Journal of Ethnopharmacology 106 (2006) 348–352

Validation of use of a traditional antimalarial remedy from

French Guiana, Zanthoxylum rhoifolium Lam
V. Jullian a , G. Bourdy b,∗ , S. Georges a , S. Maurel a , M. Sauvain a
aLaboratoire de Pharmacochimie des Substances Naturelles et Pharmacophores Redox, UMR-152 IRD - Université Paul Sabatier,
Centre IRD de Cayenne, BP 165, 97323 Cayenne, Guyane, France
b Laboratoire de Pharmacochimie des Substances Naturelles et Pharmacophores Redox, UMR-152 IRD - Université Paul Sabatier Toulouse III,

Faculté des sciences pharmaceutiques, 31062 Toulouse Cedex 09, France

Received 5 September 2005; received in revised form 10 January 2006; accepted 13 January 2006
Available online 28 February 2006

Abstract
Zanthoxylum rhoifolium bark (Rutaceae) is a medicinal plant, traditionally used in French Guiana to treat and prevent malaria. Bioassay-guided
extractions of Zanthoxylum rhoifolium bark have shown that antiplasmodial activity is concentrated in the alkaloid fraction. Further fractionation
of this extract has yielded seven benzophenanthridine alkaloids, dihydroavicine 1, dihydronitidine 2, oxyavicine 3, oxynitidine 4, fagaridine 5,
avicine 6 and nitidine 7. Antimalarial activity of the last five compounds has been evaluated, and nitidine was the most potent, displaying an
IC50 < 0.27 ␮M against Plasmodium falciparum.
Investigation of the traditional remedy, a trunk bark decoction in water, has shown that fagaridine 5, avicine 6 and nitidine 7 are also present in
the decoction, therefore justifying the traditional use of Zanthoxylum rhoifolium bark as antimalarial.
© 2006 Elsevier Ireland Ltd. All rights reserved.

Keywords: Antimalarial; Traditional remedy; Zanthoxylum rhoifolium; French Guiana; Medicinal plant; Plasmodium

1. Introduction
French Guiana is a 91,000 km2 overseas department of
France, located North East of the South American continent,
between 2◦ –6◦ North and 52◦ –54◦ West. It borders Brazil with
Oiapoque river (East) and Tumuc–Humac mountains (South),
while Maroni river delimits the western border with Suri-
name. This department is severely affected by endemic malaria
(Huguet and Clausse, 2003). During an ethnopharmacological
study aiming to detect antimalarial remedies used by the local
population, we recorded the use of Zanthoxylum rhoifolium bark
(Rutaceae) claimed to have both curative and preventive activ-
ities. As a curative remedy, Zanthoxylum rhoifolium bark is
boiled for a long time in water, alone or mixed with other ingre-
dients (Vigneron et al., 2005). In another paper, we demonstrated
this traditional preparation to be active in vivo (Bertani et al.,
2005).
∗ Corresponding author. Tel.: +33 594 299276; fax: +33 594 319855.
E-mail address: yuruma@cayenne.ird.fr (G. Bourdy).
Other therapeutic uses are also reported for this species. In
1956, Lemée (1956) was the first one to state that the bitter
bark of this species is a good tonic and febrifuge, also very
useful as a mouth rinse against toothache. Latter, from more
recent sources (Grenand et al., 2004), it was noted that the bark
decoction is used externally to cure venereal chancre, external
parasites, and pimps eruption on children, legs of children in
which case treatment should be completed by drinking some of
the bitter preparation. Pieces of roots, mixed with mango peels
are also administered in the form of a decoction used to help
placenta delivery.
Finally, it was told to us (Bourdy, unpublished data) that aque-
ous extract of Zanthoxylum rhoifolium roots would also have a
positive effect on drepanocytic erythrocytes making this species
valuable in case of sickle cell anaemia. Though this effect has
been demonstrated for Fagara zanthoxyloı̈des due its hydrox-
ymethylbenzoic acid and zanthoxylol content (Sofowora et al.,
1975), no study has been conducted in that peculiar field with
Zanthoxylum rhoifolium.
In the close-by Guiana, it is reported that inner bark and leaves
of Zanthoxylum rhoifolium are boiled and drunk as antipyretic
and antimalarial by Patamona Indians (Tiwari, 1999), and the

