Journal of Ethnopharmacology 79 (2002) 43 – 52

www.elsevier.com/locate/jethpharm

Yucatec Mayan medicinal plants:

evaluation based on indigenous uses
Anita Ankli a, Michael Heinrich b,g, Peter Bork b, Lutz Wolfram c, Peter Bauerfeind c,
Reto Brun d, Cécile Schmid d, Claudia Weiss e, Regina Bruggisser f, Jürg Gertsch a,
Michael Wasescha a, Otto Sticher a,*
a
Department of Applied Biosciences, Institute of Pharmaceutical Sciences, Swiss Federal Institute of Technology (ETH) Zurich,
Winterthurerstrasse 190, CH-8057 Zurich, Switzerland
b
Institute of Pharmaceutical Biology, Albert-Ludwigs-Uni6ersity, Schänzlestr. 1, D-79104 Freiburg, Germany
c
Di6ision of Gastroenterology, Department of Internal Medicine, Uni6ersity Hospital Zurich, Rämistr. 100, 8091 Zurich, Switzerland
d
Department of Medical Parasitology, Swiss Tropical Institute, Socinstr. 57, CH-4002 Basel, Switzerland
e
Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London, UK
f
Department of Pharmaceutical Biology, Uni6ersity of Basel, Klingelbergstr. 50, CH-4056 Basel, Switzerland
g
Centre for Pharmacognosy and Phytotherapy, The School of Pharmacy, 29 /39 Brunswick Sq., London WC1N 1AX, UK
Received 13 October 2000; received in revised form 30 August 2001; accepted 20 September 2001

Abstract

As part of an ethnopharmacological field study 48 medicinal plants were evaluated using several biological assays with the goal
to obtain information on the pharmacological effects of these plants, which may be of direct relevance to the indigenous uses.
Three species used to treat gastrointestinal disorders showed remarkable activity against Helicobacter pylori. One of them showed
activity against Giardia duodenalis. Cytotoxic effects against KB cells were found for six species. In the group of plants used for
dermatological conditions several species were active against gram-positive bacteria and Candida albicans. Two plant species of
this group were found to be active in an Nuclear Factor-kB (NF-kB) assay measuring inhibition of this pro-inflammatory
transcription factor. A species of the Solanaceae, applied in cases of pain and fever, showed a weak activity against Plasmodium
falciparum. One species traditionally used for diabetes exhibited antihyperglycemic activity. None of the six species from the group
of ‘women’s medicine’ showed relevant affinity to the D2 dopamine receptor. Based on this evaluation, plants with strong activities
should be further investigated phytochemically and pharmacologically to identify active fractions and compounds. © 2002
Elsevier Science Ireland Ltd. All rights reserved.

Keywords: Yucatec Maya; Ethnopharmacological evaluation; Medicinal plants; Traditional medicine; Antibacterial; Anti-inflammatory; Antihy-
perglycemic; Antiparasitic; D2 receptor binding

1. Introduction direct relevance to the indigenous uses (Frei et al.,

One of the numerous objectives of medical ethnob-
otany is the selection of culturally important plant
species in order to further evaluate them for pharmaco-
logical activity (Browner et al., 1988; Etkin, 1994;
Farnsworth, 1988; Frei et al., 1998a; Messner, 1978). In
order to evaluate an ethnopharmacopeia systematically,
plant extracts can be tested in bioassays, which have
* Corresponding author. Tel.: +41-1-635-6050; fax: + 41-1-635-
6882.
E-mail address: sticher@pharma.anbi.ethz.ch (O. Sticher).
1998b; Heinrich et al., 1992a,b; Lewis and Elvin-Lewis,
1994).
The knowledge of medicinal plants was a part of the
ancient Maya culture and they are still utilised by the
Yucatec Mayan inhabitants on the Peninsula of Yu-
catan, Mexico (Roys, 1933; De Landa, 1992). During
an ethnobotanical study in three Mayan communities
(February 1994 –June 1995; September 1996 –October
1996), 360 medicinal plants and 1828 reports on their
uses were documented. The uses of the plants were
divided into nine therapeutic groups (Ankli et al. 1999;
Heinrich et al. 1998). Forty-eight species were chosen

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44 A. Ankli et al. / Journal of Ethnopharmacology 79 (2002) 43–52

and evaluated in bioassays relevant to the following
groups of illnesses, gastrointestinal disorders; dermato-
logical conditions; women’s medicines as well as pain
and/or fever. Plants used to treat ‘diabetes’ were also
tested (Table 2).
In case of Staphylococcus aureus and Yersinia entero-
colitica extract C was used. Extract C was prepared by
extracting 10 g of plant material with ethanol 96%
followed by ethanol 70% and the extracts combined
and reduced under vacuum.

