Hernandulcin is the major constituent of Lippia dulcis Trev. (Verbenaceae)

Natural Product Letters

Volumn 8, Issue 2, 1996, Pages 151-158

  Souto Bachiller, Fernando A ^a   De Jesus Echevarria, Maritza ^a   Cardenas Gonzalez, Oswaldo ^a  

^a UNIVERSITY OF PUERTO RICO   (Puerto Rico)

Author keywords

Camphor; Epihernandulcin; Hernandulcin; Lippia dulcis; Verbenaceae

Indexed keywords

ESSENTIAL OIL; HERNANDULCIN; PLANT EXTRACT; SESQUITERPENOID;

ARTICLE; DRUG ISOLATION; NONHUMAN; TRADITIONAL MEDICINE;

LIPPIA DULCIS;

EID: 0029962015     PISSN: 10575634     EISSN: None     Source Type: Journal    
DOI: 10.1080/10575639608043255     Document Type: Article

References (23)

1. C. M. Compadre, J. M. Pezzuto, A. D. Kinghorn, and S. K. Kamath (1985) Science 227, 417.
2. F. Hernández (1959) Obras Completas Tomo II: Historia Natural de Nueva España, Universidad Nacional de Mexico, Mexico, D. F.
3. F. Hernández (1651) Rerum Medicarum Novae Hispaniae Thesaurus seu Plantarum, Animalium, Mineralium Mexicanorum Historia, Typographeio Vitalis Mascardi, Rome, p. 240.
4. J. Morton (1981) Atlas of Medicinal Plants of Middle America, C. C. Thomas Publishing Co., New York, N.Y., pp. 746-747; also see J. T. Roig (1988) Plantas Medicinales, Aromáticas o Venenosas de Cuba, Ministerio de Cultura, Editorial Científico-Técnica, Habana, Cuba, Second Ed., pp. 606-607.
5. J. Morton (1981) Atlas of Medicinal Plants of Middle America, C. C. Thomas Publishing Co., New York, N.Y., pp. 746-747; also see J. T. Roig (1988) Plantas Medicinales, Aromáticas o Venenosas de Cuba, Ministerio de Cultura, Editorial Científico-Técnica, Habana, Cuba, Second Ed., pp. 606-607.
6. C. M. Compadre, R. A. Hussain, R. L. Lopez de Compadre, J. M. Pezzuto, and A. D. Kinghorn, (1987) J. Agric. Food Chem. 35, 273.
7. C. M. Compadre, E. F. Robbins, and A. D. Kinghorn (1986) J. Ethnopharm. 15, 89.
8. F. A. Souto Bachiller, L. Romero Ramsey, M. De Jesús Echevarría, R. Suau Suárez, and F. Pliego Alfaro (1991) Studies on the Induction and Growth of Callus Cultures from the Verbenaceae Herb Lippia dulcis Trev, 2nd Panamerican Chemical Congress, San Juan, Puerto Rico, September 25-27.
9. N. Kaneda, I.-S. Lee, M. P. Gupta, D. D. Soejarto, and A. D. Kinghorn (1992) J. Nat. Prod. 55, 1136.
10. Quoted from F. Hernández, Ref. 3: "Huius herbae folia adeo dulcis funt, vt melipfum, faccharumque, & quaecunque alia dulcedine praeftant, longe inferiora videantur. itaque in hac vna fola planta vifa eft natura periclitary, quantam dulcedinem poflet rebus naturalibus impertiri."
11. Sterile disease free plants could be obtained adapting literature procedures developed for orchids to the in vitro germination of seeds from wild L. dulcis plants, see R. L. M. Pierik (1987) In Vitro Culture of Higher Plants, Martinus Nijhoff Publishers, Boston, pp. 149-158.
12. Upon seed germination and growth, plantlets were regenerated by single node culture, ibid. pp. 190-193.
13. A sample of dry powder from wild plant material was brought to constant weight in a desiccator and 10.0 g placed in a 250-ml Soxhlet apparatus. The extraction was performed refluxing 100 ml of the respective solvent for 1-8 h in a 200-ml round bottomed flask.
14. The TLC analysis was carried out with Analtech GHLF (normal phase) and RPSF (C-18 reversed phase) silica gel plates (250 μm, 2.5×10 cm) with fluorescent indicator. Difficult separations were performed on Analtech HLF silica gel channeled plates (250 μm, pre-adsorbent, 10×20 cm). Development of the plates after spotting (1-10 μ1) was carried out with the following mixed solvent systems (v/v). Normal phase silica: hexane-acetone, 85:15 and 70:30; chloroform-acetone, 100:0, 50:50 and 20:80; hexane-ethyl acetate, 90:10, 75:25, 70:30, 60:40 and 50:50. Reversed phase: acetone-water, 95:5 and 80:20; isopropanol-water, 95:5 and 50:50; acetonitrile-water, 100:0, 88:12, 80:20, 70:30, 60:40 and 50:50. Visualization of the separation was by UV activated fluorescence quenching.
