Biochemical Systematics and Ecology 34 (2006) 654e657www.elsevier.com/locate/biochemsyseco

Apigenin chemotypes of Matricaria chamomilla L.

Vanda Svehlıkova, Miroslav Rep�cak*

P. J. Safarik University, Faculty of Science, Institute of Biology and Ecology, Department of Botany, Manesova 23, 04154 Kosice, Slovakia

Received 26 October 2005; accepted 25 February 2006

Keywords: Matricaria chamomilla; Asteraceae; Apigenin; Malonylated derivatives; Chemotypes

1. Subject and source

Matricaria chamomilla is an important medicinal plant containing active substances possessing anti-spasmodic andanti-inflammatory properties (Achterrath-Tuckermann et al., 1980; Della Logia et al., 1986). The most important con-stituents of the chamomile drug ‘‘Chamomillae flos’’ are sesquiterpenes and flavonoids, which have been studied ex-tensively (Redaelli et al., 1980, 1981, 1982; Exner et al., 1981; Schilcher, 1987; Carle et al., 1989, 1993; Tschierschand Holzl, 1992; Franz, 1993). Plants of the diploid cv. ‘‘Novbona’’ of M. chamomilla were used as the source of fla-vonoids for this study. The plants were cultivated in the experimental field of the Botanical Garden of P.J. SafarikUniversity in Kosice (Slovakia) in the years 1997e1999 (total of 104 individuals).

2. Previous work

Chamomile plants exhibit great variation in the content of their substances; a number of chemotypes have previ-ously been described in the chamomile populations. In particular, sesquiterpene chemotypes were thoroughly exam-ined including geographical and genetic research of the types (Franz, 1993; Schilcher, 1987). Chamomile plants werealso found to differ in the spectrum of methoxylated flavonoids (Rep�cak et al., 1999).

M. chamomilla, however, also contains a range of vacuolar glycosylated flavonoids, the most important of whichare apigenin conjugates. Apigenin is accumulated in the white ligulate florets of the anthodium in the form of apigenin7-O-glucoside and its acylated derivatives [its monoacetylated derivatives, apigenin 7-O-(600-caffeoyl)-glucoside,apigenin 7-O-(600-malonyl)-glucoside, apigenin 7-O-(400-acetyl-600-malonyl)-glucoside and an unidentified apigenin7-O-acetyl-malonyl-glucoside] (Svehlıkova et al., 2004).

3. Present study

The aim of the present study was to examine quantitative and qualitative differences in the profiles of apigeninderivatives among individual plants of chamomile cultivars as well as among wild-growing plants.

* Corresponding author. Tel.: þ421 55 633 7352; fax: þ421 55 633 7353.

E-mail address: repcak@upjs.sk (M. Rep�cak).

0305-1978/$ - see front matter � 2006 Elsevier Ltd. All rights reserved.

doi:10.1016/j.bse.2006.02.002

655V. Svehlıkova, M. Rep�cak / Biochemical Systematics and Ecology 34 (2006) 654e657

Anthodia were collected from individual plants in the phase of full flowering when approximately a half of tubularflorets were in flower and dried at room temperature.

For the analysis of apigenin glucosides dry ligulate florets were homogenised and extracted with methanol for30 min at room temperature. The extract was filtered and directly analysed by HPLC (Svehlıkova and Rep�cak,2000). Identity of apigenin derivatives was previously confirmed on the basis of MS and NMR spectra (Svehlıkovaet al., 2004). Apigenin (Sigma) was used as the standard compound. t-Testing was used to evaluate differences inthe content of apigenin compounds for statistical significance.

4. Chemotaxonomic significance

Individual chamomile plants were found to differ in the spectrum of accumulated acylated substances. On the basisof these differences two chemotypes can be distinguished in the populations of both cultivated and wild-growingchamomile populations (Fig. 1). The plants of the major type synthesize all the above-mentioned compounds de-scribed in chamomile with apigenin 7-O-(400-acetyl-600-malonyl)-glucoside being the dominating substance (chemo-type 1). A number of individual plants, however, were found in the populations of M. chamomilla, which do notcontain monoacetylated derivatives of apigenin 7-O-glucoside or apigenin 7-O-(400-acetyl-600-malonyl)-glucosideand its structural isomer. These minor chemotype plants accumulate only apigenin 7-O-(600-malonyl)-glucoside alongwith a small quantity of apigenin 7-O-(600-caffeoyl)-glucoside in their ligulate flowers (chemotype 2), most likely dueto a lack of acetyltransferase activity. This suggests a biochemical connection between the chemotypes where apige-nin 7-O-(600-malonyl)-glucoside represents a precursor in the biosynthesis of the respective acetate-malonates. Bothchemotypes were also found in the wild plants from the Slovak and Czech republics (90 plants from 7 localities e datanot shown).

