METHOD VALIDATION FOR DETERMINATION OF KYNURENIC ACID CONTENT IN NATURAL PRODUCTS BY RP-HPLC-UV

Abstract

Kynurenic acid (KYNA) is a metabolite with pharmacoactive properties found primarily in chestnut honey, linden, and some other honeys. Considering the anti-inflammatory and immunosuppressive functions of KYNA, it can be seen that it has bidirectional effects on biological pathways. For this reason, determining and knowing the amount of honey, which is an important natural product, can have a high impact on health. Studies on the detection of this metabolite in both natural products and animal tissues are ongoing, and it is important to develop fast and easily applicable methods. Within the scope of this study, a new method was developed using an ultraviolet detector (RP-HPLC-UV) in reverse phase high-performance liquid chromatography to determine the KYNA content of natural products (honey, chestnut pollen, and chestnut flowers) quantitatively in a short time. According to the data obtained, LOD and LOQ values were found to be 0.030 µg/mL and 0.092 µg/mL, respectively. At the same time, solutions of KYNA prepared in ultrapure water (UPW), 70% EtOH, EtOH and MeOH solvents were analyzed in this method, and it was found that UPW was the best solvent. The findings of this research can contribute significantly, particularly in the application of measuring KYNA, to distinguishing the botanical origin of chestnut honey. Keywords; Kynurenic acid, Chestnut honey, HPLC-UV