2.3. Bioassays
2. Materials and methods

2.3.1. Antibacterial and antifungal acti6ity

2.1. Plant material
The plants were collected in the villages and sur-
roundings of Chikindzonot, Ekpedz and Xcocmil, Yu-
catan (Mexico). Authenticated voucher specimens were
deposited at the Herbarium of the Centro de Investiga-
ción Cientı́fica de Yucatán (CICY) in Mérida, the
National Herbarium of Mexico (MEXU), the Instituto
Nacional Indigenista (INI) in Valladolid, Yucatan, the
ETH Zurich (ZT) and the Centre for Pharmacognosy
and Phytotherapy, School of Pharmacy, London, UK
(AANK1-654). They were identified by comparison
with authentic specimens and in some cases with the
assistance of specialists at CICY, MEXU and The
Royal Botanical Garden, Kew, UK (K).
2.2. Extract preparation
Shade-dried and powdered plant material (20 g) was
extracted by maceration with 100 ml dichloromethane/
methanol 2:1, repeating the process two times during 36
h. The filtered solvents were combined and evaporated
under vacuum to give the non-polar extract A. The
residue of the dichloromethane/methanol mixture was
dissolved in 100 ml methanol/water 7:3 and macerated
two times during 24 h. The solvents were reduced under
vacuum and the extract was further partitioned between
n-butanol (3× 30–50 ml) and water. The n-butanol
fractions were evaporated to obtain the polar extract B.
The organisms used to test biological activities are
listed in Table 1. Antimicrobial tests were performed by
the disc diffusion technique (Rı́os et al., 1988; DIN,
1992). Overnight cultures of microorganisms were pre-
pared by transferring 30 ml of store culture to 5 ml
broth (Nutrient Broth for bacteria, Sabouraud Liquid
Medium Oxoid for fungi). Ten milliliter of Mueller–
Hinton agar or malt extract agar (for Candida albicans)
was inoculated with 50 ml of overnight culture and
poured over the agar base. Paper discs (6 mm Blanc
Discs, Oxoid) were impregnated with 200 and 600 mg
plant extracts, the solvent evaporated and the treatment
discs placed on the inoculated agar. The plates were
sprayed with methylthiazolyltetrazolium chloride
(MTT, Fluka) after 16 h of incubation at 37 °C. The
inhibition zones were measured in mm.
Susceptibility tests with Helicobacter pylori and
Campylobacter jejuni were carried out on Wilkins Chal-
gren agar plates supplemented with 5% defibrinated
sheep blood and the following antibiotics, 2 mg ampho-
tericin B per ml, 6 mg vancomycin per ml, 5 mg cefsu-
lodin per ml, and 5 mg trimethoprim per ml. Hundred
microliter of a thick H. pylori suspension (yield from
one agar plate, resuspended in 1 ml phosphate buffered
saline (PBS)) was applied onto each plate and the
treatment discs (600 mg plant extract) placed on the
plates. They were then incubated under a water-satu-
rated, micro-aerophilic atmosphere at 37 °C for 1 day
(C. jejuni ), or 3 –5 days (H. pylori ).

Table 1
Test organisms for antibacterial and antifungal activity

Microorganism Origin Clinical picture of diseasesb

Gastrointestinal problems Dermatological conditions

Bacillus cereus ATCC 10702 Diarrhea

a
Campylobacter jejuni Enteritis, diarrhea
Candida albicans H29 ATCC 26790 Mycosis
Escherichia coli ATCC 25922 Diarrhea, dysentery Infected wounds
Helicobacter pylori ATCC 43504 Gastritis, peptic ulcer
Pseudomonas aeruginosa ATCC 25922 Infected wounds
Staphylococcus aureus ATCC 25933 Diarrhea (food intoxication) Topical infection
Staphylococcus epidermidis ATCC 12228 Infection (septicemia)
Yersinia enterocolitica O3 Enterocolitis

