Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates

Sherif S. Ebada, RuAngelie Edrada-Ebel, Wenhan Lin, Peter Proksch

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) as antitumor agent for the treatment of advanced soft tissue sarcoma. In this protocol, methods for bioactivity-guided isolation, purification and identification of secondary metabolites from marine invertebrates such as sponges, tunicates, soft corals and crinoids are discussed. To achieve this goal, solvent extraction of usually freeze-dried sample of marine organisms is performed. Next, the extract obtained is fractionated by liquid-liquid partitioning followed by various chromatographic separation techniques including thin layer chromatography, vacuum liquid chromatography, column chromatography (CC) and preparative high-performance reversed-phase liquid chromatography. Isolation of bioactive secondary metabolites is usually monitored by bioactivity assays, e.g., antioxidant (2,2-diphenyl-1-picryl hydrazyl) and cytotoxicity (microculture tetrazolium) activities that ultimately yield the active principles. Special care should be taken when performing isolation procedures adapted to the physical and chemical characteristics of the compounds isolated, particularly their lipo- or hydrophilic characters. Examples of isolation of compounds of different polarities from extracts of various marine invertebrates will be presented in this protocol. Structure elucidation is achieved using recent spectroscopic techniques, especially 2D NMR and mass spectrometry analysis.
LanguageEnglish
Pages1820-1831
Number of pages12
JournalNature Protocols
Volume3
DOIs
Publication statusPublished - 6 Nov 2008

Fingerprint

trabectedin
Liquid chromatography
Invertebrates
Metabolites
Bioactivity
Biological Products
Purification
Urochordata
Thin layer chromatography
Aquatic Organisms
Anthozoa
Column chromatography
Liquids
Porifera
Solvent extraction
Reverse-Phase Chromatography
Cytotoxicity
Vacuum
Thin Layer Chromatography
Liquid Chromatography

Keywords

  • purification
  • structural elucidation
  • bioactive secondary metabolites
  • marine invertebrates

Cite this

@article{61f05306b8b3481586dcc6b5cb20a4f9,
title = "Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates",
abstract = "In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) as antitumor agent for the treatment of advanced soft tissue sarcoma. In this protocol, methods for bioactivity-guided isolation, purification and identification of secondary metabolites from marine invertebrates such as sponges, tunicates, soft corals and crinoids are discussed. To achieve this goal, solvent extraction of usually freeze-dried sample of marine organisms is performed. Next, the extract obtained is fractionated by liquid-liquid partitioning followed by various chromatographic separation techniques including thin layer chromatography, vacuum liquid chromatography, column chromatography (CC) and preparative high-performance reversed-phase liquid chromatography. Isolation of bioactive secondary metabolites is usually monitored by bioactivity assays, e.g., antioxidant (2,2-diphenyl-1-picryl hydrazyl) and cytotoxicity (microculture tetrazolium) activities that ultimately yield the active principles. Special care should be taken when performing isolation procedures adapted to the physical and chemical characteristics of the compounds isolated, particularly their lipo- or hydrophilic characters. Examples of isolation of compounds of different polarities from extracts of various marine invertebrates will be presented in this protocol. Structure elucidation is achieved using recent spectroscopic techniques, especially 2D NMR and mass spectrometry analysis.",
keywords = "purification, structural elucidation, bioactive secondary metabolites, marine invertebrates",
author = "Ebada, {Sherif S.} and RuAngelie Edrada-Ebel and Wenhan Lin and Peter Proksch",
year = "2008",
month = "11",
day = "6",
doi = "10.1038/nprot.2008.182",
language = "English",
volume = "3",
pages = "1820--1831",
journal = "Nature Protocols",
issn = "1754-2189",

}

Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates. / Ebada, Sherif S.; Edrada-Ebel, RuAngelie; Lin, Wenhan; Proksch, Peter.

In: Nature Protocols, Vol. 3, 06.11.2008, p. 1820-1831.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates

AU - Ebada, Sherif S.

AU - Edrada-Ebel, RuAngelie

AU - Lin, Wenhan

AU - Proksch, Peter

PY - 2008/11/6

Y1 - 2008/11/6

N2 - In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) as antitumor agent for the treatment of advanced soft tissue sarcoma. In this protocol, methods for bioactivity-guided isolation, purification and identification of secondary metabolites from marine invertebrates such as sponges, tunicates, soft corals and crinoids are discussed. To achieve this goal, solvent extraction of usually freeze-dried sample of marine organisms is performed. Next, the extract obtained is fractionated by liquid-liquid partitioning followed by various chromatographic separation techniques including thin layer chromatography, vacuum liquid chromatography, column chromatography (CC) and preparative high-performance reversed-phase liquid chromatography. Isolation of bioactive secondary metabolites is usually monitored by bioactivity assays, e.g., antioxidant (2,2-diphenyl-1-picryl hydrazyl) and cytotoxicity (microculture tetrazolium) activities that ultimately yield the active principles. Special care should be taken when performing isolation procedures adapted to the physical and chemical characteristics of the compounds isolated, particularly their lipo- or hydrophilic characters. Examples of isolation of compounds of different polarities from extracts of various marine invertebrates will be presented in this protocol. Structure elucidation is achieved using recent spectroscopic techniques, especially 2D NMR and mass spectrometry analysis.

AB - In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) as antitumor agent for the treatment of advanced soft tissue sarcoma. In this protocol, methods for bioactivity-guided isolation, purification and identification of secondary metabolites from marine invertebrates such as sponges, tunicates, soft corals and crinoids are discussed. To achieve this goal, solvent extraction of usually freeze-dried sample of marine organisms is performed. Next, the extract obtained is fractionated by liquid-liquid partitioning followed by various chromatographic separation techniques including thin layer chromatography, vacuum liquid chromatography, column chromatography (CC) and preparative high-performance reversed-phase liquid chromatography. Isolation of bioactive secondary metabolites is usually monitored by bioactivity assays, e.g., antioxidant (2,2-diphenyl-1-picryl hydrazyl) and cytotoxicity (microculture tetrazolium) activities that ultimately yield the active principles. Special care should be taken when performing isolation procedures adapted to the physical and chemical characteristics of the compounds isolated, particularly their lipo- or hydrophilic characters. Examples of isolation of compounds of different polarities from extracts of various marine invertebrates will be presented in this protocol. Structure elucidation is achieved using recent spectroscopic techniques, especially 2D NMR and mass spectrometry analysis.

KW - purification

KW - structural elucidation

KW - bioactive secondary metabolites

KW - marine invertebrates

U2 - 10.1038/nprot.2008.182

DO - 10.1038/nprot.2008.182

M3 - Article

VL - 3

SP - 1820

EP - 1831

JO - Nature Protocols

T2 - Nature Protocols

JF - Nature Protocols

SN - 1754-2189

ER -