Antibacterial and brine shrimp lethality

Fitoterapia 75 (2004) 360–363

Short report

Antibacterial and brine shrimp lethality tests
of biflavonoids and derivatives of
Rheedia gardneriana
Luiz G. Verdia, Moacir G. Pizzolattia,*,
ˆ M.C. Brighentea,
Ana Beatriz P. Montanhera, Ines
b
ˆ
ˆ b,
´
Artur Smania
Junior
, Elza de F.A. Smania
Edesio L. Simionattoc, Franco Delle Monached
b

a

´
´
Departamento de Quımica,
Universidade Federal de Santa Catarina, Florianopolis,
SC, Brazil
´
Departamento de Microbiologia, Universidade Federal de Santa Catarina, Florianopolis,
SC, Brazil
c
¸ ˜ Universidade Regional de Blumenau, Blumenau, SC, Brazil
´
Departamento de Quımica,
Fundacao
d
Centro Chimica dei Recettori, C.N.R., Instituto di Chimica, Largo F. Vito 1, 00168 Roma, Italy

Received 18 November 2003; accepted 11 December 2003

Abstract
The hydroalcoholic extract of the leaves from Rheedia gardneriana yielded volkensiflavone

(1), fukugetin (2), fukugiside (3), GB2a-I-7-O-glucoside (4) and epicatechin (5). Compounds 1–5, and some derivatives of 1 and 2 were evaluated for lethality to brine shrimp
larvae and for antibacterial activity.
䊚 2004 Elsevier B.V. All rights reserved.
Keywords: Rheedia gardneriana; Biflavonoids; Antimicrobial activity

Plant. Rheedia gardneriana Planch et Triana (Guttiferae), leaves collected in
February 1999 in Blumenau. A voucher specimen is deposited at the Dr Roberto
ˆ
Miguel Klein Herbarium (Departamento de Ciencias
Naturais, FURB, Blumenau)
under numbers 534–540.
*Corresponding author. Fax: q55-48-331-9711.
E-mail address: [email protected] (M.G. Pizzolatti).
0367-326X/04/$ - see front matter 䊚 2004 Elsevier B.V. All rights reserved.
doi:10.1016/j.fitote.2003.12.023

L.G. Verdi et al. / Fitoterapia 75 (2004) 360–363

361


Uses in traditional medicine. This species has been reported as folk medicine for
the treatment of inflammations of urinary tract, arthritis and to relieve pain w1x.

Previously isolated classes of constituents. Biflavonoids w2–4x, xanthones w5–8x,
triterpenes w5,9x, and 4-substituted coumarins w7,10,11x.

New-isolated
(0.004%).

constituents.

GB2a-I-7-O-glucoside (0.02%) and epicatechin

Tested material. Hydroalcoholic extract, partitioned with ethyl acetate resulting in
the EtOAc-fraction (3.42%) and aqueous fraction (5.97%). Si-gel CC of the EtOAcfraction yielded volkensiflavone (1) (0.085%), fukugetin (2) (0.15%), fukugiside
(3) (0.007%), GB-2a-I-7-O-glucoside (4) (0.02%), epicatechin (5) (0.004%).
Derivatives: volkensiflavone peracetylated (6), tetramethylated (7), tetrabenzoylated
(8); fukugetin peracetylated (9), pentamethylated (10) and pentabenzoylated (11).

