Carole Mathe

A multivariate study of the performance of an ultrasound-assisted madder dyes extraction and characterization by liquid chromatography-photodiode array detection

An extraction method of madder (Rubia tinctorum) roots dyes is established and optimized to obtain the original chemical composition. A central composite design (CCD) was developed to specify the importance of the three major factors studied (time, temperature and solvent composition) affecting the ultrasound-assisted extraction of this matrix. A preliminary granulometric study of madder roots is realized in the aim to determine the optimal particles size corresponding to the best ultrasound effects. A comparison with the classical extraction method of madder dyes by reflux is described. The identification of the constituents of R. tinctorum is carried out by liquid chromatography coupled with a photodiode array detector (LC-PDA). Anthraquinonic aglycone and heterosidic dyes compounds are characterized by retention time and UV spectrum: alizarin (1,2-dihydroxyanthraquinone), purpurin (1,2,4-trihydroxyanthraquinone), lucidin (1,3-dihydroxy-2-hydroxymethylanthraquinone), rubiadin (1,3-dihydroxy-2-methylanthraquinone), xanthopurpurin (1,3-dihydroxyanthraquinone), pseudopurpurin (1,2,4-trihydroxy-3-carboxyanthraquinone), lucidin primeveroside, ruberythric acid (alizarin primeveroside), galiosin (pseudopurpurin primeveroside) and rubiadin primeveroside. The optimal experimental conditions are 18min, 36 degrees C and 37/63 MeOH/H(2)O (v/v).

Cytohistological and phytochemical study of madder root extracts obtained by ultrasonic and classical extractions.

INTRODUCTION Madder (Rubia tinctorum) has been used since ancient times as a source of pigments for dyeing and painting. Madder dyes are localised in roots and the native chemical population is composed of glycosiled and aglycone compounds. The aim of this study is to elaborate an efficient extraction process without any chemical denaturation of dyes. OBJECTIVE To compare an optimised ultrasonic process, using for madder dye extraction, with two conventional procedures and to determine the efficiency of ultrasound on these vegetable matrix. METHODOLOGY Madder roots were extract in a methanol-water mixture in 37 : 63 (v/v) for ultrasound and 80 : 20 (v/v) for reflux and agitation. HPLC-PAD analyses showed the anthraquinone proportion for each extraction process and their denaturing effects. Finally, cytohistological observations were made to show the consequence of each process on the cell organisation in madder roots. RESULTS The results showed that the amount of extracted dyes was higher with UAE than with agitation and reflux. HPLC-PAD analysis revealed that the anthraquinone composition differed according to the extraction procedure. The UAE extracts presented an important richness in terms of anthraquinonic compounds that suggests a preserving effect. Cytohistological observations showed that the main alterations concerned the cell walls of phloem. After UAE the walls exhibited numerous pitted areas reflecting an ultrasound-induced cavitation that enhances the extraction effectiveness of this method. CONCLUSION The study has shown the improvement of madder roots extraction both quantitatively and qualitatively using the efficiency of ultrasound-assisted extraction in comparison with magnetic agitation and reflux techniques.

LIQUID CHROMATOGRAPHY OF TRITERPENIC RESINS AFTER DERIVATIZATION WITH DANSYL CHLORIDE

A chromatographic method using fluorescing derivatization was developed for the analysis of triterpenoids in natural resins. The separation was performed using a Luna-C18 column, an acetonitrile-water eluant mixture, a flow-rate of 1 mL · min−1. The fluorescent compounds were detected at 530 nm (excitation at 347 nm). A derivatization reaction of hydroxyl groups of triterpenoids using dansyl chloride is described; this process appeared unusual and provided some novelty. The detection limits were obtained in the range of 0.15–0.45 ng/mL (S/N = 3) for dansylated standard compounds. The described procedure allowed the simultaneous determination of twelve triterpenoids present in frankincense species (Boswellia carteri and B. frereana) and was successfully applied to an archaeological sample coming from an Egyptian mummy. Two other resinous materials, dammar and mastic resins employed in Cultural Heritage, were also studied, in which nine and seven triterpenoids are respectively characterized.