www.elsevier.com/locate/biochemsysecoBiochemical Systematics and Ecology 35 (2007) 533e538

Sesquiterpene lactones and other constituentsfrom Matricaria chamomilla L.

Lahcene Zaiter a, Mohamed Bouheroum a, Samir Benayache a,Fadila Benayache a, Francisco Leon b,c, Ignacio Brouard b, Jose Quintana d,

Francisco Estevez d, Jaime Bermejo b,*

a Laboratoire de Phytochimie et Analyses Physico-Chimiques et Biologiques, Equipe associee a l’A.N.D.R.S.,

Universite Mentouri, Route de A€ın El Bey, 25000 Constantine, Algeriab Instituto de Productos Naturales y Agrobiologıa (IPNA-CSIC), Instituto Universitario de Bio-Org�anica ‘‘Antonio Gonz�alez’’,

Avda. Astrofısico F. S�anchez 3, 38206 La Laguna, Tenerife, Spainc Instituto Canario de Investigacion del C�ancer (ICIC), Avda. Astrofısico F. S�anchez 2, 38206 La Laguna, Tenerife, Spain

d Departamento de Bioquımica y Biologıa Molecular, Centro de Ciencias de la Salud, Universidad de Las Palmas de Gran Canaria,Avda. San Cristobal, 35016 Las Palmas de Gran Canaria, Spain

Received 6 November 2006; accepted 11 March 2007

Abstract

An ethanolic extract of the aerial parts of Matricaria Chamomilla L. collected at Oued Tonga near El-Kala is eastern Argeliayielded two new substances named matricolone and chamolol, together with four known compounds. The structure of the new com-pounds were determined by chemical transformations and NMR spectroscopy with 1H-1H (COSY and ROESY), 1H-13C (one bondand long range correlations) two dimensional experiments. The results obtained indicate the presence of sesquiterpene lactones ofthe eudesmanolide, germacranolide and guaianolide types. These products can be considered to be of great interest from a chemo-taxonomical point of view for this wild-growing plant, because these types of lactones have not been found together in otherMatricaria species.� 2007 Elsevier Ltd. All rights reserved.

Keywords: Matricaria chamomilla; Asteraceae; Sesquiterpene lactones; Glucosyl monoterpene

1. Subject and source

The genus Matricaria (Asteraceae, tribe Anthemideae) is a frequently used source of phytopharmaceuticals ofgrowing importance (Dragland et al., 2003), employed as a component of tea-mixes and as a valuable ingredientof many galenic preparations such as tinctures and extracts. In many countries such as Argentina, Egypt, Hungary,Slovakia and Germany (Avallone et al., 2000; Miliauskas et al., 2004), Matricaria is grown as a field crop. Due totheir high content in essential oil ingredients, Matricaria flowers are of special value, despite their extreme perishabil-ity (Maia et al., 2004).

* Corresponding author. Tel.: þ34 922 318583; fax: þ34 922 318571.

E-mail address: jbermejo@ull.es (J. Bermejo).

0305-1978/$ - see front matter � 2007 Elsevier Ltd. All rights reserved.

doi:10.1016/j.bse.2007.03.008

534 L. Zaiter et al. / Biochemical Systematics and Ecology 35 (2007) 533e538

Matricaria chamomilla L. was collected at Oued Tonga near El-Kala in eastern Algeria in June 2000 and identifiedby Professor M. Kaabeche (Biology Department, University of Setif, Algeria). A voucher specimen (CMC15/06/00)has been deposited in the Herbarium of the Biology Department at Mentouri University in Constantine.

2. Previous work

A previous phytochemical study of M. chamomilla yielded a range of vacuolar glycosylated flavonoids, the mostimportant of which are apigenin conjugates (Svehlıkov�a and Repc�ak, 2006). Different types of sesquiterpenes havealso been isolated (Yamazaki et al., 1982; Gasic et al., 1983; Szoke et al., 2004).

