Continuous preparation of two opioïd peptides and recycling of organic solvent using liquid/liquid extraction coupled with aluminium oxide column during haemoglobin hydrolysis by immobilized pepsin

Ion-pairing separation of bioactive peptides using an aqueous/octan-1-ol micro-extraction system from bovine haemoglobin complex hydrolysates

“…A comparison of the haemorphin partition coefficients has been realized ( Table 1) with alkyl-sulfonic acid in the best extraction conditions, without alkyl-sulfonic acid addition and with previous studies where the extraction occurred in an aqueous/butan-2-ol-octan-1-ol biphasic medium [24,25]. It is observed a high Fig.…”

“…Previously, aluminium oxide support has allowed to haemorphin and haem separate in a butan-2-ol-octan-1-ol mixture [25]. This support could be tested to haemorphin recover in the octan-1-ol organic phase after ionpairing extraction.…”

“…[24,25] LVVh-7 partition coefficient 0.04 ± 0.01 1.64 ± 0.10 0.33 VVh-7 partition coefficient 0.03 ± 0.01 1.60 ± 0.15 0.31 partition coefficient increased when the extraction occurred in the presence of the alkyl-sulfonic acid, with an improvement of about 40 times the values for each haemorphin compared with experiments without ion-pairing agent, and a 5 time improvement compared with the previous studies [24,25]. These comparisons demonstrated the effective extraction of the opioid peptides thanks to the alkyl-sulfonic agent using and the real potential of the peptide selective separation acquired with ion-pairing extraction in controlled liquid/liquid biphasic medium.…”

“…For mixtures containing more than hundred different peptides, this strategy appears a good means to separate peptides by modifying their hydrophobicity thanks to ion pair agent-peptide complex formation and in using liquid/liquid medium to selectively extract some bioactive peptides. To allow this study, two opioid peptides, LVV-haemorphin-7 and VV-haemorphin-7, have been chosen because previous studies have shown that these peptides can be selectively extracted in a biphasic system containing butan-2-ol-octan-1-ol but a very low partition coefficient is obtained [24,25]. Therefore, ion-pairing extraction has been applied with alkyl-sulfonic acids, chosen as acidic amphipathic surface-active agents, to increase the lipophilicity of the active peptides via hydrophobic ion-pairing.…”

“…Therefore, ion-pairing extraction has been applied with alkyl-sulfonic acids, chosen as acidic amphipathic surface-active agents, to increase the lipophilicity of the active peptides via hydrophobic ion-pairing. Action of the alkyl-sulfonic acid counter anions as lipophilicity modulator at different pH and with different alkyl chain lengths, and a partition coefficient comparison with and without alkyl-sulfonic acid have been discussed in previous studies [24,25] and have demonstrated the potential of this system for selective extraction of peptides.…”