Fitoterapia 76 (2005) 718 – 721

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Short report

Biological activity of some Patagonian plants

Pedro Cuadra a,b,*, Marı́a Furrianca b, Alejandra Oyarzún b,
Erwin Yáñez a,b, Amalia Gallardo b, Vı́ctor Fajardo a,b
a
Centro de Estudios del Cuaternario (CEQUA), Punta Arenas, Chile
b
Universidad de Magallanes, Facultad de Ciencias, casilla 113-D, Punta Arenas, Chile
Received 21 October 2004; accepted in revised form 20 July 2005
Available online 17 October 2005

Abstract

Citotoxicity (inhibition of cell division in fertilized eggs of Loxechinus albus) and general
toxicity (using embryos of Artemia salina) of plants belonging to the genera Senecio, Deschampsia,
Alstroemeria, Anarthrophyllum, Chloraea and Geranium were investigated.
D 2005 Elsevier B.V. All rights reserved.

Keywords: Patagonian plants; Citotoxicity; Inhibition; Cell division

1. Plants

Plants were collected between 2002 and 2003 from several Patagonian localities.
Valeriana carnosa Sm., Alstroemeria patagonica Phil. and Senecio miser Hooker in Sierra
Baguales (Ultima Esperanza province); Berberis ilicifolia L.f., Anarthrophyllum desider-
atum Bentham ex Jackson and Chloraea magellanica Hooker in Tierra del Fuego. The
latter two species were found on the shores of Bahı́a Inútil and Geranium magellanicum
Hooker from the bushes in the steppes of Rı́o Verde, near the suburbs of Punta Arenas city.
Deschampsia antarctica Desv. in Morro Chico, 140 km to the north of Punta Arenas city.
All plant species were identified by Orlando Dollens from the Universidad de Magallanes.
Voucher specimens were kept in the Herbarium of the Universidad de Magallanes.

* Corresponding author. Centro de Estudios del Cuaternario (CEQUA), Punta Arenas, Chile. Tel.: +56 61
217315; fax: +56 61 217315.
E-mail address: pedro.cuadra@umag.cl (P. Cuadra).

0367-326X/$ - see front matter D 2005 Elsevier B.V. All rights reserved.
doi:10.1016/j.fitote.2005.07.007
 P. Cuadra et al. / Fitoterapia 76 (2005) 718–721 719

2. Use in traditional medicine

V. carnosa because of its particular medicinal properties (antioxidant, antihypertensive,

vasodilative) [1–4]. The fruits of B. ilicifolia have been used by natives as a dye. There are
no reports about the biological activity of the other plant species.

3. Previously isolated classes of constituents

Alkaloids and eremophilanolides [5] from S. miser; from V. carnosa lignan-type

compounds [6]. No reports relating to A. patagonica, also numerous alkaloids have been
isolated from the Amaryllidaceae family [7–10]. From D. antarctica, flavones [11,12].
Alkaloids in B. ilicifolia [13] and plants belonging to the Anarthrophyllum genus [14].
Phenols, pyridines and phenolic acids, in the Geranium genus [15]. C. magellanica, one of
fifty taxa of terrestrial orchids from Chile [16], no reports.

4. Tested material

Ethanol extracts. 20 mg of the extract were dissolved in 2 ml of DMSO to obtain a

solution of 10 000 ppm. From this, three different solutions of 5, 50 and 500 ppm were
prepared. Each experiment represents the average of three different determinations. A
solution of marine water was used as a control. A DMSO solution was also tested in order
to assess the solvent effects. In D. antarctica, the ethanolic extract was separated by CC
and fractions 25–35 were assessed.

5. Studied activity

5.1. Brine shrimp test (BST)

This assay uses Artemia salina larvae and was applied according to Meyer et al. [17]
and Mc Laughlin [18]. Extract solutions of 1000, 100 and 10 ppm were tested. Each LC50
value represents an average of three replicates. Ten individuals were used in each replicate.
Larvae were exposed for 24 h at 25–30 8C, and 25–35% salinity.

