Stilbene, Ferulic acid and its derivatives from the roots of Angelica sinensis

T¹p chÝ Hãa häc, T. 42 (4), Tr. 508 - 511, 2004<br /> <br /> <br /> <br /> stilbene, Ferulic acid and its derivatives from the<br /> roots of Angelica sinensis<br /> Received 17-11-2003<br /> Nguyen Thi Hong Van , Nguyen Thi Hoang Anh1, Tran Van Sung1,<br /> 1<br /> <br /> Katrin Franke2 and Ludger Wessjohann2<br /> 1<br /> Institute of Chemistry, National Center for Natural Science and Technology,<br /> 2<br /> Institute of Plant Biochemistry, Halle/S., Germany<br /> <br /> <br /> SUMMARY<br /> The rhizome of Angelica sinensis (Oliv.) Diels (Umbelliferae) is one of the most commonly<br /> used traditional drugs in China, indicated against anemia, menstrual disorders, amenorroehea<br /> and rheumatism. From the ethyl acetate extract of the roots of A. sinensis a stilbene, ferulic acid,<br /> coniferylferulate and decorticate have been isolated. Their structures were identified by MS and<br /> NMR spectroscopic methods.<br /> <br /> <br /> I - INTRODUCTION spectroscopy and by comparison with the data<br /> published in the literature to be 4,4'-dihydroxy,<br /> The roots of Angelica sinensis are also 3,3'-dimethoxystilben (1), ferulic acid (2),<br /> known as "Danggui" in China. In China it serves coniferylferulate (3) and decorticate (4).<br /> as remedy against anemia, menstrual disorders,<br /> amenorroehea and rheumatism [1]. In the II - Experimental<br /> Vietnamese folk medicine it is used for the<br /> treatment of constipation and pain [2]. In Japan, 1. Plant material<br /> Angelica acutiloba is used as a substitute for A.<br /> sinensis. Research reports on the analysis its The roots of A. sinensis were bought in the<br /> ingredients are appeared in the literature since traditional medicine mark in Hanoi, March<br /> 1924. Recently, many advances have been made 2002. A voucher specimen was deposited in the<br /> in chemical and pharmacological studies of this Herbarium of Institute of Chemistry.<br /> drug. A series of components have been found 2. Instruments and Chemicals<br /> in the roots of A. sinensis by GC, HPLC-MS<br /> analysis. 53 constituents in its essential oil and NMR: Varian Unity 300; MS: AMD 402;<br /> 34 compounds in non-volatile fraction have for analytical purposes: Merck TLC aluminum<br /> been reported by Chen Yaozu [3]. Z-ligustilide sheets silica gel 60 F254 (layer thickness 0.2<br /> is the main component and coniferyl ferulate, mm) were used. Silica gel Merck 60 (0.040 -<br /> butylphthalide etc. are the major components. 0.063 mm) is used for column chromatography.<br /> The present communication deals with the 3. Extractions and Isolation<br /> isolation and structural elucidation of four Air dried and powdered tubers of A. sinensis<br /> compounds (1 - 4). Their structures have been were extracted with EtOH : H2O (95 : 5) at<br /> determined by MS, 1H and 13C-NMR room temperature. The ethanol extract was<br /> <br /> 508<br /> concentrated under vacuum and aq. soln. was Additionally, three singlets at 6.88, 5.63 and<br /> extracted with EtOAc and n-BuOH, 3.95 with the integrals of 1, 1 and 3 protons,<br /> successively. The solvent was evaporated in respectively, were also observed. The signal at<br /> vacuum to afford 160g EtOAc and 20g n-BuOH 5.63 could be assigned to a hydroxyl group. The<br /> extracts. last signal ( 3.95) exhibited the presence of a<br /> The EtOAc extract was separated by methoxy group. The 13C-NMR and APT spectra<br /> chromatography on silica gel, eluting gradient revealed only 