Chromatography 11 Monolithic Chromatography and its Modern Applications


High-performance liquid chromatography has become the predominant global separation technique over the past four decades. However, the recent development of an alternative column packing material - a one-piece porous solid known as a ‘monolith’ - has transformed the fi eld. Monoliths have attracted a large amount of interest due to the enhanced speed and thoroughness with which they can separate complex mixtures of biological molecules.


Monolithic Chromatography and its Modern Applications is the book for any seasoned scientist already using monolithic columns, as well as those novices who are new to the area and would like to know more about this technique. It’s the ideal resource for all analytical laboratories thinking of improving their separation protocols. In this book, global experts comprehensively describe this developing fi eld, with part one covering the synthesis and characterisation of the columns and part two discussing their numerous applications.


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Innovative Filtration Technology to Help Prevent Clogging of LC Columns


Highly Stable Detector Improves GPC/SEC Results


The Differential Refractive Index (DRI) detector from Testa Analytical Solution eK offers unmatched accuracy and reliability for determination of absolute concentration and total mass balance in Gel Permeation Chromatography/Size Exclusion Chromatography (GPC/SEC) applications.


The baseline signal stability of any liquid chromatography detector is very important, as it is a limiting factor in peak determination. Detector stability gains even more importance where analysis time is long, such as is commonly encountered in high resolution GPC/SEC applications.


Use of multiple columns is common in GPC/SEC as a technique to increase resolution and also to increase the available dynamic range in terms of molecular weight. As such, high resolution GPC/SEC measurements can often take 30 to 60 minutes for a single separation. As a result, most HPLC detectors adapted for use in GPC/SEC will show a signifi cant baseline drift between the beginning and end of the chromatogram thus negatively infl uencing the ability to accurately determine the beginning and end of sample peaks and therefore to calculate the correct polydispersity.


Operating from room temperature up to 80°C with high thermal stability, Testa Analytical’s DRI detector has been designed with particular attention to eliminating drift. As a consequence of its unmatched baseline stability, the DRI detector delivers excellent performance for chromatographic separations of any duration.


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DPX Technologies developed a patent-pending Tip-on-Tip (ToT) technology as a natural progression of their INTip product lines designed to promote the simplifi cation and automation of complex sample preparation. ToT technology combines a bottom Filtration tip with a top wide bore conductive tip. The top tip aspirates a sample solution, fi ts into the Filtration tip, and dispenses the sample through the Filtration tip providing a clean, particulate-free solution. Filtration tips are available in a variety of porosities.


ToT Filtration has broad applicability, including forensic, clinical, bioanalytical, organic synthesis and cannabis sample preparation. The methodology provides an automated alternative to traditional fi ltration/centrifugation. ToT fi ltration is ideal for high-throughput online protein precipitation, beta-glucuronidase removal, or any other application requiring particulate removal. Particulates can be removed down to less than 1 µm to prevent clogging of LC columns.


ToT Filtration can be modifi ed to incorporate sorbent for more extensive purifi cation. These workfl ows can centre around binding matrix to the sorbent (ToT Cleanup) where the fi ltrate has purifi ed analyte to be further analysed or around binding the analyte (ToT SPE) to be further washed and eluted. In each workfl ow, the sorbent and sample solution are mixed in a well for binding to occur. The top tip aspirates the sorbent/sample mixture, fi ts into the Filtration tip, and dispenses through the Filtration tip where the sorbent is retained, as seen in the image provided. These workfl ows are compatible with a variety of commercially available chemistries.


ToT Cleanup or SPE methodologies are ideal for applications that require specifi c incubation conditions between sorbent and sample solution, like immunoaffi nity purifi cation. This provides an automated, cost effective alternative to workfl ows that traditionally use magnetic beads. Filtration tips are compatible with budget friendly polymer- and agarose-based resins; designed to eliminate particulates in the elution and maximise the utility of the automated liquid handling system.


The beauty in this technology is that customers can utilise their existing chemistries/workfl ow and use Filtration tips for ToT Cleanup or SPE to automate that workfl ow. DPX has application notes and a Tip-on-Tip technology guide online with images and schematics covering steps for varying workfl ows.


Download resources for INTip methods and applications. More information online: ilmt.co/PL/PaAO


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