a
Obtained from the Division of Gastroenterology, Department of Internal Medicine, University Hospital Zürich.
b
(Kayser et al., 1993).
 A. Ankli et al. / Journal of Ethnopharmacology 79 (2002) 43–52
The MIC determination for H. pylori was carried out
in a modified minimal medium according to Nedenskov
(1994). Different concentrations (in the range from 0.3
to 200 mg/ml) of the plant extracts were added to the
uninoculated medium (5 ml in a 25 ml Erlenmeyer
flask). A fresh H. pylori culture was used as a 1%
inoculum and grown under micro-aerophilic conditions
in a water-saturated atmosphere at 37 °C in a rotary
shaker (G25; New Brunswick Scientific, New Jersey,
USA). The incubation was continued for 2 days at 175
rpm. After 2 days the optical density of the cultures
were photometrically determined at 600 nm (DU-64
spectrophotometer, Beckman, UK). The MIC value
was defined as the extract concentration not allowing
visible growth (less than 0.03 in comparison with 1.2
for the control).
2.3.2. Cytotoxicity study using KB cell culture
The cytotoxicity of the plant extracts was assessed
using the KB cell line (ATCC CCL 17; human
nasopharyngeal carcinoma). The test was carried out
with some modifications according to the screening
technique of Swanson and Pezzuto (1990). The assay
was performed in 96-well plates (Falcon) with an inocu-
lum of 2.5×104 cells per ml. Total volume was 150 ml.
The dried extracts were dissolved in ethanol. Water was
added to dilute the solution 5-fold. Concentration of 50
mg/ml with maximum 1% ethanol was tested. These
solutions were diluted 20-fold by mixing it with culture
medium. For active extracts, IC50 values were deter-
mined. The quantification was performed by adding 15
ml from a 5 mg/ml solution of MTT in PBS (Mosmann,
1983). After incubation at 37 °C for 4 h, the metaboli-
cally active cells produced an insoluble formazan dye.
The medium was drawn off and the formazan dye was
dissolved using 150 ml of 10% sodium dodecylsulfate
(SDS) in water. After 24 h of incubation at room
temperature (RT), the optical density was measured at
540 nm using a microplate reader (MRX, Dynex
Technologies).
2.3.3. Inhibitory acti6ity on NF-sB
Anti-inflammatory activity of the plant extracts was
assayed in Electrophoretic Mobility Shift Assay
(EMSA) using the inhibition of nuclear factor-kB (NF-
kB) binding to a radioactive labelled oligonucleotide as
a molecular target. The bioassay was carried out as
described in Bork et al. (1996).
2.3.4. Antimalarial acti6ity
Antimalarial activity was assessed for the
chloroquine resistant K1 strain and the chloroquine
sensitive T9-96 clone of Plasmodium falciparum. The
parasites were maintained in continuous culture of in-
fected A+ human red blood cells in RPMI 1640
supplemented with 6.9 mg/ml HEPES, 2 mg/ml glucose,
45
2.33 mg/ml NaHCO3, 50 mg/ml hypoxanthine, 40 mg/ml
gentamicin (all Sigma) and 10% A+ serum (North
London Blood Transfusion Centre) (Fairlamb et al.,
1985). Antimalarial IC50 values were assessed using the
modified in vitro lactate dehydrogenase assay (Makler
et al., 1993). Extracts were tested in concentrations
from 1000 to 4.12 mg/ml (3-fold dilution). Fifty micro-
liter of a 1% parasitemia blood suspension (predomi-
nantly ring form) were added to 50 ml of drug solution
in RPMI 1640 (final hematocrit 2%). The 96-well mi-
crotiter plates were incubated for 48 h at 37 °C in an
atmosphere of 1% O2 and 3% CO2 in balanced N2.
After the incubation period, 20 ml of the parasite sus-
pension was added to 100 ml of Malstat™ reagent
(Flow Incorporated, USA) and incubated at RT for 15
min before adding 20 ml of freshly made 1:1 NBT/PES-
mixture (2 and 0.2 mg/ml, respectively) to each well.
The plates were reincubated for 20 min at RT in the
dark and subsequently read at 650 nm. All compounds
were tested twice in triplicate.
2.3.5. Giardia duodenalis
G. duodenalis trophozoites were cultivated in Dia-
mondı́s modified TYI-S-33 medium (Keister, 1983) sup-
plemented with 10% heat inactivated fetal calf serum
(FCS). The in vitro assay was performed as described
for the Alamar Blue® assay for trypanosomes by Raez
et al. (1997) with modifications for G. duodenalis WB
strain (isolated 1982 from a human in Afghanistan).
Briefly, 200 ml of a trophozoite suspension were inocu-
lated into 96-well microtiter plates (Costar, USA) at a
density of 4× 105 trophozoites per ml culture medium.
The trophozoites were incubated in the presence of
serial 3-fold dilutions of extracts for 72 h at 37 °C.
Wells without drug served as controls. Minimum in-
hibitory concentration (MIC) was determined micro-
scopically after 70 h of incubation (the lowest drug
concentration at which no trophozoite with normal
morphology could be observed). Ten microliter Alamar
Blue® were added to each well and after 2 h of incuba-
tion the fluorescence determined using a fluorometer
(Cytofluor, Millipore; excitation wavelength at 530 nm,
emission at 590 nm). IC50 values were calculated by
linear interpolation selecting values above and below
the 50% mark.
2.3.6. Dopamin D2 receptor binding assay
Two concentrations of extracts (100 and 10 mg/ml)
were tested in the dopamine receptor binding assay.
The affinity of the extracts to the dopamine receptor
was assessed according to Berger (1998).
2.3.7. h-Amylase assay
Plant extracts (1, 3, 6 mg/ml in 5 ml solvent) were