The number of analysed plants of the diploid cultivar of M. chamomilla enabled evaluation of preliminary quan-titative characteristics of the chemotypes. The total amount of apigenin aglycone bound in the conjugated form, aswell as the sum of all acylated derivatives of apigenin 7-O-glucoside is not significantly different between the types(Table 1).

Plants of the acetate-absent chemotype represent only approximately 10% of the individuals in the populations ofthe cultivated variety ‘‘Novbona’’. Such a distribution of the types suggests a possible interaction between the genescontrolling the steps of modification reactions in substance biosynthesis, in contrast to methoxylated flavonoid chemo-types, where a Mendelian inheritance of the types can be expected on the basis of 25% spreading of the minor type(Rep�cak et al., 1999).

Fig. 1. HPLC chromatogram of the methanol extract of ligulate flowers of chemotype 1 (A) and chemotype 2 (B) of Matricaria chamomilla.

1, Apigenin 7-O-glucoside; 2, apigenin 7-O-(600-malonyl)-glucoside; 3, unidentified monoacetylated derivative of apigenin 7-O-glucoside;

4, apigenin 7-O-(600-caffeoyl)-glucoside; 5, apigenin 7-O-(600-acetyl)-glucoside; 6, apigenin 7-O-(400-acetyl, 600-malonyl)-glucoside; 7, unidentified

apigenin 7-O-acetyl-malonyl-glucoside.

Table 1

Content of apigenin 7-O-glucosid aglycone in the ligulate florets of two chemotypes of the diploid cultivar ‘‘Novbona’’ of

Matricaria chamomilla (g of com

Average content of compound in

Apigenin 7-O-

glucoside

igenin 7-O-00-caffeoyl)-

coside

Apigenin 7-O-

(400-acetyl,

600-malonyl)-

glucoside

Unidentified

acetylatede

malonylated

derivative of

apigenin 7-O-

glucoside

Sum of

acylated

derivatives of

apigenin 7-O-

glucoside

Total bound

apigenin

aglycone

Chemotype 1

(92 plants)

1.63 (0.393)a 2 (0.100) 2.79 (0.713) 0.51 (0.126) 5.32 (0.971) 4.08 (0.709)

Chemotype 2

(12 plants)

2.10 (0.315) 4 (0.096) 0.10 (0.262) 0.02 (0.048) 5.01 (0.738) 4.19 (0.585)

a Standard deviations are given

65

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.Svehlıkova,

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Rep �ca

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e and its acylated derivatives and the amount of total bound apigenin

pound/100 g ligulate floret dry mass)

the ligulate florets (g of substance/100 g dry mass)

Apigenin 7-O-

(600-malonyl)-

glucoside

Unidentified

monoacetylated

derivative of

apigenin 7-O-

glucoside

Apigenin 7-O-

(600-acetyl)-

glucoside

Ap

(6

glu

1.47 (0.579) 0.20 (0.059) 0.12 (0.032) 0.2

4.62 (0.845) 0.02 (0.027) 0.01 (0.016) 0.2

in parentheses.

657V. Svehlıkova, M. Rep�cak / Biochemical Systematics and Ecology 34 (2006) 654e657

The present study shows that cultivated chamomile plants preserved the variability in the spectrum of accumulatedapigenin derivatives that originated in the wild-growing populations. Further detailed research of wild-growing cham-omile plants across the whole of Europe could bring valuable information on the geographical distribution of the typesas well as on the content of apigenin as a pharmacologically important substance in chamomile anthodia. The datacould be correlated with the distribution pattern of sesquiterpene chemotypes, as in the case of Thymus herba-baronaLoisel. (Corticchiato et al., 1995). Variation in the content of isoprenoids in the essential oil was described in the nat-ural populations from the area of origin of M. chamomilla species in South-East Europe (Gray, 1976), across CentralEurope to the Iberian Peninsula and reflects genetic differences among the populations (Franz, 1993; Schilcher, 1973).The data on flavonoid distribution could contribute to the elucidation of migration routes of the species in Europe inthe past.

Acknowledgements

The work was supported by the Grant Agency VEGA, Slovakia (grant No. 1/0444/03).

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