HPLC-DAD-MS study of polyphenols from Artemisia absinthium, A. annua, and A. vulgaris

Abstract

and then fragmentizes them, recording the MS spectra. We recorded the mass spectra of the analyzed polyphenolic compounds as standards in the analytical method. We present the MS analysis mode and the specific ions from the mass spectra that we can use to identify every compound in Table 1. Eighteen polyphenolic compounds have been studied in the aerial parts of Artemisia absinthium L., A. annua L. and A. vulgaris L. The substances were one hydroxybenzoic acid, six cinnamic acid derivatives, four quercetin glycosides, and seven aglycones of flavonol and flavone type. The results are summarized in Table 2; the polyphenolic compounds are shown in order of their retention time. Quantification of constituents was performed using UV detection at 330 nm for phenol carboxylic acids and 370 nm for flavonoids. The results indicate the presence of caffeic and chlorogenic acid in all samples. p-Coumaric acid was found in all the herbal drugs except Artemisia absinthium, in which it exist only in ester form. Ferulic acid was present in all the analyzed samples, except Artemisia vulgaris, before hydrolysis. Caftaric acid and sinapic acid are absent; gentisic acid is present only as an ester in Artemisia annua and A. vulgaris. In the case of flavonoids, rutoside and isoquercitrin were present in all three herbal drugs; hyperoside was found only in Artemisia absinthium, and quercitrin only in A. vulgaris. Fisetin was identified in A. absinthium and A. annua, after hydrolysis. Quercetin is present as the free aglycon in A. absinthium and A. annua. Patuletin was found in A. annua before and after hydrolysis and in A. absinthium as glycoside. All analyzed samples contain the flavonic aglycon luteolin. Kaempferol was found as the free aglycon in A. absinthium and A. annua, and in glycoside form in A. vulgaris. Myricetin could not be detected in any of the samples. A. absinthium and A. annua both have apigenin as the free aglycon. Apparatus and Chromatographic Conditions. Analysis was performed on an Agilent 1100 HPLC Series system equipped with a G1322A degasser, a G1311A quaternary gradient pump, and a G1313A autosampler. For the separation, a Zorbax SB-C18 reversed-phase analytical column was employed (100 3.0 mm i.d., 5 m particle), fitted with precolumn Zorbax SB-C18, both operated at 48C.