mixed with 45 ml of amylase reagent (ET-G7 PNP 1.0
mmol/l, magnesium chloride 10 mmol/l, sodium chlo-
46 A. Ankli et al. / Journal of Ethnopharmacology 79 (2002) 43–52
ride 50 mmol/l, a-glucosidase 25.000 U/l, buffer pH 7.0,
sodium azide 0.05%) obtained from Sigma Diagnostics
and incubated at 37 °C for 2– 10 min. The absorbance
was recorded at 405 nm versus water as a reference.
The incubation was continued and the absorbance was
read after exactly 1 and 2 min. The amylase activity
was calculated according to Pierre et al. (1976).
3. Results
All polar and non-polar extracts of the 48 plants
were screened for cytotoxic activity against KB cells,
Bacillus cereus, Escherichia coli, Candida and in the
NF-kB test. Additionally, extracts were evaluated in
selected test systems, which are of direct relevance to
the indigenous uses of the species (Table 2). The mi-
croorganisms chosen cause gastrointestinal problems
and/or dermatological illnesses, and generally are the
causal agents of these conditions (Table 1). The ex-
tracts, which showed noteworthy activities, are listed in
Tables 3 and 4.
3.1. Acti6e plants for gastrointestinal problems
Gram-positive and Gram-negative bacteria as well as
protozoa (G. duodenalis) were used to determine activi-
ties of species used for treating gastrointestinal disor-
ders (Table 3). Six species showed at least some activity
against G. duodenalis with IC50 values less than 100
mg/ml, three of them with MIC values less than 100
mg/ml. The most active extract was the non-polar ex-
tract A of Crossopetalum gaumeri (MIC 6.3 mg/ml),
whereas the polar extract B showed very weak antipro-
tozoal activity. The non-polar and polar extracts of
Psidium sartorianum, Piscidia piscipula, Bidens squar-
rosa and Casimiroa tetrameria and the non-polar frac-
tion of Bauhinia di6aricata showed weak activity with
IC50 values between 20 and 90 mg/ml. Several plants
were active against H. pylori (Table 3). The non-polar
and polar extracts of P. piscipula showed the highest
activities (with MIC values of 0.7 and 3 mg/ml, respec-
tively). The non-polar extracts of C. tetrameria (MIC, 3
mg/ml) and Jatropha gaumeri (MIC, 5 mg/ml) were also
active against H. pylori. Other active extracts were
Dorstenia contrajer6a (A and B) and the polar extracts
of P. sartorianum, Microgramma nitida, Chrysophyllum
mexicanum.
The non-polar root extract of J. gaumeri was found
to be the most active plant tested against B. cereus.
Extract A of C. gaumeri as well as extracts A and B of
P. sartorianum showed weak activities against this
strain. The ethanol extract of C. tetrameria weakly
inhibited the growth of S. aureus. There was no signifi-
cant activity against C. jejuni. None of the tested plant
species used for gastrointestinal problems showed activ-
ity against E. coli or Y. enterocolitica.
3.2. Acti6e plants for dermatological conditions
Selected plants, documented as medicines for derma-
tological conditions, were evaluated for anti-inflamma-
tory, antibacterial and antifungal activities (Table 4).
Cytotoxicity of the plant extracts was evaluated using
KB and the HeLa cell line (Table 4).
The non-polar fraction of C. gaumeri showed the
most potent effect in the NF-kB test with an inhibitory
concentration of 25 mg/ml. The non-polar extracts of
Diospyros anisandra and J. gaumeri inhibited NF-kB
activation (Table 4). These effects are probably due to
the potent cytotoxicity of the extracts against KB cells.
The non-polar fractions of Dalea carthagenensis and
Luehea speciosa elicited inhibition of NF-kB binding at
150 and 100 mg/ml, respectively. The former showed
cytotoxic activity against KB cells (IC50 31 mg/ml),
whereas no such effect could be detected for L.
speciosa.
Several extracts showed antibacterial effects against
S. epidermidis and E. coli (Table 4). The most active
extract against the former strain was extract B of
Aechmea bracteata. The non-polar fruit extract of
Morinda yucatanensis was active against S. epidermidis.
Other active extracts included: the non-polar one of C.
gaumeri and Casearia corymbosa as well as both ex-
tracts of P. sartorianum.
3.3. Antimalaria acti6ities
Plants used against fever and/or pain were screened
in vitro for antimalarial activity against Plasmodium
falciparum (Table 2). The non-polar extract of Cestrum
nocturnum showed some antimalarial activity (IC50
172.40 (2.80) mg/ml against chloroquine-sensitive strain
of P. falciparum HB3; IC50 283.31 (0.48) mg/ml against
chloroquine-resistant clone K1) without exhibiting
overt cytotoxicity (IC50 ] 50 mg/ml). The results of the
positive control chloroquine are the following ones:
IC50 7.96 (0.03) ng HB3 and IC50 193.96 (0.81) ng K1.
The non-polar extract of C. corymbosa showed weak
antimalarial activity (IC50 441.40 (2.26) mg/ml HB3;
IC50 494.49 (3.07) mg/ml K1). All other extracts (Ehretia
tinifolia, Caesalpinia gaumeri and Manilkara zapota)
showed any activity (IC50 ] 500 mg/ml).
3.4. D2 -receptor binding affinities
Dopamine-agonists are used in the treatment of the
premenstrual syndrome (PMS) (Steiner, 1997). For the
dopamine D2 receptor binding assay, no significant
activity was observed, however, the non-polar extracts
of Sal6ia micrantha and Aristolochia maxima showed
weak receptor affinity.
 A. Ankli et al. / Journal of Ethnopharmacology 79 (2002) 43–52 47

