Clc ods column

The content of sugars and xylitol were determined by high performance liquid chromatography (HPLC), using a Shimadzu system equipped with refractive index detector, Shimpack CLC-NH₂ column and acetonitrile/water (80/20) as the mobile phase at 1.0 mL.min^-1(30°C). Furfural and hydroxymethylfurfural (HMF) were quantified by high performance liquid chromatography (HPLC) using a Shimadzu system equipped with CLC-ODS column, kept at 40°C and with detector diode arrangement at a wavelength of 280 nm. The mobile phase was initially composed of water/methanol (95/5 p/v), andreached 100% of methanol after 13 min at a flow rate of 0.8 mL.min^-1.
Total phenolic compound content was determined using the Folin Ciocalteau reagent and expressed as g.L^-1 of ferulic acid [29 ].

The activity assay was performed by incubating the reaction mixture (1 mL) with 20 μg/mL of ZEN and the supernatant (containing approximately 147 μg Oxa) as a catalyst at 37 °C on a rotary shaker at 180 rpm for 12 h. The concentration of ZEN was analyzed using HPLC.
An HPLC analysis was performed using a Waters 600 system equipped with a four-element pump and assembled with fluorescence detectors (excitation wavelength: 275 nm and emission wavelength: 440 nm). A CLC-ODS column (Shimadzu; 250×4.6 mm i.d., particle size: 5 μg) was used to separate the mobile phase consisting of acetonitrile/water/methanol (46/46/8, v/v/v) at a flow rate of 0.9 mL/min. For HPLC, a column temperature of 30 °C and an injection volume of 20 μL were employed. The ZEN levels in the samples were calculated by comparing the area of the chromatographic peak of the sample with the standard curve, and the conversion rate was calculated using the following equation: $$\mathrm{ZEN} \mathrm{degradation} \mathrm{rate} \%=\left(1-\frac{\mathrm{remaining} \mathrm{concentration} \mathrm{of} \mathrm{ZEN}}{\mathrm{initial} \mathrm{concentration} \mathrm{of} \mathrm{ZEN}}\right)\times 100\%$$

The AX contents in mitochondrial and cytosolic fractions from Sol8 myotubes were prepared, as described previously [23 (link)]. Briefly, Sol8 myotubes were treated with AX (100 nmol) or DMSO, as control, in 10 mL culture medium/100 mm dishes and incubated at 37 °C with 5% CO₂ for 24 h. The cells were harvested to isolate the mitochondrial fraction as described in Section 2.4. After lyophilization, crude mitochondrial extracts and cytosol were solubilized in acetone and centrifuged at 12,000× g for 15 min. The supernatants were filtered using a 0.45 μm polytetrafluorethylene membrane and analyzed using HPLC and a spectrophotometer detector (JASCO, Tokyo, Japan) set at 460 nm. A Shim-pack CLC-ODS column (150 mm length and 6.0 mm internal diameter) was used.