Development and Validation of a UHPLC Method for Biogenic Amines Using Microwave-Assisted Dansyl Chloride Derivatization

Ng, Danny Kah Kit (2026) Development and Validation of a UHPLC Method for Biogenic Amines Using Microwave-Assisted Dansyl Chloride Derivatization. Final Year Project (Bachelor), Tunku Abdul Rahman of Management and Technology.

Text DANNY NG KAH KIT_Full Text.pdf Restricted to Registered users only Download (1MB)

Abstract

Determination of Biogenic Amines (BAs) is very important as their concentration levels are closely related to human health. High or excessive concentration of BAs in the human body may cause various diseases and impair normal physiological functions. Moreover, BAs can serve as an index of food freshness and as microbial spoilage indicators. The most common BAs includes tryptamine (TRP), phenylethylamine (PEA), histamine (HIS), putrescine (PUT), cadaverine (CAD), tyramine (TYR) and spermidine (SPD). In this study, the derivatization of BAs was carried out by using Dansyl Chloride as derivatization with the assist of Microwave Synthesis Reactor to increase the rate of reaction was carried out. Since BAs lack strong chromophores, this derivatization is necessary to enhance their detection sensitivity for UV-VIS spectroscopy. Dansyl Chloride was used as the derivatization reagent because it reacts with amino groups to form stable derivatives with strong UV absorbance. To speed up the derivatization process, a Microwave Synthesis Reactor was applied. Microwave-assisted derivatization was generally faster and more efficient compared to conventional methods, and it helped to improve the overall analysis. The determination of BAs was performed using Ultra-High-Performance Liquid Chromatography (UHPLC) coupled with a UV-VIS Diode Array Detector (DAD). This approach allows for the efficient separation of structurally similar BAs and provides reliable quantification. By combining dansyl chloride derivatization with UHPLC–DAD analysis, all seven targeted BAs were successfully identified and quantified.