The molecular mass and peptide sequencing were done in
positive ion mode using Electrospray ionisationemass spectrometry
(ESIeMS) and the tandem mass spectrometry (MS/MS),
respectively. ESI mass spectrometry was performed using a triple
quadrupole instrument Applied Biosystems API 3000 (PE Sciex,
Toronto, Canada) equipped with an electrospray ion source. The
system is controlled by the Analyst Software 1.4, allowing the
control of the spectrometer, the analysis and the processing data.
Interpretations of spectra MSeMS were made with the Bioanalyst
software. The freeze-dried samples from RP-HPLC were dissolved
in acetonitrile/water (20/80; v/v) containing 0.1% formic acid for
the positive mode. The solution was injected (nebulised) uninterrupted,
by a pump (Model 22, Harward Apparatus, South Natick,
USA) with a flow rate of 5 ml/min. The potential of ionisation was of
5000 V in positive mode. At the time of the recording of the
spectrum, 30 scans 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 nitrogen
generatorWhatman model 75-72 (Whatman Inc., Haverhill, MAthe
USA). The polypropylene glycol (PPG) was used for the calibration
and the optimisation of the machine. The peptide sequence was
determined from the CID spectrum of the protonated analyse
[M þ H]þ by MS/MS experiments. Peptide sequences were done
using the bioanalyst software (Applied Biosystems, USA).
The molecular mass and peptide sequencing were done inpositive ion mode using Electrospray ionisationemass spectrometry(ESIeMS) and the tandem mass spectrometry (MS/MS),respectively. ESI mass spectrometry was performed using a triplequadrupole instrument Applied Biosystems API 3000 (PE Sciex,Toronto, Canada) equipped with an electrospray ion source. Thesystem is controlled by the Analyst Software 1.4, allowing thecontrol of the spectrometer, the analysis and the processing data.Interpretations of spectra MSeMS were made with the Bioanalystsoftware. The freeze-dried samples from RP-HPLC were dissolvedin acetonitrile/water (20/80; v/v) containing 0.1% formic acid forthe positive mode. The solution was injected (nebulised) uninterrupted,by a pump (Model 22, Harward Apparatus, South Natick,USA) with a flow rate of 5 ml/min. The potential of ionisation was of5000 V in positive mode. At the time of the recording of thespectrum, 30 scans on average were added (MCA mode) for eachspectrum. The gases used (nitrogen and air) were pure (up to 99%)and produced by a compressor Jun-Air 4000-40M and a nitrogengeneratorWhatman model 75-72 (Whatman Inc., Haverhill, MAtheUSA). The polypropylene glycol (PPG) was used for the calibrationand the optimisation of the machine. The peptide sequence wasdetermined from the CID spectrum of the protonated analyse[M þ H]þ by MS/MS experiments. Peptide sequences were doneusing the bioanalyst software (Applied Biosystems, USA).
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