0378-8741/$ – see front matter © 2006 Elsevier Ireland Ltd. All rights reserved.
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 V. Jullian et al. / Journal of Ethnopharmacology 106 (2006) 348–352
same use has been reported in Bolivia (De Lucca and Zalles,
1992). Finally, in Peru, this species enjoys the reputation to have
digestive and tonic properties (Brack Egg, 1999).
The aim of our study was to carry out a scientific evaluation of
the claimed antimalarial properties of Zanthoxylum rhoifolium
bark, through isolation of antimalarial(s) compound(s), in order
to validate its use, and confirm the previously detected in vivo
activity. As far as we know, prior to this study no evaluation
of the antimalarial activity of this species had been undertaken,
though antimalarial activity has been evidenced for other Zan-
thoxylum, such as Zanthoxylum acutifolium (Arruda et al., 1992)
and Zanthoxylum chabyleum (Gessler et al., 1994).
2. Materials and methods
2.1. Plant collection
Zanthoxylum rhoifolium bark was collected in the Remire
village in 2001 (French Guiana), dried in a shade drier, and
then ground into powder. Herbarium voucher (GB2837) was
deposited in French Guiana Herbarium (CAY) and determined
by specialists.
2.2. General experimental procedure
The solvents were purchased at SDS (Peypin, France) and
were of analytical or HPLC grade. Flash column chromatog-
raphy was performed on Merck Geduran® , silica gel Si 60
(40–63 ␮m). TLC was performed on Merck TLC plates coated
with silica gel Si 60 F254. Compounds were detected with UV
light at 254 nm and by spraying Dragendorff’s reagent. HPLC
was performed with two isocratic pumps (Varian Prostar 218),
on a Waters X-Terra column (RP18, 5 ␮m, 4.6 mm × 250 mm),
with a flow rate of 1 mL/min, and compounds were detected
with a PDA detector (Varian 330). NMR spectra were recorded
on Bruker AC 250 or Bruker AMX 400 spectrometers. Mass
spectra were recorded on a Nermag R 10-10 spectrometer.
2.3. Phytochemical isolation
After Zanthoxylum rhoifolium trunk bark powder showed a
positive test to Dragendorff’s reagent, an extraction of the alka-
loids was performed.
Zanthoxylum rhoifolium trunk bark powder (520 g) was wet-
ted with an ammonium hydroxide aqueous solution (14%), and
left for 2 h in order to perform alkaloids extraction. The pow-
der was then left overnight in 1 L of dichloromethane and fil-
trate. This procedure was repeated once. The dichloromethane
layer was extracted three times with 0.1 M hydrochloric acid.
The aqueous layer was basified to pH 10 with concentrated
ammonium hydroxide solution, and then extracted twice with
dichloromethane. This organic layer was dried over anhydrous
magnesium sulfate and the solvent was removed under reduced
pressure to yield 1.2 g of a mixture of alkaloids, A1.
The A1 extract was submitted to flash column chromatogra-
phy on silica gel, using dichloromethane with increasing amount
of methanol as eluant.
349
With 1% methanol in dichloromethane, 95 mg of a mix-
ture containing dihydroavicine and dihydronitidine was eluted.
