Multifunctional-separation-mode ion chromatography method for determining major metabolites during multiple parallel fermentation of rice wine.

Facile and effective analysis methods are desirable for elucidating the behaviours of metabolites during fermentation reactions. Herein, a multifunctional-separation-mode ion chromatography (MFS-IC) method was developed for the simultaneous monitoring of major metabolites during multiple parallel fermentation, including those related to central carbon metabolism (saccharification, glycolysis, alcoholic fermentation, and the tricarboxylic acid (TCA) cycle). The use of two types of sulfo-modified size-exclusion columns and phthalic acid as the eluent allowed the separation of oligosaccharides (disaccharides, trisaccharides, and tetrasaccharides), glucose, pyruvate, and major organic acids during the TCA cycle (cis-aconitate, citrate, iso-citrate, malate, fumarate, and succinate but not α-ketoglutarate) from other non-target analytes. The MFS-IC method was successfully applied to monitoring the major metabolites in the rice wine brewing process. This approach can contribute to an improved understanding of metabolite behaviour during fermentation without requiring the use of expensive advanced instrumentation methods such as liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry.

Development of a method for the simultaneous determination of ionic nutrients in hydroponic solutions using cation-/anion-exchange chromatography with a neutral eluent.

Nutrient availability in hydroponic solutions must be accurately monitored to maintain crop productivity; however, few cost-effective, accurate, real-time, and long-term monitoring technologies have been developed. In this study, we describe the development and application of cation-/anion-exchange chromatography with a neutral eluent (20-mmol/L sodium formate, pH 7.87) for the simultaneous separation (within 50 min) of ionic nutrients, including K+, NH4+, NO2-, NO3-, and phosphate ion, in a hydroponic fertilizer solution. Using the neutral eluent avoided degradation of the separation column during precipitation of metal ion species, such as hydroxides, with an alkaline eluent and oxidation of NO2- to NO3- with an acidic eluent. The suitability of the current method for monitoring ionic components in a hydroponic fertilizer solution was confirmed. Based on our data, we propose a controlled fertilizer strategy to optimize fertilizer consumption and reduce the chemical load of drained fertilizer solutions.

Reduction of the alcohol-stimulative taste of Japanese pot-distilled spirits.

The Japanese traditional pot-distilled spirit shochu has various tastes that are produced by variations in the manufacturing processes. In this study, an alcohol-stimulative taste was organoleptically evaluated using shochu samples, and the chemical components and proton nuclear magnetic resonance (1H NMR) spectra were measured. In some shochu samples, the alcohol-stimulative taste was weaker than that of the standard 15% (v/v) EtOH-H2O mixture, and the water-ethanol hydrogen-bonding structure was stronger compared to a water-ethanol solution. However, some shochu samples filtered with ion-exchange resin had a strong alcohol-stimulative taste comparable to that of the standard 15% (v/v) EtOH-H2O mixture, and the hydrogen-bonding structure was found to be similar to that of the water-ethanol solution. We also investigated the effect of MgCl2 on reducing the alcohol-stimulative taste, and it was observed most strongly with in shochu samples filtered with ion-exchange resin. The change in chemical shift values of the 1H NMR spectra was also the largest in ion-exchange resin filtered shochu samples. The sensory reduction in the alcohol-stimulative taste could be enhanced by the strengthening of the water-ethanol hydrogen-bonding structure. Shochu samples contained many components in larger quantities compared to vodkas. It was found that MgCl2 could reduce the alcohol-stimulative taste of shochu samples. Some salts, such as MgCl2, can be introduced into spirits through the water used to dilute the ethanol content before bottling the products. Our results indicated that some components, such as MgCl2, present in water used can reduce the alcohol-stimulative taste of different spirits produced worldwide.

Dynamic sequential extraction procedure using a four-channel circulating flow system for extracting Hg from soil samples.