3. Present study

The aerial parts of the plant (0.856 kg) were macerated at room temperature with EtOHeH2O (70:30 v/v) (48 h,three times). After filtration, the filtrates were combined, concentrated, and the residue was dissolved in H2O. Thesolution was treated with Pb(OAc)4 under stirring overnight (Gonz�alez et al., 1984), filtered and extracted withCHCl3, three times. The organic phases were dried with Na2SO4, filtered and concentrated in vacuo at room temper-ature affording 15.0 g of extract. This extract was chromatographed on a silica gel column using cyclohexaneeEtOAc

with increasing polarity to yield 39 fractions obtained by combining the eluates on the basis of TLC. Fraction 11(25 mg) eluted with cyclohexaneeEtOAc (80:20) was recrystallized from hexaneeMeOH to obtain the known com-pound stigmasterol (18.3 mg). Fraction 22 (31 mg) eluted with cyclohexaneeEtOAc (65:35) was subjected to prepar-ative TLC using hexaneeacetone (1:1) followed by column chromatography using CH2Cl2eacetone (9:1) to giveethyl caffeate (17.6 mg). Fraction 24 (14.6 mg) eluted with cylclohexaneeEtOAc (60:40) gave, after purificationby successive preparative TLC using hexaneeEtOAc (7:3) and CH2Cl2eacetone (9.5:0.5), the compound 5(9 mg). Fraction 30 (71 mg) eluted with cyclohexaneeEtOAc (45:55) and recrystallized from acetoneeCHCl3gave the new sesquiterpene 1 (14 mg). Fraction 37 (91.5 mg) eluted with cyclohexaneeEtOAc (10:90) was chromato-graphed by preparative TLC and developed with hexaneeEtOAc (1:9) to afford the new compound 2 (27.5 mg) andthe known sesquiterpene 4 (34.4 mg).

In the present work on the chemical constituents of M. chamomilla L., we have isolated two new substances, namedmatricolone (1) and chamolol (2), together with four known compounds.

The structures of the known compounds were identified as dihydroridentin (4) (Abu Zarga et al., 1995); 2a-hydroxyarborescin (5) (Bohlmann et al., 1985); ethyl caffeate (Kisiel and Barszcz, 1995) and stigmasterol (Ahmadand Misra, 1997), by comparing their spectroscopic data with those reported in the literature. In the present paper,we report on the isolation and structural identification of 1 and 2 from M. chamomilla.

O

OH

AcO

O

HOH

12, R = H3, R = Ac

OHO

OH

O

ORORO

OR

OR

O

4

O

O

OHO

5

1

43 5

67 13

9

11

14

15 1

2 34

5

6

7

10

89

1`2`

3`

4` 6`

535L. Zaiter et al. / Biochemical Systematics and Ecology 35 (2007) 533e538

Matricolone (1) was obtained as a crystalline solid, mp 258e259 �C and [a]D25þ 39.0� (c 0.4 CHCl3). The

HRFABMS spectrum of 1 exhibited a quasi-molecular ion at m/z 349.1610 [MþNa]þ corresponding to the molecularformula C17H26O6 for this compound. The FABMS spectrum confirmed the molecular formula of this compound andshowed in particular ions at m/z 267 [MHeCH3CO2H]þ (21%), 249 [267�H2O]þ (19%), 231 [249�H2O]þ (6%)indicating the presence of an acetate and two hydroxyl groups in this molecule. The IR spectrum showed bands at3443 and 1732 cm�1 corresponding to these groups and a band at 1769 cm�1 typical for a five-membered lactone.

