Sequential injection analysis system for on-line monitoring of l-cysteine concentration in biological processes

A sequential injection analysis (SIA) system was developed to monitor the concentration of l-cysteine in biological processes on-line. It is based on the redox reaction of l-cysteine with iron(III) in the presence of 1,10-phenanthroline (phen) and the detection of the red-iron(II)-phen complex with a spectrophotometry. The system was fully automated using software written in the LabVIEW™ development environment. A number of system variables such as the flow rate of the carrier buffer solution, the volume ratio of the sample to the reagents, and the reaction coil length, etc., were evaluated to increase the sensitivity and performance of the SIA system. Under partially optimized operating conditions the performance of the SIA system was linear up to a concentration of l-cysteine of 1 mM (R² = 0.998) with a detection limit of 0.005 mM and a sample frequency of 15 hr⁻¹. The SIA system was employed to monitor the concentration of l-cysteine on-line in a continuously stirred reactor and a fermentation process of Saccharomyces cerevisiae. The on-line monitored data were in good agreement with the off-line data measured by a HPLC with a fluorescence detector (n = 15, R² = 09899).     

流动注射-分光光度法在线监测水中微量铝

建立了流动注射-分光光度法在线测定水中铝离子的方法,经过优化实验条件,方法的线性范围为0.01 ～0.6 mg/L,检出限为0.004 mg/L.本法试剂消耗量少,分析速度快,灵敏度高,抗干扰能力强,适宜于现场的即时检测,回收率在97.7％ ～101.7％之间.     

Sequential separation of actinides by ion chromatography coupled with on-line scintillation detection

The separation and measurement of actinides by ion chromatography (IC) coupled with on-line scintillation detection was investigated. Separation of (237)Np, (239)Pu, (241)Am, (244)Cm, (230)Th, and (233)U was accomplished using a Dionex CG5 guard column, CS5 separation column, and an elution program utilizing oxalic, diglycolic, and nitric acid eluents. Np(V) was eluted as a weak anionic oxalate complex or as free NpO(2)(+) cation, Am(III) and Cm(III) as anionic oxalate complexes, Pu(IV) and Th(IV) as nonionic diglycolate complexes, and U(VI) as a free uranyl cation. Response of the flow-cell detector was linear over the full range of activities tested (4-1500 Bq). The minimum detectable activities for the investigated analytes ranged from 3.6 Bq for (244)Cm to 5.4 Bq for (230)Th and (239)Pu. The corresponding minimum detectable concentrations based on a 1-L sample ranged from 3.6 to 5.4 KBq m(-3) (96 to 150 pCi L(-1)).     

An automated method of on-line extraction coupled with flow injection and capillary electrophoresis for phytochemical analysis

In this study, an automated system for phytochemical analysis was successfully fabricated for the first time in our laboratory. The system included on-line decocting, filtering, cooling, sample introducing, separation, and detection, which greatly simplified the sample preparation and shortened the analysis time. Samples from the decoction extract were drawn every 5 min through an on-line filter and a condenser pipe to the sample loop from which 20-μL samples were injected into the running buffer and transported into a split-flow interface coupling the flow injection and capillary electrophoresis systems. The separation of glycyrrhetinic acid (GTA) and glycyrrhizic acid (GA) took less than 5 min by using a 10 mM borate buffer (adjusted pH to 8.8) and +10 kV voltage. Calibration curves showed good linearity with correlation coefficients (R) more than 0.9991. The intra-day repeatabilities (n = 5, expressed as relative standard deviation) of the proposed system, obtained using GTA and GA standards, were 1.1% and 0.8% for migration time and 0.7% and 0.9% for peak area, respectively. The mean recoveries of GTA and GA in the off-line extract of Glycyrrhiza uralensis Fisch root were better than 99.0%. The limits of detection (signal-to-noise ratio = 3) of the proposed method were 6.2 μg/mL and 6.9 μg/mL for GTA and GA, respectively. The dynamic changes of GTA and GA on the decoction time were obtained during the on-line decoction process of Glycyrrhiza uralensis Fisch root.     

