Mci gel chp 20p

Optical rotations were measured with a Horiba SEPA-300 polarimeter (Horiba, Tokyo, Japan). UV spectra were obtained using a Shimadzu UV-2401A spectrophotometer (Shimadzu, Tokyo, Japan). IR spectra were obtained by a Tenor 27 spectrophotometer (Bruker Optics, Ettlingen, Germany) using KBr pellets. 1D and 2D NMR spectra were carried out on Bruker AM-400, DRX-500, or AVANCE III-600 spectrometers with TMS as an internal standard (Bruker, Karlsruhe, Germany). ESIMS and HRESIMS were run on an Agilent 6530 Q of spectrometer (Agilent, Palo Alto, CA, USA). Column chromatography (CC) was performed using MCI gel (CHP 20P, 75–150 m; Mitsubishi Chemical Corporation, Tokyo, Japan), Polyamide (PA, 80–100 mesh; Sinopharm Chemical Reagent Co., Ltd, Shanghai, China) and silica gel (100–200 or 200–300 mesh; Qingdao Haiyang Chemical Co., Ltd, Qingdao, China). Thin-layer chromatography (TLC) was conducted on silica gel plates GF₂₅₄ (Qingdao Haiyang Chemical Co., Ltd, Qingdao, China). Fractions were monitored by TLC using various solvent systems, and spots were visualized by spraying improved Dragendorff’s reagent to the silica gel plates or by heating silica gel plates sprayed with 10% H₂SO₄ in EtOH.

Optical rotations were recorded on a Rudolph Autopol IV-T polarimeter. UV spectra were recorded on a Hitachi U-2900 UV–Vis spectrophotometer. IR spectra were recorded on a ThermoFisher Nicolet iS5 FT-IR spectrometer. NMR spectra were acquired on Varian Mercury Plus 400 instrument and Bruker Avance III HD 600 spectrometer using CD₃OD (δ_H 3.31 and δ_C 49.0). HRESIMS were obtained on an AB SCIEX 5600+ Q-TOF mass spectrometer. MCI gel CHP-20P (75–150 μm, Mitsubishi Chemical Corporation, Tokyo, Japan), ODS gel (50 μm, YMC Co., Ltd., Japan), and Sephadex LH-20 gel (GE Healthcare Bio-Sciences, USA) were used for column chromatography. Precoated silica gel GF254 plates (Qingdao Haiyang Chemical Co., Ltd., China) were used for TLC analysis. Semi-preparative HPLC was performed on a Shimadzu Essentia LC-16 with a UV detector (210 and 254 nm) and a Kromasil C₁₈ column (150 × 10 mm, 5 μm, AkzoNobel, Co., Sweden).

UV spectra were measured using a Shimadzu UV-2401A spectrophotometer (Shimadzu, Tokyo, Japan). IR spectra were recorded on a Bruker Tensor 27 spectrophotometer (Bruker Optics, Ettlingen, Germany) with KBr pellets. 1D and 2D NMR spectra were carried out on Bruker AM-400 and DRX-500 or AVANCE III-600 and AV 800 spectrometers with TMS as an internal standard (Bruker, Karlsruhe, Germany). ESI-MS was run on an Agilent 6540 Q TOF spectrometer (Agilent, Palo Alto, CA, USA). HR-ESI-MS was performed using an Agilent G6230 TOF MS system (Agilent, Palo Alto, CA, USA). Semi-preparative HPLC was performed on an Agilent 1260 apparatus equipped with a diode-array detector and a Zorbax SB-C-18 (Agilent, 9.4 mm × 25 cm) column. Column chromatography was performed using MCI gel (CHP 20P, 75–150 mm; Mitsubishi Chemical Corporation, Tokyo, Japan), silica gel (100–200 or 200–300 mesh; Qingdao Marine Chemical Co. Ltd, Qingdao, China) and Sephadex LH-20 (GE Healthcare Bio-Sciences AB, Sala, Sweden). Thin-layer chromatography (TLC) was carried out on silica gel GF254 on glass plates (Qingdao Marine Chemical Inc.) and spots were visualized by heating silica gel plates sprayed with 10% H₂SO₄ in EtOH. All reactions sensitive to air or moisture were carried out under an argon or nitrogen atmosphere in dry and freshly distilled solvents under anhydrous conditions, unless otherwise noted.

Optical rotations were measured in MeOH with JASCO P-1020 polarimeters. 1D and 2D NMR spectra were recorded on Advance III 500 or 600 spectrometers using pyridine-d₅ as the internal standard. Chemical shifts (δ) are expressed in ppm relative to the pyridine-d₅ signals. HRESIMS was performed on an API QSTAR spectrometer. UV spectra were obtained on a Shimadzu UV-2401PC spectrophotometer. IR spectra were obtained on a Bruker Tensor-27 FT-IR spectrometer using KBr pellets. Column chromatography (CC) was performed with silica gel (100–200 mesh; Qingdao Marine Chemical, Inc., Qingdao, People’s Republic of China), MCI gel (CHP20P, 75–150 μm, Mitsubishi Chemical Corporation, Tokyo, Japan). Semi-preparative HPLC was performed on an Agilent 1200 liquid chromatograph with a Zorbax SB-C18 (9.4 mm × 250 mm) column. Fractions were monitored by thin layer chromatography, spots were visualized by UV light (254 nm and 365 nm) and by heating silica gel plates sprayed with 10% H₂SO₄ in EtOH. All solvents used in column chromatography were distilled including petroleum ether.