Another flash chromatography on silica gel eluted with cyclo-
hexane, then cyclohexane containing 10 and 20% of ethyl acetate
gave 35 mg of dihydroavicine 1 and 21 mg of dihydronitidine 2.
With 5% methanol in dichloromethane, two fractions were
obtained: the first fraction was 150 mg of a mixture containing
oxyavicine 3 and oxynitidine 4. Another flash chromatography
on silica gel eluted with dichloromethane and dichloromethane
with 0.25% methanol gave 25 mg of oxyavicine 3 and 15.5 mg
of oxynitidine 4. The second fraction was 125 mg of fagaridine
5.
With 10 and 20% methanol in dichloromethane, 500 mg of a
mixture containing avicine 6 and nitidine 7 was eluted. We failed
to separate these two compounds by classical chromatographic
methods. As we found that dihydroavicine and dihydronitidine
were easily separated, 75 mg of this mixture of avicine and niti-
dine were reduced with sodium borohydride in methanol to give
a mixture of dihydroavicine and dihydronitidine. After flash
column chromatography, 33 mg of dihydroavicine and 18 mg
of dihydronitidine were obtained and then oxidized as follows:
12 mg of dihydronitidine were dissolved in a mixture of chlo-
roform and methanol (3/1) and stirred under ambient air during
48 h. The formation of nitidine was monitored by thin layer
chromatography (TLC) and 1 H NMR. Solvents were removed
under reduced pressure and 5.6 mg of nitidine hydroxide 7 was
obtained after flash chromatography of the crude reaction mix-
ture. Under the same conditions, 20 mg of dihydroavicine gave
2.6 mg of avicine hydroxide 6.
2.4. Structural data of isolated compounds
Dihydroavicine 1: ESIMS: 332 (M–H)+; 1 H NMR
(250 MHz, CDCl3 ): 2.60 (3H, s), 4.11 (2H, s), 6.00 (2H,s), 6.05
(2H, s), 6.77 (1H, s), 7.11 (1H, s), 7.26 (1H, s), 7.49 (1H, d,
8.5 Hz), 7.60 (1H, d, 8.5 Hz), 7.64 (1H, s). Dihydronitidine 2:
ESIMS: 348 (M–H)+; 1 H NMR (250 MHz, CDCl3 ): 2.59 (3H,
s), 3.95 (3H, s), 3.99 (3H, s), 4.13 (2H, s), 6.05 (2H, s), 6.80
(1H, s), 7.11 (1H, s), 7.30 (1H, s), 7.49 (1H, d, 8.9 Hz), 7.65
(1H, s), 7.69 (1H, d, 8.9 Hz). Oxyavicine 3: EIMS: 347 (M+);
DCIMS (NH3 ): 348 (MH+); 1 H NMR (400 MHz, CDCl3 ): 3.97
(3H, s), 6.11 (2H, s), 6.13 (2H, s), 7.17 (1H, s), 7.54 (1H, d,
8.6 Hz), 7.60 (1H, s), 7.62 (1H, s), 7.90 (1H, s), 7.91 (1H,
d, 8.6 Hz); 13 C NMR (100 MHz, CDCl3 ): 41.4, 100.8, 101.7,
102.1, 102.8, 104.9, 106.7, 116.9, 118.7, 120.9, 121.0, 123.4,
131.2, 132.1, 136.0, 147.2, 147.7, 148.3, 152.6, 164.2. Oxyni-
tidine 4: EIMS: 363 (M+); DCIMS (NH3 ): 364 (MH+); 1 H
NMR (400 MHz, CDCl3 ): 4.00 (3H, s), 4.08 (3H, s), 4.12 (3H,
s), 6.12 (2H, s), 7.20 (1H, s), 7.58 (1H, d, 8.6 Hz), 7.61 (1H,
s), 7.66 (1H, s), 7.95 (1H, s), 8.01 (1H, d, 8.6 Hz); 13 C NMR
(100 MHz, CDCl3 ): 41.4, 56.3, 56.4, 101.7, 102.8, 103.0, 104.9,
108.8, 116.5, 118.5, 119.2, 121.0, 123.4, 128.5, 132.0, 136.0,
147.0, 148.2, 149.9, 154.2, 164.1. Fagaridine 5: ESIMS: 334
(M–OH)+; 1 H NMR (400 MHz, CDCl3 ): 2.70 (3H, s), 3.90 (3H,
s), 5.44 (1H, s), 6.00 (2H, s), 6.97 (1H, d, 8.5 Hz), 7.06 (1H, s),
