The protein samples were analyzed on an LC − ESI–MS platform consisting of a Surveyor MS pump system (C4 column, Higgins Analytical Proto 300 5 µm RS-0301-W045, 30 mm × 1.0 mm) coupled to an electrospray mass spectrometer (Thermo Finnigan LCQ duo, equipped with an ion trap mass analyzer). The gradient was 5% solvent B (96% acetonitrile, 2% acetic acid) in solvent A (2% acetonitrile, 2% acetic acid) for 2 min, 5–100% solvent B in solvent A in 4 min, and 100% solvent B for 8 min at a flow rate of 40 μL min−1. The ESI spectra was deconvoluted to the zero-charge domain by the ProMass Deconvolution Software (Novatia) using a standard parameter set: average mass type, mass tolerance 0.02%, minimum tolerance 2 Da and input m/z range 500–2000 units.
Finnigan lcq duo
Manufactured by Thermo Fisher Scientific
The Finnigan LCQ duo is a liquid chromatography-mass spectrometry (LC-MS) system designed for qualitative and quantitative analysis of a wide range of analytes. It features a quadrupole ion trap mass analyzer for high-sensitivity detection and accurate mass measurements.
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2 protocols using finnigan lcq duo
1
Protein Analysis by LC-ESI-MS
2
Mass Spectrometry Analysis of Lipopeptides
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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 ).
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