injection and fixed-target approaches, which allow exciting new research with XFELs. Graphical abstract.

Ultra-High Resolution Elemental/Isotopic Mass Spectrometry (m/Δm > 1,000,000): Coupling of the Liquid Sampling-Atmospheric Pressure Glow Discharge with an Orbitrap Mass Spectrometer for Applications in Biological Chemistry and Environmental Analysis.

Many fundamental questions of astrophysics, biochemistry, and geology rely on the ability to accurately and precisely measure the mass and abundance of isotopes. Taken a step further, the capacity to perform such measurements on intact molecules provides insights into processes in diverse biological systems. Described here is the coupling of a combined atomic and molecular (CAM) ionization source, the liquid sampling-atmospheric pressure glow discharge (LS-APGD) microplasma, with a commercially available ThermoScientific Fusion Lumos mass spectrometer. Demonstrated for the first time is the ionization and isotopically resolved fingerprinting of a long-postulated, but never mass-spectrometrically observed, bi-metallic complex Hg:Se-cysteine. Such a complex has been implicated as having a role in observations of Hg detoxification by selenoproteins/amino acids. Demonstrated as well is the ability to mass spectrometrically-resolve the geochronologically important isobaric 87Sr and 87Rb species (Δm ~ 0.3 mDa, mass resolution m/Δm ≈ 1,700,000). The mass difference in this case reflects the beta-decay of the 87Rb to the stable Sr isotope. These two demonstrations highlight what may be a significant change in bioinorganic and atomic mass spectrometry, with impact expected across a broad spectrum of the physical, biological, and geological sciences. Graphical Abstract "".

Automatic Mesofluidic System Combining Dynamic Gastrointestinal Bioaccessibility with Lab-on-Valve-Based Sorptive Microextraction for Risk Exposure of Organic Emerging Contaminants in Filter-Feeding Organisms.

An automatic mesofluidic system combining dynamic oral bioaccessibility with lab-on-valve (LOV)-based sorptive microextraction is herein proposed for the first time for exploring the kinetics of leaching of incurred rather than spiked organic emerging contaminants (viz., methyl paraben, butyl paraben, diclofenac, and triclosan) from exposed mussels on the basis of the Versantvoort's fed-state physiological extraction test. Our method capitalizes on programmable flow analysis, in which gastrointestinal extracts are obtained online by pumping a simulated biorelevant gastrointestinal fluid across a large-bore column (maintained at 37.0 ± 2.0 °C) loaded with 250 mg of freeze-dried and powdered mussel onto a polyvinyldiene difluoride filter membrane. The physiologically relevant extracts are then cleaned up, and the analytes are preconcentrated onto a dedicated reversed-phase solid-phase extraction (Oasis PRIME-HLB) microcolumn that is captured into the channels of an LOV mesofluidic platform. The aim behind this is to obtain analyte-laden eluates with ACN/MeOH (90:10, v/v) in unsupervised mode for direct injection into LC-MS. The LOV minicolumn (≤25 mg) is automatically disposed of and renewed for every individual fraction on account of the strong retention of (phospho)lipids by the copolymeric sorbent. The proposed dynamic bioaccessibility test features a significant shortening of the extraction time against the batch method (28 vs 240 min) while avoiding overestimation of potentially bioavailable fractions. The trueness of the online gastrointestinal extraction method was confirmed using mass-balance validation following ultrasonic-assisted solid-liquid extraction of the original mussel sample and the residual (nonbioaccessible) fraction of emerging contaminants.

Strategies for sample delivery for femtosecond crystallography.

Highly efficient data-collection methods are required for successful macromolecular crystallography (MX) experiments at X-ray free-electron lasers (XFELs). XFEL beamtime is scarce, and the high peak brightness of each XFEL pulse destroys the exposed crystal volume. It is therefore necessary to combine diffraction images from a large number of crystals (hundreds to hundreds of thousands) to obtain a final data set, bringing about sample-refreshment challenges that have previously been unknown to the MX synchrotron community. In view of this experimental complexity, a number of sample delivery methods have emerged, each with specific requirements, drawbacks and advantages. To provide useful selection criteria for future experiments, this review summarizes the currently available sample delivery methods, emphasising the basic principles and the specific sample requirements. Two main approaches to sample delivery are first covered: (i) injector methods with liquid or viscous media and (ii) fixed-target methods using large crystals or using microcrystals inside multi-crystal holders or chips. Additionally, hybrid methods such as acoustic droplet ejection and crystal extraction are covered, which combine the advantages of both fixed-target and injector approaches.