Table 2
Ethnomedical data on plants studied and chosen test systems

Family Plant name (AANKc voucher) Based on traditional use Tested for

Group of use Plant part

Acanthaceae Ruellia nudiflora (Engelm. and Gray) Urb. (115) UR ap 1–7, 14

Annonaceae Malmea depressa (Baill.) R. E. Fr. (161) UR rt 1–7, 14
Apocynaceae Tabernaemontana amygdalifolia Jacq. (190) DER lv 1–5
Araceae Anthurium schlechtendalii Kunth ssp. schlechtendalii (243) FEM lv 1–5, 15
Aristolochiaceae Aristolochia maxima Jacq. (350) GI, FEM rt 1–10, 15
Asteraceae Verbesina gigantea Jacq. (288) RES lv 1–5
Bidens squarrosa Less. (121) GI ap 1–12
Basellaceae Anredera 6esicaria C. F Gaertner (196) DER lv, tu 1–7
Bignoniaceae Parmentiera millspaughiana (L.) Williams (135) UR lv 1–5, 14
Parmentiera aculeata (Kunth) Seem. (096) UR lv, rt 1–5
Bombacaceae Pseudobombax ellipticum (Kunth) Dugand (275) RES lv 1–5
Boraginaceae Ehretia tinifolia L. (021) RES, UR, NEU lv 1–5, 13
Bromeliaceae Aechmea bracteata var. bracteata Griseb. (167) DER lv 1–7
Cactaceae Hylocereus undatus (L.) Britton & Rose (427) GI, UR lv 1–12, 14
Caesalpiniaceae Bauhinia di6aricata L. (007) RES, UR, GI lv 1–10, 14
Caesalpinia gaumeri Greenman (155) NEU lv 1–7, 13
Celastraceae Crossopetalum gaumeri (Loes.) Lundell (038) GI, DER lv 1–12
Cucurbitaceae Iber6illea millspaughii (Cogn.) C. Jeffrey (094) DER tu 1–7
Ebenaceae Diospyros anisandra Blake (134) DER lv 1–5
Diospyros cuneata Standl. (341) DER lv 1–5
Euphorbiaceae Croton reflexifolius Kunth (143) DER lv 1–5
Jatropha gaumeri Greenman (419) GI, DER rt 1–12
Flacourtiaceae Casearia corymbosa Jacq. (150) DER,NEU lv 1–7, 13
Lamiaceae Sal6ia micrantha Desf. (025) DER, FEM ap 1–7, 15
Meliaceae Cedrela mexicana L. (301) RES lv 1–5
Moraceae Brosimum alicastrum Sw. (092) RES lv 1–5
Dorstenia contrajer6a L. (330) GI, FEM rh 1–12, 15
Myrtaceae Psidium sartorianum (Berg) Nied. (211) DER, GI lv 1–10
Nyctaginaceae Neea psychotrioides F. D. Sm. (274) DER lv 1–7
Pisonia aculeata L. (154) FEM lv 1–7, 15
Papilionaceae Dalea carthagenensis var. barbata (Oerst.) Barneby (125) DER lv 1–5
Piscidia piscipula (L.) Sarg. (123) GI, RES lv 1–10
Phytolaccaceae Phytolacca icosandra Sims (388) DER fr 1–5
Ri6ina humilis L. (089) DER ap 1–5
Polygonaceae Neomillspaughia emarginata S. F. Blake (203) DER, RES lv 1–5
Polypodiaceae Microgramma nitida (J. Sm.) A. Reed Sm. (183) GI wp 1–10
Rubiaceae Borreria 6erticillata G. Meyer (276) DER ap 1–7
Morinda yucatanensis Greenman (113) DER fr 1–7
Rutaceae Casimiroa tetrameria Millsp. (049) GI, NEU lv 1–12
Sapotaceae Chrysophyllum mexicanum Brandegee (386) GI rt 1–10
Manilkara zapota (L.) Royen, Achras zapota L. (234) GI, NEU ba 1–10, 13
Selaginellaceae Selaginella longispicata Underw. (214) UR, RES ap 1–5
Simaroubaceae Al6aradoa amorphoides Liebm. (136) DER lv 1–7
Solanaceae Cestrum nocturnum L. (050) DER, NEU lv 1–5, 13
Solanum erianthum G. Don f. (334) DER lv 1–5
Solanum nigrum L. (267) DER lv 1–5
Sterculiaceae Helicteres baruensis Jacq. (176) FEM lv 1–5, 15
Tiliaceae Luehea speciosa Willd. (347) DER lv 1–7, 11, 12

