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Validation and evaluation of an HPLC methodology for the quantification of the potent antimitotic compound (+)‐discodermolide in the Caribbean marine sponge Discodermia dissoluta
Author(s) -
Valderrama Katherine,
Castellanos Leonardo,
Zea Sven
Publication year - 2010
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200900852
Subject(s) - sponge , chromatography , chemistry , high performance liquid chromatography , extraction (chemistry) , calibration curve , fraction (chemistry) , dichloromethane , methanol , detection limit , solvent , biology , botany , organic chemistry
The sponge Discodermia dissoluta is the source of the potent antimitotic compound (+)‐discodermolide. The relatively abundant and shallow populations of this sponge in Santa Marta, Colombia, allow for studies to evaluate the natural and biotechnological supply options of (+)‐discodermolide. In this work, an RP‐HPLC‐UV methodology for the quantification of (+)‐discodermolide from sponge samples was tested and validated. Our protocol for extracting this compound from the sponge included lyophilization, exhaustive methanol extraction, partitioning using water and dichloromethane, purification of the organic fraction in RP‐18 cartridges and then finally retrieving the (+)‐discodermolide in the methanol–water (80:20 v/v) fraction. This fraction was injected into an HPLC system with an Xterra RP‐18 column and a detection wavelength of 235 nm. The calibration curve was linear, making it possible to calculate the LODs and quantification in these experiments. The intra‐day and inter‐day precision showed relative standard deviations lower than 5%. The accuracy, determined as the percentage recovery, was 99.4%. Nine samples of the sponge from the Bahamas, Bonaire, Curaçao and Santa Marta had concentrations of (+)‐discodermolide ranging from 5.3 to 29.3 μg/g −1 of wet sponge. This methodology is quick and simple, allowing for the quantification in sponges from natural environments, in situ cultures or dissociated cells.