期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1043, 期 1, 页码 9-18出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2004.03.070
关键词
mobile phase composition; trifluoroacetic acid; peptides
资金
- NIGMS NIH HHS [R01 GM061855-05, R01 GM61855] Funding Source: Medline
Trifluoroacetic acid (TFA) remains the dominant mobile phase additive for reversed-phase high-performance liquid chromatography (RP-HPLC) of peptides after more than two decades since its introduction to this field. Generally, TFA has been employed in a concentration range of 0.05-0.1% (6.5-13 mM) for the majority of peptide separations. In order to revisit the question as to whether such a concentration range is optimum for separations of peptide mixtures containing peptides of varying net positive charge, the present study examined the effect of varying TEN concentration on RP-HPLC at 25 and 70degreesC of three groups of synthetic 10-residue synthetic peptides containing either one (+ 1) or multiple (+3, +5) positively charged groups. The results show that the traditional range of TEN concentrations employed for peptide studies is not optimum for many, perhaps the majority, of peptide applications. For efficient resolution of peptide mixtures, particularly those containing peptides with multiple positive charges, our results show that 0.2-0.25% TFA in the mobile phase will achieve optimum resolution. In addition, the use of high temperature as a complement to such TFA concentration levels is also effective in maximizing peptide resolution. (C) 2004 Elsevier B.V. All rights reserved.
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