Cuaderno 7 Cytotoxic Screening of Tropical Plants Using Brine Shrimp Lethality Test

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Cytotoxic Screening of Tropical Plants Using Brine Shrimp Lethality Test

Abstract

In recent years the study of plants has been the subject of renewed interest as a source of natural

products with medicinal value. This study aims at the development and application of bioassays

in order to detect potential sources of cytotoxic, and antitumour compounds from endemic and

native plants from Puerto Rico. For this purpose, as a strategy for primary evaluation, the brine

shrimp lethality test was chosen. The native and endemic species selected for the study were:

Canella winterana, Croton Discolor, Goetzea elegans, Guaiacum officinale, Pimenta racemosa ,

Simarouba tulae and Thouinia striata. These species were dried and extracted with a mixture of

CH2Cl2-MeOH (1:1). The crude extract was analyzed by 1H-NMR spectroscopy and afterwards

it was suspended in water and extracted with solvents of different polarities. Fifteen of the

twenty-eight extracts were active exhibited a LC50 ≤ 200 µg/mL. The most promising activity

was displayed by extracts of Simarouba tulae and Guaiacum officinale with lethality values of 2

and 4, respectively.

Key words: Brine shrimp lethality test, endemic and native plants, cytotoxic activity

Introduction

Natural products are organic compounds that are present in living organisms, animals and

plants. These are divided in two groups: primary and secondary metabolites. Primary metabolites

are those compounds which occur in all cells and play a central role in the metabolism and

reproduction of those cells. However, secondary metabolites such as polyketides, fatty acids,

terpenoids, phenylpropanoids and alkaloids are characteristic of a limited range of species and do

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not have a direct role in plant growth and development. Secondary metabolites have evolved as

defense agents against predators and the surrounding environment.1 In addition, these

compounds exert their biological effect within the organism that is responsible for their

production and can affect functions in other organisms. For example, some plant metabolites can

suppress cell division, a trait that makes them useful in treating cancer in human patients.

Moreover, these plant products also have a wide variety of different uses in health human and

industrial applications. It has been estimated that over 40% of medicines have their origins in

these secondary metabolites. 2

Due to the wide application at pharmacological level, medicinal plants have gain

importance worldwide. The work in this area begins with an assessment of the biological activity

of crude (total lipid) extracts from plants. These preliminary results provide further access for an

exhaustive analysis of its principal constituents. The lack of information related to the specific

compounds and its biological effect in plants and other organisms, provides alternative lines of

investigations, drug discoveries, and chemical modifications.

Endemic plants are species unique to a particular geographic location whereas native

plants are species belonging to a region if its presence in this region is the result of only natural

resources. Endemic and native Caribbean plants have been less studied that those from Africa,

India and Europe thus, a preliminary biological screening and subsequent isolation of the

secondary metabolites from these plants would be a great contribution to document and expand

the chemotaxonomic knowledge of these species.

During the last two decades, there have been reports of the biological screenings of the

molluscicidal (2002)3, antimycobacterial (1998 and 2001)4,5, antiplasmodial (2001)5, and

antibacterial (1996, 2006) 6,7 activities of some tropical plants including Musaceae, Labiatae,

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Rutaceae, Myrtaceae, Apiaceae, Rubiaceae, and Malvaceae from Puerto Rico. Additionally, two

studies about the cytotoxic activity of some Puerto Rican plants has been documented. The first

was reported by Guerrero and Robledo in 1993.8 In this work, six crude extracts of the

Euphorbiaceae, Solanaceae, Myrsinaceae, Polygonaceae, and Polygalaceae families showed

LC50 values below 200 µg/mL in the Artemia Salina Test, indicating the potential presence of

bioactive compounds. No isolation of bioactive compounds was reported later. In the second

report by Chavez et al. tested the dichloromethane portion of the ethanol extract in a

concentration of 1000 µg/mL using the brine shrimp assay.9 The extracts with a LC50 ≤ 1000

µg/mL were further tested against Hela and CHO cells. The extracts from Annona glabra,

Simarouba tulae, Tithonia diversifolia, Dendropanax arboreous, Piper jacquemontanium,

Annona montana, Polygala hecatantha were active in both assays.

These previous reports document the biological importance of secondary metabolites

from Puerto Rican plants as potential antitumour agents. The isolation and characterization of

the metabolites responsible for the biological activities is needed, as well as the study of some

other species from families that have produced bioactive compounds.

In the present study, seven species of native and endemic plants present in Puerto Rico

were studied. These were: Canella winterana, Pimenta racemosa, Guaiacum officinale, Croton

discolor, Gotzea elegans, Thouinia striata and Simarouba tulae,

Canella winterana is a tree found naturally in Florida and the Caribbean. Some important

constituents of this plant are monoterpenes like canellal (1)10, eugenol (2)11, eucalyptol (3)11, and

drimane sesquiterpenoids like 9α-hydrocinnamolide (4)12 (Figure 1). The sesquiterpenoids

exhibited phototoxic activity in a Lemna minor bioassay13. The drimane sesquiterpenoids

isolated from other species of the Canellaceae family are known for their broad antifeedant,

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antifungal, molluscicidal, and cytotoxic activities, however, the bioactivity of the Canella

winterana secondary metabolites have not been reported.

