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Family: Zygophyllaceae Genus: Guaiacum Species: Officinale Common name: Guayacn, Palo Santo, Lignum Vitae Distribution: Native to West Indies and South America Historical uses: Gout, syphilis, arthritis, resin used to detect blood in human stool.
Simarouba tulae
Family:
Simaroubaceae
Commonly
Trees
Distribution:
Simarouba tulae
Simaroubacea family
Traditional Uses: Anti-malaria Anti-feedant Anti-inflammatory Anti-leukemic Anti-viral
To expand to the phytomedicinal knowledge of native and endemic plants of Puerto Rico and to discover their chemotaxonomy. Isolate, purify and identify chemical compounds of Guaiacum officinale and Simarouba tulae leaves. Evaluate the cytotoxic activity of pure compounds of Guaiacum officinale and Simarouba tulae leaves.
Biological test
Plant Collection
Jayuya
Guaiacum Officinale
1.12kg de hojas
98.5 g extracto crudo
Extraction
1H
Alyphatic
1H
Guaiacum oficinale
Crude
Hexane
30.765
Chloroform
0.692
91
76
Ethyl Acetate
4.479
* All extracts were evaluated at a single dose of 100 g/mL. ** Results performed by Dr. Marianela Prez Torres in the UPR-MSC.
*** LC values greater than 200 g/ml are not considered cytotoxic 50
First
collection
Table 2. Dry weight for crude extracts and solvent used for each extraction in the first collection
Plant Extract Extract dry weight
(g) 0.02 Simarouba tulae Crude Hexane Chloroform Ethyl acetate 15.0 2.33 9.21 0.90
Table 3. Cytotoxicity results for Simarouba tulae leaves extracts against Artemia Salina test
Simarouba tulae Extract Artemia Salina Test LC50 value in g/mL
Crude Hexane
2 > 200
Chloroform
Ethyl acetate
161
35
Extraction: Chloroform
Aqueous layer
Organic layer
Drying at Room Temp.
Rotoevaporation
Extract
TLC
Chloroform /Methanol (9.8:0.2) Hexane/Ethyl Acetate (9:1)
Second
collection
Table 6. Dry weight for crude extracts in the second collection of Simarouba tulae
Extract
Simarouba tulae Crude
Hexane
Chloroform Ethyl Acetate
30.88
39.64 6.08
Simarouba
tulae Extract
MCF-7
ZR-75-1
T47D
Crude Hexane
82 87
< 80 < 80
94 92
Chloroform
Ethyl acetate
157.141
26.243
95
< 80
< 80
< 80
97
92
tulae
Extract
Growth inhibition % *
MCF-7
ZR-75-1
T47D
Crude
Hexane Chloroform
23.703
> 200 157.141
82
87 95
< 80
< 80 < 80
94
92 97
Ethyl acetate
26.243
< 80
< 80
92
Simarouba
tulae Extract
MCF-7
ZR-75-1
T47D
Crude Hexane
82 87
< 80 < 80
94 92
Chloroform
Ethyl acetate
157.141
26.243
95
< 80
< 80
< 80
97
92
28 fractions
28 fractions
SH2C3
28 fractions
SH2C3C
SH2C3
28 fractions
SH2C3C-C
SH2C3C
SH2C3
Allylic
Vinylic
-Heteroatoms
Vinylic
Carbonyls
Oxygenated
Aliphatic
CC Si gel (CHCl3/Acetona)
CC Si gel (CHCl3/Acetona)
23 fractions
1H
NMR Spectrum (400 MHz) for crude extract of the leaves of S. tulae
Alkenes
Every extract of Guaiacum officinale presented activity in the Artemia Salina bioassay. From spectroscopic chloroform extracts of Guaiacum officinale are rich in metabolites.
The chloroform extract of Simarouba tulae leaves showed high cytotoxic activity against Artemia Salina and two breast cancer cell lines. From spectroscopic data SH2C3C-C is rich in metabolites The hexane extract of Simarouba tulae leaves showed high cytotoxic activity against two breast cancer cell lines.
Identification of the main compound of the Chloroform extract using NMR spectroscopy. Perform a Chromatographic analysis of Chloroform To evaluate the cytotoxicity of pure compounds against cancer cell lines.
Batista, J.; Braz, R.; Curcino, I.; Da Silva, M.; Rodrigues E.; Vireira, P. 20(R)- and 20(S)- Simarolide Epimers Isolated from Simaba cuneata Chemical Shifts Assignment of Carbon and Hydrogen Atoms. J. Braz. Chem. Soc., 1999, 10, 76-84. Beutler, J.; Clement, J.; Goncharova, E.; et al. Quassinoid Inhibition of AP-1 Function does not Correlate with Cytotoxicity or Protein Synthesis Inhibition. Journal of Natural Products, 2009 Anderson, M.; Gupta, M.; Phillipson, D.; Solis, P.; Colin, W. A Microwell Cytotoxicity Assay using Artemia Salina (Brine Shrimp). Planta Med, 1993, 59, 250-252.
Guo, Z.; Sindelar, R.D.; Sindelar, R.W.; Vangapandu, S.; Walker, L.A. Biological Actives Quassinoids and Their Chemistry: Potential Leads for Drug Design. Curr. Med. Chem. 2005, 12, 173-190.
Rhodes, M.; Robins, R. High-performance liquid chromatographic methods for the analysis and purification of quassinoids from Quassia amara L. J. Chromato. 1984, 283, 436-440. Ospina, C. A.; Pagn, M.; Carvajal, A.; Claudio, K; Rivera, J.; Ortiz, I.; Hernndez, J. In Cytotoxic Screening of Tropical Plants Using Brine Shrimp Lethality Test.; Montes, E. L.; Eds.; Cuadernos de Investigacin Number 7; Instituto de Investigaciones Interdisciplinarias: Cayey, 2009; 1-20.
Advising
Technical Support
Chemistry Department
Melvin De Jesus, UPR Humacao
Financial Support
Collaborators
Augusto Carvajal, M.S Marianela Prez School of Pharmacy UPR Karla Claudio- Graduate student
Undergraduate students
Reynaldo J. Morales Rodriguez Yadira D. Cora Amaro Elsa M. Luciano Nunez Claudia A. Ospina, Ph.D. Mayra Pagn Ortiz, Ph.D.
Saline Solution
Yeast
Marine salt
Distilled water
Samples Preparation
Positive Control: Berberine Chloride Negative Control: Saline Solution Plants extractions dissolved in DMSO
Bioassay
Prepare the concentration assigned to each line Add 10-15 brine shrimps by pippeting
Data Collection Count the death shrimps Add MeOH Count the total shrimps