Improving Signal Intensity in the Analysis of Peptides with Trifluoroacetic Acid-containing Mobile Phase by Reversed-Phase Capillary Liquid Chromatography-electrospray Mass Spectrometry
Date Issued
2007
Date
2007
Author(s)
Lee, Wan-Li
DOI
zh-TW
Abstract
Trifluoroacetic acid (TFA) is a commonly used ion-pair reagent in RP-HPLC to improve peak shape and enhance separation efficiency of peptides. However, TFA forms very strong ion pairs with peptides that are not broken apart in the process of electrospray ionization and consequently reduces the ionization efficiency of peptides. Moreover, TFA also leads to spray instability and signal reduction due to high conductivity and high surface tension of the mobile phase.
In this work, a low flow-liquid junction interface was used to alleviate the adverse effect of TFA on the peptide signal in ESI/MS. Different voltages are applied to the make-up solution and sheath liquid to create an electrical field between liquid junction and low flow interface. Under acidic conditions, EOF velocity will be lower than the velocity of TFA anion. TFA anions thus turn out to move away from ESI/MS and the sensitivity loss caused by TFA can be reduced. By using this low flow-liquid junction interface, it is possible to employ TFA in LC/MS for peptide separation.
Subjects
三氟醋酸
液相層析
電灑法質譜
訊號抑制
胜肽
液體結合式界面
低流速鞘流界面
TFA
LC-ESI/MS
signal suppression
peptide
liquid junction interface
low flow interface
Type
thesis
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