A Densitometric Method for Determination of Mangiferin, an Antioxidant Compound, with Thin Layer Chromatography in the Leaf Extracts of Coffee (Coffea Arabica L.)

Abstract

Mangiferin is one of the antioxidants in Coffea arabica L. leaves that has many pharmacological effects. The content of secondary metabolites in the leaves including mangiferin can be affected by age. A Thin Layer Chromatography (TLC) method for the quantitative analysis of mangiferin in Coffea arabica L. leaves extract was developed and validated. The method was developed using a mobile phase prepared with analytical grade solvents: ethyl acetate, methanol, formic acid, and deionized water (8:2:1:1 v/v/v/v). Regression functions were established over the 199.98-600.00 ng/spot range with r=0.999. The limit of detection (LOD) and limit of quantification (LOQ) were 13.87 and 41.61 ng, respectively. The method was selective with a resolution value of more than 1.5 and specific with the spectra correlation value for purity and identity check of more than 0.99. The percentage RSD was found to be 2.43% for repeatability precision and 2.05% for intermediate precision. The method's accuracy was determined through the standard addition method by adding known quantities of standard mangiferin to the pre-analyzed test solution and the mean recovery was 101.69± 1,21%. This TLC Densitometry method was linear, sensitive, selective and specific, precise, accurate, and can be used for routine analysis of mangiferin. On the young Coffea arabica L. leaves, the concentration of mangiferin ± RSD was 0.830 ± 1.71% w/w, and on the old Coffea arabica L. leaves was 1.128± 1.59% w/w.