Optical rotation indices were determined in methanol on a Rudolph Autopol II digital polarimeter (Rudolph, Hackettstown, NJ, USA). ESI-MS analysis was performed on a Quatro Premier instrument (Waters, Milford, MA, USA). HR-ESI-MS spectra were recorded on an Agilent Technologies 6550 Q-TOF (Santa Clara, CA, USA). 1D- and 2D-NMR spectra were recorded on Bruker-AVANCE 400 (Bruker, Rheinstetten, Germany) and Bruker-AVANCE 600 instrument (Bruker, Rheinstetten, Germany) using TMS as an internal standard. Analytical HPLC was performed on a Waters e2695 Separations Module system coupled with a 2998 Photodiode Array Detector and an Accurasil C-18 column (4.6 mm × 250 mm, 5 μm particles, Ameritech, Chicago, IL, USA). Semipreparative HPLC was performed on a system comprising an LC-6AD pump equipped with an SPD-20A UV detector (Shimadzu, Kyoto, Japan) and an Ultimate XB-C18 (10 mm × 250 mm, 5 μm particles) or YMS-Pack-ODS-A (10mm × 250 mm, 5 μm particles) column. Silica gel was purchased from Qingdao Haiyang Chemical Group Corporation (Qingdao, China).
E2695 separations module system
Manufactured by Waters Corporation
Sourced in United States
The E2695 Separations Module system is a laboratory equipment product manufactured by Waters Corporation. It is designed to perform chromatographic separations, which is a core function of the system. The E2695 Separations Module system is used for the analysis and purification of chemical samples in various applications.
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2 protocols using e2695 separations module system
1
Analytical Techniques for Natural Product Characterization
2
Protein Quantification via HPLC
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Separation method used was adapted from Ee, Zhao, Rehman, and Agboola (2009) and was carried out for protein quantification. Two different mobile phases were applied, mobile phase Awater and acid trifluoroacetic (TFA) (99.9:0.1, v/v) -and mobile phase Bacetonitrile and TFA (99.9:0.1, v/v),under the following conditions: gradient elution starts at 100% mobile phase A and ends at 45% after 45 min, at a continuous flow of 0.5 mL min -1 . Between 45 and 50 min the mobile phase A returns to 100% and remains at this percentage for 2 min (until 52 min). Detection was performed at a wavelength of 280 nm, peaks were analyzed and quantified using a calibration curve of pure protein (HPLC gradient) comparing retention time and spectra. The chromatographic analysis was performed using a Waters e2695 separations module system interfaced with a Photodiode array UV/Vis detector . Separation was performed in a reverse phase column (COSMOSIL 5C1 8-AR-II Packed Column -4.6 mm I.D. × 250 mm; Dartford, UK). Data acquisition and analysis were accomplished using Software Empower 3. Three independent analysis were performed for each experiment.
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