Sequential injection system for on-line analysis of total nitrogen with UV-mineralization

An automatic method for the determination of total nitrogen in wastewater by sequential injection analysis and mineralization with UV radiation has been developed. The method is based on the mineralization of the samples with sodium persulphate in basic medium under UV radiation. Small volumes of sample and reagents are firstly aspirated into a single channel and then propelled by flow reversal to the UV reactor and then to the detector. The organic and inorganic nitrogen compounds are oxidized to nitrate that is then measured at 226 nm. The sequential injection procedure has been optimized and the factors affecting the efficiency of the oxidation have been studied with a number of test substances with different chemical structures and properties. Solutions in the concentration range 1-56 g l⁻¹ of nitrogen can be analyzed with the described procedure. The sample rate is of 30-40 samples h⁻¹. The LOD is 0.6 mg l⁻¹ N and the reproducibility is 1.8% (28 mg l⁻¹ N). Organic carbon in the form of glucose was added to a number of test solutions to study the potential interference of organic matter.The method was compared with the Kjeldahl digestion method by analyzing 15 wastewater samples with both methods. The nitrate and nitrite content of the non-oxidized samples were subtracted from the corresponding nitrogen content determined after photo-oxidation and the value compared with the Kjeldahl nitrogen content.     

Biomass estimation using on-line glucose monitoring by flow injection analysis

Applied Biochemistry and Biotechnology - A new Flow Injection Analysis (FIA) system was developed that permits the on-line monitoring of glucose. This information may be used to identify the...     

Dielectrophoresis microsystem with integrated flow cytometers for on-line monitoring of sorting efficiency

Dielectrophoresis (DEP) and flow cytometry are powerful technologies and widely applied in microfluidic systems for handling and measuring cells and particles. Here, we present a novel microchip with a DEP selective filter integrated with two microchip flow cytometers (FCs) for on-line monitoring of cell sorting processes. On the microchip, the DEP filter is integrated in a microfluidic channel network to sort yeast cells by positive DEP. The two FCs detection windows are set upstream and downstream of the DEP filter. When a cell passes through the detection windows, the light scattered by the cell is measured by integrated polymer optical elements (waveguide, lens, and fiber coupler). By comparing the cell counting rates measured by the two FCs, the collection efficiency of the DEP filter can be determined. The chips were used for quantitative determination of the effect of flow rate, applied voltage, conductivity of the sample, and frequency of the electric field on the sorting efficiency. A theoretical model for the capture efficiency was developed and a reasonable agreement with the experimental results observed. Viable and non-viable yeast cells showed different frequency dependencies and were sorted with high efficiency. At 2 MHz, more than 90% of the viable and less than 10% of the non-viable cells were captured on the DEP filter. The presented approach provides quantitative real-time data for sorting a large number of cells and will allow optimization of the conditions for, e.g., collecting cancer cells on a DEP filter while normal cells pass through the system. Furthermore, the microstructure is simple to fabricate and can easily be integrated with other microstructures for lab-on-a-chip applications.     

Sequential injection lab-on-valve system for the on-line monitoring of hydrogen peroxide in lens care solutions

A sequential injection lab-on valve (SI-LOV) method for the enzymatic determination of hydrogen peroxide was developed. The spectrophotometric assay is based on the reaction between hydrogen peroxide and ABTS (2,2′-Azino-bis(3-Ethylbenzothiazoline 6-sulfonic acid)) in the presence of the enzyme HRP (horseradish peroxidase). The produced oxidized ABTS is measured at 410 nm. The sample consumption was 15 µL/assay and the consumption of HRP and ABTS was 34.6 mg L^{− 1} and 0.06 g L^{− 1}, respectively with a determination rate of 45 h^{− 1}.Relative deviations lower than 9.0% were found when the results were compared to those obtained by the reference procedure in the analysis of bleaches and disinfection solutions for contact lenses. With the incorporation of an in-line dilution (dialysis) process, was possible to attain a response range up to 342 mg L^{− 1} of hydrogen peroxide. The developed method was applied to monitor on-line of the disinfection–neutralization process of contact lenses. The study of two different one-step systems for cleaning contact lenses demonstrated that the neutralization of the hydrogen peroxide is completed within 6 h as recommended by the manufactures. The developed flow method was proved to be a useful tool for monitoring the dynamic process of disinfection–neutralization.     