CHOH3C

H

OH

O H

canellal (1)

OH

OCH3

eugenol (2)

O

eucalyptol (3)

CHOH3C

H

OH

O H

9-hydrocinnamolide (4)

Figure 1. Some Compounds Isolated from Canella winterana.

Pimenta racemosa is a native tree to the islands of the Caribbean, that is used in folk

medicine for the treatment of different diseases, as tooth ache, abdominal pain, fever, influenza,

rheumatism and pneumonia. This three is well known for the essential oil present in leaves and

stems. This oil is composed of eugenol (5) and terpenes, such as α-terpineol (6) among other of

its respective derivatives (Figure 2). In 2001, García et al. reported the isolation of the

triterpene lupeol (8) and the anti-inflammatory effect of the methanol extract of this species. 14

In 2004, Saenz et al reported the antibacterial and antinociceptive activity of the essential oils

and the aqueous extract of the plant. 15

eugenol (5)

OH

OCH3

OH

terpineol (6)

HO

lupeol (7)

Figure 2. Some Compounds Isolated from Pimenta racemosa

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Guaiacum officinale is a tree common in the Antilles and tropical zones of America. The

bark and the resin are used to treat rheumatism, tooth ache and skin disorders. The fruits and

leaves contain triterpene saponins such as guaianin and officigenin (8).16 The resin contains

lignans such as α-guaiaconic acid (9). (Figure 3).17

HO

officigenin (8)

COOH

HO HO

COOH

O

O

O

H3C CH3

OH

H3CO

HO

OCH3

guaiaconic acid (9)

Figure 3. Some Compounds Isolated from Guaiacum officinale

Croton discolor is a shrub of the floral region of Puerto Rico and the Virgen Islands. Folk

medicine has used the tea from the leaves to treat rheumatism. Young leaves and branch tips

have been used to treat coughs. Two aporphine alkaloids, crotonosine (10) and discolorine (11)

had been isolated from this plant (Figure 4).18

NH

HO

H3CO

O

crotonosine (10)

NCH3

HO

H3CO

HO

discolorine (11)

Figure 4. Alkaloids Isolated from Croton discolor

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Simarouba tulae, is an endemic tree of Puerto Rico belonging to the Simaroubaceae

family. Some species have been used in folk medicine as fever reducer, tonic and antimalarial. In

the last years, its possible therapeutic purposes have increased because of its antimalarial, anti-

inflammatory, antileukemic, antifeedant and antiviral activities.19 In 1997, Chavez reported the

cytotoxic activity of Simarouba tulae against Hela and CHO cells.9 No studies about the chemical

constituents of this species had been reported.

Finally, Thouinia striata (Sapindaceae) and Gotzea elegans (Solanaceae) are also

endemic plants of Puerto Rico. To our knowledge, no references about the secondary metabolites

and biological activity of these species had been reported.

An efficient, rapid, and general test for evaluate the cytotoxic activity of extracts and

compounds from plants is the brine shrimp lethality bioassay. This bioassay has a good

correlation with cytotoxic activity in some human solid tumors and pesticide activity. 20,21 We

decided to study the cytotoxic activity of selected tropical plants from Puerto Rico using the

brine shrimp lethality test, and in the future use a bioassay-guide fractionation to guide the

purification, isolation and subsequent characterization of the active constituents.

Materials and Methods General Experimental Procedures

The NMR spectra were recorded on an Anasazi 60 MHz FT-NMR or in a Bruker (400

MHz). The chemical shifts are given in δ (ppm). Samples were dissolved in CDCl3 or (CD3)2SO

and tetramethyl silane (TMS) was used as internal reference. The alive shrimp in the Artemia

salina bioassay were counted by inspection of the well with the aid of a Stereo zoom microscope

(10X widefield eyepieces 45 degree inclined body, 7X to 45X magnification). Silica Gel (70-

230 mesh, 60A) was used for column chromatography. The deuterated solvents (CDCl3 and

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(CD3)2SO), TMS, berberine chloride, hexane, dichloromethane and methanol were purchased

from Sigma Aldrich. Celite (521), chloroform and ethyl acetate were purchased from Thermo

Fisher Scientific. Brine shrimp eggs were acquired in a local pet shop (San Juan, Puerto Rico)

and the yeast was purchased in a local grocery store (Caguas, Puerto Rico).

