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Development of an online capillary comprehensive 2D‐LC system for the analysis of proteome samples
Author(s) -
Sommella Eduardo,
Cacciola Francesco,
Donato Paola,
Dugo Paola,
Campiglia Pietro,
Mondello Luigi
Publication year - 2012
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100877
Subject(s) - proteome , chromatography , chemistry , capillary electrophoresis , capillary action , materials science , biochemistry , composite material
In the present contribution, a fully automated capillary comprehensive two‐dimensional LC (LC×LC) method, for proteomic analysis, was developed for the first time. The investigated platform was characterized by the coupling of high‐pH RP with low‐pH RP separations thus ensuring the generation of high peak capacity despite the employment of identical stationary phases. The use of capillary columns in both dimensions allowed to reduce mobile‐phase consumption and enhance sensitivity. Fraction transfer from the first to the second dimension was performed by means of two 2‐position 6‐port nano‐switching valves, under stop‐flow conditions. Values as high as 1208 and 955 were obtained for the theoretical and practical peak capacity, respectively. The investigated LC×LC system showed good retention time repeatibility with RSD values ranging from 0.8 to 6.0% for the first dimension and from 1.0 to 3.0% for the second dimension, respectively. RSD peak area values below 9.5% were also attained, thus demonstrating the precision of the LC×LC method employed.