Cto 20a column oven

NIL was measured by a HPLC method consisting of a LC-20AT Shimadzu pump, a DGU-20A Shimadzu degasser, an SIL-10AF auto sampler, a CTO-20A column oven, and an SPD-20A UV detector (HPLC, Shimadzu Corp., Kyoto, Japan). A quantity of 50 mM phosphate buffer/acetonitrile/methanol (50/25/25, v/v%) was applied as a mobile phase and flowed at 0.25 mL/min. An Inertsil^® ODS-3 column (2.1 × 50 mm) was used (GL Science Co., Inc., Tokyo, Japan), and the wavelength for detection was selected as 242 nm. In this study, 1 mg/mL propyl p-hydroxybenzoate was used as an internal standard. The measurement was performed at 35 °C using a column oven, and the samples (10 μL) were injected using a SIL-10AF. The measuring time was set at 16.5 min.

HPLC was performed on a Shimadzu LC-20AD separations module connected to a SIL-20A autosampler, a CTO-20A column oven, and an SPD-20AUV/visible detector (Shimadzu Corporation, Kyoto, Japan). A constant temperature shaker (Shanghai Tiancheng Experimental Instrument Manufacuring Co., Ltd., Shanghai, China) was used for static adsorption of macroporous resins. The HSCCC systems were comprised of a TBE-300A module, a TBP5002 constant-flow pump, and an ultraviolet (UV) monitor (TautoBiotechnique Co., Ltd., Shanghai, China). The TBE-300 Amodule was equipped with three polytetrafluoroethylene preparative coils (2.6 mm, i.d.; total volume, 300 mL) and a 20 mL sample injection loop, and the revolution speed could be adjusted in the range from 0 to 1000 rpm. A constant temperature regulator (HX-105, Beijing Changliu Science Implement, Beijing, China) was used to control the temperature of the separation coils. A CBS-A program-controlled automatic fraction collector (Shanghai Huxi analysis instrument factory Co., Ltd., Shanghai, China) was applied for collecting the effluent from HSCCC.

Residual glucose and xylose concentrations were determined using a high-performance liquid chromatograph (Shimadzu) equipped with an Aminex Fermentation Monitoring Column (Bio-Rad Laboratories, Hercules, CA, USA) and Micro-Guard Cation H Refill Cartridges in a Standard Cartridge Holder (Bio-Rad Laboratories). The detector was an RID 10A refractive index detector (Shimadzu). The column was held at 60°C using a CTO 20A column oven (Shimadzu). A sulfuric acid solution (5 mM) served as the mobile phase at a constant flow rate of 0.6 mL/min.

The GC analysis was carried out on a Shimadzu GC-2010 chromatography (Kyoto, Japan) with a flame ionization detector on a SUPELCOWAX 10 (Supelco, Bellefonte, PA, USA) capillary column (30 m × 0.25 mm × 0.25 μm). Carrier gas was He at 30 cm/s. The GC–MS analysis was performed with a Shimadzu CMS-QP5050A instrument (Kyoto, Japan) (electron impact at 70 eV) with a MDN-5s (Supelco, Bellefonte, PA, USA) capillary column (30 m × 0.25 mm ID). Carrier gas was He at 30 cm/s.
The HPLC–HRMS analysis of polar lipids was performed with a Shimadzu Prominence liquid chromatograph equipped with two LC-20AD pump units, a high pressure gradient forming module, CTO-20A column oven, SIL-20A auto sampler, CBM-20A communications bus module, DGU-20A3 degasser, and a Shim-Pack diol column (50 mm × 4.6 mm ID, 5 μm particle size) (Shimadzu, Kyoto, Japan). Lipids were detected by a high resolution tandem ion trap–time of flight mass spectrometry with a Shimadzu LCMS-IT-TOF instrument (Kyoto, Japan) operating both at positive and negative ion mode during each analysis at electrospray ionization (ESI) conditions. Ion source temperature was 200 °C, the range of detection was m/z 200–1600, and potential in the ion source was −3.5 and 4.5 kV for negative and positive modes, respectively. The drying gas (N₂) pressure was 200 kPa. The nebulizer gas (N₂) flow was 1.5 L/min.

Fifty microliters of sample was added to 100 µL of methanol containing 0.3 µg of benzophenone (internal standard) and filtered through a Chromatodisk 4A (pore size: 0.45 µm, Kurabo Industries Ltd., Osaka, Japan). Ten microliters of the filtrated solution was injected into an LC-20AT HPLC system using an SIL-10AF auto sampler (Shimadzu Corp., Kyoto, Japan). We used an Inertsil^® ODS-3 column (2.1 mm × 50 mm, GL Science Co., Inc., Tokyo, Japan), and the column temperature was set to 35 °C using a CTO-20A column oven (Shimadzu Corp., Kyoto, Japan). Here, 40% acetonitrile containing 0.1% trifluoroacetic acid was used as a mobile phase and the flow rate was adjusted to 0.25 mL/min, and the levels of DIS and DDC were detected at 215 nm in an SPD-20A UV detector (Shimadzu Corp., Kyoto, Japan).

HPLC was performed using an HPLC series Nexera X2 equipped with a SPD-M20A DAD set at 276 nm and CTO 20A column oven (Shimadzu, Japan). Chromatographic separation was carried out with an Atlantis^TM T3 column (3 μm, 4.6 × 150 mm) (Waters Co. Milford, CT, USA). The column temperature was set at 20 °C. The injection volume was set at 100 μL and the flow rate at 0.8 mL/min. Solvent A was H₂O and solvent B was acetonitrile (ACN). Gradient elution program was as follows: 0–13 min 5% B, 13–15 min 100% B, 15–20 min 5% B.

The LC-20AD UPLC system (Shimadzu Co., Kyoto, Japan) consisted of a LCMS-8030 (Shimadzu Co., Kyoto, Japan) triple quadrupole mass spectrometer equipped with LC-20AD pump (Shimadzu Co., Kyoto, Japan), CTO-20A column oven (Shimadzu Co., Kyoto, Japan), DGU-20A3 online degasser (Shimadzu Co., Kyoto, Japan), and SIL-20ACXR autosampler (Shimadzu Co., Kyoto, Japan). Furthermore, the tandem quadrupole mass spectrometer equipped with electrospray ionization (ESI) turbo ion interface was used with the following parameters: interface voltage (kV): 4.5; DL temperature (°C): 250; heat block temperature (°C): 400; nebulizing gas flow (L/min): 3; and drying gas flow (L/min): 15 and nitrogen was used in all cases.
A C18 column (ACQUITY BEH, 100 mm × 2.1 mm i.d., particle size 1.7 mm, Waters, Ireland) was used for UPLC separation. The mobile phase consisted of (A) methanol with 0.1% formic acid and (B) 5 mM NH₄OAC. The gradient elution program of mobile phase was as follows: 0-1 min: 25–70% A; 1–3.5 min: 70–90% A; 3.5–7.5 min: 90-90% A; 7.5–8.5 min: 90–25% A; and 8.5–13 min: 25-25% A, v/v. The flow rate for the mobile phase was set at 0.2 mL/min and the analytical volume was 5 μL of JWXYS extract.