Plant Material

The leaves of Canella winterana, Thouinia striata, Croton discolor and Guaiacum

officinale were collected at the dry forest in Guánica, PR in February 2008. The leaves of

Pimenta racemosa were collected in Cidra, PR in February 2008. The leaves of Goetzea elegans

were colleted at the Botanical Garden of the University of Puerto Rico in Río Piedras. The

leaves of Simarouba tulae were collected in Patillas, PR. The plants were identified by professor

Augusto Carvajal of the Department of Biology at the University of Puerto Rico at Cayey. A

voucher specimen was reserved to be deposited in the herbarium of the Botanical Garden of the

University of Puerto Rico.

Extraction and Isolation

The air-dried leaves of Canella winterana (283.80 g) were crushed in a blender with

three portions of 1L of CH2Cl2/CH3OH (1:1, v/v). The solid debris was removed from the

combined extracts by a Celite filtration and the extract was concentrated under vacuum to yield

65.09 g of crude. The extract was suspended in 500 mL of water and extracted with solvents of

increasing polarity, hexane (3 x 150 mL), chloroform (3 x 200 mL) and ethyl acetate (3 x 150

mL). Each of the extracts was concentrated to dryness by rotoevaporation. The weights of the

extracts were hexane 25.01 g, chloroform 15.76 g and ethyl acetate 2.10 g.

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The air-dried leaves of Croton discolor (46.59 g) were crushed in a blender with three

portions of 500 mL of CH2Cl2/CH3OH (1:1, v/v). The solid debris was removed from the




mL). Each of the extracts was concentrated to dryness by rotoevaporation. The weights of the

extracts were hexane 0.75 g, chloroform 3.09 g and ethyl acetate 0.19 g.

The air-dried leaves of Goetzea elegans (270.11 g) were crushed in a blender with three

portions of 1L of CH2Cl2/CH3OH (1:1, v/v). The solid debris was removed from the combined

extracts by a Celite filtration and the extract was concentrated under vacuum to yield 25.10 g of

crude. The extract was suspended in 500 mL of water and extracted with solvents of increasing

polarity, hexane (3 x 150 mL), chloroform (3 x 150 mL) and ethyl acetate (3 x 100 mL). After

removal of the solvents, the weights of the dry extracts were hexane 5.88 g, chloroform 1.06 g

and ethyl acetate 0.52 g.

The air-dried leaves of Guaiacum officinale (270.14 g) were crushed in a blender with





mL). After removal of the solvents, the weights of the dry extracts were hexane 5.05 g,

chloroform 4.96 g and ethyl acetate 2.06 g.

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The air-dried leaves of Pimenta racemosa (279.66 g) were crushed in a blender with


combined extracts by a Celite filtration and the extract was concentrated in vacuum to yield





The air-dried leaves of Simarouba tulae (65.71 g) were crushed in a blender with three

portions of 500 mL of CH2Cl2/CH3OH (1:1, v/v). The solid debris was removed from the



increasing polarity, hexane (3 x 100 mL), chloroform (3 x 100 mL) and ethyl acetate (3x 100



The air-dried leaves of Thouinia striata (106.39 g) were crushed in a blender with three

portions of 1L of CH2Cl2/CH3OH (1:1, v/v). The solid debris was removed from the combined

extracts by a Celite filtration and the extract was concentrated under vacuum to yield 15.84 g of

crude. The extract was suspended in 500 mL of water and extracted with solvents of increasing

polarity, hexane (3 x 150 mL), dichloromethane (3 x 100 mL) and ethyl acetate (3 x 100 mL).

Each of the extracts was concentrated to dryness by rotoevaporation. The weights of the extracts

were hexane 3.61 g, dichloromethane 6.03 g and ethyl acetate 0.13 g.

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Brine shrimp lethality bioassay

Brine shrimp lethality bioassay was performed to assess the cytotoxicity of the crude and

extracts of the plants.20 The bioassay was performed as reported earlier with some minor

modifications. Brine shrimp eggs were hatched in artificial seawater (0.5 g eggs per liter) at the

dark portion of a divided chamber. The artificial seawater was prepared using sea salt 30 g/L,

containing 0.006 g of yeast as food source. After approximately 48 h the phototropic nauplii

move through a hole in the division to the portion of the camber kept under continuous light.

The nauplii were collected with a pipette and concentrated in a beaker. The concentration of the

nauplii was adjusted adding seawater to the beaker until approximately 10-15 nauplii were found

in 100 µL of solution measured with an automatic micropipette.

The sample (0.002g) to be tested was dissolved in 100 µL of DMSO and diluted with

seawater (1900 µL) to a concentration of 1 mg/mL. Berberine chloride was used as a positive

control and prepared according to the sample. The blank solution (negative control) was prepared

diluting 50 µL of DMSO in 950 µL of seawater. The bioassay was conducted in a 96 microwell

plate. Several solutions of each sample ranging from 1.95 µg/mL to 500 µg/mL were prepared

in triplicate. To each solution 100 µL of seawater containing 10-15 nauplii were added. The

microwell plate was incubated at room temperature during 24 h. with constant lighting.