8 carbons, including 3 quaternary<br /> with n-hexan, EtOAc and MeOH to furnish 345 signals, four protonated and one methoxyl<br /> fractions, which were combined according to signals. The analysis of spectroscopic data<br /> TLC monitoring. suggested that, compound 1 consisted of two<br /> The fraction containing compound 1 was equivalent moieties. The singlet at 6.88 and<br /> chromatographed on Sephadex LH-20, washing one protonated carbon more in the aromatic<br /> with MeOH, and further purified by preparative range led to conclude that two moieties of 1<br /> TLC eluted with CHCl3 : MeOH (9:1) to afford were connected with each other by a double<br /> compound 1, an amorphous powder, EI-MS m/z bond. Its structure was confirmed by<br /> (rel. int.): 272 [M]+ (30), 242 [M-2xCH3]+ (4), comparison with the data described in the<br /> 212 [M-CH3-CO-H2O+H]+ (9), 208 (8). literature [4] to be 4,4'-dihydroxy-3,3'-<br /> dimethoxy-stilben. This compound was isolated<br /> The fraction containing compound 2 was for the first time from the roots of Leuzea<br /> subjected on a Sephadex LH-20 column, eluting carthamoides [4].<br /> with MeOH and following by preparative TLC,<br /> developing with CHCl3 : MeOH (9 : 1) to yield Compound 2 was isolated as an amorphous<br /> compound 2, an amorphous powder, negative powder and gave the molecular ion peak at m/z<br /> ESI-MS: 193 [M-H]+, HRESIMS m/z 217.04734 194 [M]+ in the EI-MS spectrum. Its molecular<br /> [M+Na]+, C10H10O4 cal. 217.04713. formula was obtained from the HRESIMS as<br /> C10H10O4. The 1H-NMR spectrum indicated a<br /> Chromatography of fraction 35 on Sephadex trisubstituted aromatic ring [7.17 d (2.0), 7.05<br /> LH-20 with MeOH as solvent afforded dd (2.0, 8.2) and 6.30 d (8.2)], two trans ethenyl<br /> compounds 3 and 4, compounds 3 and 4 were protons with coupling constant of 15.92 Hz.<br /> isolated. Compound 3: colourless oil, EIMS: Furthermore, only methoxyl group was<br /> 356 [M]+ (2.8), 220 [M - C8H9O2 + H]+ (3.5), observed. 13C-NMR spectrum revealed 10<br /> 194 [M - C10H11O2 + H]+ (ferulic acid) (64), 176 carbons. Among them, 8 signals could be<br /> [M - C10H11O2- H2O + H]+ (100). Compound 4: assigned for aromatic ring and one double bond<br /> yellow powder, positive ESI-MS: 353 [M+Na]+. (111.5 - 150.2 ppm). Besides these carbons, a<br /> carbonyl (171.1 ppm) and a methoxyl groups<br /> III - RESULTS AND DISCUSSION (56.4 ppm) have been seen. Comparison of the<br /> spectroscopic data of 2 with the data described<br /> The roots of Angelica sinensis were in the literature [5] could conclude that<br /> extracted with ethanol at room temperature. The compound 2 is ferulic acid. But it seems that<br /> aqueous ethanol extract has been treated further compound 2 existed as salt because of the large<br /> by extraction with ethyl acetate and n-butanol, difference of chemical shifts of carbonyl signal<br /> successively. Chromatographic separation of the ( 2.2), when compared with that of ferulic<br /> ethyl acetate extract yielded compounds 1 - 4. acid. Some studies [6, 7] reported that ferulic<br /> Compound 1 was obtained as an amorphous acid showed anti-inflammatory and inhibitory<br /> powder. Its EI-MS spectrum indicated the effects on uterine movement.<br /> molecular ion peak at m/z 272 [M]+. The 1H- The EI-MS spectrum of compound 3, an<br /> NMR of 1 contained 3 protons at 6.89 d amorphous powder, showed the molecular ion<br /> (8.06), 6.99 dd (1.95, 8.06) and 7.01 d (1.95) peak at m/z 356 [M]+. 