Doğal Ürünlerdeki Kynurenik Asit İçeriğinin RP-HPLC-UV ile Belirlenmesine Yönelik Yöntem Doğrulama

Year 2024, , 188 - 196, 18.11.2024

Yakup Kara , Sevgi Kolaylı

https://doi.org/10.31467/uluaricilik.1468749

Abstract

Kynurenik asit (KYNA) başta kestane balı olmak üzere ıhlamur ve diğer bazı ballarda bulunan farmakoaktif özelliğe sahip bir metabolittir. KYNA’nın antiinflamatuar ve immünosüpresif fonksiyonu düşünüldüğünde biyolojik yolaklar için çift taraflı bir etkiye sahip olduğu görülebilir. Bu sebeple de önemli bir doğal ürün olan ballardaki miktarının belirlenmesi ve bilinmesi sağlık üzerinde yüksek bir etkiye sahip olabilir. Bu metabolitin hem doğal ürünlerde hem de hayvan dokularında tespitine yönelik çalışmalar devam etmekte olup, hızlı ve kolay uygulanabilir metotların geliştirilmesi önem arz etmektedir. Bu çalışma kapsamında doğal ürünlerin (bal, kestane poleni ve kestane çiçeği) KYNA içeriğinin kantitatif olarak kısa sürede belirlenmesi amacıyla ters faz yüksek performanslı sıvı kromatografisinde ultraviyole dedektörü (RP-HPLC-UV) kullanılarak yeni bir metot geliştirilmiştir. Elde edilen verilere göre LOD ve LOQ değerleri sırasıyla 0,030 µg/mL ve 0,092 µg/mL olarak bulunmuştur. Aynı zamanda KYNA’nın ultra saf su (UPW), %70 EtOH, EtOH ve MeOH çözücülerinde hazırlanmış çözeltileri bu metotta analiz edilmiş ve UPW ‘un en uygun çözücüsü olduğu görülmüştür. Bu araştırmanın bulguları, özellikle kestane balının botanik kökeninin ayırt edilmesinde uygulanması açısından KYNA'nın ölçülmesine önemli ölçüde katkıda bulunabilir.

Keywords

Kynurenik asit, Kestane balı, HPLC-UV

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 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, 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 created 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

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