A Protocol for Determination of Proteinogenic Amino Acids in Biological Fluids by the High-Speed UHPLC-MS Method: Application on Transgenic Spontaneously Hypertensive Rat-24 Plasma and Cerebrospinal Fluid Samples

Abstract

Recently, proteinogenic amino acids have become very interesting molecules, accompanied by a large variety of metabolic processes in humans and are associated with various diseases. In the era of system biology, including a broad spectrum of associated disciplines (e.g., metabolomics, lipidomics, proteomics, etc.), the possibility of identifying trustworthy biomarkers of diseases becomes much more likely. Changes in amino acid levels in plasma, serum, or cerebrospinal fluid reflect physiological or pathological conditions and, therefore, their regular monitoring can lead to early detection of the occurrence of a disease. Therefore, the exact determination of amino acids in biological fluids is of great importance. However, it is necessary to dispose with an effective, accurate, precise, selective, and robust analytical method. This protocol describes the complex procedure of amino acid analysis based on a combination of UHPLC with single quadrupole MS. The protocol presents a highly reproducible and robust methodology that has already been established in the quality control of biopharmaceuticals and determination of proteinogenic amino acids in urine in our laboratory. Here, the application potential is extended to the most frequently investigated biological fluid, that is, plasma and to the cerebrospinal fluid, which is investigated in many neurological conditions.