7.41 (1H, d, 8.5 Hz), 7.49 (1H, d, 8.5 Hz), 7.62 (1H, s), 7.69
(1H, d, 8.5 Hz); 13 C NMR (100 MHz, CDCl3 ): 40.5, 62.0, 86.3,
350 V. Jullian et al. / Journal of Ethnopharmacology 106 (2006) 348–352
100.5, 101.2, 104.7, 117.2, 119.7, 120.0, 122.9, 123.7, 124.2,
125.1, 126.8, 131.0, 137.9, 145.2, 147.4, 148.1, 149.2. Avicine
hydroxide 6: EIMS: 317 (M–CH3)+; DCIMS (NH3 ): 318; 1 H
NMR (250 MHz, CDCl3 /CD3 OD): 4.87 (3H, s), 6.17 (2H, s),
6.28 (2H, s), 7.35 (1H, s), 7.80 (1H, s), 7.96 (1H, s), 7.99 (1H,
s), 8.00 (1H, d, 9.1 Hz), 8.26 (1H, d, 9.1 Hz), 9.99 (1H, s). Niti-
dine hydroxide 7: EIMS: 333 (M–CH3)+; 1 H NMR (250 MHz,
CDCl3 /CD3 OD): 4.05 (3H, s), 4.16 (3H, s), 4.87 (3H, s), 6.15
(2H, s), 7.35 (1H, s), 7.88 (1H, s), 7.90 (1H, s), 7.97 (1H, s),
7.99 (1H, d, 8.8 Hz), 8.35 (1H, d, 8.8 Hz), 9.98 (1H, s).
2.5. Preparation of the traditional remedy and alkaloid
extraction
Fifty grams of bark powder was left to boil in 1 L of water
until it reduces by half, and the mixture was left overnight to
cool. The resulting decoction was then filtered.
The previous decoction was acidified to pH 1 with hydrochlo-
ric acid and extracted with dichloromethane. The aqueous layer
was basified to pH 10 with ammonium hydroxide solution, and
then extracted twice with dichloromethane. The organic layer
was dried over anhydrous magnesium sulfate and the solvent
removed under reduced pressure to yield 22 mg of an alkaloid
mixture (A2).
2.6. Detection of compounds in the alkaloid mixture A2
Compounds were detected and identified by means of HPLC
and TLC, in comparison with fagaridine 5, avicine 6 and nitidine
7 previously isolated from A1.
Nitidine and avicine were detected with HPLC using the fol-
lowing solvants: solvant A was a phosphate buffer, c = 0.01 M,
pH 11.2 and solvant B was methanol. Eluants were 40% of A
and 60% of B, or 35% of A and 65% of B.
Nitidine, avicine and fagaridine were detected on TLC.
Fagaridine was eluted with a mixture of dichloromethane and
methanol (95:5). Nitidine and avicine were eluted with a mix-
ture of ethyl acetate, methanol and 25% ammonium hydroxide
(50:1:0.5).
2.7. In vitro antiplasmodial drug assay
In vitro activity: Plasmodium falciparum (chloroquine-
resistant strain FCB1 kindly given by A. Valentin from the
laboratory of Parasitology, Faculty of Pharmacy, Montpellier,
France) was evaluated by two methods: a micromethod using
the lactate dehydrogenase (LDH) enzyme of Plasmodium fal-
ciparum (Delhaes et al., 1999; Makler and Hinrichs, 1993),
and a micromethod mesuring [3 H]-hypoxanthine incorporation
(Desjardins et al., 1979).
Erythrocytes were infected with Plasmodium falciparum,
from parasite cultures obtained using the method of Tragger
and Jensen (1976) and synchronized by 5% D-sorbitol (Merck,
Germany) lysis (Lambros and Vanderberg, 1979).
LDH micromethod: Infected erythrocythes were resuspended
in the complete culture medium at a haematocrit of 1.5%.
The suspension was distributed in 96-well, microtitre plates
(200 ␮L/well). Drug testing was performed in triplicate with
