Chemical analysis and biological evaluation of tropical plants: goetzea elegans and pimienta...

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7/30/2019 Chemical analysis and biological evaluation of tropical plants: goetzea elegans and pimienta racemosa

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Chemical Analysis andhemical Analysis and

Biological Evaluation of Tropicaliological Evaluation of TropicalPlants:lants: Goetzeaoetzea eleganslegans andnd

Pimentaimenta racemosaacemosaJaymie M. Rivera Gutiérrez

Dr. Claudia Ospina1, Dr. Mayra Pagán1 andProf. Augusto Carvajal2

Chemistry Department University of Puerto Rico at Cayey2Biology Department, University of Puerto Rico at Cayey

Aimsims:{ Main:

z The development and application of bioassay for theresearch of antitumor activity on tropical plant’s

extract.

Specifics:z Studying cytotoxic properties of tropical species.

z Chemotaxonomic study of tropical plants in Puerto

Introductionntroduction• Since ancient times, in Puerto Rico someplants have been used to treat diseases likecolds, coughs, diarrhea, respiratory infections,

skin lesions and others. Unfortunately, it isestimated that only a few group of endemicand native plants have been studied around

entire world.

• In Puerto Rico, we are at risk of losing the

ancestral knowledge of the healing propertiesof plants.Meléndez, 2006

Plant-based anticancer moleculesO

P o d o p h y l l o t o x i n

L ig n a n is o l a t e d f r o m P o d o p h y llu m p e l ta t u m

E t o p o s i d e

O G l u

S y n t h e t ic D r u g

C a m p t o t h e c i n

A lk a lo id e x t ra c te d fro m C a m p o th e c a a c u m in a ta

T o p o t e c a n

S y n t h e tic D r u g

Srivastava, 2005

GENERAL STRATEGY:

Selection of the Organism

GENERAL STRATEGY:

Selection of the OrganismThe species that we chose for this study were:

•Pimenta Racemosa (P.R.) – antiseptic and astringentproperties; against headache, sore throat, stomach painreliever and it helps to regulate the digestion.

•Goetzea Elegans (G.E. - endemic) – Has not been reportedany healing power yet.

Alain, 2006

Goetzea Elegans

Pimenta Racemosa

Cidra, Puerto Rico

Río Piedras, Puerto Rico

February 23, 2008:

A collection expeditionwas organized takingin consideration the

distribution andabundance of the

organisms in aspecific area

GENERAL STRATEGY:ENERAL STRATEGY:

Collectionollection

MethodologyethodologyP l a n t

D r y i n g a n d M a c e r a t i o nw i t h a m i x t u r e o f C H 2 C l 2 / M e O H ( 1 : 1 )

C r u d e E x t r a c t

S u s p e n d e d i n W a t e r a n dE x t r a c t e d w i t hs o l v e n t s o f d i f f e r e n t p o l a r i t y

H e x a n e C h l o r o f o r m E t h y l A c e t a t e B u t a n o l W a t e r*

S o m e t i m e s , b u t a n o l e x t r a c t i o n i s r e u i r e d

Methodologyethodology{ Rotovaporation of the samples

Results and Discussionesults and Discussion

-Generaleneral-

Leaves dry

Weight (g)

±0.02g

Solvent name Solvent

Crude extract

dry weight (g)

±0.02

Hexane 400

Chloroform 300

Ethyl Acetate 300

Buthanol 300

P.R. 279.66

Aqueous 175

Hexane 400

Chloroform 450

Ethyl Acetate 300

G.E. 270.10 25.10

Aqueous 200

Methodologyethodology• Biological evaluation:

•Bioassay

• Serial dilutions of extracts are made in wells of96-wells microplates in triplicate in 100μL seawater. A suspension of nauplii containing 10-15

brine shrimp larvae are added to each microwellplate incubated at room temperature for 24 hours.

Methodologyethodology

Bioassay

-Biological evaluationiological evaluation-Extract LC50 (µg/mL)

P.R. Crude 1.6x104

P.R. Hexane 68.9

P.R. CHCl3

G.E. Crude 6.2x104

G.E. Hexane 171.0

G.E. CHCl3 4.1x102

Three of the six extracts were active with a LC50 ≤200µg/mL. These extracts were obtained after partition with

solvents of different polarities. The crude extracts of thesespecies were inactive with a LC50 ≥200 µg/mL.

-Chemical analysishemical analysis-1H NMR spectrum of Pimenta Racemosa in CD3OD

The functional groups for the crude extract of P.R. are

aliphatic carbons under the 1-2 ppm region, aromatics in 6-7.1ppm, and double bonds at 5pmm. The high peaks under3.5-4.8ppm is the methanol-d used as solvent.

Results and Discussionesults and Discussion-Chemical analysishemical analysis-

1H NMR of chloroform extract of Pimenta Racemosa

The corresponding compound for this NMR is eugenol

and the corresponding molecule is:

-Chemical analysishemical analysis-1H NMR spectrum of Goetzea Elegans in CDCl3

The functional groups for the crude extract of G.E.

are aliphatic protons under the 0.6-2.4 ppm region,and double bonds at 5pmm.

Methodologyethodology•Chemical analysis:

•TLC Analysis:

•A mixture of different concentrations ofsolvents was prepared as the liquid phase.We took drops of the corresponding samples

of G.E. crude, G.E. hexane extract and G.E.chloroform extract and settled in thestationary phase.

Results and Discussionesults and Discussion-Chemical analysishemical analysis-

•TLC analysis:

Where A is the

crude extract, Bis the hexaneextract and C isCHCl3 extract

for G.E.

•Better mixture of

solvents to purify thecompounds 9.0mLHexane with 1.0mLAcoEt

Methodologyethodology{ Chemical analysis:

z Separation by column:{

We made a separation by columns using the bestmixture of solvents with different polarities basedon the results of the TLC.

Column and the six fractions.

-Chemical analysishemical analysis-•TLC analysis:

A B C D

Where A is thecrude extract, B

is fraction #1 , Cis the fraction #2and D is thefraction #3 for

the hexane extractfor G.E.

Mixture of solvents:9.5mL Hexane with0.5mL AcoEt

-Chemical analysishemical analysis-1H NMR of the third fraction of G.E. hexane extract.

The possible functional groups for the hexane extract ofG.E. are aliphatic carbons under the 1-2 ppm region,

aromatics in 7-8ppm, and double bonds at 5pmm.

Future Work:

{ Extracts purification and separation will be basedon the bioactivity results.

{13C NMR for the third fraction of G.E. hexane

extract.

Sam, et Al, 2007

Acknowledgments:{ Dr. Claudia Ospina

{ Isamar Ortiz, Karla Claudio andJanibeth Hernandez

Biominds Program{ Chemistry and Technicians

Department

{ Interdisciplinary InvestigationInstitute

{ RISE Program

Chemical Analysis andhemical Analysis and

Biological Evaluation of Tropicaliological Evaluation of TropicalPlants:lants: Goetzeaoetzea eleganslegans andnd

Pimentaimenta racemosaacemosaJaymie M. Rivera Gutiérrez

Dr. Claudia Ospina1, Dr. Mayra Pagán1 andProf. Augusto Carvajal2

1Chemistry Department University of Puerto Rico at Cayey2Biology Department, University of Puerto Rico at Cayey

Chemical analysis and biological evaluation of tropical plants: goetzea elegans and pimienta...

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