An approach to high throughput measurement of accurate retention data in liquid chromatography

Stoll, Dwight R.; Kainz, Gudrun; Dahlseid, Tina A.; Kempen, Trevor; Brau, Tyler; Pirok, Bob W.J.

doi:10.1016/j.chroma.2022.463350

Cited by 9 publications

(24 citation statements)

References 33 publications

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“…A number of other small adjustments have been made to the protocol as described in the following sections. The major reasons for moving to the "feed injection" approach for sample introduction were: 1) to avoid flow rate variation during sample introduction due to compression of the sample, and using a simpler valve technology than was used in our previous work [11]; and 2) to enable a series of rapid sample introductions from one sample draw without the time overhead associated with the robotic motions of a conventional autosampler. Chromatograms from absorbance at 254 nm were exported to .csv files for processing outside of ChemStation as described previously [11].…”

Section: Methodsmentioning

confidence: 99%

“…The data treatment and calculations used in this work are the same as described previously [11]; the key aspects are repeated here due to their central importance to the work, and for clarity. Please see the prior publication for details not mentioned here.…”

Section: Calculations and Full Datasetmentioning

confidence: 99%

“…Please see the prior publication for details not mentioned here. The peaks of interest in chromatograms from short columns are first fit to the modified Pearson VII function [11], and then the first moment of the fit function is calculated numerically and used as a measure of "retention time". Next, a selectivity value is calculated for a compound of interest based on measurements made using a short column (αi,S) as follows, where the retention of the reference compounds (toluene in this case; tr,ref,S) is measured in a mobile phase of 50/50…”

Section: Calculations and Full Datasetmentioning

confidence: 99%

“…), can be used to accurately predict retention factor for a compound of interest on the longer column, which can in turn be used to guide method development, for example. This approach can overcome inaccuracies in retention factors measured using short columns that result from the volumes of the frits at the column ends that are difficult to account for experimentally [11].…”

Section: Introductionmentioning

confidence: 99%

“…In each case a series of retention measurements was made starting with the mobile phase with the highest ACN level, and working systematically toward the mobile phase with the lowest ACN level. The rationale for this approach was described in the preceding paper in this series [11].…”

Section: Feed Injection Approach To Sample Introductionmentioning

confidence: 99%

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Characterization of a High Throughput Approach for Large Scale Retention Measurement in Liquid Chromatography

Kempen

Dahlseid

Lauer

et al. 2023

Preprint

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Many contemporary challenges in liquid chromatography - such as the need for “smarter” method development tools, and deeper understanding of chromatographic phenomena - could be addressed more efficiently and effectively with larger volumes of experimental retention data than we have been accustomed to historically. The paucity of publicly accessible, high-quality measurements has been due, at least in large part, to the high cost in time and resources associated with traditional retention measurement approaches. Recently we described an approach to improve the throughput of such measurements by using very short columns (typically 5 mm), while maintaining measurement accuracy. In this paper we present a perspective on the characteristics of results obtained using this approach using a dataset containing about 13,000 retention measurements, and describe an approach to sample introduction that improves upon the prior work. The dataset is comprised of results for 35 different small molecules, nine different stationary phases, and several mobile phase compositions for each analyte/phase combination. During the acquisition of these data, we have interspersed repeated measurements of a small number of compounds for quality control purposes. The data from these measurements not only enable detection of “out of control” measurements, but also assessment of the repeatability and reproducibility of retention measurements over time. For retention factors greater than 1, the mean relative standard deviation (RSD) of replicate (typically n=5) measurements is 0.4%, and the standard deviation of RSDs is 0.4%. Most differences between selectivity values measured six months apart for 15 non-ionogenic compounds were in the range of +/- 1%, indicating good reproducibility. A critically important observation from these analyses is that selectivity (defined here as retention of a given analyte relative to the retention of a reference compound; kx/kref) is a much more consistent measure of retention over time (months) compared to the retention factor alone. While this work and dataset also highlights the importance of stationary phase stability over time for achieving reliable retention measurements, we are nevertheless optimistic that this approach will enable the compilation of large databases (>> 10,000 measurements) of retention values over long time periods (years), which can in turn be leveraged to address some of the most important contemporary challenges in liquid chromatography. All of the data discussed in the manuscript are provided as Supplemental Information.

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Section: Methodsmentioning

confidence: 99%

Section: Calculations and Full Datasetmentioning

confidence: 99%

Section: Calculations and Full Datasetmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Feed Injection Approach To Sample Introductionmentioning

confidence: 99%

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