Size-Exclusion Chromatography (SEC)

At HPLC 2023, Matthias Pursch presented his findings on using UHPLC and SEC to monitor small molecule and polymer reactions in real time.

Characterizing lipid nanoparticles (LNPs) is a developing delivery modality in biotherapeutic analysis. We address some of the current challenges and opportunities in this field, including an examination of the most common tools used to characterize LNPs.

The latest developments and challenges of using liquid chromatography (LC) for copolymer analysis are discussed.

What parameters can influence an siRNA separation?

Determining a theoretical number of plates is an effective tool to ensure that the column is still applicable for the desired separation.

Determination of molar mass distributions and the molar mass averages derived therefrom are the main objectives of gel permeation chromatography/size-exclusion chromatography (GPC/SEC) analysis. But what is the meaning of these averages and how are they influenced by the setting of baselines and integration limits? This instalment of Tips and Tricks in GPC/SEC will try to provide a better understanding.

This installment of Tips & Tricks will provide some helpful pointers to keep your GPC/SEC system in good shape.

The state of protein-derived self-associated, aggregated, and fragmented impurities in therapeutics are critical quality attributes (CQAs) and are widely monitored using non-denaturing size-exclusion chromatography (SEC).

Developing and building a talent base in macromolecular separations should be a priority for employers. Here’s why.

What experimental details need to be considered when analyzing PEGs?

In 2015, the American Chemical Society’s Committee on Professional Training added a requirement to the ACS degree certification program that undergraduates learn about macromolecules, supramolecular aggregates, and nanomaterials (MSN). This requirement can be met by a specialized course in these topics, but many programs are also choosing the distribute these topics across the curriculum.

This exploration into commercially available light scattering detection systems for size-exclusion chromatography (SEC) emphasizes new technologies and will help users understand the options.

This instalment describes the interplay between column length, pore size distribution, and particle size to optimize GPC/SEC separations.

Hydrolyzed collagens (collagen peptides) are water-soluble products obtained by hydrolysis of natural proteins and used for dietary supplements. A simple GPC/SEC method is described for molar mass determination of collagen peptides, allowing reliable molar mass determination using ultraviolet (UV) detection.

Analytical size-exclusion chromatography (SEC) often suffers from several limitations. This article shows how some of these limitations can be partly (or even completely) resolved by a multi-angle light scattering (MALS) detector for the determination of molar mass distributions of synthetic and natural polymers.

How to create a calibration curve when no chemically-matching reference materials with narrow molar mass distribution are available.

A review of alternative approaches to narrow standard calibration.

Light scattering detectors are ideally suited for size-exclusion chromatography (SEC) because they provide the molecular weight and radius of gyration information of polymeric samples without column calibration. In this article, the fundamentals of light scattering as applied to SEC are introduced, with emphasis on the origin of the Rayleigh equation. This tutorial is geared to those new to the field or who already utilize light scattering and seek clarification regarding the multitude of equations associated with light scattering. Part X of this series will present a brief summary of commercially available light scattering instrumentation with emphasis on new detector technology. Owing to the complexity of data analysis and the many equations involved with light scattering measurements, part X will also include a glossary of principal symbols and a summary of relevant equations.

The macroscopic properties of material based on poly(D,L-lactic-co-glycolic acid) (PLGA) polymers are tunable by molar mass distribution and degree of branching, enabling optimization for applications in the pharmaceutical and medical industries. Size-exclusion chromatography followed by online multi-angle light scattering with intrinsic viscosity detection (SEC–MALS–IV) is an advanced analytical method for determining absolute molar mass distributions, identifying polymer conformation and quantifying branching. SEC–MALS–IV overcomes the errors that can be encountered in molar mass determined by conventional SEC, which arise from chemical composition and molecular structure, and provides comprehensive characterization of PLGA to facilitate the targeted development of optimized polymer.

Water-soluble macromolecules require water as a mobile phase, and method development therefore seems to be straightforward. At second glance, however, aqueous gel permeation chromatography/size-exclusion chromatography (GPC/SEC) is complex and the choice of the stationary phase and pH are crucial. In addition, mobile phase additives are often required to allow for interaction-free separations.

The chemistry of samples analyzed using gel permeation chromatography/size-exclusion chromatography/gel filtration chromatography (GPC/SEC/GFC) is very diverse. Different chemistries of stationary phases are required to allow for true size separation. Several types of materials are available, all of which have their advantages and limitations. While silica‑based stationary phases are most common in high performance liquid chromatography (HPLC), for macromolecules polymer-based phases are popular.

The “greenest solution” is certainly using no solvent but gel permeation chromatography/size-exclusion chromatography (GPC/SEC) as a liquid chromatography (LC) technique requires the use of a mobile phase. The growing awareness of the need for more sustainable (greener) solutions has focused attention on environmentally- and health-friendly solvents and solutions.

The accuracy of a measurement describes how close the measured result is to the true value. Accuracy is influenced by systematic errors, which are difficult to detect even for experienced scientists.

One of the major problems with plastics is recycling. Only a few materials can be recycled and the acceptance of recyclates is sometimes low. GPC/SEC can be applied to investigate the quality of materials containing recycled portions.