cultures mostly at ring stage at 1% parasitaemia. For each assay,
parasite culture was incubated with drug for 48 h in 5% CO2 at
95% relative humidity, and frozen until the biochemical assay
could be run.
A 20 ␮L subsample of the contents of each well was mixed
with 100 ␮L of a substrate solution containing 20 mg of sodium
l-lactate (Sigma), 5.5 mg Tris (Sigma) and 3.7 mg of 3-acetyl
pyridine adenine dinucleotide (APAD; Sigma)/mL, in the well
of another microtitre plate. After incubation for 30 min at 37 ◦ C,
25 ␮L of a mixture of NBT (1.6 mg/mL; Sigma) and PES
(0.1 mg/mL; Sigma) were added to each well. After another
35 min of incubation at the same temperature, the reaction was
stopped by the addition of 25% acetic acid (25 ␮L/well). The for-
mation of the reduced form of APAD was measured at 650 nm,
using a spectrophotometer (microplate reader, Polarstar BMG).
The reference product was artemisinine.
[3 H]-hypoxanthine micromethod: Infected erythrocythes
were resuspended in the complete culture medium at a haema-
tocrit of 1%. The suspension was distributed in 96-well,
microtitre plates (200 ␮L/well). Drug testing was performed in
triplicate with cultures mostly at ring stage at 1% parasitaemia.
For each assay, the parasite culture was incubated with drug for
48 h in 5% CO2 at 95% relative humidity. After 36 h of incuba-
tion, 20 ␮L of a solution of [3 H]-hypoxanthine (12.5 mCi/mL,
Perkin-Elmer) was added. Twelve hours later, cultures were
frozen at −20 ◦ C, then defrozen and each well was harvested
onto a glass fiber filter. The incorporated [3 H]-hypoxanthine
was determined with a ␤-counter (145 Microbeta Trilux, Wallac
Perkin-Elmer). The reference product was chloroquine.
3. Results and discussion
In vitro antiplasmodial and cytotoxicity activities of extracts
and pure compounds are given in Table 1. Fig. 1 displays the
structure of the compounds.
3.1. Zanthoxylum rhoifolium bark
The antiplasmodial activity of Zanthoxylum rhoifolium bark
was concentrated in the alkaloid fraction (A1) (44% inhibi-
tion at 10 ␮g/mL, using LDH micromethod) and was similar to
A2. From A1, we isolated seven benzophenanthridine alkaloids.
The compounds were identified by mass and NMR spectro-
scopies and by comparison with published data: dihydroavicine
1 (Joshi et al., 1991), dihydronitidine 2 (Hanaoka et al., 1987),
oxyavicine 3 (Le et al., 2004), oxynitidine 4 (Hanaoka et al.,
1987; Le et al., 2004) and fagaridine 5 (Nakanishi and Suzuki,
1998). For avicine 6 and nitidine 7, some chemical shifts on the
1 H NMR spectra were different, certainly because previously
reported values (Ishii et al., 1984; Hanaoka et al., 1987) were
from spectra recorded for the hydrochloride salts in dimethylsul-
foxide. However, the structure of avicine and nitidine obtained
in this work was confirmed by their reduction in dihydronitidine
and dihydroavicine with sodium borohydride.
The following alkaloids have been already described
in Zanthoxylum rhoifolium: berberine, allocryptopine, can-
 V. Jullian et al. / Journal of Ethnopharmacology 106 (2006) 348–352 351

