Comparison of commercial organic polymer‐based and silica‐based monolithic columns using mixtures of analytes differing in size and chemistry

Abstract

Commercially available silica-based monolithic columns Chromolith RP-8e, Chromolith RP-18, and Chromolith HR RP-18, and polymer-based monolithic columns ProSwift RP-1S, ProSwift RP-2H, and ProSwift RP-3U varying in pore size and bonded phase have been tested for the fast separation of selected sets of analytes. These mixtures of analytes included small molecules (uracil, caffeine, 1-phenylethanol, butyl paraben, and anthracene), acylated insulins, and intact proteins (ribonuclease A, cytochrome C, transferrin, apomyoglobin, and thyroglobulin), and covered wide range of chemistries and sizes. Small molecules were well separated with a height equivalent to theoretical plate of 11-26 μm using silica-based monolithic columns, while organic polymer-based monoliths excelled in the fast sub 1 min baseline separations of large molecules. A peak capacity of 37 was found for separation of acylated insulins on Chromolith columns using a 3 min gradient at a flow rate of 3 ml/min. Poor recovery of proteins from Chromolith columns and significant peak tailing of small molecules using ProSwift columns were the major obstacles in using monolithic columns in those applications.

High-Throughput and Ultrafast Liquid Chromatography

“…UHPLC is regularly being applied for HTS applications (typically with subminute separation time) and is often coupled to MS for detection . To rapidly separate peptides and proteins, SPPs, , monolithic columns, , and novel sphere-on-sphere particles , have all been used (Figure ). SPPs with very large pores (1000 Å) have been especially useful for the successful separation of large biomolecules with shorter method times, − partially due to unique perfusion-like flow through these particles that has been predicted by flow simulations .…”

“…Copyright 2017. Panel b is adapted from ref : Comparison of Commercial Organic Polymer-based and Silica-based Monolithic Columns Using Mixtures of Analytes Differing in Size and Chemistry, Vyviurska, O.; Lv, Y.; Mann, B. F.; Svec, F. J. Sep. Sci. , Vol.…”

Silica-Based Monolithic Columns as a Tool in HPLC—An Overview of Application in Analysis of Active Compounds in Biological Samples

“…Considering the type of material used for synthesis, monolithic columns can be divided into two groups; the first is based on silica gel and the second is based on polymeric materials [ 6 ]. Vyviurska et al presented an exhaustive comparison of both types of commercially available monoliths [ 7 ]. The major disadvantage of most polymeric monolithic fillings is their inability to separate small molecules [ 8 ]; hence, their significance in the analysis of samples with a complex matrix such as plant material is low.…”

Preparation of restricted‐access material precolumns by grafting δ‐gluconolactone onto a hybrid silica monolithic column for on‐line solid‐phase extraction of tetracycline residues from milk

“…The column has low column pressure and high mass transfer efficiency owing to unique pore structure . With the advantages of facile preparation, abundant surface functional groups, and controlled pore sizes distribution, the monolithic columns have been numerous reports such as organic polymer‐based, silica‐based and organic‐silica hybrid monolithic columns . Compard to the former monoliths, the hybrid monolithic column combines the merits of the former two monoliths and overcomes the inherent drawbacks like the instability under extreme pH and harsh solvents .…”