UR, urological problems including ‘diabetes’ (based on traditional knowledge), DER, dermatological conditions including injuries caused by
venomous animals; FEM, women’s medicines; GI, gastrointestinal disorders; NEU, fever and/or pain; RES, respiratory illnesses; ap, aerial parts;
ba, bark; fr, fruits; lv, leaves; rh, rhizome; rt, root; tu, tuber; wh, whole plant; 1, B. cereus; 2, E. coli; 3, C. albicans; 4, KB-cell line; 5, NF-kB;
6, P. aeruginosa; 7, S. epidermidis; 8, H. pylori; 9, C. jejuni; 10, G. duodenalis; 11, S. aureus; 12, Y. enterocolitica; 13, P. falciparum; 14, a-amylase;
15, D2-receptor binding assay.

3.5. Hypoglycemic effects diabetes were tested for their ability to inhibit a-amy-
lase (Table 2). The inhibition of the enzyme leads to a
The polar extracts of five plant species, used against reduced splitting of food based poly- and disaccharides
48 A. Ankli et al. / Journal of Ethnopharmacology 79 (2002) 43–52

in the colon resulting in a delayed resorption (Keller
and Berger, 1983). B. di6aricata was the most potent
inhibitor of the enzyme (0 U/l for extract 1 [1 mg/ml]),
2 [3 mg/ml]) and 3 [6 mg/ml]) followed by Hylocereus
undatus (255 U/l for extract 1; 0 U/l for extracts 2 and
3, respectively). Polyphenols, known to interfere with
enzymes, do not occur very frequently in these genera
or families (Hegnauer, 1989; Hegnauer and Hegnauer,
1994). So the inhibition of the enzyme seems not to be
due polyphenols. The following species were inactive,
Ruellia nudifora, Malmea depressa, Parmentiera
millspaughiana. The negative control resulted in a value
of 620 U/l, the control of methanol gave 548 U/l.
3.6. Other acti6ities
Some extracts showed activity in assays, which are
not directly related to an indigenous use. The antibiotic
effects against S. epidermidis and B. cereus of the
non-polar fractions of Caesalpinia gaumeri used for
pain of the body and headache are particularly note-
worthy (2 mm using a concentration of 200 mg).
4. Discussion
The species evaluated in this paper were selected
because of their cultural importance to the Yucatec
Maya and their use(s) for specific syndromes. In this
section the relevance of these findings to interpreting
such indigenous uses is discussed using selected
examples.
The roots of Crossopetalum gaumeri are used orally
for diarrhea and snake-bites and topically to prevent
inflammation after a snake-bite. In a detailed phyto-
chemical study of C. gaumeri, the non-polar fraction

Table 3
Screening of plant species used for gastrointestinal disorders by the Yucatec Maya

Plant name Extract KB Giardia duodenalis Helicobacter pylori Bacillus cereus Staphylococcus
aureus
IC50 (mg/ml) MIC (mg/ml) IC50 (mg/ml) MIC (mg/ml) 600 mg (mm) 200 mg (mm) 600 mg (mm) 200 mg (mm)

Bauhinia A – – 51 nt – – – nt
di6aricata
Bidens B – – 70 nt – – – –
squarrosa
Casimiroa A – – 72 3 4 – – C: 1
tetrameria
Chrysophyllum B – – – nt 3 – – nt
mexicanum
Crossopetalum A 0.7 6.3 2.1 nt – 1 2 –
gaumeri
B 10.2 – 90 nt – – – nt
Dorstenia A – – – 10 4 – – –
contrajer6a
B – – – nt 3 – – nt
Jatropha A 7.8 – – 5 8 2 3 –
gaumeri
Microgramma B – – – nt 3 – – nt
nitida
Piscidia A – 41 27 0.7 11 – – nt
piscipula
B – – 70 3 12 – – nt
Psidium A – 69.2 51 nt – B1 1 nt
sartorianum
B – – 65 nt 4 – 1 nt
Metronidazole 8.5 5
Ornidazole 1.1 0.5
Tetracycline 20 (10 mg)
Ampicillin 23 (100 mg)
Kanamycin 19 (40 mg)
Streptomycin 19 (50 mg)
Chloramphenic 22 (25 mg) 8 (10 mg) 8 (10 mg)
ol
Ciprofloxacin 1 (0.01)

