Stability and repeatability of capillary columns based on porous monoliths of poly(butyl methacrylate-co-ethylene dimethacrylate)

被引:106
作者
Geiser, Laurent
Eeltink, Sebastiaan
Svec, Frantisek
Frechet, Jean M. J. [1 ]
机构
[1] Univ Calif Berkeley, Coll Chem, Berkeley, CA 94720 USA
[2] Univ Calif Berkeley, Lawrence Berkeley Lab, Mol Foundry, Berkeley, CA 94720 USA
关键词
nanoHPLC; monolithic columns; column technology; capillary; mass spectrometry; proteins; repeatability; reproducibility; validation;
D O I
10.1016/j.chroma.2006.11.079
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns have been prepared via either thermally or photochemically initiated polymerization of the corresponding monomers and the repeatability of their preparation has been explored. Three separate batches of 5 columns each were prepared using thermal and photochemical initiation for a total of 30 columns. All 30 capillary columns were tested in liquid chromatography-electrospray ionisation mass spectrometry mode for the separation of a model mixture of three proteins-ribonuclease A, cytochrome c and myoglobin. Excellent repeatability of retention times was observed for the proteins as evidenced by relative standard deviation (RSD) values of less than 1.5%. Somewhat broader variations with RSD values of up to 10% were observed for the pressure drop in the columns. The stability of retention times was also monitored using a single monolithic column and no significant shifts in either retention times or back pressure was observed in a series of almost 2200 consecutive protein separations. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:140 / 146
页数:7
相关论文
共 37 条
[1]   Determination of the porosities of monolithic columns by inverse size-exclusion chromatography [J].
Al-Bokari, M ;
Cherrak, D ;
Guiochon, G .
JOURNAL OF CHROMATOGRAPHY A, 2002, 975 (02) :275-284
[2]   Studies on the long-term thermal stability of particulate and monolithic poly(styrene-divinylbenzene) capillaries in reversed-phase liquid chromatography [J].
Andersen, T ;
Nguyen, QNT ;
Trones, R ;
Greibrokk, T .
ANALYST, 2004, 129 (03) :191-197
[3]  
[Anonymous], STP PHARM PRATIQUES
[4]  
Caporal-Gautier J, 1992, STP PHARM PRATIQUES, V2, P227
[5]   Novel monolithic materials using poly(ethylene glycol) as porogen for protein separation [J].
Courtois, J ;
Byström, E ;
Irgum, K .
POLYMER, 2006, 47 (08) :2603-2611
[6]   Evaluation of ring-opening metathesis polymerization (ROMP)-derived monolithic capillary high performance liquid chromatography columns [J].
Gatschelhofer, C ;
Magnes, C ;
Pieber, TR ;
Buchmeiser, MR ;
Sinner, FM .
JOURNAL OF CHROMATOGRAPHY A, 2005, 1090 (1-2) :81-89
[7]   Preparation and evaluation of poly (polyethylene glycol methyl ether acrylate-co-polyethylene glycol diacrylate) monolith for protein analysis [J].
Gu, BH ;
Armenta, JM ;
Lee, ML .
JOURNAL OF CHROMATOGRAPHY A, 2005, 1079 (1-2) :382-391
[8]  
Hancock WS, 2002, PROTEOMICS, V2, P352, DOI 10.1002/1615-9861(200204)2:4<352::AID-PROT352>3.0.CO
[9]  
2-U
[10]   Surface-alkylated polystyrene monolithic columns for peptide analysis in capillary liquid chromatography-electrospray ionization mass spectrometry [J].
Huang, XA ;
Zhang, S ;
Schultz, GA ;
Henion, J .
ANALYTICAL CHEMISTRY, 2002, 74 (10) :2336-2344