Reducing pressure and increasing peak capacity by incorporating an adaptable flow stream splitting platform to high-performance liquid chromatography columns: A study on C18 silica-based monoliths.

In this study, an adaptable end-column platform was fitted to a commercially available monolith, which enabled the column to be fitted with a flow-splitting device. A variety of flow-splitting adapters could be incorporated into the platform, and in this study, a radial flow stream splitter was utilized. The advantage of the radial flow stream spitter was that it overcame issues relating bed density variations that could cause bands to distort in the radial cross-section of the column. Using propylbenzene as a test standard in isocratic elution mode, height equivalent to a theoretical plate curves were constructed across ten flow rates, and it was found that the column efficiency improved by as much as 73%. Furthermore, the dual outlet flow splitter enabled a very substantial reduction in column back pressure, with the decrease being consistently between 20 to 30% depending on the column length. Additionally, sensitivity increased by 45%, consistent with the observed increase in efficiency. The adaptable end-column platform could be retrofitted to almost any commercial column with the expectation of gaining efficiency, sensitivity, and reducing back pressure.

Using HPLC with In-Column Derivatization to Authenticate Coffee Samples.

Coffee is one of the world's most popular beverages, with the global coffee capsule market worth over USD 4 billion and growing. The incidence of coffee fraud is estimated to be up to one in five coffees being contaminated with cheaper blends of coffee. Given the worsening extent of climate change, coffee crop yields are harder to maintain, while demand is increasing. The 2021 Brazil frost delaying or destroying many coffee crops is an example. Hence, the incidence of coffee fraud is expected to increase, and as the market becomes more complex, there needs to be faster, easier, and more robust means of real-time coffee authentication. In this study, we propose the use of novel approaches to postcolumn derivatization (termed herein as in-column derivatization) to visualize the antioxidant profiles of coffee samples, to be later used as indicators for authentication purposes. We propose three simple mathematical similarity metrics for the real-time identification of unknown coffee samples from a sample library. Using the CUPRAC assay, and these metrics, we demonstrate the capabilities of the technique to identify unknown coffee samples from within our library of thirty.

A Rapid Screening Analysis of Antioxidant Compounds in Native Australian Food Plants Using Multiplexed Detection with Active Flow Technology Columns.

Conventional techniques for identifying antioxidant and phenolic compounds in native Australian food plants are laborious and time-consuming. Here, we present a multiplexed detection technique that reduces analysis time without compromising separation performance. This technique is achieved using Active Flow Technology-Parallel Segmented Flow (AFT-PSF) columns. Extracts from cinnamon myrtle (Backhousia myrtifolia) and lemon myrtle (Backhousia citriodora) leaves were analysed via multiplexed detection using an AFT-PSF column with underivatised UV-VIS, mass spectroscopy (MS), and the 2,2-diphenyl-1-picrylhydrazyl (DPPH(•)) derivatisation for antioxidants as detection methods. A number of antioxidant compounds were detected in the extracts of each leaf extract.

Regeneration of a silica monolithic rod column using harsh methods followed by firm thermodynamic and kinetic validation.

In this study, a numerical tool is introduced--based on thermodynamic and kinetic separation theory--for validating the regeneration of monolithic rod columns after cutting their inlet sections. A long-used RP-18e monolithic column was deemed to be unfit for further coffee analysis because of poor separation performance. The columns brownish inlet section was physically removed with a lathe, leaving a clean white inlet section. The original and regenerated columns were extensively analyzed and compared using numerical tools for processing adsorption data. The perturbation peak method was used to measure the adsorption isotherm of phenol on the original and regenerated monolith and the adsorption energy distributions were calculated for identifying any change in the degree of heterogeneity. Although peak shapes improved considerably after regeneration, no significant differences were found in the detailed characterization of the processed adsorption data between the original column and the regenerated one. This indicates that the removal of a section of the monolithic bed can be undertaken without damaging the column and columns in which their inlet head sections are contaminated may still function with normal adsorption behavior. In addition, the combined thermodynamic and kinetic methodology could accurately be used to evaluate any regeneration method of columns.

Segmented flow and curtain flow chromatography: overcoming the wall effect and heterogeneous bed structures.

The variation in mobile phase velocity as a function of the column radius has been shown to be a major limitation in the efficiency of HPLC columns. One contributing factor to the variability in the flow velocity stems from the heterogeneity in the radial packing density, leading to what has been described as the 'wall-effect'. The wall-effect generates parabolic-type elution profiles, which dilutes the sample and creates tailing bands. In this communication a new column technology is discussed that has been designed to overcome the wall effect, minimising the limitations associated with packing heterogeneity. This technology has been referred to as active flow technology and consists of two types of column designs, parallel segmented flow and curtain flow. In both these column designs sample that elutes through the column in the radial central region of the bed is separated from the flow that elutes along the wall region. Hence, the sample that elutes through the most efficiently packed region of the bed is collected to the detector. As a consequence more theoretical plates are obtained, and sensitivity is increased since the sample is not diluted by the diffuse tail. Sensitivity is enhanced further in the curtain flow design. The benefits of these new columns are discussed.

Two-dimensional HPLC analysis of oligostyrenes: comprehensive and online heart-cutting techniques.

2-D HPLC incorporating two reversed phase (RP) environments was employed for the isolation of oligomers and their diastereomers of low molecular weight oligostyrenes. The operation of a comprehensive method of analysis was compared to a heart-cutting approach. The comprehensive approach employed a high resolution diastereomer separation in the first dimension and a low peak capacity C18, high speed separation according to molecular weight. Because of solvent incompatibility between the dimensions in the comprehensive method, successful separation of the diastereomers of the oligomers was not possible. The heart-cutting approach used a C18 monolith in the first dimension, which was selective only for molecular weight. Entire molecular weight fractions were then transported to the second dimension in an online heart-cutting process for the separation of diastereomers. The heart-cutting process was more successful in that 228 components of the 511 within the sample were recognized. This series of separations was undertaken in less than 6 h.