Mercury (Hg) is harmful to the human body. Its physical properties and toxicity differ greatly depending on its chemical form. The quantification of soluble Hg in soils or sediments is crucial for preventing further environmental contamination by Hg sources such as products, processes, and storage. In this study, the risk of leachable Hg that release from soil to the hydrosphere was evaluated by the sequential extraction procedure (SEP), a speciation method of a targeted element based on the solubility of its various compounds. The SEP, which consists of a four-channel circulating flow system (FCFS), was developed to reduce the time and amount of liquid required for SEP of leachable Hg in the Bloom method, which is a conventional SEP for Hg in a solid sample. The SEP with FCFS was optimized by considering the impact of extractant volume, circulation extraction time, and flushing of the extraction line. In the three-step (Fraction 1-3) sequential extraction of leachable Hg in soil samples, the optimized SEP with FCFS required 45 mL of extractant and 2.5 h, while the common batch SEP required 75 mL of extractant and 49 h. Furthermore, the combination of the SEP with FCFS for the leachable forms (Fraction 1-3) and the batch method for the insoluble forms (Fraction 4-5) was applied to the five-step SEP of Hg from CRM-JSAC0403. The sum of Hg concentrations that were classified into five fractions was in good agreement with that obtained from the batch method for all extraction, indicating that there are no significant differences in the concentrations extracted from the four channels.

Retention of inorganic anions using mesoporous zirconia spheres modified with anion-exchange groups as the stationary phase for ion chromatography.

A zirconia (ZrO2) stationary phase with a chemically fixed silane coupling agent (trimethyl [3-(trimethoxysilyl)propyl]ammonium chloride; TMA), which possesses quaternary ammonium functional groups, is evaluated as a separation column for ion chromatography (IC) of anions. The selectivity for anions varies depending on the amount of TMA immobilized on the ZrO2. The TMA-ZrO2, with an anion-exchange capacity of 17 ± 3 µeq/g, shows an anion-exchange reaction that involves the specific retention of fluoride ion on ZrO2. The TMA-ZrO2 exhibits a decrease of the anion resolution with an increase of the eluent pH and an enhancement of the selective separation of fluoride ion with an increase of the column temperature. Through this study, the TMA-ZrO2 stationary phase shows potential as a new medium for ion separation.

Implementation of a conductivity cell electrode as an ion chromatography detector.

This technical note illustrates the possibility of using a conductivity cell electrode (CCE) as an ion chromatography (IC) detector to extend the application fields of this analytical technique. A conventional non-suppressed IC system consists of an eluent delivery pump, a separation column, column oven, and conductivity detector (CD). In this study, the conventional CD, which is one of the expensive parts of the instrument, is replaced with a relatively inexpensive CCE, leading to comparable peak resolution, detection sensitivity, and relative standard deviation. The separation effectiveness was retained and the developed IC-CCE system was successfully applied to the simultaneous separation of inorganic anions (SO42-, Cl-, and NO3-) and cations (Na+, NH4+, K+, Mg2+, and Ca2+) in three natural mineral water samples, with good accordance between the monitored values obtained using the CCE and CD. The commercially available CCE is potentially suitable for application as an IC detector for monitoring ionic components with overall IC cost reduction.

Simple mercury determination using an enclosed quartz cell with cold vapour-atomic absorption spectrometry.

A simple method for determining total inorganic mercury (Hg) in solution using an enclosed quartz cell applying cold vapour-atomic absorption spectrometry (CV-AAS) can reduce sample volume, reductant, and cost compared with those of official methods. The Hg determination under the optimised conditions achieves precision similar to that of conventional CV-AAS.

Simultaneous capillary electrophoresis of anions and cations in a single injection using an anion exchanger-modified capillary for determination of salivary ions in combination with statistical analyses.

This paper describes the simultaneous capillary electrophoresis (CE) of anions and cations using an anion exchange-modified capillary, which was prepared by chemical coating with a cationic silylating reagent, and its application to saliva analysis. The CE method provides high-throughput (5 min for a single sample injection) analysis by generating a high-velocity electroosmotic flow at pH 3.0-3.5. The detection limits at a signal-to-noise ratio of 3 ranged from 1.2 to 18 µM for anions and 1.0 to 2.7 µM for cations. The relative standard deviations for the migration times and peak areas of analytes (n = 4) ranged from 0.05% to 0.40% and 0.94% to 4.7%, respectively. The CE system was used to analyze 11 common ions in saliva samples collected from long-distance runners and sedentary university students before and after running for a set distance or a set time. Interestingly, the SCN- concentrations decreased in the saliva samples of all 14 athletes and 16 sedentary students after running. Furthermore, when the concentrations of the analyzed ions were compared with that of cortisol as a typical stress marker by multiple regression analysis, SCN- and NO3- in saliva samples from the two subject groups strongly correlated with cortisol levels, as determined by an electrochemiluminescence immunoassay. This study improves our knowledge of both the analytical methodology for CE and statistical methods for identifying common ions that could be used as physical stress markers.

