Fast isolation of the environmentally relevant halogenated natural product MHC-1 by means of countercurrent chromatography.

Environmentally relevant halogenated natural products (HNPs) are frequently similarly high concentrated in marine biota as major anthropogenic persistent organic pollutants (POPs). The lack of widely available reference standards, however, hampers the in-depth research of several HNPs. For instance, (1R,2S,4R,5R,1'E)-2-bromo-1-bromomethyl-1,4-dichloro-5-(2'-chloroethenyl)-5-methylcyclohexane (MHC-1), which is produced by species referred to the red seaweed Plocamium cartilagineum has not yet been synthesized due to its complex structure and stereochemistry. For this reason, we aimed to establish a method for fast isolation of mg-amounts of MHC-1 from its natural producer based on countercurrent chromatography (CCC). Five biphasic solvent systems were tested and finally, the solvent system acetonitrile/n-hexane/toluene (9:9:2, v/v/v) was selected for the separations due to its suitable partition coefficient of MHC-1 (K = 0.52). n-Hexane extracts of dried P. cartilagineum were directly injected into the CCC system. Four subsequent CCC runs from three samples of Plocamium cartilagineum (two from Heligoland, Germany and one from Brittany, France) could be performed with high reproducibility. Together, the main fraction provided ~16 mg MHC-1 in a purity of >97% according to GC/FID, GC/ECNI-MS and NMR analysis. This amount could be used to prepare ~1600 quantitative standard solutions of MHC-1. The high MHC-1 content in the seaweed sample collected at Brittany indicated that this area was another hotspot of MHC-1.