KB, cytotoxicity, (–), IC50]50 mg/ml) I, G. duodenalis, (–), IC50]100 mg/ml; H. pylori, (–), 52 mm; antibacterial activity was measured as
inhibition zone in (mm) (–), no activity; A, non-polar extract; B, polar extract; C, ethanol extract; nt, not tested.
 A. Ankli et al. / Journal of Ethnopharmacology 000 (2002) 000–000 49

Table 4
Screening of plant species used for dermatological conditions by the Yucatec Maya

Plant name Extract KB, IC50 NF-kB Staphylococcus epidermidis Escherichia coli Candida albicans
(mg/ml) (mg/ml)
200 mg (mm) 600 mg (mm) 600 mg (mm) 200 mg (mm) 600 mg (mm)

Aechmea bracteata B – – 3 5 – – –
Al6aradoa A 10 – – – – – –
amorphoides
B 14 – – – – – –
Casearia A – – 1 1 – – –
corymbosa
Crossopetalum A 0.7 25 2 3 – – –
gaumeri
B 10.2 * – – – – –
Croton reflexifolius A 39 – nt nt – – –
Dalea A 31 150 – – – – –
carthagenensis
Diospyros A 14 100 nt nt 1 – 2
anisandra
Diospyros cuneata A – 75 nt nt – – 1
Jatropha gaumeri A 7.8 * – – – – –
B – – – – – 1 2
Luehea speciosa A – 100 – – – – –
Morinda A – – 2 4 – – –
yucatanensis
Psidium A – – – 1 – – –
sartorianum
B – – 1.5 2 – – –
Podophyllotoxin 0.006
Parthenolide 10 mM
PDTL 100 mM
Chloramphenicol 8 (10 mg) 8 (10 mg)
Tetracycline 6 (10 mg)
Miconazole 5 (1 mg) 5 (1 mg)

KB, cytotoxicity; (–), IC50 ]50 mg/ml; NF-kB (HeLa cell line); (–), \150 mg/ml; *, cytotoxic at 100 mg/ml during the period of the test.
Antimicrobial activity was measured as inhibition zone in (mm); nt, not tested.

