High-Sensitivity Simultaneous Analysis of Chiral Amino Acids in 10 Minutes without Derivatization
Release of the LC/MS/MS Method Package for D/L Amino Acids
Contributing to Various Fields Such as Fermented Foods, Clinical and Antiaging Research
Shimadzu announces the release of its LC/MS/MS Method Package for D/L Amino Acids, for use with triple quadrupole liquid chromatograph mass spectrometers.
Background to the Development
The 20 amino acids that form the structure of proteins exist as mirror image optical isomers: the D-amino acids (D-form) and the L-amino acids (L-form) (chiral amino acids)*1). The L-form of amino acids exists in large quantities in foods and in the body as the structural elements of proteins and as nutritional sources. As food ripens or the body ages, a portion of these is known to change to the D-form. Assessing the amounts of the D- and L-forms is a focus of interest in a variety of areas. Examples include the evaluation of fermented foods, physiological functional analysis of the cranial nerve system, searches for biomarkers, and the health and beauty fields. However, the amino acids must be derivatized before being analyzed, and there have been calls for faster and more convenient analysis techniques.
In this context, the Fukusaki Laboratory at Osaka University’s Graduate School of Engineering successfully developed a technique for simultaneously analyzing chiral amino acids at high sensitivity in 10 minutes*2, *3). This technique uses dedicated chiral columns rather than performing derivatization. The LC/MS/MS Method Package for D/L Amino Acids was commercialized based on the results of the analyses using this technique by the Osaka University Shimadzu Analytical Innovation Research Laboratory.
Using this package can dramatically accelerate product development and research at food companies dealing with fermented foods, and at universities and research laboratories performing physiological functional analysis of D-amino acids.
*1: Excluding glycine
*2: Reference literature:
Nakano, Y., Konya, Y., Taniguchi, M., Fukusaki, E.
Journal of Bioscience and Bioengineering, 123, 134-138 (2016)
*3: With this technique, both D/L- forms of the secondary amino acid proline are co-eluted.
1. Simultaneous Analysis of Chiral Amino Acids in 10 Minutes
This technique allows chiral amino acids to be analyzed simultaneously in 10 minutes by using the CROWNPAK CR-I(+) and CR-I(-), dedicated columns with a chiral stationary phase.
2. Quantitatively Determine All D/L-Amino Acids Using Two Types of Columns
With the CROWNPAK CR-I(+) dedicated column, elution of amino acids is in the sequence from D- to L-, and with the CROWNPAK CR-I(-), the elution sequence is reversed. Extremely similar physicochemical properties are seen between glutamine and lysine, isoleucine and allo-isoleucine, and threonine and allo-threonine, so they may not separate properly. If amino acids with similar properties are found to overlap when using the CR-I(+), switching to the CR-I(-) makes it possible to separate and confirm the two overlapping amino acids.
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LC/MS/MS Method Package for D/L Amino Acids