Primaide

Optical rotations were recorded using a Perkine Elmer 341 polarimeter (Hertford, UK). ECD spectra were recorded with a Chirascan circular dichroism spectrometer (Applied Photophysics, Surrey, UK). UV spectra were recorded on a UV-2600 UV–vis spectrophotometer (Shimadzu, Kyoto, Japan). The 1D and 2D NMR spectra were recorded on a Bruker AC 500 and 700 NMR (Broker, Fallanden, Switzerland) spectrometer with TMS as the internal standard. HRESIMS spectra were measured with a Bruker micro TOF-QII (Bruker, Fallanden, Switzerland) mass spectrometer in positive/negative ion mode. silica gel GF-254 (10–40 mm) was used for thin-layer chromatography (TLC) (Qingdao Marine Chemical Factory, Qingdao, China). Sephadex LH-20 (Amersham Biosciences, Uppsala, Sweden) and silica gel (200–300 mesh, 100–200 mesh) (Qingdao Marine Chemical Factory, Qingdao, China) were applied in column chromatography (CC). HPLC was carried out on a Hitachi Primaide with a YMC ODS Series column (YMC-Pack ODS-A, YMC Co. Ltd. (Kyoto, Japan), 250 × 10 mm i.d., S-5 μm, 12 nm). All solvents were analytical grade (Tianjin Fuyu Chemical and Industry Factory). The fermentation culture medium and reagents were obtained from Guangzhou Haili Aquarium Technology Company, Guangzhou, China.

The UV spectrum was recorded on a Shimadzu UV-2600 PC spectrometer (Shimadzu, Beijing, China). The IR spectrum was obtained using an IR Affinity-1 spectrometer (Shimadzu). Optical rotations were determined with an Anton Paar MPC 500 polarimeter. HRESIMS spectra were recorded with a Bruker maXis Q-TOF mass spectrometer. The NMR spectra were recorded on a Bruker Avance-500 spectrometer (Bruker BioSpin International AG, Fällanden, Switzerland), and chemical shifts were recorded as δ-values. Semipreparative high-performance liquid chromatography (HPLC) was performed on the Hitachi Primaide with a DAD detector, using an ODS column (YMC-pack ODS-A, 10 × 250 mm, 5 μm). Thin-layer chromatography analysis (TLC) and column chromatography (CC) were carried out on plates precoated with silica gel GF254 (10–40 μm) and over silica gel (200–300 mesh) (Qingdao Marine Chemical Factory, Qingdao, China) and Sephadex LH-20 (Amersham Biosciences, Uppsala, Sweden), respectively. Spots were detected on TLC (Qingdao Marine Chemical Factory) under 254 nm UV light. All solvents employed were of analytical grade (Tianjin Fuyu Chemical and Industry Factory, Tianjin, China).

The toxin peptide (MQKCQLFDYGGCRGNDNRFDSEEECLELCNLD) was synthesized by solid-phase polypeptide synthesis according to previously reported methods [52 (link),53 (link)]. Ct-kunitzin (10 mg) was dissolved in buffer containing (5 mM, GSH; 0.5 Mm GSSH; 0.1 M Tris-HCL; 0.1 M Nacl; pH = 9.0; 25 °C) for 24 h. Then the semipreparative high-performance liquid chromatography (HPLC) was performed on the Hitachi Primaide with a DAD detector, using an ODS column (YMC-pack ODS-A, 10 × 250 mm, 5 μm). On a C18 column using linear gradients of buffer A (0.1% trifluoroacetic acid; TFA; water) and buffer B (0.1% trifluoroacetic acid; TFA; acetonitrile). The mobile phase used a gradient of 5–50% buffer B in 45 min. The flow rate was 1 ml/min and the absorbance was monitored at 215 nm.

The photocatalytic degradation activity of the samples was measured using a 500 W Xenon lamp (CHF-XM500, PerfectLight, Beijing, China) with a 420 nm cutoff filter at room temperature. The target pollutant was 10 mg/L of BPA solution. In a typical degradation process for each experiment, 10 mg of photocatalyst powder were added into 50 mL of the aforementioned BPA solution, and the mixture was kept being stirred for 30 min, in order to achieve the adsorption–desorption equilibrium. At a specific time interval, 0.5 mL of solution was taken out from the reaction system and immediately centrifuged. After that, BPA concentrations of the obtained samples were measured using a high-performance liquid chromatography (HPLC, Primaide, Hitachi, Dalian, China). The temperature of the chromatographic column was 30 °C. The mobile phase was composed of deionized water, acetonitrile and formic acid with a volume ratio of 1000:1000:1, and the flow rate was set as 0.5 mL/min. All experiments were conducted twice. The intermediate compounds of BPA degradation were determined through Liquid Chromatograph Mass Spectrometer (LC-MS; LC-MS-2020, Shimadzu, Kyoto, Japan). The preliminary preparation of catalyst cyclic stability test was the same as the above. Centrifugation was performed after each degradation, and the obtained catalyst was used for the next degradation test, with a total of 5 cycles.

The roots of Gan and the flower of Ju were used in this study as herbal material. All herbs used in this study were of Chinese origin and identified by Prof. Mo Wang of the College of Plant Sciences and Technology of Huazhong Agricultural University, Wuhan, China. The standard contents of some compounds in herbal material used in this study were quantified by liquid chromatography (HPLC, Primaide, Hitachi Limited). The methods for detecting Gan and Ju by HPLC are shown in Tables 1,2. The injection volume was 20 μl, and the flow rate was kept at 0.8 ml/min. The C18 column (250 × 4.6 mm, 5.0 µm, Agilent, Santa Clara, CA, USA) was used for the chromatographic separations. The contents of the main components in the medicinal materials and aqueous extract of Gan and Ju are shown in Tables 4,5 and the HPLC profiles are shown in Supplementary Figures S1-S4. Compound standards liquiritin (99.5%), chlorogenic acid (3-CQA) (98%), 3,5-dicaffeoylquinic acid (3,5-diCQA) (98.9%), luteoloside (LUS) (99%), and glycyrrhizic acid ammonium salt (99.3%) were purchased from Chengdu Pusi Biotechnology Co., Ltd.

All solvents and reagents were purchased from commercial sources and used directly without further purification. The starting plant material was provided as a fine powder of two types of Chenopodium quinoa Willd. saponins (red and white quinoa), obtained by the company JIWRA SAC (Arequipa, Peru). High-Performance Liquid Chromatography (HPLC) was conducted on a Hitachi Primaide with a DAD detector Primaide 1430 and a Thermo Scientific™ Hypersil GOLD™ C18 Reversed Phase HPLC Column, 5 μm, 2.1 mm × 20 mm. The purity of all compounds was >95% as determined by HPLC. Methanol : acid water (89 : 11) was used as a mobile phase with a flow rate of 1.5 mL/min. The Thin Layer Chromatography (TLC) analysis was carried out on silica gel plates that were visualized under UV at 366 nm and sprayed with Liebermann–Burchard reagent before heating.

HPLC analysis GSSG formation was performed using
a Hitachi Primaide instrument equipped with a C18 column (Xbridge
Peptide BEH C18 column from Waters, 4.6 mm × 150 mm, pore size
300 Å, particle size 3.5 μm), using 0.1% aqueous TFA (solvent
A) and 90% CH3CN/0.1% TFA in water (solvent B) with a linear gradient
from 5% to 10% solvent B in 7 min.