showed the presence of terpenoids, whereas the extract
B consisted of several cardenolides (Ankli et al., 1999,
2000). The study indicated that the terpenoids possess
antibacterial activity that may be relevant to the plant’s
traditional use as a treatment for diarrhea. The potent
antiprotozoal effect against G. duodenalis is probably
due to the high non-specific cytotoxicity of the extracts
(Table 3). The cytotoxicity of the polar and non-polar
extracts may also be responsible for the potent in-
hibitory activity against NF-kB (Table 4).
The leaves of P. piscipula are used as a medicine for
treating gastrointestinal disorders (especially diarrhea
and cramps) and for cough. The remarkable activity of
this species against H. pylori and to some extent against
G. duodenalis may be a reason why Yucatec Maya
value this plant for treating gastrointestinal problems.
In a study by Cáceres et al. (1991), P. piscipula was
shown to have antimycotic effects. Another species,
Piscidia erythrina L., has been widely investigated yield-
ing spasmolytic isoflavones (Della Loggia et al., 1988).
Among other ethnic groups, P. piscipula is used as a
fish poison (Acevedo-Rodrı́guez, 1990).
B. di6aricata is used for a variety of illnesses such as
gastrointestinal problems, but more frequently for dia-
betes and respiratory problems. Its activity against G.
duodenalis may be one of the reasons for its indigenous
use. In a separate study the leaf extract of Bauhinia
purpurea L. was reported to have significant antidi-
arrheal activity in vivo (Mukherjee et al., 1998). A
possible hypoglycemic activity was also found in our
a-amylase test which supports the reported hypo-
glycemic effects in a previous study (Roman et al.,
1992).
The roots of J. gaumeri are used for treating diarrhea
and the resin is used as a medicine for herpes labialis.
The therapeutic application of this plant as an anti-di-
arrhetic may be the result of its antibacterial activity as
observed in this study against B. cereus. A variety of
pharmacological effects are reported in the literature
concerning Jatropha spp., including antimicrobial ef-
50 A. Ankli et al. / Journal of Ethnopharmacology 79 (2002) 43–52
fects (Odebiyi, 1980). The potent cytotocitity against
the KB-cells is probably due to the cytotoxic phorbol
esters, which are known for many taxa of the Euphor-
biaceae (Hänsel et al., 1999).
P. sartorianum is employed internally (diarrhea) and
externally (mostly for measles or any kind of pimples).
The antibacterial activities against H. pylori, B. cereus
and S. epidermidis observed in this study might be of
interest in light of its traditional use as a treatment for
diarrhea and skin problems. No pharmacological data
for P. sartorianum are available, but P. guaja6a L. was
shown to have antibacterial effects, especially against
the enterobacterium Shigella.
C. tetrameria is used as a medicine for treating
gastrointestinal problems including diarrhea as men-
tioned in the Ethno-Botany of the Maya (Roys, 1976
[orig. 1931]). The antiprotozoal effect as well as the
antibacterial activities against H. pylori and S. aureus
may be of relevance to the internal use. The antidi-
arrheal and antispasmodic effects are currently investi-
gated (Heneka, 2000).
The two Diospyros species, D. anisandra and D.
cuneata, were reported to be ‘strong’ medicines. They
are used topically for treating dermatological problems
(pimples, scabies, inflammation). The antibiotic and the
antifungal activities, shown in the assays, are probably
responsible for the growth inhibition of the bacterial
and fungal secondary infection of scabies and pimples.
The NF-kB inhibiting activity, probably a result of the
non-specific cytotoxicity of the extract, could be the
reason of its anti-inflammatory use among the Mayas.
These results are strengthened by reports of antimicro-
bial activity of Diospyros lycioides Desf. (Li et al.,
1998), as well as in vivo anti-inflammatory activity of
Diospyros leucomelas Poir (Recio et al., 1995).
Various species of the genus Croton are used in the
medical system of the Yucatec Maya for dermatological
problems but also for fever and respiratory illnesses.
The resin of Croton reflexifolius is used for pimples in
the mouth (herpes) and eye problems. The possible
occurrence of phorbol esters in this species could be a
reason for its medicinal use as an antiviral drug (Hänsel
et al., 1999). The application of the resin into the eyes
could have severe side effects due to a high level of
cytotoxicity (Table 4).
L. speciosa is an effective inhibitor of the transcrip-
tion factor NF-kB. The Yucatec Maya value this plant
for treating skin diseases and toothache, and apply the
medicine in the form of plasters. The inhibitory effect
on NF-kB may explain the use of this plant. The weak
antibacterial activity against Y. enterocolitica can not
substantiate the traditional use of the plant. To our
knowledge no pharmacological or phytochemical data
have been published yet concerning this plant.
C. nocturnum, applied for children with night fever
and cold bodies, shows a weak effect against P. falci-
parum. Further testing of fractions of this plant would
be of interest. Phytochemically the species is well inves-
tigated but no published data has been found for this
antimalarial effect.
5. Conclusion
One goal of this evaluation is to better understand
the use of plants by the Yucatec Maya. In this paper we
show some correlations between uses of the medicinal
plants and relevant biological activities. Other indige-
nous uses currently cannot be explained in a bio-scien-
tific manner because the bioassays applied are not
appropriate. In other cases the symbolic aspects might
be more important to the Maya. We hope that the
combination of a detailed documentation of ethnomed-
ical use and the study of selected species in relevant
bioassays correlating with their application may lead to
a better understanding of the ethnopharmacopoeia of
the Yucatec Maya (and other indigenous groups).
Phytochemical and further pharmacological studies
are important tasks for the future in order to better
understand the effects of these important pharmaceuti-
cal resources. Organizations like World Health Organi-
zation (WHO) and TRAMIL (Central America)
encourage the use of remedies provided that they are
safe and that some scientific evidence on their biological
and pharmacological effects exists. Hopefully this study
contributes to a selection of the most appropriate spe-
cies of the indigenous medicine of the Yucatec Maya.
Herbal medicines are a valuable and readily available
resource for primary health care and complementary
health care systems.
Acknowledgements
The authors wish to thank all persons who have
helped in the field study and especially the healers,
midwives and the inhabitants of Chikindzonot, Ekpedz
and Xcocmil, Yucatan, for their collaboration, for their
friendship and hospitality. The botanical identification
at CICY and MEXU (National Herbarium of Mexico)
was performed in collaboration with the numerous
specialists of these institutions. Particularly we would
like to thank Dr I. Olmsted, J. Granados, P. Simá, J.C.
Trejo, Dr R. Durán of CICY as well as O. Tellez, Dr
R. Lira, Dr J. Villaseñor and Dr M. Sousa of MEXU.
This research owes a lot to the help of Dr J. Heilmann
(Zürich), Dr J. Orjala (Davis), Dr B. Frei Haller
(Zernez), Professor Dr W. Schaffner (Basel) and Profes-
sor Dr H. Rimpler (Freiburg). Financial support by
Swiss Agency for Development and Cooperation (SDC,
Berne, Switzerland) and the Swiss Academy of Natural
Sciences (SANW) is gratefully acknowledged. We are
 A. Ankli et al. / Journal of Ethnopharmacology 79 (2002) 43–52
grateful to Dr Paul Bremner (ULSOP) for carefully
checking the MS.
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