The 1H NMR spectral data of 1 (Table 1) showed two singlets at dH 1.37 and 1.01, and a doublet at dH 1.23 revealingthe presence of three methyls, while a singlet at dH 2.10 confirmed the presence of an acetoxy group. The samespectrum showed a typical triplet for H-6 at dH 4.18. This proton correlated with the carbon at dC 79.8 in theHSQC spectrum experiment, indicating a C-6 lactonized sesquiterpene lactone. These signals were in part similarto those described for 1a,3b,4b-trihydroxy-(5a,7a,11bH-10a-methyl)-eudesman-12, 6a-olide (Abdel-Mogib et al.,1989). However, a hydroxyl group was replaced by an acetoxy group. To determine the position of the acetoxy group,the HMBC spectrum of 1 was useful. For this purpose, this spectrum showed correlation between the signal at dH 4.81(H-3) and the signal at dC 18.9 (CH3-15) and with the carbonyl at dC 170.4 of the acetate group, thus situating thisgroup at C-3. The coupling patterns and the magnitude of the coupling constants of H-1, H-5 to H-7 in the 1HNMR spectrum of compound 1 were in full agreement with an H-1a configuration for C-1, and a trans disposition

H

O

H

O

H

H

AcO

H

H

OH

HO

13

4 5 611

7

14

Fig. 1. Selected ROESY correlations for compound 1.

Table 11H and 13C NMR data for compound 1

Position dC dH

1 75.7, CH 3.58, dd (11.7, 3.6)

2 33.5, CH2 1.62, ma

2.05, ddd (12.6, 4.5, 3.6)

3 75.9, CH 4.81, dd (12.5, 4.5)

4 73.4, qC

5 54.0, CH 1.70, d (11.4)

6 79.8, CH 4.18, t (10.8)

7 53.2, CH 1.65, ma

8 23.3, CH2 1.46, qd (12.9, 3.3)

1.91, dq (12.9, 3.3)

9 39.2, CH2 1.23, ma

2.00, dt (12.9, 3.3)

10 42.1, qC

11 40.4, CH 2.29, dq (12.6, 6.9)

12 177.8, qC

13 12.4, CH3 1.23, d (6.9)

14 13.7, CH3 1.01, s

15 18.9, CH3 1.37, s

OAc 170.4, qC 2.10, s

21.4 CH3

400 MHz, CDCl3, J values (Hz) are given in parentheses.a Overlapped.

536 L. Zaiter et al. / Biochemical Systematics and Ecology 35 (2007) 533e538

of H-5/H-6 and H-6/H-7. The configuration at C-11 was deduced to be 11bH by the trans diaxial coupling for H-7 andH-11 (J¼ 12.6 Hz) and the chemical shifts in the 13C NMR spectrum (Table 1) at dC 40.4 (C-11) and dC 12.4 (C-13).Thus, the relative configuration of 1 was confirmed by a ROESY experiment (Fig. 1) in which correlations observedbetween dH 3.58 (H-1), 4.81 (H-3), 1.70 (H-5) and 1.65 (H-7) clearly showed that these protons were on the same face.Correlations were also observed between dH 1.01 (CH3-14) and 1.37 (CH3-15), and dH 4.18 (H-6) and 2.29 (H-11).Thus, the C-3 acetoxy and the C-1 hydroxyl groups were deduced to be b-oriented in an equatorial disposition. There-fore, compound 1 was identified as 3b-acetoxy-1b,4a-dihydroxy-(5a,6b,7a,11bH-10b-methyl)-eudesman-12,6a-olide named matricolone.

Matricolone (1): colorless needles; mp 258e259 �C; [a]D25þ 39.0� (c 0.4 CHCl3); IR (KBr) nmax 3443, 2931, 2874,

1769, 1732, 1454, 1380, 1309, 1248, 1152, 1127, 1054, 1015, 928, 905, 752, 647 cm�1; 1H and 13C NMR, see Table 1;FABMS m/z 349 [MþNa]þ (16), 327 [MþH]þ (12), 309 [(MþH)�H2O]þ (19), 279 [(MþH)�H2OeCH2O]þ

(9), 267 [(MþH)eCH3CO2H]þ (21), 249 [267�H2O]þ (19), 231 [249�H2O]þ (6), 167 (17), 154 (22); HRFABMSm/z 349.1610 [MþNa]þ (Calcd for C17H26O6Na, 349.1627), 327.1819 [MþH]þ (Calcd for C17H27O6, 327.1819).

