3rd meeting of young researchers at UP 1 - IJUP - Universidade do ...
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Screening <strong>of</strong> is<strong>of</strong>lavones levels in several wild Portuguese<br />

Leguminosae using m<strong>at</strong>rix solid-phase dispersion (MSPD) and<br />

HPLC/DAD<br />

T. Sousa 1 , I. Sousa 1 , M.A. Faria 2 , S.C. Cunha 2 , M. E. Nunes 1<br />

1 CIBIO-ICETA/Faculty <strong>of</strong> Sciences, University <strong>of</strong> Porto, R. Padre Arman<strong>do</strong> Quintas 4485-661 Vairão,<br />

Portugal<br />

2 REQUIMTE/Department <strong>of</strong> Brom<strong>at</strong>ology, Faculty <strong>of</strong> Pharmacy, University <strong>of</strong> Porto, Rua Aníbal<br />

Cunha 164, 4099-030 Porto, Portugal<br />

Email: sara.cunha@ff.up.pt<br />

Is<strong>of</strong>lavones (3-phenyl chromones) are plant secondary metabolites displaying numerous<br />

biological activities in vertebr<strong>at</strong>es among which estrogen-like activity is one <strong>of</strong> most described<br />

in liter<strong>at</strong>ure [1]. They are found primary in Leguminosae such as soybean and red clover.<br />

However the pr<strong>of</strong>ile <strong>of</strong> is<strong>of</strong>lavones amongst species is quite variable (e.g. red clover contains<br />

mainly formononetin and biochanin A whereas soybean contains primarily daidzein, glycitein<br />

and genistein). Thus, the analysis <strong>of</strong> these compounds in plants is still a challenge for analytical<br />

chemists due to the wide range <strong>of</strong> amounts and type <strong>of</strong> molecules with different polarities<br />

which difficult their simultaneous extraction from the m<strong>at</strong>rix. In a previous work a simple<br />

procedure which permits the simultaneous extraction <strong>of</strong> 12 is<strong>of</strong>lavones from the leaves <strong>of</strong> red<br />

clover was optimized [2]. This procedure is based on a m<strong>at</strong>rix solid-phase dispersion (MSPD)<br />

metho<strong>do</strong>logy consisting on the simultaneously sample homogeniz<strong>at</strong>ion, extraction and cleanup,<br />

requiring a small sample size and minimal amounts <strong>of</strong> organic solvents. The is<strong>of</strong>lavones<br />

extracted were then identified and quantified by high-performance liquid chrom<strong>at</strong>ography<br />

(HPLC) coupled to a diode array detector.<br />

The aim <strong>of</strong> this work was to explore the applicability <strong>of</strong> the referred MSPD HPLC/DAD<br />

method for the quantific<strong>at</strong>ion <strong>of</strong> 12 is<strong>of</strong>lavones in several wild Portuguese Leguminosae<br />

species collected in several loc<strong>at</strong>ions in the Portuguese territory. Samples <strong>of</strong> the species Vicia<br />

s<strong>at</strong>iva, Genista spp, Cytisus scoparius, Ornithopus compressus, Ornithopus pin<strong>at</strong>us, Medicago<br />

lupulina, Lotus edulis, Medicago Arabica, Lotus cornicul<strong>at</strong>us, Pterospartum trident<strong>at</strong>um and<br />

Ulex spp were collected, morphologically identified, stored on desicc<strong>at</strong>ing silica gel and<br />

submitted to the overall procedure for is<strong>of</strong>lavones quantific<strong>at</strong>ion. The is<strong>of</strong>lavone pr<strong>of</strong>iles <strong>of</strong> the<br />

referred plants are presented.<br />

References:<br />

[1] Rostagno, M.A., Villares A., Guillamón E., García-Lafuente A. and Martínez, J.A. (2009), Sample<br />

prepar<strong>at</strong>ion for the analysis <strong>of</strong> is<strong>of</strong>lavones from soybeans and soy foods, Journal <strong>of</strong> Chrom<strong>at</strong>ography A,<br />

1216, 2-29.<br />

[2] Visnevschi-Necrasov T., Cunha S.C., Nunes E. and Oliveira M.B.P.P. (2009), Optimiz<strong>at</strong>ion <strong>of</strong><br />

m<strong>at</strong>rix solid-phase dispersion extraction method for the analysis <strong>of</strong> is<strong>of</strong>lavones in Trifolium pr<strong>at</strong>ense<br />

Journal <strong>of</strong> Chrom<strong>at</strong>ography A,1216, 3720-3724.<br />

Acknowledgments:<br />

This research was supported by U.Porto /Santander Totta “Projectos Pluridisciplinares 2009”. S.C.C. is<br />

gr<strong>at</strong>eful to “POPH-QREN- Tipologia 4.2, Fun<strong>do</strong> Social Europeu e Fun<strong>do</strong> Nacional MCTES”. M.A.<br />

Faria is gr<strong>at</strong>eful to FTC for the Grant SFRH/BPD/20725/2004.<br />

130 3 rd <strong>meeting</strong> <strong>of</strong> <strong>young</strong> <strong>researchers</strong> <strong>at</strong> <strong>UP</strong>

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