After 24 h the dead nauplii were counted with the aid of a microscope and the lethal

concentration (LC50 value) was calculated by probit analysis. Larvae were considered dead if

they did not move during the observation. The LC50 value was obtained by regression analysis of

the data of percentage lethality versus concentration.

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Results and Discussion

The selection of the plant species for this project was based primarily on the little

information regarding their chemical composition in spite of the fact that some of the species

belong to families well recognized for the bioactivity of its compounds. In addition, most of the

selected species are native, meaning that they occur naturally in a region (the Caribbean) and

two of them are endemic species, meaning that they are prevalent in a particular locality (the

island of Puerto Rico). Two important aspects of this project are worth to be mentioned, first

the importance of identifying secondary metabolites that may be unique to endemic species. The

identification of such chemotaxonomic markers may aid in the identification of species by its

chemical composition and it demonstrates the multidisciplinary character of the project. In

addition, the isolation and characterization is guided by the bioactivity found in the extracts of

the species.

The solid-liquid extraction that occurs in the blender with dichloromethane/methanol

produces a crude mixture of secondary metabolites. The complex mixture is further simplified

by liquid-liquid extraction suspending the crude in water and using solvents of increasing

polarity to separate each class of metabolites based on its polarity. The crude extract of each

species was analyzed by 1H-NMR Spectroscopy and its response to the Artemia salina bioassay

was assessed. The extracts of the crude were also screened by the Artemia salina bioassay.

The most difficult challenge in natural products chemistry is the separation and

purification of a compound from a complex mixture and its further identification. NMR

spectroscopy is the most important tool for structure elucidation and it is extensively used for

that purpose. However, when working with complex mixtures the interpretation of the spectra is

difficult because of the crowding and overlapping of signals. The 1H-NMR spectra of the total

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lipid extraction shows the occurrence of functional groups according to the chemical shift of the

protons. The area of the signal is proportional to the abundance of that type of proton in the

mixture. The absence or low intensity of any signal does not necessarily indicate the absence of

the corresponding proton, because the signal may be diminished by strong signals in the

spectrum. The results form the 1H-NMR spectra for the total extract is summarized in Table 1

and individual spectrum are shown in Figures 5-10.

All of the species show absorbance in the region corresponding to aliphatic and allylic

protons being one of these signals the predominant in the spectrum of Canella winterana,

Thouinia striata, Guaiacum officinale, and Goetzea elegans. Pimenta racemosa has the most

predominant signal in the aromatic region. In the spectrum of Pimenta racemosa the strong

signals in 3.5 and 4.4 ppm correspond to methanol and water respectively. In the spectrum of

Guaiacum officinale the signal at 3.5 ppm correspond also to residual methanol from the

extraction.

The occurrence of most of the proton types in Canella winterana is consistent with the

variedty of functional groups as seen in Figure 1. In the Goetzea elegans spectrum the most

predominant signals correspond to aliphatic protons although there are minor resonances in the

vynilic and allylic regions of the spectrum. In Pimenta racemosa there is evidence in the

spectrum of the occurrence in the species of aromatic and olefinic functional groups

corresponding to compounds such as eugenol and its derivatives (Figure 2). The spectrum of

Simarouba tulae has signals across the different chemical shifts. It is worth mentioning that the

region from 3.0 to 4.8 has several signals corresponding to protons adjacent to heteroatoms,

which could be due to the presence of highly oxygenated species as the quassinoids. The

aliphatic protons are dominant in the 1H- NMR spectrum of Goetzea elegans with minor signals

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in the aromatic, vynilic and α-heteroatom regions. Thouinia striata has its principal proton

signals in the aliphatic and allylic regions. None of these last two species has literature precedent

regarding their chemical composition.

Table 1. Types of Proton in the 1H-NMR of the Total Extract

Species Aliphatic

(0.2-1.5)

Allylic

X=C-CH3

(1.5-3.0)

α-Heteroatoms

H-C-Y

(3-4)

Vinylic

C=C-H

(4.5-5.5)

Aromatic

Ar-H

(6.5-8.0)

Canella winterana √ √ √ √ √

Thouinia striata √ √ √

Guaiacum officinale √ √ √ √

Pimenta racemosa √ √ √ √ √

Goetzea elegans √ √ √ √

Simarouba tulae √ √ √ √

X= C, O, N

Y= X, O, N, S

Figure 5. 1H NMR spectrum (60 MHz) of Canella Winterana in CDCl3

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Figure 6. 1H NMR spectrum (60 MHz) of Thouinia striata in CDCl3

Figure 7. 1H NMR spectrum (60 MHz) of Guaiacum officinale in CDCl3

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Figure 8. 1H NMR spectrum (60 MHz) of Pimenta racemosa in CD3OD

Figure 9. 1H NMR spectrum (60 MHz) of Goetzea elegans in CDCl3

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Figure 10. 1H NMR spectrum (400 MHz) in CDCl3

Seven plants were collected, dried and extracted with a mixture of CH2Cl2-MeOH (1:1).

