Molecules 2–3 and 15 were isolated from extracts of P. luteoviolacea 2ta16. 10×100 mL cultures were grown as before and incubated for 24 h at 30 °C with shaking at 200 rpm. Cultures were pooled and extracted with an equal volume of ethyl acetate. The organic layer was dried with anhydrous MgSO4, filtered and solvent was removed in vacuo. The residue was dissolved in 5 mL MeOH and filtered through a 0.2 µM vacuum filter. 2–3 , and 15 were purified by semi-preparative scale HPLC using a reverse phase C18 column (Phenomenex, 250 mm). Solvents used were water + 0.1% TFA (A) and MeCN + 0.1% TFA (B). An identical elution profile as described before for analytical scale experiments but at an increased flow rate of 2 mL/min was used for all semi-preparative purification experiments in this study. Guided by LC-MS/MS, fractions containing 2–3 , and 15 were lyophilized. 500 µg of each product was dissolved in 50 µL CDCl3. 1H NMR, and 13C NMR spectra for 2 were obtained using a 600 MHz Varian NMR microprobe. Spectra of 3 and 15 were verified with literature values40 (link).
Nmr microprobe
Manufactured by Agilent Technologies
The NMR microprobe is a specialized analytical instrument used in nuclear magnetic resonance (NMR) spectroscopy. It is designed to analyze small sample volumes with high sensitivity and resolution. The core function of the NMR microprobe is to detect and measure the magnetic properties of atomic nuclei within a sample, providing valuable information about its chemical composition and structure.
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2 protocols using nmr microprobe
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Isolation and Purification of Bioactive Molecules from P. luteoviolacea
2
Isolation and Purification of Bioactive Molecules from P. luteoviolacea
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Molecules 2–3 and 15 were isolated from extracts of P. luteoviolacea 2ta16. 10×100 mL cultures were grown as before and incubated for 24 h at 30 °C with shaking at 200 rpm. Cultures were pooled and extracted with an equal volume of ethyl acetate. The organic layer was dried with anhydrous MgSO4, filtered and solvent was removed in vacuo. The residue was dissolved in 5 mL MeOH and filtered through a 0.2 µM vacuum filter. 2–3 , and 15 were purified by semi-preparative scale HPLC using a reverse phase C18 column (Phenomenex, 250 mm). Solvents used were water + 0.1% TFA (A) and MeCN + 0.1% TFA (B). An identical elution profile as described before for analytical scale experiments but at an increased flow rate of 2 mL/min was used for all semi-preparative purification experiments in this study. Guided by LC-MS/MS, fractions containing 2–3 , and 15 were lyophilized. 500 µg of each product was dissolved in 50 µL CDCl3. 1H NMR, and 13C NMR spectra for 2 were obtained using a 600 MHz Varian NMR microprobe. Spectra of 3 and 15 were verified with literature values40 (link).
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