Extracts found to contain nuisance or pan-toxic metabolites were subjected to preparative mass-guided chromatography and edited to selectively remove such compounds. A standard, reverse phase elution gradient was used for each crude extract. A portion of the LC output was diverted to a fraction collector, and separated based on retention time and UV absorbance (220 nm). A portion of the eluent was simultaneously subjected to HRESIMS mass analysis to identify compounds in each fraction based on the molecular ion (M+H). Fractions containing previously identified nuisance compounds were maintained in separate collection vessels, while the remaining fractions and elution waste were recombined to form a 'nuisance compound subtracted' edited extract.
Preparative HPLC was performed on a Shimadzu LC-20 AT system equipped with an ultraviolet detector. Reverse Phase HPLC samples were dissolved in MeOH and eluted with a gradient of H2O to either ACN or MeOH on a semi-preparative Phenomenex C18 column (10 μm, 100 Å, 250× 10 mm). HRESIMS was obtained on an Agilent 6230A LC/TOF with positive mode ionization.
Preparative HPLC was performed on a Shimadzu LC-20 AT system equipped with an ultraviolet detector. Reverse Phase HPLC samples were dissolved in MeOH and eluted with a gradient of H2O to either ACN or MeOH on a semi-preparative Phenomenex C18 column (10 μm, 100 Å, 250× 10 mm). HRESIMS was obtained on an Agilent 6230A LC/TOF with positive mode ionization.