Field measurement of nitrate in marine and estuarine waters with a flow analysis system utilizing on-line zinc reduction

A sensitive reagent-injection flow analysis method for the spectrophotometric determination of nitrate in marine, estuarine and fresh water samples is described. The method is based on the reduction of nitrate in a micro column containing zinc granules at pH 6.5. The nitrite formed is reacted with sulfanilamide and N-(1-naphthyl)ethylene diamine (Griess reagent), and the resulting azo compound is quantified spectrophotometrically at 520 nm. Water samples in the range of 3-700 μg L⁻¹ NO₃⁻-N can be processed with a throughput of up to 40 samples per hour, a detection limit of 1.3 μg L⁻¹ and reproducibility of 1.2% RSD (50 μg L⁻¹ NO₃⁻-N, n = 10). The proposed method was successfully applied for the determination of nitrate in estuarine waters and the reliability was assessed by the analyses of certified reference materials and recovery experiments. The method is suitable for waters with a wide range of salinities, and was successfully used for more than 3200 underway nitrate measurements aboard SV Pelican1 in the “Two Bays” cruise in January 2010.     

Monitoring of vitamin C species in aqueous solution by flow injection analysis coupled with an on-line separation with reversed-phase column

Two vitamin C species of ascorbic acid and dehydroascorbic acid in aqueous solution were monitored by flow injection analysis. Ascorbic acid and dehydroascorbic acid were resolved by a reversed-phase column, and dehydroascorbic acid was reduced to ascorbic acid by an on-line post-column reaction with dithiothreitol. Both natural and reduced ascorbic acids were photometrically detected at 260 nm, and the two vitamin C species were simultaneously determined. The determination range was from 0 to 8 × 10⁻⁵ M with a limit of detection of 1.7 × 10⁻⁶ M. The proposed method was applied to the conversion monitoring of ascorbic acid and dehydroascorbic acid in weakly acidic to weakly alkaline aqueous solutions, as well as to the determination of the vitamin C in some beverage samples.     

On-column polymer-imbedded graphite inlet electrode for capillary electrophoresis coupled on-line with flow injection analysis in a poly(dimethylsiloxane) interface

A method for coupling an electrophoretic driven separation to a liquid flow, using conventional fused-silica capillaries and a soft polymeric interface is presented. A novel design of the electrode providing high voltage to the electrophoretic separation was also developed. The electrode consisted of a conductive polyimide/graphite imbedded coating immobilized onto the capillary electrophoresis (CE) column inlet. This integrated electrode gave the same separation performance as a commonly used platinum electrode. The on-column electrode also showed good electrochemical stability in chronoamperometric experiments. In addition, with this electrode design, the electrode position relative to the inlet end of the CE column will always be constant and well defined. The on-line flow injection analysis (FIA)-CE system was used with electrospray ionization (ESI)-time of flight (TOF)-mass spectrometry detection. The preparation of the PDMS (poly(dimethylsiloxane)) interface for FIA-CE is described in detail and used for initial tests of the on-column polymer-imbedded graphite inlet electrode. In this interface, a pressure-driven liquid flow, a make up CE electrolyte and a CE column inlet meet in a two-level cross (95 microm ID) in the PDMS structure, enabling independent flow characterization.     