Chamolol (2) was isolated as colorless needles, mp 58e59 �C and [a]D25�120.0� (c 0.58 CHCl3), and its molecular

formula was determined to be C16H28O6 by HRFABMS m/z 339.1798 [MþNa]þ. The IR spectrum of 2 showed ab-sorption bands at 3344, and 1657 and 913 cm�1 assigned to hydroxyl and olefinic functions, respectively. Its 1H NMRspectrum (Table 2) revealed the presence of four olefinic protons including an ABX system of a vinyl group at dH 4.97(Ha-9), 5.02 (Hb-9), and 5.96 (H-8), and a separate olefinic proton at dH 5.11 (H-4). An anomeric proton signal at dH

4.49 (1H, d, J¼ 7.6 Hz) suggested the presence of a sugar residue with a b-configuration.The 13C and DEPT NMR spectra revealed the presence of four methyl carbons at (dC 18.2, 22.8, 26.0, and 26.1),

one aliphatic methine (dC 52.3), four olefinic carbons (dC 115.4, 122.6, 134.2, and 138.2), and a quaternary oxygenatedcarbon (dC 80.2). All these findings, in addition to the values of the chemical shifts of the carbon atoms of the sugarmoiety (Agrawal, 1992), indicated that the structure of compound 2 and its peracetylated derivative 3 proved to bea glucosidic natural derivative of santolina alcohol (Poulter et al., 1972). The HMBC spectrum experiment of 2(Fig. 2) showed a correlation between H-10 (dH 4.49) and C-2 (dC 80.2), supporting the attachment of b-glucopyra-nosyl moiety at the C-2 position. Thus, the structure of compound 2 was deduced to be 2,5-dimethyl-3-vinyl-hex-4-en-2-O-b-glucopyranoside.

Chamolol (2): colorless needles; mp 58e59 �C; [a]D25 �120.0� (c 0.58 CHCl3); IR (KBr) nmax 3570, 3344, 3072,

2978, 2922, 2860, 1726, 1657, 1629, 1607, 1446, 1404, 1375, 1361, 1271, 1168, 1134, 1103, 1015, 932, 913, 874, 848,778, 662 cm�1; 1H and 13C NMR, see Table 2; FABMS m/z 339 [MþNa]þ (100), 317 [MþH]þ (3), 176 (5), 154 (6),137 (54), 132 (6), 121 (4), 109 (6); HRFABMS m/z 339.1798 [MþNa]þ (Calcd for C16H28O6Na, 339.1784).

Table 21H and 13C NMR data for compound 2

Position dC dH

1 26.1, CH3 1.20, s

2 80.2, qC

3 52.3, CH 3.09, dd (9.6, 6.5)

4 122.6, CH 5.11, d (9.6)

5 134.2, qC

6 26.0, CH3 1.73, s

7 18.2, CH3 1.60, s

8 138.2, CH 5.96, ddd (16.5, 11.0, 6.5)

9 115.4, CH2 4.97, br d (16.5)

5.02, br d (11.0)

10 22.8, CH3 1.16, s

10 97.1, CH 4.49, d (7.6)

20 73.6, CH 3.37, dd (8.1, 7.8)

30 76.4, CH 3.57, ma

40 69.9, CH 3.57, ma

50 75.4, CH 3.28, br d (8.0)