Optical rotations were measured on Jasco P-1020 or SGW_®-3 (INESA Instrument Co., Ltd., Shanghai, China) automatic digital polarimeter. UV data were obtained from HPLC online analysis. IR spectra (KBr) were obtained on a Bruker Tensor-27 infrared spectrophotometer. NMR spectra were carried out on a Bruker Avance III 600 or Bruker AV-400 (Bruker BioSpin GmbH, Rheinstetten, Germany) spectrometer with deuterated solvent signals used as internal standards. ESIMS and HRESIMS were measured using Bruker HCT Esquire 3000 and API QSTAR time-of-flight mass spectrometers, respectively. Silica gel (200–300 mesh, Qingdao Marine Chemical Inc., China), MCI gel CHP-20P (75–150 μm, Mitsubishi Chemical Corporation, Japan), Chromatorex C-18 (40–75 μm, Fuji Silysia Chemical Ltd., Japan) and Sephadex LH-20 (Amersham Biosciences, Sweden) were used for column chromatography. Fractions were monitored and analyzed using TLC, in combination with Agilent 1200 series HPLC system equipped by Extend-C18 column (5 μm, 4.6 × 150 mm). A TCI Chiral MB-S column (5 μm, 4.6 × 250 mm, Tokyo Chemical Industry Co., Ltd., Tokyo, Japan) was applied for determination of acid hydrolyzates using Agilent 1200 series HPLC system with an external Alltech 3300 ELSD detector (Grace, Deerfield, USA).

Optical rotations were measured on a Jasco P-1020 automatic digital polarimeter. UV data were obtained from HPLC online analysis. IR spectra (KBr) were obtained on a Bruker Tensor-27 infrared spectrophotometer. NMR spectra were carried out on a Bruker Avance III 600 or Bruker DRX-500 spectrometer with deuterated solvent signals used as internal standards. ESIMS and HRESIMS were measured using an Agilent G6230 time-of-flight mass spectrometer. MPLC was performed on a Büchi apparatus equipped with Büchi fraction collector C-660, Büchi pump module C-605 and manager C-615. Preparative HPLC separation was performed using an Agilent 1260 series HPLC system equipped with a Zorbax SB-C₁₈ column (5 μm, 21.2 × 150 mm). Silica gel (200–300 mesh, Qingdao Marine Chemical Inc., China), MCI gel CHP-20P (75–150 μm, Mitsubishi Chemical Corporation, Japan), Chromatorex C₁₈ (40–75 μm, Fuji Silysia Chemical Ltd., Japan) and Sephadex LH-20 (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) were used for column chromatography. Fractions were monitored and analyzed using TLC, in combination with Agilent 1200 series HPLC system equipped by an Extend-C₁₈ column (5 μm, 4.6 × 150 mm).

NMR spectra were performed on Bruker ascend 600 MHz and/or Bruker ascend 500 MHz NMR spectrometer (Bruker BioSpin AG Industriestrasse 26 CH-8117) with tetramethylsilane as an internal standard. Negative-ion mode ESI-Q-Orbitrap MS were determined on a Thermo ESI-Q-Orbitrap MS mass spectrometer connected to the UltiMate 3000 UHPLC instrument via ESI interface (Thermo Scientific). Optical rotations, UV, IR, and ECD spectra were run on a Rudolph Autopol^® IV automatic polarimeter (l = 50 mm) (Rudolph Research Analytical, Hackettstown), Varian Cary 50 UV-Vis (Varian, Inc.), Varian 640-IR FT-IR spectrophotometer (Varian Australia Pty Ltd.), and Circular dichroism spectrum (J-815, JASCO company), respectively.
Column chromatorgtaphies (CC) were accomplished on macroporous resin D101 (Haiguang Chemical Co., Ltd.), silica gel (48–75 μm, Qingdao Haiyang Chemical Co., Ltd.), ODS (50 μm, YMC Co., Ltd.), MCI gel CHP 20P (Mitsubishi Chemical Corporation, CHP20/P120), and Sephadex LH-20 (Ge Healthcare Bio-Sciences). HPLC column: Cosmosil 5C18-MS-II (4.6 mm i. d. × 250 and 20 mm i. d. × 250 mm) (5 μm, Nakalai Tesque, Inc.) were used to analysis and separate the constituents, respectively.
Dichloromethane (CH₂Cl₂), methanol (MeOH), acetonitrile (CH₃CN), acetic acid (HAc), and other reagents (chromatographically pure or analytical pure) were purchased from Tianjin Concord Technology Co., Ltd.