Sample delivery for serial crystallography at free-electron lasers and synchrotrons.

The high peak brilliance and femtosecond pulse duration of X-ray free-electron lasers (XFELs) provide new scientific opportunities for experiments in physics, chemistry and biology. In structural biology, one of the major applications is serial femtosecond crystallography. The intense XFEL pulse results in the destruction of any exposed microcrystal, making serial data collection mandatory. This requires a high-throughput serial approach to sample delivery. To this end, a number of such sample-delivery techniques have been developed, some of which have been ported to synchrotron sources, where they allow convenient low-dose data collection at room temperature. Here, the current sample-delivery techniques used at XFEL and synchrotron sources are reviewed, with an emphasis on liquid injection and high-viscosity extrusion, including their application for time-resolved experiments. The challenges associated with sample delivery at megahertz repetition-rate XFELs are also outlined.

Flow injection tyrosinase biosensor for direct determination of acetaminophen in human urine.

An amperometric biosensor compatible with a flow injection analysis (FIA) for highly selective determination of acetaminophen (APAP) in a sample of human urine was developed. This biosensor is also suitable for use in the routine pharmaceutical practice. To prove this statement, two different commercially available pharmaceutical formulations were analyzed. This nano-(bio)electroanalytical device was made from a commercially available screen-printed carbon electrode covered by a thin layer of non-functionalized graphene (NFG) as amperometric transducer. A biorecognition layer was prepared from mushroom (Agaricus bisporus) tyrosinase (EC 1.14.18.1) cross-linked using glutaraldehyde, where resulting aggregates were covered by Nafion®, a known ion exchange membrane. Owing to the use of tyrosinase and presence of NFG, the developed analytical instrument is able to measure even at potentials of 0 V. Linear ranges differ according to choice of detection potential, namely up to 130 µmol L-1 at 0 V, up to 90 µmol L-1 at -0.1 V, and up to 70 µmol L-1 at -0.15 V. The first mentioned linear range is described by the equation Ip [µA] = 0.236 - 0.1984c [µmol L-1] and correlation coefficient r = 0.9987; this equation was used to quantify the content of APAP in each sample. The limit of detection of APAP was estimated to be 1.1 µmol L-1. A recovery of 96.8% (c = 25 µmol L-1, n = 5 measurements) was calculated. The obtained results show that FIA is a very selective method for APAP determination, being comparable to the chosen reference method of reversed-phase high-performance liquid chromatography.

Automatic determination of arsenate in drinking water by flow analysis with dual membrane-based separation.

The sequential application of a polymer inclusion membrane (PIM), composed of poly(vinylidenefluoride-co-hexafluoropropylene) and the anionic extractant Aliquat 336, and a microporous polytetrafluoroethylene (PTFE) gas-permeable membrane was utilized for the first time to develop a flow analysis (FA) system, for the automatic determination of trace levels of arsenate (As(V)) in drinking water as arsine. The system incorporated a flow-through extraction cell for separation and preconcentration of arsenate and a gas-diffusion cell for the separation of arsine prior to its spectrophotometric determination based on the discoloration of a potassium permanganate solution. Under optimal conditions the FA system is characterized by a limit of detection of 3.0 µg L-1 As(V) and repeatability of 1.8% (n = 5, 25 µg L-1 As(V)) and 2.8% (n = 5, 50 µg L-1 As(V)). The newly developed FA method was successfully applied to the determination of arsenate in drinking water samples in the µg L-1 concentration range.

Integration of Flow Injection Capillary Liquid Electrode Discharge Optical Emission Spectrometry and Microplasma-Induced Vapor Generation: A System for Detection of Ultratrace Hg and Cd in a Single Drop of Human Whole Blood.

A simple and miniature analytical system was developed to determine Hg and Cd in small amounts of samples by integrating flow injection capillary liquid electrode discharge (CLED) optical emission spectrometry (OES) and microplasma-induced vapor generation (PIVG) atomic fluorescence spectrometry (AFS). With the assistance of the inherent capillary driving force and the force arising from the solution vaporization in the microplasma, the sample solution was automatically transported into the discharge chamber wherein analytes were simultaneously excited to generate their atomic emission lines and converted to their volatile species. Subsequently, the volatile species were further swept into AFS for their further determination. Therefore, the same sample could be successively analyzed by OES and AFS. Owing to the unique independent linear-range and sensitivity of CLED-OES and PIVG-AFS, the developed system not only significantly extended its linear range to 6 orders of magnitude but also remarkably reduced the sample consumption to several microliters. Thus, wide linear-range and ultrasensitive determination of Hg and Cd in limited amounts of samples were accomplished simply by sharing one single capillary liquid electrode discharge source. Under the optimized conditions, limits of detection (LODs) of 10 µg L-1 were obtained for both Hg and Cd when CLED-OES was used as a detector, whereas the LODs for Hg and Cd were improved to 0.03 µg L-1 and 0.04 µg L-1 with AFS detector, respectively. In addition, the extremely wide linear-range of 0.001-100 mg L-1 and 0.001-40 mg L-1 were obtained for Hg and Cd, respectively. The potential application of this method was validated by successfully analyzing three Certified Reference Materials (ZK021-1, GBW(E)090033, and GBW(E)090034) and six human blood samples.