Table 1
In vitro antiplasmodial activity of Zanthoxylum rhoifolium extracts and alkaloids
Plasmodium falciparum FCB1 (LDH) Plasmodium falciparum FCB1 ([3 H]-hypoxanthine)

IC50 (␮g/mL) IC50 (␮M) IC50 (␮g/mL) IC50 (␮M)

A2 15.3 3.3
Nitidine (hydroxide) 1.8 4.9 <0.1 <0.27
Avicine (hydroxide) 11.7 33 4.3 12.3
Fagaridine 13.6 38 NT NT
Oxynitidine NA NA NA NA
Oxyavicine NA NA NA NA
Artémisinine 0.069
Chloroquine 0.175

NT, not tested; NA, not active.

dicine, magnoflorine, N-methylcanadine, N-methylisocorydine,
N-methylthalicmidine, tembetarine, chelerythrine and niti-
dine (Mester, 1973), dihydronitidine, oxynitidine, skimmi-
anine (De Moura et al., 1997), rhoifoline A, rhoifoline
B, 6-acetonyldihydroavicine, 6-acetonyldihydronitidine, 6-
acetonyldihydrochelerythrine and xanthoxyline (Gonzaga et al.,
2003). So, dihydroavicine, oxyavicine, fagaridine and avicine
are described for the first time in this species.
Three of the A1 isolated compounds displayed antiplas-
modial activity, ranging from good (nitidine, the most potent
compound) to moderate (avicine and fagaridine). Oxyavicine
and oxynitidine were inactive. Dihydronitidine and dihy-
droavicine have not been tested because of their insta-
bility in solution being oxydized in nitidine and avicine,
respectively.
Gakunju et al. (1995) and Nyangulu et al. (2005) have already
reported the antimalarial activity of nitidine with an IC50 value
of 0.05 ␮g/mL on Plasmodium falciparum FCB strain, using
the [3 H]-hypoxanthine micromethod. The value we found using
the [3 H]-hypoxanthine micromethod is in agreement with these
results. However, the antimalarial activity of nitidine displayed
by using the LDH micromethod was much lower (the same for
avicine and A2 extract), certainly due to the difference in sensi-
bility between both tests.
3.2. Traditional remedy
The traditional preparation displayed a moderate in vivo
activity (78% of inhibition at 715 mg/kg) (Bertani et al., 2005).
As observed for A1, the in vitro activity was concentrated in the
alkaloidic fraction, A2.
From A2, were detected and identified by means of HPLC
and TLC three compounds, previously isolated from A1: avicine,
nitidine and fagaridine, all three compounds displaying antiplas-
modial activity.
These findings indicate that the traditional way of preparing
the remedy allows active compounds extraction, and therefore,
corroborate its antimalarial reputation. Still, this does not pre-
clude that other components present in the aqueous phase are
not involved in the claimed antimalarial activity.

