Sodium dodecyl sulphate removal from tryptic digest samples for on-line capillary liquid chromatography/electrospray mass spectrometry.
An on-line microcolumn switching method was developed for the removal of sodium dodecyl sulphate (SDS) from tryptic digest samples. The system includes two micro-precolumns: a specific ionic detergent trapping column and a preconcentration column. Characterization of the proteinaceous samples, after isolation from the SDS, was performed by capillary liquid chromatography (LC) with UV absorption detection and electrospay mass spectrometry (ESI-MS). Loading and clean-up of the samples and regeneration of the detergent trapping column were performed at 50 microl min(-1), resulting in sample clean-up times of only 30 s. SDS-containing tryptic digested protein samples were directly applied to the micro-precolumns without any previous sample pretreatment. The developed microcolumn switching method permits the on-line analysis of small tryptic digest samples by capillary LC/ESI-MS in the presence of SDS. The method is completely automated and can be performed unattended. The maximum amount of SDS, in terms of loadability and breakthrough, were determined. Also studied were the selection of the loading and clean-up solvents and the recovery of the peptides. Chromatographic separations and mass spectral data confirmed the removal of SDS.
Vissers
J P
JP
LC Packings, Amsterdam, The Netherlands.
Chervet
J P
JP
Salzmann
J P
JP
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009862:Online Systems; D010455:Peptides / Q000737:chemistry*; D012967:Sodium Dodecyl Sulfate; D013056:Spectrophotometry, Ultraviolet; D014357:Trypsin / Q000737:chemistry*
D010455:Peptides; D012967:Sodium Dodecyl Sulfate; D014357:Trypsin