Polygoprep c18

The fermentation and extraction conditions were the same as described in the feeding experiments, except that H. aurantiacus 114-95^T was grown on a 20 L scale in VNY medium and that the cultures were only supplemented with non-labeled D-glucose. The extract that was obtained after the XAD-2 elution was dissolved in 20% (v/v) aqueous MeOH and fractionated by flash column chromatography over Polygoprep C18 (Macherey-Nagel) using an increasing concentration of MeOH in water. Fractions containing 1 were identified by LC–MS analysis, pooled and purified by reversed-phase HPLC on a Shimadzu UFLC liquid chromatography system equipped with a Nucleodur C18 HTec column (VP 250 × 10 mm, 5 μm; Macherey-Nagel) using a gradient from 10% (v/v) MeOH in H₂O (+ 0.1% trifluoroacetic acid) to 100% (v/v) MeOH over 30 min. The purity of the isolated herpetopanone determined by HPLC was >95% at all wavelengths (compared to sum of total peak areas).

The particle sizes vary from 40 to 1,000 μm, the specific surface area from 10 to 550 m² g⁻¹ and the cost per weight from 111 to 18,600 € kg⁻¹. Adsorbents were chosen based on their potential for terpene purification. Those include mostly hydrophobic polymeric or inorganic materials with possible functionalizations (see Table 1). The Bio-Beads™ S-X12 and SM-2 were both sponsored by Bio-Rad Laboratories, Germany. The former is a polymeric size exclusion material, which gives defined porosities in a swollen state. Polygoprep™ C18 was purchased from Macherey Nagel, Germany. Rensa^® PY and RP were purchased from Biotage, Sweden. Silica gel 60, Amberlite^® XAD-2 and XAD-7 from Sigma-Aldrich, USA. The dimensions of the chromatographic glass columns (Omnifit^®) were 0.66 cm and 1 cm in diameter, with a variable height from 0 to 15 cm. Frits with a mean pore size of 10 μm and made of PTFE were used. Ethanol was >99% pure, acetonitrile >98% and both were purchased from VWR, Germany. β-caryophyllene (CAS-Nr. 87-44-5, see Figure 1) was purchased from Molekula GmbH with a purity of >98%.