Spd 20a 20av series prominence hplc uv vis detectors

YH (25 g) was sequentially fractionated using solvents of different polarities. In particular, YH (25 g) was subjected to solvent fractionation using ethyl acetate (3 × 800 mL) and butanol (3 × 800 mL), and each fraction was concentrated and lyophilized. For the butanol fraction, the active compound was purified using reverse-phase (RP)-C18 silica gel (Merck, 230–400 mesh, 10% MeOH → 100% MeOH) and Sephadex LH-20 (Pharmacia, Uppsala, Sweden, 10% MeOH isocratic system, 100% MeOH for washing). The fraction obtained using the Sephadex LH-20 process was purified using an Agilent Eclipse C18 column (250 × 21.2 mm) and semi-preparative HPLC (SPD-20A/20AV Series Prominence HPLC UV-Vis Detectors; Shimadzu, Tokyo, Japan). Each fraction was confirmed using LC–MS and thin-layer chromatography (TLC). LC–MS was run using an analytical Kinetex^® 5 µm C18100 Å column (5 µm, 2.1 × 100 mm, Phenomenex, Torrance, CA, USA), while TLC was performed using silica gel F254 and RP-18 F254s plates from Merck. The results were detected either under UV light or by heating after spraying with anisaldehyde–sulfuric acid.

Optical rotations were calculated using a P-1020 polarimeter (Jasco, Easton, MD, USA). IR spectra were acquired on an IFS-66/S FT-IR spectrometer (Bruker, Madison, WI, USA). LC/MS analysis was performed on an Agilent 1200 Series HPLC system (Santa Clara, CA, USA) equipped with a diode array detector and a 6130 Series ESI mass spectrometer using a Kinetex C18 100 Å analytical column (100 mm × 2.1 mm i.d., 5 μm). NMR spectra were recorded on a UNITY INOVA 800 NMR spectrometer (Varian, Palo Alto, CA, USA) operating at 800 MHz (¹H) and 200 MHz (¹³C), with chemical shifts given in ppm (δ). Preparative high-performance liquid chromatography (HPLC) used a Waters 1525 Binary HPLC pump with a Waters 996 Photodiode Array Detector (Waters Corporation, Milford, CT, USA). Semi-preparative HPLC utilized a Prominence HPLC System with SPD-20A/20AV Series Prominence HPLC UV-Vis Detectors (Shimadzu, Tokyo, Japan). silica gel 60 (70–230 mesh and 230–400 mesh, Merck, Darmstadt, Germany) and RP-C18 silica gel (Merck, 40–63 μm) were used for column chromatography. Merck precoated silica gel F254 plates and RP-18 F254s plates were used for TLC. Spots were detected on TLC under UV light or by heating after spraying with anisaldehyde-sulfuric acid.

Specific rotations were measured on a Jasco P-1020 polarimeter (Jasco, Tokyo, Japan). IR spectra were recorded on a Bruker IFS-66/S FT-IR spectrometer (Bruker, Karlsruhe, Germany). UV spectra were acquired on an Agilent 8453 UV-visible spectrophotometer (Agilent Technologies, Milford, MA, USA). Liquid chromatography/mass spectrometry (LC/MS) analysis was performed on an Agilent 1200 Series HPLC System (Agilent Technologies) equipped with a diode array detector and a 6130 Series ESI mass spectrometer using an analytical Kinetex C18 100 Å column (100 mm × 2.1 mm i.d., 5 μm). NMR spectra were recorded on a Bruker AVANCE III 700 NMR spectrometer operating at 700 MHz (¹H) and 175 MHz (¹³C). A Shimadzu Prominence HPLC System with SPD-20A/20AV Series Prominence HPLC UV-Vis detectors (Shimadzu, Tokyo, Japan) was used for semi-preparative HPLC. Column chromatography was performed with silica gel 60 (Merck, 230–400 mesh, Frankfurt, Germany) and RP-C18 silica gel (Merck, 230–400 mesh). The packing material for molecular sieve column chromatography was Sephadex LH-20. Merck pre-coated silica gel F254 plates and reversed-phase (RP)-18 F254s plates were used for thin layer chromatography (TLC). Spots on TLC plates were detected under UV light or by heating the plates after spraying with anisaldehyde-sulfuric acid.

NMR spectra were recorded on a Varian UNITY INOVA 700 NMR spectrometer operating at 700 MHz (¹H) and 175 MHz (¹³C), with chemical shifts given in ppm (δ). Preparative high performance liquid chromatography (HPLC) using a Waters 1525 Binary HPLC pump with a Waters 996 Photodiode Array Detector (Waters Corporation, Milford, CT, USA) was also performed. Semi-preparative HPLC was conducted using a Shimadzu Prominence HPLC System with SPD-20A/20AV Series Prominence HPLC UV-Vis Detectors (Shimadzu, Tokyo, Japan). Silica gel 60 (Merck, 70–230 mesh and 230–400 mesh) and RP-C18 silica gel (Merck, 40–63 μm) were used for column chromatography. The packing material for molecular sieve column chromatography was Sephadex LH-20 (Pharmacia, Uppsala, Sweden).

Optical rotations were measured using a Jasco P-1020 polarimeter (Jasco, Easton, MD, USA). Infrared (IR) spectra were recorded using a Bruker IFS-66/S FT-IR spectrometer (Bruker, Karlsruhe, Germany). Electrospray ionization (ESI) mass spectra were recorded using a Waters Micromass Q-Tof Ultima ESI-TOF mass spectrometer (Waters, New York, NY, USA). Nuclear magnetic resonance (NMR) spectra were recorded using a Bruker AVANCE III 700 NMR spectrometer operating at 700 MHz (¹H) and 175 MHz (¹³C) (Bruker, Karlsruhe, Germany) with chemical shifts reported in parts per million (δ). Preparative HPLC used a Waters 1525 Binary HPLC pump with a Waters 996 Photodiode Array Detector (Waters Corporation, Milford, CT, USA). Semi-preparative HPLC was performed using a Shimadzu Prominence HPLC System with SPD-20A/20AV Series Prominence HPLC UV-Vis Detectors (Shimadzu, Tokyo, Japan). Silica gel 60 (Merck, 70–230 mesh and 230–400 mesh) and RP-C18 silica gel (Merck, 40–63 μm) were used for column chromatography. Merck precoated silica gel F254 plates and RP-18 F254s plates (Merck, Darmstadt, Germany) were used for thin layer chromatography (TLC). Spots were detected on TLC under UV light or by heating after spraying with anisaldehyde-sulfuric acid.