Quantitative Analysis of Eugenol in Clove Extract and Nanoparticle Formulation by a Validated High-Performance Thin-layer Chromatographic Method

Abstract

Eugenol (4-allyl-2-methoxyphenol) is the aromatic compound of the essential oil with the potential to modulate neuronal excitability. Nose to brain route of drug delivery via nanoparticle formulation has emerged as an alternative route drug delivery system for treatment of epilepsy. A selective and sensitive analytical method is required for evaluation of eugenol-based novel drug delivery systems. The objective of present study is to develop and validate a high-performance thin-layer chromatographic (HPTLC) method for the quantitative analysis of eugenol as bulk, in clove extract and in developed eugenol-loaded nanoparticle formulation. Chromatographic separation was achieved on silica gel pre-coated TLC aluminium plates 60F254, using methanol:distilled water (6:4, v/v) as the mobile phase. Quantitative analysis was carried out by densitometry at a wavelength of 280 nm. The method was validated as per ICH guidelines, to analyse eugenol in clove extract and to evaluate eugenol-loaded nanoparticles. Eugenol spots were observed at Rf value 0.58 ± 0.02. The detector response was linear (r = 0.9991) between 0.5 and 5.0 ng/spot. The intra- and inter-day precisions were 1.08–2.17 and 1.95-3.86 %, respectively. The limit of detection was 50 ng/spot and the limit of quantification was 150 ng/spot. The method proved to be simple, accurate, reproducible and rugged for eugenol. Evaluation of eugenol-loaded nanoparticle formulation demonstrated drug loading of 35.0%, encapsulation efficiency of 47.0% and sustained drug release following biphasic pattern. The present method is useful for the quantitative and qualitative analysis of eugenol and eugenol-loaded nanoparticle formulation. It provides significant advantages in terms of greater specificity and rapid analysis. © 2020 iGlobal Research and Publishing Foundation. All rights reserved.