A label-free, direct solid-phase fluorimetric analysis of ochratoxin A in agricultural products with monoclonal antibody-immobilized monolith.

We established a method for directly measuring mycotoxin ochratoxin A (OTA) in foods by solid-phase fluorescence of monolith-immobilized antibodies. The antibody was introduced onto only one side of an 8 mm-diameter, 3 mm-thick monolith via covalently immobilized protein G. 4 µg (2.7 × 10-11 mol) of antibody was immobilized per one monolith. A maximum of 10 µg (2.4 × 10-11 mol) OTA adsorbed to the activated side of each monolith. The amount of OTA adsorbed and the fluorescence intensity showed good linearity in the range of 0.5-3 ng OTA. Loading the sample solution onto the non-antibody side on the monolith blocked the hydrophobic fluorescent matrices from reaching the immobilized surface of the antibody. The proposed method was able to detect 1 ng OTA/g in solid samples with complex matrices. Mean recoveries obtained at spiked concentration of 3 ng g-1 OTA/g were 78-90% with relative standard deviations of <7.9%.

Effects of solutes on the alcohol-stimulative taste of vodkas.

Quantitative analyses of chemical components and sensory tests were carried out on vodka samples to test for stimulative taste of ethanol. Proton nuclear magnetic resonance (1H NMR) spectra of the vodkas were measured with 600 MHz NMR. The effects of salts on the alcohol-stimulative taste were investigated for 15% (v/v) EtOH-H2O solution. 1H NMR spectroscopy results showed that a magnesium salt could reduce the stimulative taste of ethanol organoleptically and, at the same time, strengthen the hydrogen-bonding structure of water-ethanol, although the effect of the magnesium could not be clarified quantitatively in the products of vodka. It was also suggested that a change in the water-ethanol hydrogen-bonding structure could lead to a reduction in the stimulative taste of ethanol in vodka.

Selective Recovery of Copper(II) from Incineration Ash Produced from a Municipal Waste Incineration Facility Using a Flow-through Type Electrolysis Method.

Recovery of copper was attempted from municipal waste incineration ashes using acid extraction and electrodeposition by a flow-through type electrolysis cell (FE). Efficient extraction of copper from the incineration ash was obtained using 0.5 mol L-1 sulfuric acid, i.e., copper extraction rate: 78% from bottom ash and 88% from fly ash. The copper in the 10-fold diluted extractant injected into the FE was quantitatively adsorbed by applying -0.4 V and was quantitatively desorbed by eluting with nitric acid.

Utilization of Anion-exchange Guard Column as an Ion Chromatographic Column of Anions Including Application to Simultaneous Separation of Anions and Cations.

This study demonstrated that a guard column containing anion-exchange resin has the potential for use as a separation column for acid eluent. Specifically, a 1-cm long anion-exchange guard column with a 4.6-mm internal diameter provided good separation of monovalent inorganic anions, by elution of 8 mM tartaric acid or 4 mM malic acid. Using the guard column with acid eluent could be applied to evaluation of nitrite and nitrate ions in mountain and urban river water samples. When the guard column was connected in front of a cation-exchange separation column (15 cm long × 4.6 mm internal diameter) in a series, the system provided simultaneous separation of anions and cations in eluent of 8 mM tartaric acid and 0.5 mM 18-crown-6 ether by a single injection.

Dynamic Four-step Sequential Extraction Procedure Using a Four-channel Circulating Flow System for Extracting Cd, Cu, Pb, and Zn from Solid Environmental Samples.