362


Table 1
Antibacterial activity and Brine Shrimp Lethality Test (BST) of extract, fraction, isolated compounds and derivatives of R. gardneriana
Antibacterial activitya

compounds

Escherichia
coli

Bacilli
cereus

Pseudomonas
aeruginosa

Staphylococcus
aureus

Staphylococcus

saprophyticus

10
8
NI
NI
NI
NI
NI
NI
NI
NI
9
NI

30
22
12.5
9
12

25
21
10
NI
12.5
17
18

NI
9.5
9
NI
9
15
11
NI
NI
10
9
11


30
18
9
9
8.5
20
21
15
NI
14
15
19

29
14.5
NI
NI
NI
18

15
NI
NI
9
10.5
12

Crude extract
EtOAc fraction
1
6
7
8
2
9
10
11
3
4
5

a

BST
Artemia salina
LC50, ppmb
977 (751–1258)
)1000
)1000

)1000
851 (681–1071)
933 (724–1207)
)1000
)1000
)1000

Values (mean of three replicates) are diameter of zone of inhibition (mm): -9, inactive; 9–12, partially active; 13–18, active; )18, very active w13x.
wCompoundsxs1 mgy50 ml; wFractionxs3 mgy50 ml; NIsno inhibition.
b
Confidence intervals 95%.


L.G. Verdi et al. / Fitoterapia 75 (2004) 360–363

Fractiony

L.G. Verdi et al. / Fitoterapia 75 (2004) 360–363

363

Table 2
Minimum inhibitory (MIC) and minimum bactericidal concentrations (MBC) from compounds of R.
gardnerianaa
Compound

Volkensiflavone
Fukugetin
Fukugiside
a

B. cereus


E. coli

P. aeruginosa

S. aureus

MIC

MBC

MIC

MBC

MIC

MBC

MIC

MBC

0.15
0.12
1.0

0.62
)1.25
)1.25

1.0
1.0
1.0

)1.25
)1.25
)1.25

1.0
1.0
1.0

)1.25
)1.25
)1.25

0.25
0.09
1.0

)1.25
)1.25
)1.25

Concentrations in mg mly1.

Studied activity. Brine shrimp lethality test w12x, antimicrobial activity by the agarwell diffusion method and determination of minimum inhibitory and minimum
bactericidal concentrations w13x.

Used microorganisms. Listed in Table 1.

Results. Reported in Tables 1 and 2.

Conclusions. The extract and methylated and benzoylated fukugetin showed toxicity
to brine shrimp. Compounds 1–5 were active against all Gram-positive bacteria
tested. Derivatives of compound 1 and 2 were less active than the starting material.
References
w1x Cordeiro R, Nunes V do A, Almeida CR de. Plantas que Curam. Ed. Tres,
ˆ 1996.
w2x Botta B, Mac-Quhae M, Delle Monache F, Delle Monache G. J Natl Prod 1984;47:1053.
w3x Luzzi R, Guimaraes
˜ CL, Verdi LG, Simionatto EL, Delle Monache F, Yunes RA, Floriani AEO,
Cechinel Filho V. Phytomedicine 1997;4:141.
w4x Cechinel Filho V, Da Silva KL, De Souza MM. Z Naturforsch 2000;55c:820.
w5x Braz Filho R, Magalhaes
˜ GC, Gottlieb OR. Phytochemistry 1970;9:673.
w6x Delle Monache F, Delle Monache G, Marini-Bettolo GB. J Natl Prod 1983;46:655.
w7x Delle Monache F, Delle Monache G, Waterman PG, Crichton EG, Alves de Lima R. Phytochemistry 1984;23:1757.
w8x Delle Monache F, Botta B, De Mello JF, Coelho JSB, Menichini F. J Natl Prod 1984;47:620.
w9x Santos MH, Nagen TJ, Oliveira TT, Braz Filho R. Quimica Nova 1999;22:654.
w10x Hegnauer R. Chemotaxonomie der Pflanzen 1966;4:216.
w11x Basle BV, Rezende CMA, Gottlieb OR. Biochem Syst Ecol 1973;1:111.
w12x Meyer BN, Ferrigni NR, Putman JE, Jacobson LB, Nichols DE, McLaughlin JL. Planta Med
1982;45:31.
w13x Smania
ˆ
ˆ
´
Junior
A, Delle Monache F, Smania
EFA, Gil ML, Benchetrit LC, Cruz FS. J
Ethnopharmacol 1995;45:177.

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