HPLC Fluorescence Method for Eugenols in Basil Products Derivatized with DIBI

Area of Science:

• Analytical Chemistry
• Food Chemistry
• Pharmacognosy

Background:

• Eugenols (Eugs), including eugenol (Eug), methyleugenol (MeEug), and linalool (Lin), are key bioactive compounds in basil products.
• These compounds possess a terminal double-bond, crucial for their chemical reactivity and biological activity.

Purpose of the Study:

• To develop a sensitive high-performance liquid chromatography (HPLC)-fluorescence method for determining Eugs in basil products.
• To validate the method's performance for accurate and reliable quantification of Eugs.

Main Methods:

• Derivatization of Eugs using 4-(4,5-diphenyl-1H-imidazol-2-yl)iodobenzene (DIBI).
• Separation of derivatized Eugs (DIB-Eugs) using an Atlantis T3 column with a methanol-water mobile phase.
• Detection via HPLC-fluorescence with optimized parameters for sensitivity and linearity.

Main Results:

• The method achieved good separation of DIB-Eugs within 20 minutes.
• Excellent linearity (r ≥ 0.999) was observed for Eug standards from 0.1-50 µM.
• Low limits of detection (1.0-6.0 nM) and quantitation (<19.9 nM) were achieved.
• High accuracy (96.8-104.6%) and precision (RSD < 1.2% intra-day, < 9.6% inter-day) were demonstrated.
• Recoveries ranged from 96.0% to 99.0% for the Eugs.
• DIB-Eugs exhibited stability at room temperature and 4°C for extended periods.

Conclusions:

• The developed HPLC-fluorescence method is sensitive, accurate, and precise for the simultaneous determination of Eugs in basil products.
• This method serves as a valuable tool for the quality control and evaluation of commercially available dried basil products.