Simultaneous Colorimetric Measurements of Antioxidant Capacity by Flow Injection Analysis with Paired Emitter Detector Diode.

An effective flow injection analysis (FIA) system employing paired emitter detector diode (PEDD) for simultaneous dual antioxidant assays is proposed. The total antioxidant capacity (TAC) was measured using the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay in parallel with the ferric reducing antioxidant power (FRAP) assay. Ascorbic acid was used as the reference antioxidant and the TAC value reported as ascorbic acid equivalent (AAE). A 10-port valve furnished with two injection loops allowed the sample from one loop to react with the ABTS reagent and be detected by one PEDD. At the same time, the sample from the second loop reacted with the FRAP reagent and is detected at the second PEDD. A pair of red light emitting diodes (634 nm) was employed for both PEDDs. The linearity range was 10 to 50 µM ascorbic acid, with limit of detection of 3.18 and 4.37 µM, and precision of 2.41 and 2.15% RSD (for 30 µM ascorbic acid, n = 10) for the ABTS and FRAP assay, respectively. Sample throughput of 90 samples/h was achieved. The method was applied to the measurement of TAC of commercial fruit juice, instant tea products and vitamin C tablets. The observed AAE values were in good agreement with those obtained using batch methods with a spectrophotometer.

Fully automated method based on on-line molecularly imprinted polymer solid-phase extraction for determination of lovastatin in dietary supplements containing red yeast rice.

A fully automated method for the determination of lovastatin in dietary supplements containing red yeast rice has been developed. It uses a sequential injection analysis system combined with solid-phase extraction applying highly selective molecularly imprinted polymer sorbent. A miniaturized column for on-line extraction was prepared by packing 4.5 mg of the sorbent in a 5.0 × 2.5-mm-i.d. cartridge, which was used in the flow manifold. Sequential injection analysis manifold enabled all steps of lovastatin extraction and continuous spectrophotometric detection at 240 nm. A limit of detection of 60 µg g-1, a limit of quantitation of 200 µg g-1, and a linear calibration range of 200-2000 µg g-1 were achieved. Intra-day and inter-day precision values (RSD) were ≤ 6.7% and ≤ 4.9%, respectively, and method recovery values of spiked red yeast rice extracts at 200, 1000, and 2000 µg g-1 concentration levels were 82.9, 95.2, and 87.7%. Our method was used for determination of lovastatin lactone in four dietary supplements containing red yeast rice as a natural source of lovastatin, also known as monacolin K. The extracted samples were subsequently analyzed by the reference UHPLC-MS/MS method. Statistical comparison of results (F test, t test, α = 0.05) obtained by both methods did not reveal significant difference. A substantial advantage of the new automated approach is high sample throughput thanks to the analysis time of 7.5 min, miniaturization via down-scaling the extraction column, and smaller sample and solvent consumption, as well as reduced generation of waste. Graphical abstract ᅟ.

Multisyringe flow injection analysis (MSFIA) for the automatic determination of total iron in wines.

This work presents a multisyringe flow injection analysis (MSFIA) system for the automatic spectrophotometric determination of total iron in wine. The reaction is based on the complexation of Fe(II) with 2-(5-bromo-2-pyridylazo)-5-(diethylamino)-phenol (Br-PADAP). Ascorbic acid was used as reducing reagent for Fe(III) to Fe(II) and, in this way, to determine the total iron content in wine. The absorbance of the Fe(II)-(Br-PADAP)2 complex was measured at 748 nm. The proposed method provided a working rage from 0.36 to 5 mg L-1 of Fe(II), with a detection limit of 0.11 mg L-1 of Fe(II), a relative standard deviation of 0.42% (3 mg L-1 of Fe(II), n = 10), and a 46 h-1 injection throughput. The system is very simple, rapid and selective, and has been successfully applied to determine total iron in red, rosé and white wine without any need for sample pre-treatment steps. The results agree well with ICP-AES, which used as a reference method.