The resulting crude extract was suspended in water and extracted with solvents of different

polarities. We used the bioassay-guided fractionation and tested the extracts for the cytotoxic

activity using the brine shrimp lethality test. Table 2 show that 15 extracts of 6 species exhibited

LC50 values below 200 µg/mL. The most promising activity was displayed by crude extracts of

Simarouba tulae, Guaiacum officinale and Croton discolor with lethality values of 2, 21 and 111

µg/mL, respectively. These species belong to families of plants whose genera have demonstrated

contain compounds with anticancer activity such as quassinoids, lignans and cembranes. In

addition the crude extracts also show activity, which infers that the cytotoxic activity can be

attributed to a particular class of compounds. In contrast, the crude extracts of Canella, Pimenta

and Gotzea elegans did not show activity against Artemia salina, but some extracts resulting

from solvent extraction showed activity. This may be due to antagonistic effects of the complex

mixture of compounds present in the crude extract.

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From this preliminary screening we identified extracts of Puerto Rican plants with

cytotoxic activity. Subsequent isolation and identification of the active constituents is needed, as

well as the determination of other possible bioactivities as antimicrobial and the testing against

specific cancer cell lines.

Table 2. Brine Shrimp Lethality Data of Puerto Rican Plants

Plant Extract LC50 value in µg/mL Canella winterana Crude >200

Hexane 78 Chloroform >200

Ethyl Acetate >200 Croton discolor Crude 111

Hexane 132 Chloroform >200

Ethyl Acetate >200 Gotzea elegans Crude >200

Hexane 91 Chloroform 188

Ethyl Acetate >200 Guaiacum officinale Crude 21


Ethyl Acetate 4 Pimenta racemosa Crude >200


Ethyl Acetate 189 Simarouba tulae Crude 2

Hexane >200 Chloroform 161

Ethyl Acetate 35 Thouinia striata Crude >200

Hexane >200 Dichloromethane >200

Ethyl Acetate >200

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Aknowledgements

This work was supported by Institutional funds provided by the “Instituto de

Investigaciones Interdisciplinarias” at UPR-Cayey. Jaymie Rivera Gutierrez and Isamar Ortiz

Rivera thanks the Amgen Bio-Minds Program and the PR-LSAMP Program for their financial

support. We are grateful to personnel of the Chemistry and Biology Departments for the

technical assistance. We also acknowledge Melvin De Jesus and the NMR facilities from the

Department of Chemistry at UPR-Humacao.

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6 Robineau, L.; Soejarto, D. D. “Tramil: A Research Project on the Medicinal Plant Resources of

the Caribbean” Medicinal Resources of the Tropical Forest. Columbia University Press, New

York. 1996.

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7 Meléndez, P. A.; Capriles, V. A. “Antibacterial Properties of Tropical Plants from Puerto Rico”

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Procedure” Pharmaceutical Biology, 1997, 35, 222-226.

10 Farouk S.; El-Feraly.; McPhail, A. T.; Onan, K. D. “X-Ray crystal structure of canellal, a

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221.

12 Claus, E. P. “Pharmacognosy” 3rd ed, Henry Kimpton, London, 1956, 326.

13 Ying, B-P.; Peiser, G. D.; Ji, Y-Y.; Mathias, K. M.; Karasina,F.; Hwang, Y-S.”Structure

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15 Saenz, M. T.; Tornos, M. P.; Alvarez, A.; Fernández, M. A.; García, M. D. “Antibacterial

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grisea” Fitoterapia, 2004, 75, 599-602.

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16 Ahmad, V. U.; Bano, N.; Bano, S. “Officigenin, a New Sapogenin of Guaiacum officinale” J.

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Canella winterana

Canella winterana it is also known as canella alba, wild cinnamon and barbasco

C

c

s

r

d

s

a

c

General Information

Venezuela.

Traditional Uses sesquiterpenes, and warburganal

among other non identified

compounds.

Canellal, isolated from bark is

antifungal, antimicrobial, cytotoxic,

and inhibits insect feeding.

The bark and leaves are still used in

the West Indies to spice beverages

and season food.

The volatile oils found in the leaves

have also been used as additives in

perfumes.

In folk medicine, the inner bark tea is

used to treat fevers, relieve indigestions

and is gargled to treat inflamed tonsils.

Externally, the leaves are appli to

relieve rheumatism and headache.

The rum extract of the bark is used as a

liniment to relieve rheumatism and

other pains, and its liquor is consumed

to treat stomach pains.

On distillation the bark yields a volatile

oil containing benzoyleugenol,

caryophyllene resins, canellal, cineol,

clovanidiol, helicid, mannitol,

myristicin, 1-pinene, drimane

The generic name is reputed

to originate from the Latin

word for cinnamon as it is

used in Spanish “canela”.