A method for extracting metals from solid samples was developed. The system had four flow channels (each containing a column packed with a solid sample), a single-flow peristaltic pump, and eight six-port valves. An extractant was passed into each channel for a specified period, then the valves were closed. We evaluated the system by performing a four-step sequential extraction procedure to extract heavy metals from a lake sediment sample. The four extractants were, in order of use, magnesium chloride, ammonium acetate, hydroxylamine hydrochloride, and hydrochloric acid. The concentrations of the analytes extracted agreed well with the concentrations determined using the batch method. The system was also successfully used to analyze heavy metals in a soil sample from a parking lot and fly ash from a domestic waste disposal facility. The total amount of extractant required per sample using the system was two-thirds of the amount required using the batch method.

Single injection ion-exclusion/cation-exchange chromatography for simultaneous determination of organic/inorganic anions, inorganic cations, and ethanol in beer samples.

Multicomponent simultaneous analysis is important for management programs, which are required in beer industries because the beer constituents measure by combining several methods in the present. In response to a requirement, our research group developed single sample injection ion chromatography systems, which comprise ion-exclusion/cation-exchange chromatography (IEC/CEC) and post-column derivatization and show promise for simultaneously determining concentrations of organic and inorganic species and alcohol commonly found in beer. Optimal chromatographic resolutions for determining 17 different species in beer samples by IEC/CEC were obtained on a H+-formed weakly acidic cation-exchange resin column with an eluent comprising 2 mM phthalic acid and 1 mM 18-crown-6. Consequently, the usefulness of developed method for monitoring beer samples was demonstrated in terms of beneficial information such as dependency of K+ concentration on the malt amount, influences of organic anion concentrations on different types of bottling methods, and validation of ethanol concentrations displayed in the ingredient table.

Direct quantitation of the preservatives benzoic and sorbic acid in processed foods using derivative spectrophotometry combined with micro dialysis.

The preservatives benzoic acid and sorbic acid are generally quantified with separation techniques, such as HPLC or GC. Here we describe a new method for determining these compounds in processed food samples based on a narrowness of the UV-visible spectral band width with derivative processing. It permits more selective identification and determination of target analytes in matrices. After a sample is purified by micro dialysis, UV spectra of sample solutions were measured and fourth order derivatives of the spectrum were calculated. The amplitude between the maximum and minimum values in a high-order derivative spectrum was used for the determination of benzoic acid and sorbic acid. Benzoic acid and sorbic acid levels in several commercially available processed foods were measured by HPLC and the proposed spectrometry method. The levels obtained by the two methods were highly correlated (r2>0.97) for both preservatives.

Simultaneous analysis of silicon and boron dissolved in water by combination of electrodialytic salt removal and ion-exclusion chromatography with corona charged aerosol detection.

Selective separation and sensitive detection of dissolved silicon and boron (DSi and DB) in aqueous solution was achieved by combining an electrodialytic ion isolation device (EID) as a salt remover, an ion-exclusion chromatography (IEC) column, and a corona charged aerosol detector (CCAD) in sequence. DSi and DB were separated by IEC on the H(+)-form of a cation exchange resin column using pure water eluent. DSi and DB were detected after IEC separation by the CCAD with much greater sensitivity than by conductimetric detection. The five-channel EID, which consisted of anion and cation acceptors, cathode and anode isolators, and a sample channel, removed salt from the sample prior to the IEC-CCAD. DSi and DB were scarcely attracted to the anion accepter in the EID and passed almost quantitatively through the sample channel. Thus, the coupled EID-IEC-CCAD device can isolate DSi and DB from artificial seawater and hot spring water by efficiently removing high concentrations of Cl(-) and SO4(2-) (e.g., 98% and 80% at 0.10molL(-1) each, respectively). The detection limits at a signal-to-noise ratio of 3 were 0.52µmolL(-1) for DSi and 7.1µmolL(-1) for DB. The relative standard deviations (RSD, n=5) of peak areas were 0.12% for DSi and 4.3% for DB.

Indirect UV detection-ion-exclusion/cation-exchange chromatography of common inorganic ions with sulfosalicylic acid eluent.

