HPLC was performed using cecal contents to determine the presence of 5-ASA and its primary metabolite, N-acetyl-5-ASA as previously described58 (link). The mobile phase consisted of 0.1% formic acid and acetonitrile containing 0.1% formic acid (50:50). Then, 20 mg of cecal contents were weighed and dissolved in 400 μL of the mobile phase. The samples were centrifuged at 17,800×g for 5 min at room temperature and filtered through a Cosmonice filter W (pore size 0.45 μm) (Nacalai Tesque, Kyoto, Japan). Chromaster 5430 (Hitachi High-Tech Corporation, Tokyo, Japan), Lichrospher 100 RP-18 endcapped (5 μm) LichroCART 150–4.6 (Merck KGaA, Darmstadt, Germany), and Manu-CART NT cartridge holders (Merck KGaA) were used for HPLC. As controls, 1 mg/ml solutions of 5-ASA (Sigma-Aldrich) and N-acetyl-5-ASA (Sigma-Aldrich) were used. The detection wavelength was set at 298 nm for 5-ASA and 313 nm for N-acetyl-5-ASA59 (link).
Chromaster 5430
Manufactured by Hitachi
Sourced in Japan
The Chromaster 5430 is a high-performance liquid chromatography (HPLC) system designed for analytical and preparative applications. It features a modular design, allowing for the integration of various components such as a pump, autosampler, column oven, and detector. The Chromaster 5430 is capable of handling a wide range of sample types and separation techniques.
Automatically generated - may contain errors
Lab products found in correlation
5 asa, by Merck Group (1 mentions)
Cosmonice filter w, by Nacalai Tesque (1 mentions)
R 220, by Büchi (1 mentions)
Cm 5000 series, by Hitachi (1 mentions)
Silica gel, by Cytiva (1 mentions)
Agilent 500 mhz dd2 spectrometer, by Agilent Technologies (1 mentions)
Sephadex lh 20, by Cytiva (1 mentions)
P 1020, by Jasco (1 mentions)
Pack ods a, by YMC (1 mentions)
J 815 spectropolarimeter, by Jasco (1 mentions)
4 protocols using chromaster 5430
1
Quantifying Gut 5-ASA and Metabolites
2
Quantifying Soy Isoflavone Composition
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The composition of soy isoflavones was carried out using an Agilent 1200 reversed phase High-performance liquid chromatography coupled with a diode-array detector (Hitachi, Chiyoda City, Japan, Chromaster 5430). A HIQ Sil C18W reversed-phase column was used (4.6 mm × 250 mm, 5 μm). The results were expressed in mg/100 g. Soy isoflavones (daidzin, daidzein, genistin, genistein, and cajanol) were measured according to the method presented in Vo et al. [11 (link)].
Biochanin A was detected as described by Lee et al. [23 (link)]. The mobile phase was: solvent A: 1% acetic acid in distilled water, and solvent B: acetonitrile. The flow rate was 1 mL/min. The gradients were: 0–35 min, 80–60% (A); 35–45 min, 65–0% (A); 50–51 min, 0–80% (A); and 51–60 min, 80% (A). The absorbance was measured at 278 nm, and the injection volume was 20 μL.
Biochanin A was detected as described by Lee et al. [23 (link)]. The mobile phase was: solvent A: 1% acetic acid in distilled water, and solvent B: acetonitrile. The flow rate was 1 mL/min. The gradients were: 0–35 min, 80–60% (A); 35–45 min, 65–0% (A); 50–51 min, 0–80% (A); and 51–60 min, 80% (A). The absorbance was measured at 278 nm, and the injection volume was 20 μL.
3
Hispidin Analysis in Sanghuang Mycelia
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Hispidin analysis was performed according to a previously reported method with slight modifications (Jang et al., 2010 ). In brief, 20 g powder was placed into 1,000 ml of absolute alcohol, filtered through Whatman filter paper No. 4, and concentrated through a rotary evaporator (R‐220, Büchi Labortechnik AG) to obtain dried ethanolic extract. The qualitative determination of hispidin in S. sanghuang mycelia ethanolic extract was carried out by a HPLC equipment (CM‐5000 series, Hitachi) connected to a Chromaster 5430 diode array detector (DAD). Hispidin was separated on a Kinetex column (4.6 × 150 mm, 5 μm), using a linear gradient of methanol and 0.1% formic acid, from 30:70 to 100:0 in 13 min followed by 30:70 in 2 min. The column temperature was set at 40°C, the flow rate was set at 1.0 ml/min, and chromatograms were processed at 370 nm.
4
Comprehensive Analytical Characterization of Natural Products
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Optical rotations were recorded on a JASCO P-1020 (JASCO Corporation, Tokyo, Japan) digital polarimeter. UV spectra were obtained on HITACHI Chromaster 5430 (HITACHI Corporation, Tokyo, Japan), while the ECD spectra were obtained on JASCO J-815 spectropolarimeter (JASCO Corporation, Tokyo, Japan). 1H NMR, 13C NMR, DEPT, and 2D NMR spectra were recorded on an Agilent 500 MHz DD2 spectrometer (Agilent Technologies Inc., Santa Clara, CA, USA). HRESIMS spectra were obtained using a Thermo Scientific LTQ Orbitrap XL mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). Column chromatography (CC) was achieved with silica gel (200–300 mesh, Qingdao Marine Chemical Inc., Qingdao, China) and Sephadex LH-20 (Amersham Biosciences, San Francisco, CA, USA). MPLC was done on a Bona-Agela CHEETAHTM HP100 (Beijing Agela Technologies Co., Ltd., Beijing, China). RP-HPLC was accomplished on an ODS column (HPLC (YMC-Pack ODS-A, 10 × 250 mm, 5 µm, 3 mL/min)) (YMC Co., Ltd., Kyoto, Japan).
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