60 61.8, CH2 3.80, m

400 MHz, CDCl3, J values (Hz) are given in parentheses.a Overlapped.

537L. Zaiter et al. / Biochemical Systematics and Ecology 35 (2007) 533e538

Acetylation of 2: compound 2 (8 mg) was dissolved in pyridine (2 mL) and acetic anhydride (2 mL) and thesolution was stirred for 24 h at room temperature. The product was dried under vacuum to furnish 3 (8 mg) as an amor-phous solid; IR (KBr) nmax 2976, 2927, 1741, 1437, 1367, 1254, 1222, 1172, 1126, 1084, 1033, 980, 911, 838, 690,651, 619 cm�1; 1H NMR (CDCl3, 300 MHz) d 5.88 (1H, ddd, J¼ 7.2, 10.0, 17.0 Hz, H-8), 5.19 (1H, br dd, J¼ 7.8,9.4 Hz, H-20), 5.06e4.94 (5H, m, H-4, Ha-9, Hb-9, H-30, H-40), 4.65 (1H, d, J¼ 7.8 Hz, H-10), 4.19 (1H, dd, J¼ 5.9,12.0 Hz, Ha-6

0), 4.07 (1H, dd, J¼ 2.5, 12.0 Hz, Hb-60), 3.67 (1H, m, H-50), 2.95 (1H, dd, J¼ 7.2, 9.6 Hz, H-3), 2.06(3H, s, OAc), 2.03 (3H, s, OAc), 2.00 (3H, s, OAc), 1.98 (3H, s, OAc), 1.71 (3H, s, CH3-6), 1.58 (3H, s, CH3-7), 1.17(3H, s, CH3-1), 1.09 (3H, s, CH3-10); 13C NMR (CDCl3, 75 MHz) d 170.5 (C, OCOCH3), 170.3 (C, OCOCH3), 169.4(C, OCOCH3), 169.0 (C, OCOCH3), 137.6 (CH, C-8), 133.4 (C, C-5), 122.6 (CH, C-4), 115.6 (CH2, C-9), 95.3 (CH,C-10), 80.4 (C, C-2), 73.0 (CH, C-30), 71.5 (CH, C-20), 71.4 (CH, C-50), 69.0 (CH, C-40), 62.1 (CH2, C-60), 52.9 (CH,C-3), 26.2 (CH3, C-1), 26.0 (CH3, C-6), 22.4 (CH3, C-10), 20.8 (CH3, OCOCH3), 20.7 (CH3, OCOCH3), 20.6 (CH3,OCOCH3), 20.5 (CH3, OCOCH3), 18.1 (CH3, C-7); FABMS m/z 507 [MþNa]þ (14), 331 (27), 169 (36), 138 (11),137 (100); HRFABMS m/z 507.2243 [MþNa]þ (Calcd for C24H36O10Na, 507.2206).

4. Chemotaxonomic significance

About 16 of more than 80 species of the genus Matricaria have been studied chemically. The most important con-stituents were sesquiterpenes and flavonoids. The results obtained in the study of M. chamomilla collected in Algeriaindicate the presence of sesquiterpene lactones of the eudesmanolide, germacranolide and guaianolide types and a glu-cosyl monoterpene. These products can be considered to be of great interest from a chemotaxonomical point of viewfor this wild-growing plant, because these types of lactones have not been found together in other Matricaria species(Baer and Schultze, 1996; Bohlmann and Zdero, 1975). Compounds 1 and 2 were new and compounds 4 and 5 wereisolated from this genus for the first time.

Acknowledgments

This work was supported in part by a grant from the Programa de Iniciativa Comunitaria INTERREG IIIB AzoreseMadeiraeCanarias (04/MAC/3.5/C5), ICIC (G-04-09) and A.N.D.R.S. The authors wish to thank Prof. M. Kaabeche(Biology Department, University of Setif, Algeria) for the identification of the plant material.

Appendix A. Supplementary information

Supplementary data associated with this article can be found, in the online version, at doi:10.1016/j.bse.2007.03.008.

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OHO

HOOH

OH

O

Fig. 2. Selected HMBC (H / C) correlations for compound 2.

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