Mass spectrometry analysis of RRC101 fractions was performed using the aforementioned column and solvent system, at a flow rate of 200 µl/min. HPLC-MS analyses of extracts were conducted using a Finnigan micro AS, Finnigan surveyor MS pump plus, and Finnigan LCQ duo (Thermo Scientific). This analysis was conducted in positive ion mode, utilizing an ESI source with 5-kV spray voltage and a capillary temperature of 190°C. Other instrument conditions were maximized for sensitivity by "tuning" to a direct injection of surfactin A (Sigma-Aldrich). Samples were also analyzed on a Dionex UltiMate 3000 AS attached to a Dionex UltiMate 3000 UHPLC + pump, in tandem with a Thermo Scientific LTQ XL. Conditions were the same as above, except the instrument was "tuned" by direct injection of iturin A (Sigma-Aldrich). Commercially obtained surfactin A (Sigma-Aldrich) was employed as a standard for retention and fragmentation patterns of surfactins, with previously published data used to guide fengycin characterization (Bie et al. 2009; Pathak et al. 2012 ).
Surfactin a
Manufactured by Merck Group
Sourced in United States
Surfactin A is a cyclic lipopeptide produced by the bacterium Bacillus subtilis. It is a naturally occurring compound with surface-active properties.
Automatically generated - may contain errors
Lab products found in correlation
Ltq xl, by Thermo Fisher Scientific (1 mentions)
Iturin a, by Merck Group (1 mentions)
Finnigan lcq duo, by Thermo Fisher Scientific (1 mentions)
Finnigan surveyor ms pump plus, by Thermo Fisher Scientific (1 mentions)
Ultimate 3000, by Thermo Fisher Scientific (1 mentions)
Hplc grade, by Thermo Fisher Scientific (1 mentions)
717 autosampler, by Waters Corporation (1 mentions)
Millenium 32, by Waters Corporation (1 mentions)
2 protocols using surfactin a
1
Mass Spectrometry Analysis of Lipopeptides
2
Lipopeptide Extraction and Analysis from B. subtilis
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Samples of B. subtilis cultures were centrifuged at 10,000× g for 10 min. A volume of 1 mL of the supernatant was purified through C18 Maxi-Clean cartridges 1 g (Alltech, Deerfield, IL, USA). The column was washed with 8 mL of water. The lipopeptides were then eluted with 8 mL of 100% methanol (HPLC grade, Acros Organics, Geel, Belgium). The extract was dried before dissolution in 200 µL of methanol. The sample was then injected and analyzed by high-performance liquid chromatography (Online Degaser, 717 Autosampler, 660S Controller, 626 Pump, 996 PhotoDiodeArray, Waters Corp., Milford, MA, USA) using a C18 column (5 µm, 250 × 4.6 mm, 218 TP, VYDAC). The mycosubtilins were separated with an acetonitrile/water/trifluoroacetic acid solvent, 40:60:0.1, v/v/v. The flow rate was 1 mL/min and the detection wavelength was 214 nm. Purified iturin A and surfactin A, used as standards, were from Sigma (Sigma-Aldrich, Saint Louis, MI, USA). The retention time and second derivative of the absorption spectrum between 200 and 400 nm (Diode Array PDA 996, Waters) were used to identify the eluted molecules (Millenium 32 Software, Waters).
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