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Capillary LC Columns : Packing Techniques and Applications Capillary LC columns : packing techniques and applications Citation for published version (APA): Vissers, J. P. C. (1998). Capillary LC columns : packing techniques and applications. Technische Universiteit Eindhoven. https://doi.org/10.6100/IR515594 DOI: 10.6100/IR515594 Document status and date: Published: 01/01/1998 Document Version: Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers) Please check the document version of this publication: • A submitted manuscript is the version of the article upon submission and before peer-review. There can be important differences between the submitted version and the official published version of record. People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website. • The final author version and the galley proof are versions of the publication after peer review. • The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal. If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license above, please follow below link for the End User Agreement: www.tue.nl/taverne Take down policy If you believe that this document breaches copyright please contact us at: [email protected] providing details and we will investigate your claim. Download date: 11. Oct. 2021 CAPILLARY LC COLUMNS Packing Techniques and Applications Proefsch rift ter verkrijging van de graad van doctor aan de Technische Universiteit Eindhoven, op gezag van de rector Magnificus, prof.dr. M. Rem, voor een commissie aangewezen door het College voor Promoties in het openbaar te verdedigen op vrijdag 18 december 1998, om 16.00 uur door JOHANNES PETRUS CORNELIS VISSERS geboren te Zevenbergen Dit proefschrift is goedgekeurd door de promotoren: prof.dr.ir. C.A.M.G. Cramers en prof.dr. P..I.F. Sandra copromotor: dr. J. Laven if you compare yourself with others, you may become vain and bitter; for always there will be greater and lesser persons than yourself be yourself Max Ehrmann "Desiderata" (1927) in herrinering aan mijn grootmoeder Janssen-Akkermans aan mijn ouders Vissers, Johannes Petrus Cornelis Capillary LC Columns; packing techniques and applications I Johannes Petrus Cornelis Vissers. - Eindhoven: Eindhoven University of Technology Thesis Eindhoven University of Technology. -with ref. - With summary in Dutch. ISBN 90-386-0608-7 NUGI: 813 Subjects headings: liquid chromatographic columns CONTENTS CONTENTS 1. GENERAL INTRODUCTION AND SCOPE 1.1 Introductory Remarks 1.2 Theoretical Aspects of Column Liquid Chromatography 2 1.3 Stationary Phases and Filling Techniques 8 1.4 Scope 17 1.5 References 18 2. MICROCOLUMN LIQUID CHROMATOGRAPHY: AN OVERVIEW OF 21 INSTRUMENTATION, DETECTION AND APPLICATIONS 3. SEDIMENTATION BEHAVIOUR AND COLLOIDAL PROPERTIES OF POROUS, 61 CHEMICALLY MODIFIED SILICAS IN NON-AQUEOUS SOLVENTS 4. COLLOIDAL ASPECTS OF SLURRY PACKING CAPILLARY LC COLUMNS 4.1 Colloid Chemical Aspects of Slurry Packing Techniques in 79 Microcolumn Liquid Chromatography 4.2 Comparison of Spherically and Irregularly Shaped Stationary Phase 97 Packings in Microcolumn Liquid Chromatography 5. HYDRODYNAMIC ASPECTS OF SLURRY PACKING PROCESSES IN 109 MICROCOLUMN LIQUID CHROMATOGRAPHY 6. APPLICATIONS OF CAPILLARY LC COLUMNS 6.1 Calculation of Retention in Capillary Electrochromatography Based on 133 Chromatographic and Electrophoretic Data 6.2 On-Line Ionic Detergent Removal from Minute Protein/Peptide Samples 143 Prior to Capillary Liquid Chromatography-Eiectrospray Mass Spectrometry 6.3 Two-dimensional Capillary Liquid Chromatography Based on 157 M icro-Fractionation APPENDIX A The Van Der Waals Interaction Between Particles of Composite Materials 171 in a Medium CHAPTER 1 GENERAL INTRODUCTION AND SCOPE 1.1 INTRODUCTORY REMARKS Analytical chemistry is involved with the separation, identification and quantitation of target compounds in complex matrices. With today's analysis techniques, the identification and quantitation of reasonably pure compounds does not pose serious problems. There is how­ ever an increasing demand for the analysis of small sample quantities in which the com­ pounds of interest are present as minor constituents. Hence, there is a growing need for sensitive, reliable analytical procedures that provide qualitative and quantitative information. In practice this involves the development of both sample preparation and separation methods -as well as suitable detection techniques. Gas chromatography (GC) and column liquid chromatography (LC) have found wide spread acceptance as the separation technique in numerous application areas. Environmental chem­ istry, food and polymer chemistry and the bioanalysis field are just a few areas to which they have been applied. GC remains the method of choice for volatile and thermally-stable com­ pounds because of its high separation power and favorable limits of detection for many classes of compounds. Moreover, the on-line coupling with mass spectrometry (MS) is well estab­ lished allowing separation and subsequent identification on a routine bases. Ionic, polar, non­ volatile, thermally-label and high-molecular weight compounds are typically separated by LC since they are not directly amenable to GC. the reduced separation efficiency and detectability obtainable with LC compared to the range of compounds that can be di- rectly analyzed is much greater and as a matter of fact still expanding. In virtue of this wide­ spread applicability of LC. much effort has been put into the development of sensitive identi­ fication techniques for LC. Especially LC-MS has gained in popularity after the recent introduc­ tion of atmospheric pressure ionization based interfaces and has found its way into routine applications in the biochemical and environmental field. Although other "liquid-based" sepa­ ration techniques have become available among them electrophoresis, supercritical fluid chromatography and electrochromatography they have not found a broad acceptance yet, which is mainly due the versatility, robustness and ease of use of LC. CHAPTER 1 Despite that sensitive detection and identification techniques have become available for LC, there is a continuous need for detecting lower concentrations and analyzing increasingly smaller sample quantities. Although miniaturized LC has the potential to fulfill these require­ ments, these techniques are not yet generally accepted and still in development. Miniaturization in separation sciences has been evident for some time [4-7]. The introduc­ tion of open tubular capillary columns for gas chromatography by Golay in the late SO's is generally regarded as one of the first steps towards miniaturization in chromatography. De­ creasing the particle size has been the main stimulus towards miniaturization in column liquid chromatography, and still is nowadays [8]. Consistent efforts towards decreasing the column diameter of packed columns were not started before the early 80's. The groups of Kucera, lshii and Novotny [9-1 ·11 were among the first who recognized the advantages of reducing the column size in LC. Their work attracted the attention of a significant number of groups, each pursuing significantly different aims, e.g. increased detection sensitivity due to the reduced chromatographic dilution, the ability to work with small quantities of sample in case of limited availability, easier interfacing to flame based detectors and mass spectrometry, hyphenation to other separation techniques, and other inherent advantages of reducing the column i.d. Other directions towards miniaturization in LC are the use of coated open tubular columns with an i.d. of 5-50 IJm and etched channels on a micro-chip as separation columns. In poten­ tial, open tubular LC is a very promising technique since it can produce large theoretical plate numbers in short analysis times [12]. However, the technique has been applied with limited success so far, which is mainly due to the demanding instrumental requirements. Micro-chip LC is still in its development stage and seems to move towards "sensor-type" of applications. Generally this type of separation devices lack the resolving power of more conventional LC techniques due to the limited column length. Furthermore, the difficulty with techniques such as open tubular LC and micro-chip LC remains sample preparation and introduction. Packed LC columns with an i.d. of 50-300 IJm haven proven to provide the best analytical results so far. Although such columns may not be the ultimate goal in miniaturization in col­ umn liquid chromatography, they already provide the possibility to separate complex mixtures. More importantly, the advantages of miniaturization with such columns can be achieved rela­ tively easy, allowing them ultimately to be used in routine
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