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Effect of trifluoroacetic acid on the reduction of disulfide bridges in peptides analyzed by fast‐atom bombardment mass spectrometry
Author(s) -
Visentini J.,
Gauthier J.,
Bertrand M. J.
Publication year - 1989
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1290031105
Subject(s) - chemistry , trifluoroacetic acid , fast atom bombardment , matrix (chemical analysis) , mass spectrometry , cystine , dithiothreitol , thiol , chromatography , inorganic chemistry , cysteine , organic chemistry , enzyme
Some of the factors that influence the reduction of disulfide‐containing peptides under fast‐atom bombardment have been investigated using two neurohormonal peptides that include disulfide bridges in their structures. Deaminoarginine‐vasopressin (DAVP) and arginine‐vasopressin (AVP) have been analyzed as their acetate and trifluoroacetate salts. Results obtained in a thioglycerol matrix indicate that the peptides analyzed as their acetate salts are completely reduced under bombardment, whereas the trifluoroacetate salts show little evidence of reduction. Addition of trifluoroacetic acid to the acetate sample prior to bombardment inhibits reduction whereas addition after bombardment shows no effect on the reduction, thereby indicating the irreversibility of the process. Time‐monitoring experiments conducted with the acetate salts of DAVP and AVP in common matrices such as thioglycerol, dithiothreitol + diethioerythritol, glycerol, hydroxyethyldisulfide and nitrobenzyl‐alcohol demonstrate an important effect of the chemical nature of the matrix on reduction. In matrices containing thiol groups, the reduction is extensive, whereas it is almost suppressed in matrices such as hydroxyethyldisulfide and nitrobenzylalcohol. However, the addition of trifluoroacetic acid to all of these matrices essentially eliminates reduction and provides measured isotopic peak ratios that are in agreement with theoretically calculated values for these peptides.