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Fitoterapia 73 (2002) 727729

Short report

Hyaluronidase inhibitory activity from the

polyphenols in the fruit of blackberry (Rubus
fruticosus B.)
M.A. Marquina, G.M. Corao*, L. Araujo, D. Buitrago, M. Sosa
en Cultivos Celulares, Laboratorio de Bioqumica,
Grupo de Investigacion Facultad de Farmacia,

Universidad de los Andes, Merida 5101, Venezuela

Received 14 May 2002; accepted in revised form 8 August 2002


The hyaluronidase inhibitory activity of different fractions obtained from aqueous extract
of blackberry fruits was evaluated. We found that only fractions F3 and F7 inhibit the tested
2002 Elsevier Science B.V. All rights reserved.

Keywords: Blackberry; Rubus fruticosus B.; Anti-inflammatory activity; Polyphenols

Plant. Blackberry (Rubus fruticosus B.), from rosaseae family, is a small tree with
red-blackish fruit containig polyphenols w1x.

Uses in traditional medicine. There is a large number of biological activities due

to polyphenols in various plants w27x. In particular, anti-inflammatory activity was
described for blackberry fruit w8x.

Previously isolated constituents. Flavonoids, flavonols, flavonones, catechin, antho-

cyanidins and polyflavanoid w8,9x.
*Corresponding author. Tel.: q58-274-2493476; fax: q58-274-2493445.
E-mail address: (G.M. Corao).

0367-326X/02/$ - see front matter 2002 Elsevier Science B.V. All rights reserved.
PII: S 0 3 6 7 - 3 2 6 X 0 2 . 0 0 2 2 2 - 8
728 M.A. Marquina et al. / Fitoterapia 73 (2002) 727729

Table 1
Summary of results on polyphenols present in aqueous extraction of the blackberry fruit (Rubus fruti-
cosus B.)
Paper chromatography (R f =100) Polyphenol test
Sample UV FeCl3 FeCl3 pH


6% 6%
F1 81 91
69 16 q 3.0
F2 75 91
14 q 4.0
F3 80
q 4.1
F4 80 88
18 16 q 3.1
F5 q 4.6
F6 79
18 q 2.9
F7 81 91
16 q 3.0
Key: aCrude extract; bfractions; c polyphenols testsFeCl3 2% pyv in methanol; running solventss
BAW (butanolyacetic acidywater; 4:1:5), AcOH 6% (acetic acid 6% vyv); detectionsUV (ultraviolet),
spray FeCl3 (FeCl3 al 2% wyv in methanol).

Tested material. Blackberry fruits were obtained from a local shop and extracted
as follows: the fruits were blended with methanol and filtered through two layers
of gauze. The methanolic filtrate (F1) was partitioned (Si-gel CC, increasing
gradient MeOHywater). The obtained fractions were running in PTLC wBuOHy
Acetic acidywater (4:1:5)x and acetic acid 6% wyv. The corresponding fractions
(Table 1), pooled, evaporated to dryness, weight and re-dissolved in 1 ml of water,
were tested for anti-inflammatory activity.

Studied activity. The hyaluronidase inhibitory activity was evaluated according to

Lee et al. w10x. Potassium hyaluronate, p-dimethylaminobenzaldehyde and hyaluron-
idase were purchased from Sigma Chemical Co.

Results. Reported in Fig. 1.

Conclusion. We found that the fractions F3 and F7 obtained from blackberry inhibit
the enzyme hyaluronidase and showed better anti-inflammatory activity when
compared with aspirin. Aspirin in this assay has shown a low response, so it is
advisable to continue research using a different drug.
M.A. Marquina et al. / Fitoterapia 73 (2002) 727729 729

Fig. 1. Histogram of inhibition of hyaluronidase by crude extract and fractions from chromatographic
separation of blackberry fruit.


This work was supported by a grant from the Consejo de Desarrollo Cientfico,

y Tecnologico (C.D.C.H.T) ULA, Merida-Venezuela.


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