Recent developments in microcolumn liquid chromatography

Vissers, Johannes P.C.

doi:10.1016/s0021-9673(99)00692-5

Cited by 174 publications

(105 citation statements)

References 270 publications

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“…Recent MALDI-TOF research improvements to overcome limitations associated with the identification of mixtures or the presence of contaminates include in post source decay and MALDI MS/MS [8,9]. LC column and interface miniaturization [10][11][12], the utility of alternative fragmentation techniques to collision -5 -The most common approach to protein identification, searching of tandem LC-MS data against a non-redundant protein database, relies upon the availability of a protein or translated nucleotide databases to match the experimental precursor and fragment ion data. It is widely believed that this approach cannot be used for the identification of proteins from unknown or partially sequenced genomes, due to the lack of available sequence information.…”

Section: Introductionmentioning

confidence: 99%

The use of proteome similarity for the qualitative and quantitative profiling of reperfused myocardium☆

Vissers

Pons

Hulin

et al. 2009

Journal of Chromatography B

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An LC-MS based approach is presented for the identification and quantification of proteins from unsequenced organisms. The method relies on the preservation of homology across species and the similarity in detection characteristics of proteomes in general. Species related proteomes share similarity that progresses from the amino acid frequency distribution to the complete amino sequence of matured proteins. Moreover, the comparative analysis between theoretical and experimental proteome distributions can be used as a measure for the correctness of detection and identification obtained through LC-MS based schemes.Presented are means to the identification and quantification of rabbit myocardium proteins, immediately after inducing cardiac arrest, using a data-independent LC-MS acquisition strategy. The employed method of acquisition affords accurate mass information on both the precursor and associated product ions, whilst preserving and recording the intensities of the ions. The latter facilitates label-free quantification. The experimental ion density observations obtained for the rabbit sub proteome were found to share great similarity with five other mammalian samples, including human heart, human breast tissue, human plasma, rat liver and a mouse cell line. Redundant, species-homologues peptide identifications from other mammalian organisms were used for initial protein identification, which were complemented with peptide identifications of translated gene sequences. The feasibility and accuracy of label-free quantification of the identified peptides and proteins utilizing above mentioned strategy is demonstrated for selected cardiac rabbit proteins.

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Section: Introductionmentioning

confidence: 99%

The use of proteome similarity for the qualitative and quantitative profiling of reperfused myocardium☆

Vissers

Pons

Hulin

et al. 2009

Journal of Chromatography B

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“…Advances in microseparations, include techniques such as capillary liquid chromatography (cLC), 1 capillary electrophoresis (CE), 2 and capillary electrochromatography 3 have played a large role in improving the separation efficiency. The chromatographic dilution (D) of cLC, for example, is given by (1) where C 0 is the initial concentration of the analyte, C max is the final compound concentration at the peak maximum, r is the column radius, ϵ is the column porosity, k is the retention factor, L is the column length, H is the column plate height, and V inj is the injected sample volume. Equation 1 shows that a smaller column radius results in less chromatographic dilution and higher concentration elution peaks.…”

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confidence: 99%

Magnetic Resonance Microimaging and Numerical Simulations of Velocity Fields Inside Enlarged Flow Cells Used for Coupled NMR Microseparations

Zhang

Webb

2005

Anal. Chem.

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The coupling of various chemical microseparation methods with small-scale NMR detection is a growing area in analytical chemistry. The formation of enlarged flow cells within the active volume of the NMR detector can significantly increase the coil filling factor and hence the signal-to-noise ratio of the NMR spectra. However, flow cells can also lead to deterioration of the separation efficiency due to the development of complex flow patterns, the form of which depend on the particular geometry of the flow cell and the flow rate used. In this study, we investigated the flow characteristics in different flow cell geometries relevant to the coupling of capillary liquid chromatography and NMR. Computational fluid dynamics was used to simulate fluid flow inside flow cells with a volume of ~ 1 µL. Magnetic resonance microimaging was used to measure experimentally the velocity fields inside these flow cells. The results showed good agreement between experiment and simulation and demonstrated that a relatively gradual expansion and contraction is necessary to avoid areas of weak recirculation and strong radial velocities, both of which can potentially compromise separation efficiency.An essential component in the purification and analysis of unknown compounds is efficient separation of individual components from an often complex chemical or biological system. Advances in microseparations, include techniques such as capillary liquid chromatography (cLC), 1 capillary electrophoresis (CE), 2 and capillary electrochromatography 3 have played a large role in improving the separation efficiency. The chromatographic dilution (D) of cLC, for example, is given by (1) where C 0 is the initial concentration of the analyte, C max is the final compound concentration at the peak maximum, r is the column radius, ϵ is the column porosity, k is the retention factor, L is the column length, H is the column plate height, and V inj is the injected sample volume. Equation 1 shows that a smaller column radius results in less chromatographic dilution and higher concentration elution peaks. The smaller scale is also much more amenable to very rapid separations. In addition to these general benefits of microseparations, in electrophoretic separations the high surface area-to-volume ratio of the small capillaries used in CE systems,

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“…O completo entendimento da origem e da magnitude destes diferentes processos de alargamento extracoluna é de fundamental importância para o desenvolvimento de sistemas cromatográficos capilares de alta eficiência e, uma vez que -por motivos óbvios -tais processos de dispersão sempre estarão presentes em maior ou menor grau [23] , a principal preocupação no desenvolvimento de equipamentos dedicados a c-LC é o quanto esses efeitos podem influenciar a separação.…”

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Cromatografia líquida capilar: 2. Alargamento das bandas por efeitos extracoluna

Coutinho¹,

Lanças

2011

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ResumoO alargamento da banda é um fator altamente indesejável em todas as técnicas cromatográficas, uma vez que diminui a eficiência da coluna (menor número de pratos, N) e deteriora a resolução (menor Rs). Este efeito é particularmente crítico na cromatografia líquida capilar (c-LC) devido aos pequenos volumes da instrumentação, conexões, colunas, etc., empregados na técnica. Como parte de uma série de artigos a respeito da c-LC, neste trabalho serão abordados os principais fatores que ocasionam o alargamento das bandas cromatográficas em c-LC, assim como a maneira de evitá-lo. Palavras-chaveCromatografia líquida capilar; alargamento de bandas; fatores extra coluna; micro LC; LC capilar. Capillary liquid chromatography: 2. Band broadening due to extracolumn effects AbstractBand broadening is one of the major factors to be avoided in chromatographic techniques, once it decreases the column efficiency (the number of plates, N) and deteriorates the Resolution (Rs). This effect is particularly critical in capillary liquid chromatography due to the very low volumes, instrumentation, connections and columns employed in this technique. As a part in a series of papers dealing with c-LC, this work describes the major factors responsible for band broadening in chromatography, as well as the ways to minimize it.

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Recent developments in microcolumn liquid chromatography

Cited by 174 publications

References 270 publications

The use of proteome similarity for the qualitative and quantitative profiling of reperfused myocardium☆

The use of proteome similarity for the qualitative and quantitative profiling of reperfused myocardium☆

Magnetic Resonance Microimaging and Numerical Simulations of Velocity Fields Inside Enlarged Flow Cells Used for Coupled NMR Microseparations

Cromatografia líquida capilar: 2. Alargamento das bandas por efeitos extracoluna

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