The species epithet honors a

captain Winter who first

introduced its bark to

Europe.

Classification

Kingdom Plantae

Subkingdom Tracheobionta

Superdivision Spermatophyta

Division Magnoliophyta

Class Magnoliopsida

Subclass Magnoliidae

Order Magnoliales

Family Canellaceae

Genus Canella

Species Winterana

ssed June 2009

ss Inc, 1997

ed

anella winterana, also known as wild

innamon is a salt tolerant evergreen

hrub. It has an open canopy that

eaches a height of 10 meters with a

iameter of 8 inches. Purple and white

howy flowers cover the tree in summer

nd fall followed by bright red berries

lustered near the tips of the branches.

While the outer bark is gray the inner

bark is yellow and aromatic with the

smell of mixed spices.

Distribution This species occurs naturally from south

Florida through the West Indies and

has been introduced in Brazil and

1) Plants Profile for Canella winterana http://plants.usda.gov/java/profile?symbol=CAWI acce2) Canella winterana hort.ufl.edu/shrubs/CANWINA.PDF accessed June 2009 3) Nellis, D. W. Poisonous Plants and Animals of Florida and the Caribbean 1st Ed. Pineapple Pre

r

©MPO/CO

General

C

s

a

u

f

I

T

a

d

Tradition

F

t

l

u

c

d

l

s

d

gastroenterit

Croton discolo

M

Classification

Kingdom Plantae




Class Magnoliopsida

Subclass Rosidae

Order Euphorbiales

Family Euphorbiaceae

Genus Croton L.

Species Discolor

Variety willd (lechecillo)

This plant is also called white maran.

Information m

d

f

The species name is Latin and it refers to the fact that the top and the bottom of its leaves are of different

color.

Distribution

The genus of over 600 species of is

distributed worldwide and includes

aromatic herbs, shrubs and trees.

al Uses

is.

Externally, the oil is an irritant and

may cause blistering of the skin

The esters of theteracyclic

diterpenoid phorbol have been

found to promote tumors in mouse

skin previously treated with

subcarcinogenic doses of

car
Croton discolor fromPuerto R
cinogens.

roton discolor is an aromatic 8 ft. tall

hrub. It has alternate leaves which

re dark on the top lighter on the

ndersides. It has male an female

lowers which born on separate stalks.

ts seeds are 0.25 inches long.

he plant toxicity is attributed to the

lkaloids crotonosine, 8,14-

ihydrosalutadirine,

ethylcrotonosine, linearisine and

iscolorine, which have been isolated

rom the plant.

Isla de Mona

olk medicine has used the tea from

he leaves to treat rheumatism. Young

eaves and branch tips have been

sed to treat coughs. A component of

roton oil (phorbol 12-tiglate 13

ecanoate) has been found to inhibit

eukemia. Croton oil found in leaves,

tem and seeds is effective in small

oses but in high doses causes severe

ico

1) Plant Profile for Croton discolor USDA http://plants.usda.gov/gallery/thumbs/crdi8_001_tvp.jpg accessed June 2009 2) Nellis, D. W. Poisonous Plants and Animals of Florida and the Caribbean 1st Ed. Pineapple Press Inc, 1997

©MPO/CO

General

2J3

pertains to th

Solanac

r

w

c

wheel-shape

M

Goetzea elegans

The Solanaceae family includes the potatoes,

tomatoes, peppers, tobacco and petunias.

Wydler

Goetzea elegans is an endemic plant of Puerto Rico also known as beautiful, Matabuey and Manzanilla.

Information

k

t

s

h

a

l

S

f

c

eggplant, potato and chili pepper. e Solanaceae family also

Atropine has been isolated from

several species of the Solanaceae

family. It is a secondary metabolite

that serves as a drug with a wide

variety of effects. It is a competitive

anta

acet

eae family

d or cylinder-shaped.

Goetzea elegans is characterized by its

trumpet shaped orange flowers. It is an

evergreen shrubby tree that can

grow

up to 9 meters in height. This endemic

species is only found in the semi-

evergreen forests in northern Puerto

Rico. In 2005 fewer than fifty individuals

of the species remained. It is very rare,

it can be found in moist limestone and

moist coastal forests. Goetzea elegans

nown as the potato or nightshade family

hat has 75 genera and over 3000

pecies. Several species of this family

ave provided many poisons as atropine

nd scopolamine that can be deadly in

arge amounts.

everal vegetables of the Solanaceae

amily are important for their nutrient

omponents these include the tomato,

gonist for the muscarinic

ylcholine receptor.

Classification Kingdom Plantae




Class Magnoliopsida

Subclass Asteridae

Order Solanales

Family Solanaceae

Genus Goetzea Wydler.