High-throughput dynamic microwave-assisted extraction on-line coupled with solid-phase extraction for analysis of nicotine in mushroom

A simple and low-cost high-throughput dynamic microwave-assisted extraction (HTDMAE) device was firstly assembled and validated by the extraction of nicotine in mushroom samples. In this device, a household microwave oven was applied to provide the microwave energy; a vacuum pump was used to deliver the solvent. Compared with traditional dynamic microwave-assisted extraction method, the sample throughput and microwave energy utilization were improved by the HTDMAE, up to 20 samples could be treated simultaneously in 9 min. Taking extraction of nicotine in mushroom sample as an example, a method was established with extraction, separation and enrichment of nicotine in a single step by the device on-line coupled with solid-phase extraction (SPE). Nicotine was first extracted from the mushroom samples with water under the action of microwave energy, and then directly introduced into the SPE column which was packed with cation-exchange resins. Subsequently, the nicotine trapped on the resins was eluted with methanol-ammonia (95:5, v/v) and determined by high-performance liquid chromatography. The limit of detection of nicotine obtained is 5.6 μg kg(-1) in fresh mushroom sample. The recovery of nicotine in mushroom samples is in the range of 87.4-104.0%. The proposed method which significantly reduced the overall analysis time and increased sample throughput should be favored for routine analyse of complex solid sample.     

Capillary electrophoresis on-line coupled with hydride generation-atomic fluorescence spectrometry for speciation analysis of selenium

A new method for speciation analysis of two inorganic selenium species was developed by on-line coupling of capillary electrophoresis (CE) with hydride generation-atomic fluorescence spectrometry (HG-AFS) and on-line conversion of Se(VI) to Se(IV). Baseline separation of Se(VI) and Se(IV) was achieved by CE in a 50 cm x 75 microm inside diameter (ID) fused-silica capillary at -20 kV using a mixture of 15 mmol.L(-1) NaH2PO4 and 0.5 mmol.L(-1) cetyltrimethylammonium bromide (pH 7.5) as electrolyte buffer. Se(VI) was on-line reduced to Se(IV) by mixing the CE effluent with concentrated HCl. The precision (relative standard deviation, RSD, n=7) ranged from 0.7 to 1.3% for migration time, 6.4 to 3.7% for peak height response, and 5.9 to 6.1% for peak area for the two selenium species at the 500 microg.L(-1) (as Se) level. The detection limits were 33 and 25 microg.L(-1) (as Se) for Se(VI) and Se(IV), respectively. The recoveries of the two selenium species in five locally collected water samples ranged from 88 to 114%. The developed method was applied to speciation analysis of inorganic selenium species in spiked natural water samples.     

Lab-on-valve (LOV) system coupled to irreversible biamperometric detection for the on-line monitoring of catechol

The analytical performance of lab-on-valve (LOV) system using irreversible biamperometry for the determination of catechol was evaluated. By integrating miniaturized electrochemical flow cell (EFC) designed and processed which is furnished with two identical polarized platinum electrodes, into the LOV unit, the lab-on-valve system combines sampling with analysis, realizing automated on-line analysis for catechol in a closed system. The biamperometric detection system was established to record the relationship between oxidation current and time by coupling the irreversible oxidation of catechol at one pretreated platinum electrode with the irreversible reduction of platinum oxide at the other pretreated platinum electrode. Factors influencing the analytical performance were optimized, including the potential difference (ΔE), buffer solution and pH, and flow variables in the LOV. A linear calibration curve was obtained within the range of 1.0 × 10⁻⁶-5.0 × 10⁻⁴ mol L⁻¹ of catechol with the detection limit (3σ) of 5.09 × 10⁻⁷ mol L⁻¹. The relative standard deviation (R.S.D.) was 2.39% for 11 successive determinations of 1 × 10⁻⁵ mol L⁻¹ catechol and the sample throughput was 35 h⁻¹. Moreover, this proposed method was applied to the analysis of catechol in beer sample, which was testified by high-performance liquid chromatography (HPLC).     