5.2. Inhibition of cell division test (ICDT)

Samples of Loxechinus albus (Molina) were collected during 2004 in Punta Carrera and
Punta Santa Ana, in the south of the Magallanes Region. These sea urchins are individuals
with separate sex, without sexual dimorphism. Male and female can be distinguished only
at the reproductive stage when the male gonads are covered by a white liquid (sperms) and
female gonads become yellow-orange, just like the gonad pigmentation [19]. In order to
induce the oviposition, sea urchins were injected through their peristomatic membrane
with 2–6 ml of KCl 0.5 M. To evaluate the fecundity percentage, 1.0 ml of unfertilized egg
720 P. Cuadra et al. / Fitoterapia 76 (2005) 718–721

Table 1
Effect of Patagonian plants ethanolic extracts on BST and ICDT a
Plants Plant organ BST b (LC50) ICDT c % inhibition Main constituents
D. antarctica Whole plant 7.0 (1.5–12.7) 4.0 (500) Flavones
S. miser Leaves 92 (500) Eremophilanolides
B. ilicifolia Roots 16 (500) Alkaloids
V. carnosa Leaves and stems 23 (500) Lignans
A. patagonica Roots 26 (1–135) 96 (500) Alkaloids
A. desideratum Whole plant 96 (500) Alkaloids
C. magellanica Whole plant 270 (47–975) 97 (500) Not reported
G. magellanicum Whole plant 65 (500) Phenolics
a
All values represent the average of 3 three replications (N = 10).
b
BST, Brine Shrimp Test (95% confidence limits in brackets).
c
ICDT, Inhibition of Cell Division Test with fertilized sea urchin eggs; extract concentration in ppm is given in
brackets.

solution plus three drops of sperm suspension was made up to 100 ml with filtered marine
water, according to Jacobs et al. [20]. Samples were kept at 7 8C until fecundation had
occurred and then 1.0 ml of each plant extract solution of 5, 50 and 500 ppm was added to
1.0 ml of fertilized egg solution and mixed thoroughly. After 2 h at 20 8C, the fertilization
process was evaluated (once the first cell division step was complete).

6. Organisms used

A. salina eggs were purchased from Aqua Tecno Co. and kept in marine water in a
closed vial, illuminated with a 60 W lamp. They were kept at 25–30 8C with 25–35%
salinity for 48 h. After this time, the eggs were collected with a Pasteur pipette and tested.
Samples of L. albus were collected in Punta Carrera and Punta Santa Ana and transported
to the biology laboratory in a fridge containing marine water, together with Macrosistys
sp. to avoid stress and oviposition. In the laboratory, samples were moved to an aquarium
filled with filtered marine water and aeration.

Table 2
Effect of D. antarctica ethanolic extracts fractions, tricin and luteolin on BST and ICDT a
BST b (LC50) ICDT c % inhibition
Fraction 25 53.6 (30.6–123.1)
Fraction 26 43.8 (15.3–93.5)
Fraction 34 72 (500)
Fraction 46 100 (500)
Tricin 62.0 (59.6–103.2)
Luteolin 16.6 (19.5–69.7)
a
All values represent the average of three replications (N = 10).
b
BST, Brine Shrimp Test (95% confidence limits in brackets).
c
ICDT, Inhibition of Cell Division Test with fertilized sea urchin eggs; concentration of samples in ppm is
given in brackets.
 P. Cuadra et al. / Fitoterapia 76 (2005) 718–721 721

7. Results

Shown in Tables 1 and 2.

8. Conclusions

The ethanolic extract of D. antarctica was highly toxic for A. salina in the BST test. S.
miser, A. patagonica, A. desideratum and G. magellanicum demonstrated a high cell
division inhibitory effect against L. albus as indicated in the ICDT test. D. antarctica was
further analyzed by means of a bioassay-guided fractionation. The most active fractions
were fraction 26 (LC50 43.8 ppm in the BST test) and fraction 46 (100% inhibition at 500
ppm in the ICDT test). From fraction 26, the following were identified: 5,7,4V-trihydroxy-
3V,5V-dimethoxyflavone and 5,7,3V,4V-tetrahydroxyflavone.

Acknowledgements

We thank the Dirección de Investigación of the Universidad de Magallanes for financial

support given (PR-F4-01RN-03 and PR-F4-02RN-03) and Miss Stephanie Whitfield for
her kindness in revising this manuscript.

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