Species Goetzea elegans

In the solanaceae family the leaves

can be opposite, clustered or

alternate. There is also variety in the

arrangement and the form of the

flowers, but generally the corolla is

adially symmetrical and five lobed

ith five stamens. The calyx and

orolla can be either bell-shaped,
1) Liogier, A. H.; Liogier, H. A.; Martorell, L. F. Flora of Puerto Rico and adjacent islands 2nd Ed. UPR Editorial, 2000.
) National Collection of Imperiled Plants http://www.centerforplantconservation.org/ASP/CPC_ViewProfile.asp?CPCNum=2041 accessedune 2009 ) Plants Profile for Goetzea elegans http://plants.usda.gov/java/profile?symbol=GOEL accessed June 2009

4) Kirkpatrick, Z. M. Wildflowers of the western plains 1st Ed. University of Texas Press, 1992

©MPO/CO

2

4

General G

e

f

c

a

a

t

a

p

o

h

acrid taste

Tradition

Guaiacum re

has been u

chronic skin

prevent go

venereal di

was successf

its introducti

three thousa

from its thera

M

Guaiacum officinale

R1O

COOR2

R1= azúcaresR2= H o azucares

Saponins form G. officinale

Guaiacum officinale is also known as the tree of life, it is the source of the true lignum vitae, a trade wood also called guayacan.

Information

and is odorless, unl

al Uses ess

It is believed that the guaiacum did not

had power to cure the syphilis but

rather alleviate the symptoms.

A number of saponins have been

reported from different plant parts.

Guaicins A to G have been found in

leav

Hea

furo

sin is an acrid stimulant. It

sed to treat rheumatism,

diseases, scrofula and to

ut. As a remedy for

seases guaiacum wood

ully used. Nine years after

on to Europe more than

nd persons had benefit

py

uaiacum officinale is an ornamental

ver

green tree with pretty rich blue

lowers, the trunk is a greenish-brown

olor, the wood of slow growth but

ttains a height of 40 to 60 feet, stem

lmost always crooked, bark furrowed;

he wood is extraordinarily heavy, solid

nd dense, fibers cross-grained;

innate leaves, oval obtuse; fruit

bcordate capsule; seeds solitary,

ard, oblong. The wood has a slight

heated, when it emits an agreeable

scent. This wood was once very important

for uses requiring strength, weight, and

hardness. All species of the genus are

now listed in CITES (Convention on

International Trade in Endangered

Species of Wild Fauna and Flora) as

potentially endangered species. The bark

yields 1 per cent volatile oil of delicious

fragrance.

Distribution This species occurs naturally in Florida

Puerto Rico and the Virgin Islands.

es, stem bark and fruits.

rtwood contains lignans such as

guaiacidin and furoguaiaodin.

Classification

Kingdom Plantae




Class Magnoliopsida

Subclass Rosidae

Order Sapindales

Family Zygophylaceae

Genus Guaiacum L.

Species Guaicaum

officinale L

Guaiacum officinale is the national flower of

Jamaica.

1) Pereira, J. The Elements of Materia Medica and Therapeutics 4th Edition, Longman, 1857. ) A Modern Herbal/Guaiacum http://www.botanical.com/botanical/mgmh/g/guaiac42.html accessed June 2009 accessed June 2009

3) Magical Flower http://www.flickr.com/photos/kam74/2462585674/ accessed June 2009 ) Plants Profile for guaiacum officinale http://plants.usda.gov/java/profile?symbol=GUOF accessed June 2009

5) Daniel, M. Medicinal Plants Science Publishers 2005.

©MPO/COM

General In

2

varieties that ha

Traditiona

T

c

t

t

b

D

e

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u

a

Pimenta racemosa

Pimenta racemosa is also known as malagueta, guayabita, wild cinnamon and bay rum tree.

formation

v

c

c

w

i

Puerto Rico and the Virgin Islands.

ve been described is the

l Uses

Eugenol is the main constituent of bay leaf oil

w

f

Pimenta racemosa

(Bay rum tree) is

native to Puerto Rico and perhaps to the

Virgin Islands and the other West Indian

islands. The tree is identified by its

opposite, smooth-margined, dark green

leaves, which when crushed have a

strong bay rum (clove-cinnamon)

fragrance. Its bark is gray to light brown,

white flowers borne in round clusters and

the fruits are small, elongated green fruits

that turn black in maturity. One of the

grisea with its oil rich in geraniol, methyl

eugenol and isoeugenol. The citriodora

ariety has strongly lemon-scented oil

onsisting mostly of geranial. A believed

chemovariety produces an anise-

scented oil consisting mostly of methyl

havicol and methyl eugenol.

Harvesting and extraction of any variety

ith true bay rum produces an oil of

nferior quality.

Distribution Pimenta racemosa can be found in

Classification

Kingdom Plantae




Class Magnoliopsida

Subclass Rosidae

Order Myrtales

Family Myrtaceae

Genus Pimenta Lindl.

