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Elution behavior of short ds DNA strands in silicon micropillar array columns in ion pair reversed‐phase chromatography mode
Author(s) -
Zhang Lei,
Majeed Bivragh,
Lynen Frederic,
Van Hoof Chris,
De Malsche Wim
Publication year - 2012
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200226
Subject(s) - chromatography , materials science , analytical chemistry (journal) , high performance liquid chromatography , silicon , phase (matter) , elution , chemistry , optoelectronics , organic chemistry
In the present paper, ds DNA separation has been studied in a silicon micropillar array column using ion‐pair RP‐ HPLC ( IP ‐ RP ‐ HPLC ). The deep‐etched (32.0 μm) silicon micropillar array was fabricated by advanced deep UV lithography and by a dedicated B osch etch process and then sealed by anodic bonding to a P yrex glass. The pillar surface was subsequently conditioned hydrophobic. Working in isocratic mode under nonretained conditions, van D eemter curves of ds DNA and coumarin were established to assess the performance of the micropillar array column, resulting in plate heights of only a few micrometers. Working in gradient mode, separations of ds DNA fragments were evaluated. The relevant gradient operation parameters were studied to understand their influence on ds DNA separations. The correlation between DNA length and retention was measured and theoretically described in a length range of 50–500 bp, promising for the determination of DNA of an unknown length. Finally, a separation example demonstrated the excellent separation power of on‐chip IP ‐ RP chromatography by achieving a large operation range of DNA length (10–300 bp) with a 5‐bp difference among 11 ds DNA fragments.