1H-NMR spectrum<br /> corresponding to an ABX aromatic ring. indicated the signals of 2 aromatic rings [7.02 d<br /> 509<br /> (2.01), 7.07 dd (2.01, 8.32), 6.88 d (8.32) and officinale. Compound 4, an amorphous powder,<br /> 6.94 d (2.01), 6.92 d (8.24), 6.92 dd (2.01, has the molecular ion peak at m/z 330 [M]+ in<br /> 8.24)], each corresponding to an ABX system, the ESI-MS spectrum. The NMR spectra of 4<br /> two double bonds with coupling constant of were similar to those of 3.<br /> 15.92 and 15.73 Hz, two methoxyl signals (3.90 Compound 4 consisted also trans-feruloyl<br /> s and 3.91 s), two hydroxyl groups (5.72 s and moiety [ 7.17 d (1.83), 7.06 dd (1.83, 8.32)<br /> 5.94 s) and one oxygenated methylene (4.84 6.78 d (8.3)] and a 1,4-disubstituted aromatic<br /> dd). Besides the carbons, which according to ring [7.24 d (8.0), 6.80 d (8.0)]. Additionally,<br /> above mentioned proton signals, a carbonyl the signals resonanced at 4.86 (1H, dd, J = 4.8<br /> signal at 166.9 and 6 quaternary aromatic and 7.3) and 4.23 m (2H) showed the existence<br /> carbons have been seen in the 13C-NMR. These of a -CH(OH)CH2O- subunit. Consequently, the<br /> spectroscopic data suggested that, 3 could be an structure 4 was elucidated to be 2-[4'-<br /> ester of ferulic acid. This suggestion was hydroxyphenyl]-glycol mono trans-ferulate by<br /> confirmed by comparison with the spectroscopic detailed comparison its spectroscopic data with<br /> data of coniferylferulate, published in the published literature [8]. This compound has<br /> literature [1]. Thus, the structure of 3 has been been isolated for the first time from<br /> determined to be coniferylferulate. It was Peucedanum decursivum. Its trivial name was<br /> described for the first time as a constituent of A. decorticated. The assignments of all protons and<br /> sinensis, Ligusticum porteri and Levisticum carbons of compound 1 - 4 were listed in the<br /> tables 1 and 2.<br /> <br /> Table 1: 1H-NMR data of compounds 1 - 4<br /> (300 MHz, 1 measured in CDCl3 and 2, 3, 4 in CD3OD)<br /> <br /> H 1 2 3 4<br /> 2 7.01 d (1.95) 7.17 d (2.01) 7.02 d (2.01) 7.17 d (1.83)<br /> 5 6.89 d (8.06) 6.30 d (8.23) 6.88 d (8.32) 6.78 d (8.3)<br /> 6 6.99 dd (1.95, 8.06) 7.05 dd (2.01, 8.23) 7.07 dd (2.01, 8.32) 7.06 dd (1.83, 8.32)<br /> 7 6.88 s 7.57 d (15.92) 7.65 d (15.92) 7.63 d (15.92)<br /> 8 6.35 d (15.92) 6.33 d (15.92) 6.37 d (15.92)<br /> 3-OMe 3.95 s 3.89 s 3.90* s 3.88 s<br /> 4-OH 5.63 s 5.72+ s<br /> 1'<br /> 2' 6.94 d (2.01) 7.24 d (8.0)<br /> 3' 6.80 d (8.0)<br /> 3'-OMe 3.91* s<br /> 4'-OH 5.94+ s<br /> 5' 6.92 d (8.24) 6.80 d (8.0)<br /> 6' 6.92 dd (8.24, 2.01) 7.24 d (8.0)<br /> 7' 6.62 d (15.73) 4.86 dd (4.76, 7.32)<br /> 8' 6.21 dt (6.59, 15.73) 4.23 m<br /> 9' 4.84 dd (1.28, 6.59)<br /> +, *<br /> : Could be exchanged in the column.<br /> 510<br />  O<br /> 5'<br /> 4' OH 2<br /> 6' MeO 3 1 7<br /> 9<br /> OH<br /> 7'<br /> <br /> 2 1' 3'<br /> MeO 3 1 4<br /> 6<br /> 7 7' 2' OMe 5<br /> HO<br /> 6<br /> 4<br /> 5<br /> HO 2: Ferulic acid<br /> 1: 4,4'-dihydroxy, 3,3'-dimethoxystilben OH<br /> 4'<br /> O<br /> O<br /> 7' OMe 1'<br /> 9'<br /> 6 7 1' 3' O<br /> 1 9<br /> O 8' OH<br /> 6' HO<br /> 4 4'<br /> HO 3 OH OMe<br /> OMe<br /> 3: Coniferylferulate 4: Decursidate<br /> <br /> <br /> Table 2: 13C-NMR chemical shift values of Acknowledgements: We thank the Bundesmin-<br /> compounds 1 - 4 isterium fuer Bildung und Forschung (BMBF),<br /> (75 MHz, 1 measured in CDCl3 and 2, 3, 4 Bonn, Germany for financial support, Dr. A.<br /> in CD3OD) Porzel and Dr. J. 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