Figure 5.
Separation of oligo‐isoprenoid mixtures by FPLC, HPLC and HPLC‐MS. 200 μl oligo‐isoprenoid solution in methanol (prepared by digestion of polyisoprene with LcpK30 as described in the main text) was applied to a reversed‐phase FPLC column (Pep RPC HR 5/5, 1 ml bed volume) and separated by a step gradient from 50% water: methanol to 100% methanol (green line) (left image). Absorption at 210 nm (red line) was used to fractionate peaks representing different oligo‐isoprenoids (A1‐A11, corresponding to n = 1–11). Aliquots of each separately collected fraction (A1 to A11) were applied to analysis via HPLC. An overlay image consisting of all eleven HPLC chromatograms is shown on the right. The superposition of the chromatograms shows the high resolution power of the used FPLC column. The masses (m/z values) of each isolated compound were confirmed by HPLC‐ESI‐MS and are provided in Table 1.