Species Pimenta

racemosa

Bay leaf oil or myrcia oil is yellow to darBay Leaf Oil

k

brown. The crude oil has a sweet

penetrating odor. Approximately 80

compounds have been detected

including monoterpenes, sesquiterpenes,

aromatic and aliphatic compounds.

The major use for bay oil is in hair lotions

ith minor uses in perfumery and as a

ood flavoring.

he leaves have been used as a spice in

ooking, as a perfume, and as a liniment

o ease muscle aches, P. racemosa var.

erebinthina is used in the Caribbean

asin for different afflictions. In the

ominican Republic, the essential oil

xtracted from the leaves is used for the

reatment of rheumatism or for

toothache. P. racemosa var. grisea is

sed for its anti-inflammatory and

nalgesic properties.
1) E. A. Weiss Spice Crops CABI Publishing, 2002
) Plants Profile Pimenta racemosa http://plants.usda.gov/java/profile?symbol=PIRA accessed June 20093) Kirk, T. K. Tropical Trees of Florida and the Virgin Islands Pineapple Press Inc, 2009.

©MPO/CO

This family

genera an

by their

includes o

Simar

the quassi

slender, wi

top of a s

little berries

Quass

The ma

Simaroub

belong

family an

bioactivi

chemica

Simarouba tulae

Simarouba tulae is an endemic plant of Puerto Rico commonly known as “aceitillo falso”

M

consists of approximately 25

d 150 species that are known

chemical composition that

xy

oubaceae Family

genated triterpenes such as

noids. Usually their trees are

th long pinnate leaves on the

lender hole and racemes of

.

ci

T

t

u

inoids in active compounds of

a are the quassinoids, which

to the triterpene chemical

d which are responsible for its

ty and are considered

l markers of the family.

There are about 100 quassinoids

known and some like isobruceine A

posses a strong antileukemic activity.

Some other quassinoids have antiviral,

antimalarial, antifeedant and

insecticidal properties.

Classification

Kingdom Plantae




Class Magnoliopsida

Subclass Rosidae

Order Sapindales

Family Simaroubaceae

Genus Simarouba Aubl.

Species Simarouba

tulae Urb
Traditional Uses
In the early twentieth entury quassinoids were ntroduced in Europe as

nicotine substituent.

he bark, wood and leaves from

he genus Simarouba have been

sed for their analgesic,

antimicrobial, antimalarial,

astringent, sudorific,

antidysenteric, amoebicidic and

antibacterial properties. Its bark

has been used as a remedy for the

treatment of diarrhea, dysentery

and dyspeptic affections.

O O

Basic skeleton of a Quassinoid C-20

1) Plants Profile for Simarouba tulae http://plants.usda.gov/java/profile?symbol=SITU accessed June 2009

2) Savory, J. A compendium of domestic medicine 7th Edition 1865.

3) Daniel, M. Medicinal Plants Science Publishers 2005.

4) Wiart, C. Medicinal Plants of Asia and the Pacific CRC Press, 2006.

5) Memory, P. F. Medicinal botany 2nd Ed. John Wiley and Sons 2003.

©MPO/CO

3) Nelson, G. Th

General T

c

a

P

M

k

a

c

Sapinda

M

Thouinia striata

e trees of Florida Pineapple Press

Classification

Kingdom Plantae



Class Magnoliopsida

Subclass Rosidae

Order Sapindales

Family Sapindaceae

Genus Thouinia

Species striata

Variety portoricensis

striata

Thouinia striata is an endemic plant found only in the island of Puerto Rico

Information

“The common name of the genus refers to the

presence of saponins in the soft pulps of the trees fruits. Solutions made from this pulp produce lather very much like the one

produced by manufactured soap. Some species of the

family have been used as detergents in tropical

regions.”

ceae family M

m

t

t

S

o

e

p

b

Inc. 2004.

houinia striata is also known as

eboruquillo and serrasuela. This tree

nd shrub is an endemic plant of

uerto Rico. Ceboruquillo is part of the

agnoliophyta division otherwise

nown as flowering plants. This plant is

lso a member of the magnoliopsida

lass usually referred to as

dicotyledons. Thouinia striata pertains

to the Sapindaceae botanical family,

otherwise known as the Soapberry

family.

Sapindaceae, also known as the

soapberry family, is a family of

flowering plants in the order

Sapindales. There are about 140-150

genera with 1400-2000 species,

including maple, horse chestnut and

lychee. Sapindaceae members occur

in temperate to tropical regions

throughout the world.

any are lactiferous, i.e. they contain

ilky sap, and many contain mildly

oxic saponins with soap-like qualities in

he foliage and/or the seeds, or roots.

ome of the sapindaceae species are

f economic importance including

dible fruits, medicinal plants, fish

oisons, soaps and stimulating

everages.

1) ITIS Standard Report Page Thouinia striata http://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=28708 accessed June 2009

2) Simpson, M. G. Plant Systematics Academic Press 2006

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