4. Conclusion

We demonstrated in this study that a widely used traditional

Fig. 1. Alkaloids isolated from Zanthoxylum rhoifolium trunk bark.
remedy made out of Zanthoxylum rhoifolium bark watery decoc-
tion, used in French Guiana against malaria contains three active
compounds, also present in the trunk bark, therefore justifying its
use. The most active compound identified in this traditional rem-
edy is nitidine, which, like other benzophenanthridine alkaloids
is a well known cytotoxic agent, inhibitor of DNA Topoiso-
merase I (Fang et al., 1993) and therefore has been widely studied
for its in vivo antitumor properties (Ishii et al., 1985).
Nitidine was also found to be the major antimalarial com-
ponent of Toddalia asiatica, a traditional remedy used in Kenya
against malaria and fever, active on a chloroquino-resistant strain
of Plasmodium falciparum (Gakunju et al., 1995). Moreover,
recently, nitidine was used as a lead compound to perform syn-
thesis of various benzophenanthridines with antimalarial activity
(Nyangulu et al., 2005).
352 V. Jullian et al. / Journal of Ethnopharmacology 106 (2006) 348–352
We consider that one of the main interest lying in a prepa-
ration containing nitidine is that, beyond its antimalarial prop-
erties, it could be useful for treating cases of malaria due to
chloroquine-resistant strains, a case of interest in French Guiana,
since a high prevalence of Plasmodium falciparum, with a high
chemoresistance level has been observed over the past few years
(Carme, 2001).
Also, it might prove fruitful to perform tests measuring the
possible synergism of action of a Zanthoxylum rhoifolium prepa-
ration with standard molecules since quite often, people from
French Guiana mix plants and remedies from the pharmacy when
treating malaria.
Acknowledgment
Thanks are due to M.F. Prévost, for the determination of
Herbarium vouchers.
References
Arruda, M.S.P., Fernandes, J.B., Da Silva, M.G.F., Vieira, P.C., Pirana, J.R.,
1992. Quinolone alkaloids from Zanthoxylum acutifolium. Phytochemistry
31, 3617–3619.
Bertani, S., Bourdy, G., Landau, I., Robinson, J.C., Esterre, P., Deharo, E.,
2005. Evaluation of French Guiana traditional antimalarial remedies. Jour-
nal of Ethnopharmacology 98, 45–54.
Brack Egg, A., 1999. Diccionario enciclopédico de plantas utiles del Perú.
PNUD, CBC, Cuzco, Peru, p. 556.
Carme, B., 2001. Les parasitoses humaines en Guyane Française. Presse
Médicale 30, 1601–1608.
De Lucca, M., Zalles, J., 1992. Flora Medicinal Boliviana. Los Amigos del
libro Editions, La Paz, Cochabamba, Bolivia, p. 498.
De Moura, N.F., Ribeiro, H.B., Machado, E.C.S., Ethur, E.M., Zanatta,
N., Morel, A., 1997. Benzophenanthridine alkaloids from Zanthoxylum
rhoifolium. Phytochemistry 46, 1443–1446.
Delhaes, B.J., Lazaro, J.E., Gay, F., Thellier, M., Danis, M., 1999. The micro-
culture tetrazolium assay (MTA): another colorimetric method of testing
Plasmodium falciparum chemosensitivity. Annals of Tropical Medicine
and Parasitology 93, 31–40.
Desjardins, R.E., Canfield, C.J., Haynes, J.D., Chulay, J.D., 1979. Quantita-
tive assesment of antimalarial activity in vitro by a semi microdilution
technique. Antimicrobial Agents and Chemotherapy 16, 710–718.
Fang, S.D., Wang, L.K., Hecht, S.M., 1993. Inhibitors of DNA Topoisomerase
I isolated from the roots of Zanthoxylum nitidum. Journal of Organic
Chemistry 58, 5025–5027.
Gakunju, D.M.N., Mberu, E.K., Dossaji, S.F., Gray, A.I., Waigh, R.D.,
Waterman, P.G., Watkins, W.M., 1995. Potent antimalarial activity of the
alkaloid nitidine, isolated from a Kenyan herbal remedy. Antimicrobial
Agents and Chemotherapy 39, 2606–2609.
