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Novel Method for HPLC Analysis of Triterpenic Acids Using 9-Anthryldiazomethane Derivatization and Fluorescence Detection

Woźniak, Łukasz, Marszałek, Krystian, Skąpska, Sylwia, Jędrzejczak, Renata
Chromatographia 2017 v.80 no.10 pp. 1527-1533
alkaline hydrolysis, derivatization, epicuticular wax, esters, flowers, fluorescence, fruits, high performance liquid chromatography, leaves, mass spectrometry, metabolites, quantitative analysis
Triterpenic acids are a group of secondary plant metabolites which are part of the cuticular waxes covering fruits, leaves, and flowers. To date, quantitative analysis of these compounds has often been conducted using high-performance liquid chromatography coupled with spectrophotometric detection or mass spectrometry; however, these methods have some major drawbacks. This paper reports a new method of analysis implementing derivatization with 9-anthryldiazomethane and fluorescence detection. The method consists of the extraction of analytes from a matrix, purification with anion exchanging SPE columns, and an optional step of the alkaline hydrolysis of triterpenic acid esters. The paper also describes a fast and easy method for the synthesis of the derivatization agent. The detection limits of the method presented are approximately 100-fold lower than in a similar method using ultraviolet spectrophotometry as the mode of detection. The recovery and repeatability of the method are at satisfactory levels.