Synthesis of a molecularly imprinted polymer for the solid-phase extraction of betulin and betulinic acid from plane bark.
BACKGROUND
Plant extracts are usually complex mixtures of various polarity compounds and their study often includes a purification step, such as solid-phase extraction (SPE), to isolate interest compounds prior analytical investigations. Molecularly imprinted polymers (MIPs) are a new promising type of SPE material which offer tailor-made selectivity for the extraction of trace active components in complex matrices. Numerous specific cavities that are sterically and chemically complementary of the target molecules, are formed in imprinted polymers. A molecularly imprinted polymer (MIP) was synthesised in order to trap a specific class of triterpene, including betulin and betulinic acid from a methanolic extract of plane bark.


METHODS
Imprinted polymers were synthesised by thermal polymerisation of betulin as template, methacrylic acid (MAA) or acrylamide (AA) as functional monomer, ethylene glycol dimethacrylate as crosslinking agent and chloroform as porogen. Afterwards, MAA- and AA-MIPs were compared with their non-imprinted polymers (NIPs) in order to assess the selectivity vs betulin and its derivatives. Recovered triterpenes were analysed by HPLC during MIP-SPE protocol.


RESULTS
After SPE optimisation, the MAA-imprinted polymer exhibited highest selectivity and recovery (better than 70%) for betulin and best affinity for its structural analogues. Thus, a selective washing step (chloroform, acetonitrile) removed unwanted matrix compounds (fatty acids) from the SPE cartridge. The elution solvent was methanol. Finally, the MAA-MIP was applied to fractionate a plane bark methanolic extract containing betulin and betulinic acid.


CONCLUSIONS
This study demonstrated the possibility of direct extraction of betulin and its structural analogues from plant extracts by MIP technology.
Claude
Berengere
B
Institut de Chimie Organique et Analytique, Université d'Orléans, CNRS FR 2708, UMR 6005, 45060 Orléans, France. berengere.claude@univ-orleans.fr
Viron-Lamy
Cecile
C
Haupt
Karsten
K
Morin
Philippe
P
D029401:Betulaceae / Q000737:chemistry*; D054802:Molecular Imprinting / Q000379:methods*; D053978:Pentacyclic Triterpenes; D024301:Plant Bark / Q000737:chemistry; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D052616:Solid Phase Extraction / Q000295:instrumentation / Q000379:methods*; D014315:Triterpenes / Q000302:isolation & purification*; D000094062:Betulinic Acid
D053978:Pentacyclic Triterpenes; D011108:Polymers; D014315:Triterpenes; C002503:betulin; D000094062:Betulinic Acid