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MS and MS/MS measurements were performed in positive ion mode using electrospray ionization (ESI) and ESI/MS/MS, respectively. ESI mass spectrometry was performed using a triple quadrupole instrument from Applied Biosystems API 3000 (PE Sciex, Toronto,Canada). The system is controlled by the Analyst Software 1.4, allowing the control of the spectrometer, the analysis, and the processing of the data. Interpretations of MS–MS spectra were made with the Bioanalyst software. The freeze-dried samples were dissolved in acetonitrile/water (20/80v/v) solvent containing formic acid 0.1% for the positive mode. The solution was injected (nebulized) uninterrupted, thanks to a pump (Model 22, Harward Apparatus, South Natick, USA) with a flow rate of 5 μL/min. The potential of ionizationwas of 5,000 V (volt) in positive mode. At the time of the recording of the spectrum, 30scans on average were added (MCA mode) for each spectrum. The gases used (nitrogen and air) were pure (up to 99%) and produced by a compressor Jun-Air 4000-40M and a Whatman model 75-72 nitrogen generator (Whatman Inc Haverhill, MA, the USA). The polypropylene glycol was used for the calibration and theoptimization of the machine system. The peptide sequence was determined from the CID spectrum of the protonated analyze [M+H]+by MS/MS experiments. |
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