Simultaneous flow injection analysis for two components with a double on-line dialyser

Summary An on-line double membrane dialyser was designed and incorporated into the manifold of a flow injection system for the simultaneous determination of more than one component in a single sample injection. A fast and reliable fully automated two-component flow injection procedure is described in which a single sample (50 l of industrial effluent water) is injected into a carrier stream and simultaneously dialysed for the determination of chloride and calcium. The results obtained for the chloride and calcium in industrial effluent water at a sampling rate of 90 per hour compared well with those obtained by standard methods.Dedicated to Professor Dr. Wilhelm Fresenius on the occasion of his 80th birthday     

High-performance liquid chromatographic determination of phenolic compounds in natural water coupled with on-line flow injection membrane extraction-preconcentration

A liquid chromatographic method for determination of trace phenolic compounds has been established, coupled with an on-line supported liquid membrane extraction-preconcentration flow-injection system. Tributyl-phosphate dissolved in kerosene was used as the carrier of the supported liquid membrane. Four phenolic compounds (phenol, catechol, resorcinol and hydroquinone) were chosen as the model compounds and the experiment conditions were optimized. Under the optimum conditions, calibrations were linear in the range of 1-500 microg/L, with good correlation coefficients (r > 0.999). The total analysis time of the system was 22 min, including the membrane extraction, liquid chromatographic separation and equilibration times.     

New membraneless vaporization unit coupled with flow systems for analysis of ethanol

This work presents the development of a new design for a membraneless vaporization (MBL-VP) unit, called dual chamber MBL-VP for measurement of volatile compounds. With this unit, exact volumes of sample and reagent are introduced into their respective cone-shaped chambers from the base of the cones. Diffusion of volatile analyte then takes place. After an appropriate time interval, the acceptor solution is withdrawn from the chamber into the detector flow-cell, while the sample solution is withdrawn to waste. Unlike the previous MBL-VP design, problems with overflow of solutions are eliminated by precise control of the input volume to be less than the volume of the chamber. The developed flow system with the dual chamber MBL-VP unit was applied to the determination of the ethanol content of various liquid samples, using the oxidation reaction between potassium dichromate and the diffused ethanol. In addition, in order to accelerate the gas diffusion process, the donor chamber was aerated. As the result, relatively short analysis time of 144 s was achieved for ethanol content in the range of 5–50% (v/v). The proposed method was successfully validated against a gas chromatographic method for 17 alcoholic samples. Percentage recovery was in the range of 96–109%.     

A continuous liquid-liquid extraction system coupled on-line with capillary gas chromatography for environmental and ecotoxicological analysis

The applicability of a liquid-liquid extraction system which is coupled on-line with a capillary gas chromatograph was studied for environmental and ecotoxicological analyses. The optimized and automated system was used for the determination of apolar and rather non-volatile organic compounds in aqueous samples. Relevant aspects of sample introduction, phase separation and selection of extraction solvent are discussed. Three routine-tipe applications are described, viz. an improved method for the determination of hexachlorocyclohexanes in ground water and the determination of the so-called NCC-ether and ACC-ether in ecotoxicological studies. Depending on the application studied, the concentration levels varied from 0.1 to 6000 μg/I, using ECD and/or FID detection. Typical coefficients of variation obtained with the total extraction–GC procedure were 2–25%. The system was found to be rugged, it saves time compared with set-ups involving off-line liquid-liquid extraction and considerably reduces the manual work load.     

Performance of capillary gel electrophoretic analysis of oligonucleotides coupled on-line with electrospray mass spectrometry

Synthetic oligonucleotides (ODNs) are routinely analyzed using capillary gel electrophoresis (CGE) for size-sieving based separations as well as electrospray mass spectrometry (ESI-MS) for identification. On-line coupling of these methods is therefore desired in order to combine the analytical capabilities provided by both methods. Performance of on-line CGE-ESI-MS systems is influenced by various parameters, and choice of optimal conditions is crucial for successful coupling experiments. In this study, we explore characteristics of the on-line coupled CGE-ESI-MS system for ODN analysis. Effects of CGE buffer concentration, capillary length, separation and orifice voltage on CGE separation and MS detection of a phosphodiester ODN mixture were examined. Attention was paid to the influence of the interface, such as geometry of capillary alignment, sheath liquid flow-rate and sheath liquid composition on performance of the system.