Herein, we describe indirect UV detection-ion-exclusion/cation-exchange chromatography (IEC/CEC) on a weakly acidic cation-exchange resin in the H(+)-form (TSKgel Super IC-A/C) using sulfosalicylic acid as the eluent. The goal of the study was to characterize the peaks detected by UV detector. The peak directions of analyte ions in UV at 315 nm were negative because the molar absorbance coefficients of analyte anions and cations were lower than that of the sulfosalicylic acid eluent. Good chromatographic resolution and high signal-to-noise ratios of analyte ions were obtained for the separations performed using 1.1 mM sulfosalicylic acid and 1.5 mM 18-crown-6 as the eluent. The relative standard deviations (RSDs) of the peak areas ranged from 0.6 to 4.9%. Lower detection limits of the analytes were achieved using indirect UV detection at 315 nm (0.23 - 0.98 µM) than those obtained with conductometric detection (CD) (0.61 - 2.1 µM) under the optimized elution conditions. The calibration curves were linear in the range from 0.01 to 1.0 mM except for Cl(-), which was from 0.02 to 2.0 mM. The present method was successfully applied to determine common inorganic ions in a pond water sample.

Utilization of a diol-stationary phase column in ion chromatographic separation of inorganic anions.

We describe the ion chromatographic separation of inorganic anions using a diol-stationary phase column (-CH(OH)CH(2)OH; diol-column) without charged functional groups. Anions were separated using acidic eluent as in typical anion-exchange chromatography. The retention volumes of anions on the diol-column increased with increasing H(+) concentration in the eluent. The anion-exchange capacities of diol-columns in the acidic eluent (pH 2.8) were larger than that of zwitterionic stationary phase column but smaller than that of an anion-exchange column. The separation of anions using the diol-column was strongly affected by the interaction of H(+) ions with the diol-functional groups and by the types of the eluents. In particular, the selection of the eluent was very important for controlling the retention time and resolution. Good separation was obtained using a diol-column (HILIC-10) with 5 mM phthalic acid as eluent. The limits of detection at a signal-to-noise ratio of 3 ranged from 1.2 to 2.7 µM with relative standard deviations (RSD, n=5) of 0.04-0.07% for the retention time and 0.4-2.0% for the peak areas. This method was successfully applied to the determination of H(2)PO(4)(-), Cl(-), and NO(3)(-) in a liquid fertilizer sample.

Water quality monitoring system for determination of ionic nutrients by ion-exclusion chromatography with spectrophotometric detection on cation- and anion-exchange resin columns using water eluent.

A unified ion-exclusion chromatography (IEC) system for monitoring anionic and cationic nutrients like NH4+, NO2-, NO3-, phosphate ion, silicate ion and HCO3- was developed and applied to several environmental waters. The IEC system consisted of four IEC methodologies, including the IEC with ultraviolet (UV) form connected with detection at 210 nm for determining NH4+ on anion-exchange separation column in OH anion-exchange UV-conversion column in I- form in tandem, the IEC with UV-detection at 210 nm for determining simultaneously NO3- and NO3- on cation-exchange separation column in H+ form, the IEC with UV-detection at 210 nm for determining HCO3- on cation-exchange separation column in H+ form connected with anion-exchange UV-conversion column in I- form in tandem, and the IEC with visible-detection based on molybdenum-blue reaction for determining simultaneously silicate and phosphate ions on cation-exchange separation column in H+ form. These IEC systems were combined through three manually-driven 6-port column selection valves to select each separation column to determine selectively the ionic nutrients. Using this sequential water quality monitoring system, the analytical performances such as calibration linearity, reproducibility, detection limit and recovery were also tested under the optimized chromatographic conditions. This novel water quality monitoring system has been applied successfully for the determination of the ionic eutrophication components in sub-urban river waters.

Recent progress and applications of ion-exclusion/ion-exchange chromatography for simultaneous determination of inorganic anions and cations.

One of the ultimate goals of ion chromatography is to determine both anions and cations found in samples with a single chromatographic run. In the present article, recent progress in ion-exclusion/ion-exchange chromatography for the simultaneous determinations of inorganic anions and cations are reviewed. Firstly, the principle and the control for the simultaneous separation and detection of analyte ions using ion-exclusion/cation-exchange chromatography with a weakly acidic cation-exchange column are outlined. Then, advanced chromatographic techniques in terms of analytical time, selectively and sensitivity are summarized. As a related method, ion-exclusion/anion-exchange chromatography with an anion-exchange column could be used for the simultaneous determination of inorganic nitrogen species, such as ammonium, nitrite and nitrate ions. Their usefulness and applications to water-quality monitoring and related techniques are also described.