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Type of publication: research article
Type of publication (PDB): Straipsnis Clarivate Analytics Web of Science / Article in Clarivate Analytics Web of Science (S1)
Field of Science: Biochemija / Biochemistry (N004)
Author(s): Škudas, Romas;Grimes, Brian A;Machtejevas, Egidijus;Kudirkaitė, Vilma;Kornyšova, Olga;Hennessy, Tom P;Lubda, Dieter;Unger, Klaus K
Title: Impact of pore structural parameters on column performance and resolution of reversed-phase monolithic silica columns for peptides and proteins
Other Title: Porų struktūrinių parametrų įtaka kolonėlių efektyvumui ir skiriamajai gebai tiriant atvirkščių fazių monolitines silikagelio kolonėles, skirtas peptidams ir baltymams
Is part of: Journal of chromatography. A. Amsterdam : Elsevier B.V, 2007, Vol. 1144, iss. 1
Extent: p. 72-84
Date: 2007
Keywords: Silica monolith;Efficiency;Resolution;Gradient elution
Abstract: In this work, monolithic silica columns with the C4, C8, and C18 chemistry and having various macropore diameters and two different mesopore diameters are studied to access the differences in the column efficiency under isocratic elution conditions and the resolution of selected peptide pairs under reversed-phase gradient elution conditions for the separation of peptides and proteins. The columns with the pore structural characteristics that provided the most efficient separations are then employed to optimize the conditions of a gradient separation of a model mixture of peptides and proteins based on surface chemistry, gradient time, volumetric flow rate, and acetonitrile concentration. Both the mesopore and macropore diameters of the monolithic column are decisive for the column efficiency. As the diameter of the through-pores decreases, the column efficiency increases. The large set of mesopores studied with a nominal diameter of 25 nm provided the most efficient column performance. The efficiency of the monolithic silica columns increase with decreasing n-alkyl chain length in the sequence of C18 < C8 < C4. The resolution of proteins and peptides by reversed-phase gradient liquid chromatography on n-octadecyl, n-octyl, and n-butyl bonded monolithic silica columns is optimized. The results obtained imply the use of acetonitrile concentration gradient up to 75% for n-octadecyl and n-octyl bonded monolithic silica columns, and the use of acetonitrile concentration gradient up to 85% for n-butyl bonded monolithic silica columns. With the respect to the gradient times and flow rates, the optimum conditions are the best with n-octyl and n-butyl bonded monolithic silica columns, where the range of optimum gradient times is up to 30 min and mobile phase flow rates in the range of 0.5–1 ml/min [...]
Affiliation(s): Vytauto Didžiojo universitetas
Appears in Collections:Universiteto mokslo publikacijos / University Research Publications

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