15. Capillary GC was carried out in a Hewlett-Packard 5790A GC with an FID detector set at 250°C. Oil components were preliminary identified by comparison with the relative retention times of authentic samples. The columns used were a Hewlett Packard Ultra-2 fused silica capillary column, 5% phenylmethylsilicone cross-linked film (25 m long × 0.2 mm internal diameter × 0.33 μm film thickness) and a Supelco SPB-5 capillary column with the same film (60 m long × 0.25 mm internal diameter × 1 μm film thickness). Helium was used as carrier gas with a column head pressure of 10-15 psi. The injector temperature was set at 80°C when using the SPB-5 column and at 225°C for the Ultra-2 column. The temperature of the Ultra-2 column was maintained isothermal at 200°C or programmed at 70°C - 4 min, 10°C/min to 125°C, 125°C - 5 min, 2°C/min to 250°C, and 250°C - 2 min. The Chromatographic conditions for the SPB-5 60-m column were programmed at 25°C - 4 min, 4°C/min to 250°C, and 250°C - 10 min. Chromatography was performed by injecting 0.20-1.0 μ1 of 10% solutions of the oil or tissue extract in chloroform or hexane-acetone 70:30. Samples were injected using direct mode and split mode with ratios of 40:1, Ultra-2 column, and 15:1, SPB-5 column.
16. Selected samples were analyzed by gas chromatography/mass spectrometry. The essential oil components were identified by visual or computerized matching with published mass spectral compilations. The experiments were carried out under electron impact ionization (EI) mode, with electron energies of 70 eV, in a Hewlett Packard 5970B mass selective detector. Other experimental parameters were electron multiplier voltage, 1600-1800 V; scan rate, 1.21 scan/s; mass range scanned, 45-400 uma; unit mass resolution over entire range; ion source temperature, 200°C; and transfer line temperature, 180°C.
17. f 0.5 in normal phase silica developed with hexane-acetone 85:15.
18. 3 with TMS as internal standard, using either a QE-300 General Electric instrument or a Gemini-300 Varian spectrometer. Low resolution mass spectra were obtained injecting the pure compounds in the GC-MSD system described above.
19. D. W. Mayo, R. M. Pike, and S. S. Butcher (1989) Microscale Organic Chemistry Laboratory, Wiley, New York, N.Y., Second Ed., pp. 105, 106.
20. 2 and analyzed by TLC and capillary GC.
21. A. A. Craveiro, J. W. Alencar, F. J. A. Matos, C. H. S. Andrade, and M. I. L. Machado (1981) J. Nat. Prod. 44, 598.
22. Since oils obtained from different varieties of the same plant species may differ in composition, while oils from plants of different families may have similar compositions, it is difficult to utilize the chemical composition for the classification of plants in the botanical system. In addition, essential oils from seemingly identical plants may differ in composition due to environmental pressures that lead to adaptation to new conditions by spontaneous mutation. For a complete discussion, see K. Bauer and D. Garbe (1985) Common Fragrance and Flavor Materials: Preparation, Properties and Uses, VCH Publishers, Deerfield Beach, Florida, Chap. 3.
23. Acknowledgements. - Prof. Miguel Vives for his field work and invaluable assistance in the identification of the diversity of Lippia species growing in Aguadilla. Mr. Benigno Rodriguez for his kind gift of three specimens of Lippia dulcisand one each of the other Lippiae. Dr. Henri A. Liogier, taxonomist, for his invaluable contribution to this work through his systematic studies of the flora of Puerto Rico and adjacent islands. Mrs. Perfa Liogier for her kindness in providing access to all the specimens of Lippiae collected in Puerto Rico and deposited at the herbarium of the Botanical Garden of Puerto Rico, Rio Piedras. Mr. Francisco Lopez for access to his collection of medicinal plants and the supply of two specimens of L. dulcis. Mrs. Maria Ortolaza for allowing us to collect abundant samples of this species from a natural population at her country estate. The Chemistry Department, University of Puerto Rico, Mayagüez Campus for very generous financial assistance, support and encouragement to carry out this research project. The National Science Foundation, RIMI and EPSCoR Programs, for making it possible to develop the intellectual climate and research infrastructure essential to undertake a demanding project with interdisciplinary scope.