An aliquot of the CHI–OA–CAP complex particle suspension was collected and diluted with methanol. The sample was centrifuged at 2100× g for 5 min (Micro Six MS-1, As One Corporation, Tokyo, Japan), followed by filtration using a syringe-connected membrane filter (0.45 µm pores; Dismic 13HP045AN, Advantec Toyo Kaisha, Ltd., Tokyo, Japan) for the removal of precipitate. The absorption spectra of the filtered solution were measured using an ultraviolet–visible (UV–Vis) spectrophotometer (UV-1800, Shimadzu Corporation, Kyoto, Japan). The CAP content was determined using the absorbance recorded at 280 nm by plotting the values on a standard curve preliminarily obtained using the amount of CAP dissolved in methanol. To determine the free CAP content in the matrix surrounding the CHI–OA–CAP complex particles, the matrix was separated from the suspension via centrifugal ultrafiltration at 10,000× g (Centrisart® I, molecular weight cut-off = 300,000, Sartrius AG, Göttingen, Germany).
Dismic 13hp045an
Manufactured by Advantec
Sourced in Japan
The Dismic 13HP045AN is a lab equipment product designed for specific tasks. It features core functionality to perform essential laboratory operations. The description of its intended use or performance capabilities is not available at this time.
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3 protocols using dismic 13hp045an
1
Quantitative Determination of CAP in CHI-OA-CAP Particles
2
HPLC Quantification of Taro Oxalates
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Oxalic acid was quantified using high‐performance liquid chromatography (HPLC).23 To extract soluble and total oxalates, taro powder was mixed with distilled water and 0.5 mol L–1 hydrochloride solution (20 mg mL−1). Samples were vortexed (Se‐08, Taitec, Saitama, Japan) and sonicated for 30 min each and centrifuged at 1600×g for 10 min (Model 3500, Kubota, Tokyo, Japan). The supernatant was collected and filtered through a 0.45 μm filter (DISMIC®‐13HP045AN, Advantec) and used as the sample for analysis. The HPLC unit comprised a pump (PU‐4180, Jasco, Japan), a detector (UV‐4075, Jasco, Tokyo, Japan), an autosampler (AS‐4050, Jasco, Tokyo, Japan), and a column oven (SLC‐25A, MEE, Japan). The HPLC analysis was performed using a C18 reversed‐phase column (Cosmosil 5C18‐PAQ, 4.6 mm I.D. × 250 mm, Nacalai Tesque, Kyoto, Japan) maintained at 40 °C. The eluent used was 20 mM phosphate buffer (pH 2.5) with a flow rate of 1.0 mL min−1, a detection wavelength at 254 nm, and an injection volume of 10 μL. The calibration curve was obtained using oxalic acid anhydride (Wako, Japan) as standards (0, 31.3, 62.5, 125, 250, 500, and 1000 μg mL−1, n = 3). Insoluble oxalate was calculated as the difference between the total and soluble oxalates.
3
Measurement of N2O Emissions in Bioreactors
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A sample was filtered through a 0.45-µm pore size membrane (DISMIC-13HP045AN; Advantec, Tokyo, Japan). DOC and total dissolved nitrogen (TDN) concentrations were measured by a TOC analyzer (TOC 5000A, Shimadzu, Kyoto, Japan). Nitrate (NO 3 -), nitrite (NO 2 -) and ammonium (NH 4 + ) concentrations were measured using a flow injection analyzer (PE-230, Human Manufacture Engineering, Japan). t-test was employed in SPSS 13.0 (IBM Co., New York, USA) to compare DOC and dissolved nitrogen concentrations in the effluents of the two reactors. Dissolved N 2 O concentration was measured using a Clark-type microelectrode (Unisense, Aarhus, Denmark). The N 2 O emission factor was estimated as the ratio of dissolved N 2 O concentration in the bulk liquid to the influent TDN concentration. This factor assumes negligible N 2 O exhaustion to the gas phase due to bubbleless aeration, which was confirmed from the gaseous N 2 O concentration in the ports for microelectrode insertion, in the gas compartment of the MABR, and in lumens of the hollow-fibers in the CBR.
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