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Instrument considerations for reliable amino acid analysis

Waters
5 Dec 2023

Amino acid analysis (AAA) is performed regularly for a wide range of samples including hydrolyzed proteins, cell culture, as well as and food and feeds samples. However, traditional HPLC methods for these analyses can have long run times and lower sensitivity. To address these challenges, Waters created an ACQUITY™ UPLC™ AAA Solution which includes a pre-column derivatization kit, sub-2 µm columns and pre-packaged mobile phases suitable for UPLC analyses. By scaling and adapting these reliable and reproducible methods from sub-2 µm particles to HPLC particle sizes (i.e., 2.5 µm), the AccQ•Tag™ Ultra solution chemistry can be used at lower pressures with conditions compatible for a wider range of systems.

In this application note, Waters Corporation details a study that presents the considerations for adapting this pre-column derivatization method from UPLC to UHPLC/HPLC systems. After adaptation and optimization of the separation method, performance is demonstrated, including linearity, repeatability, reproducibility, limit of quantitation, and limit of detection.

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UHPLC and HPLCHigh performance liquid chromatography (HPLC) and ultra high performance liquid chromatography (UHPLC), also known as UPLC, are analytical techniques used to separate, identify and quantitate components of complex mixtures including biological samples such as proteins and lipids as well as chemical mixtures of pesticides, drugs and oils. Both techniques are liquid chromatographic methods but differ by operating pressures (HPLC < 6000 psi < UHPLC ). Components of HPLC and UHPLC systems include columns, detectors, pumps, autosamplers and column heaters. Explore a range of UHPLC and HPLC columns for your specific sample needs including reverse phase, normal phase, ion exchange, HILIC, ion exclusion and size exclusion columns. For more specialized HPLC, explore FPLC, countercurrent LC and simulated moving bed systems. Find the best UHPLC and HPLC equipment in our peer reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Sample PreparationSample preparation can improve the quality and speed of separation techniques. Products to assist sample preparation include filtration equipment, evaporators, membranes and sieves.Preparative ChromatographyPreparative chromatography equipment is used for purification purposes, separating and collecting compounds eluted from a mixture which can then be subsequently used for further purposes. This can be done in large quantities or small. Preparative chromatography systems are usually low to medium pressure liquid chromatographs that are semi-automated. Explore a range of preparative columns and fraction collectors. Find the best preparative chromatography equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Amino AcidsUVUltraviolet (UV) light is used in a variety of applications, including disinfection, spectrophotometry, and molecular biology. UV light can damage DNA, making it a powerful tool in sterilization and in studies of mutagenesis and gene repair. Explore UV-related products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Cell CultureCell culture involves growing cells under controlled conditions, typically outside of their natural environment, to study cellular behavior, drug response, and disease models. Cell culture is crucial in biotechnology and biomedical research. Explore cell culture products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.

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Instrument considerations for reliable amino acid analysis