Development of an ultra-sensitive laser stimulated fluorescence system for simultaneous detection of amino acids

Abstract

An ultra-sensitive, high-performance liquid chromatography-based laser-stimulated fluorescence detection system was developed and validated for the simultaneous detection of 20 derivatized amino acids. Dansyl chloride was used as a derivatizing agent, and key derivatization parameters, such as reaction time and temperature, were optimized to enhance sensitivity and reproducibility. The majority of amino acids showed a relative standard deviation of less than 5%, indicating the reliability of the approach. The method demonstrated excellent sensitivity for all 20 amino acids, with detection limits ranging from 4.32 to 85.34 femtomoles. It also exhibited good linearity, with regression (R²) values greater than 0.98 for the amino acids. The system's performance was tested on human serum, and the eluted amino acids were identified. This method has great potential for analyzing amino acids in various body fluids and can be used in various clinical applications. It is ultra-sensitive, reliable, user-friendly, and cost-effective, offering a valuable tool for diagnosing and managing amino acid-related disorders.