Ascend tm 600 spectrometer

Bioactive compounds were isolated and identified following specific and distinctive steps, including bioassay-guided fractionation and isolation of the compounds from the primary methanol extract of S. hillii. Primary fractions were eluted with a solvent gradient consisting of 600 mL volume fractions with 10% (v/v) increments in each progressive step with dichloromethane (DCM) (Thermo Fisher Scientific, Scoresby, Victoria, Australia) and methanol using a glass column containing 50 g of silica gel (Merck, Darmstadt, Germany) 60 Å conditioned with hexane. Testing for bioactivity was carried out at each primary fraction processing step. Active primary fractions collected from S. hillii methanol extracts were separated by preparative and analytical HPLC C18 columns. The mobile phase consisted of (A) 0.05% v/v formic acid in milli-Q water and (B) methanol. Compound structures were identified by ¹H, ¹³C, and 2D NMR spectra (COSY, HSQC, and HMBC) recorded at 600 MHz on an Ascend^TM 600 spectrometer (Bruker, Thebarton, Australia). TopSpin^TM 4.0.6 software was used to analyze the NMR data. Finally, samples were analyzed on positive and negative ion modes with a range of m/z 100–800, at a scan rate of 0.5 Hz on a LTQ XLTM ion trap mass spectrometer (Thermo Scientific, Victoria, Australia) to determine and confirm the molecular weights of the structures.

The TBE-200V type-J high-speed counter-current chromatographic apparatus (Tautochem, Shanghai, China) was utilized for this study. The separation column was comprised of a 1.6 mm internal diameter polytetrafluoroethylene tube with a 190 mL capacity. For this experiment, the optimal coil speed was 800 rpm. The sample loop had a volume of 20 mL. An MP-0106 constant flow pump was used to pump the stationary and mobile phase into the coil. Chromatograms were recorded using a 21C-B detector (Shanghai Kanghua Biochemical Instrument Manufacturing Factory, Shanghai, China) and SEPU 3010 workstation (Dell, Round Rock, TX, USA). The temperature of the column was regulated with the use of an SDC-6 thermostatic bath (Hinotek, Ningbo, China), whilst sample components were collected utilizing a BSZ-100 (Jiapeng, Shanghai, China) automatic component collector.
High-performance liquid chromatography (HPLC) was conducted using a Shimadzu LC-20AT HPLC Labsolution system (Shimadzu, Kyoto, Japan), consisting of a LC-20AD quaternary pump, a CBM-20A system controller, an SIL-20A autosampler, a CTO-20AC temperature controller, and an SPD-20A detector.
Mass spectrometry (MS) was conducted using an AB SCIEX Triple TOF 5600+ system (AB SCIEX Pte. Ltd., Los Angeles, CA, USA). ¹H NMR and ¹³C NMR spectra were recorded on a Bruker Ascend TM 600 spectrometer (Bruker BioSpin, Rheinstetten, Germany).

The TBE-200V type-J high-speed counter-current chromatographic apparatus (Tautochem, Shanghai, China) was utilized for this study. The separation column was comprised of a 1.6 mm internal diameter polytetrafluoroethylene tube with a 190 mL capacity. For this experiment, the optimal coil speed was 800 rpm. The sample loop had a volume of 20 mL. An MP-0106 constant flow pump was used to pump the stationary and mobile phase into the coil. Chromatograms were recorded using a 21C-B detector (Shanghai Kanghua Biochemical Instrument Manufacturing Factory, Shanghai, China) and SEPU 3010 workstation (Dell, Round Rock, TX, USA). The temperature of the column was regulated with the use of an SDC-6 thermostatic bath (Hinotek, Ningbo, China), whilst sample components were collected utilizing a BSZ-100 (Jiapeng, Shanghai, China) automatic component collector.
High-performance liquid chromatography (HPLC) was conducted using a Shimadzu LC-20AT HPLC Labsolution system (Shimadzu, Kyoto, Japan), consisting of a LC-20AD quaternary pump, a CBM-20A system controller, an SIL-20A autosampler, a CTO-20AC temperature controller, and an SPD-20A detector.
Mass spectrometry (MS) was conducted using an AB SCIEX Triple TOF 5600+ system (AB SCIEX Pte. Ltd., Los Angeles, CA, USA). ¹H NMR and ¹³C NMR spectra were recorded on a Bruker Ascend TM 600 spectrometer (Bruker BioSpin, Rheinstetten, Germany).