Gessler, M.C., Nkunya, M.H.H., Mwasumbi, L.B., Heinrich, M., Tannera,
M., 1994. Screening Tanzanian medicinal plants for antimalarial activity.
Acta Tropica 56, 65–77.
Gonzaga, W.d.A., Weber, A.D., Giacomelli, S.R., Dalcol, I.I., Hoelzel, S.C.S.,
Morel, A.F., 2003. Antibacterial alkaloids from Zanthoxylum rhoifolium.
Planta Medica 69, 371–374.
Grenand, P., Moretti, C., Jacquemin, H., Prévost, M.F., 2004. Pharmacopées
Traditionnelles en Guyane. Créoles, Palikur, Wayãpi. IRD Editions, Paris,
p. 816.
Hanaoka, M., Yamagishi, H., Marutani, M., Mukai, C., 1987. Chemical
transformation of protoberberines. XIII. A novel and efficient synthesis
of antitumor benzo[c]phenanthridine alkaloids, nitidine, and fagaronine.
Chemical and Pharmaceutical Bulletin 35, 2348–2354.
Huguet, P., Clausse, J., 2003. Profil Épidémiologique du Paludisme en
Guyane 2000–2002. Cellule de veille épidémiologique (DSDS), Cayenne,
Guyane, France.
Ishii, H., Ichikawa, Y.I., Kawanabe, E., Ishikawa, M., Ishikawa, T., Kuretani,
K., Inomata, M., Hoshi, A., 1985. Studies on the chemical constituent
of rutaceous plant. LX. Development of a versatile method for syn-
these of the antitumor benzo[c]phenanthridine alkaloids. (9). Efficient
syntheses and antitumor activities of nitidine and related nonphenolic
benzo[c]phenanthridine alkaloids. Chemical and Pharmaceutical Bulletin
33, 4139–4151.
Ishii, H., Ishikawa, T., Ichikawa, Y.I., Sakamoto, M., Ishikawa, M., Taka-
hashi, T., 1984. Studies on the chemical constituent of rutaceous plant.
LV. The development of a versatile method for synthese of the antitumor
benzo[c]phenanthridine alkaloids. (5). A new method for quaternarization
of the benzo[c]phenanthridine nucleus. Chemical and Pharmaceutical Bul-
letin 32, 2984–2994.
Joshi, B.S., Puar, M.S., Moore, K.M., Pelletier, S.W., 1991. Isolation of
dihydroavicine and rhetsisnine from Zanthoxyllum budrunga. The revision
of 1H and 13C NMR spectral assignments for sanguinarine. Heterocycles
32, 1365–1370.
Lambros, C., Vanderberg, J.P., 1979. Synchronization of Plasmodium falci-
parum erythrocytic stages in culture. Journal of Parasitology 65, 418–
420.
Le, T.N., Gang, S.G., Cho, W.J., 2004. A versatile total synthesis
of benzo[c]phenanthridine and protoberberine alkaloids using lithiated
toluamide-benzonitrile cycloaddition. Journal of Organic Chemistry 69,
2768–2772.
Lemée, A., 1956. Flore de la Guyane Française. Paul Chevalier Editions,
Paris, p. 124.
Makler, M.T., Hinrichs, D.J., 1993. Measurement of the lactate deshydroge-
nase activity of Plasmodium falciparum as an assessment of parasitemia.
American Journal of Tropical Medicine and Hygiene 48, 205–210.
Mester, I., 1973. The occurence of the alkaloids in Rutaceae. Fitoterapia 44,
123–152.
Nakanishi, T., Suzuki, M., 1998. Revision of the structure of fagaridine based
on the comparison of UV and NMR data of synthetic compounds. Journal
of Natural Products 61, 1263–1267.
Nyangulu, J.M., Hargreaves, S.L., Sharples, S.L., Mackay, S.P., Waigh,
R.D., Duval, O., Mberu, E.K., Watkins, W.M., 2005. Antimalarial
benzo[c]phenanthridines. Bioorganic and Medical Chemistry Letters 15,
2007–2010.
Sofowora, E.A., Isaac-Sodeye, W.A., Ogunkoya, L.O., 1975. Isolation and
characterisation of anti-sickling agent from Fagara zanthoxylloides root.
Llodya 38, 387–390.
Tiwari, S., 1999. Ethnomedecine of the Patamona Indians of Guyana. Lehman
College, City University of New York, New York, p. 560.
Tragger, W., Jensen, J.B., 1976. Human malaria in continuous culture. Science
193, 673–675.
Vigneron, M., Deparis, X., Deharo, E., Bourdy, G., 2005. Antimalarial reme-
dies in French Guiana: a knowledge attitudes and practices study. Journal
of Ethnopharmacology 98, 351–360.