Inside amber: the structural role of succinic acid in Class Ia and Class Id resinite.

For the first time, molecular evidence of the structural role played by succinic acid within the macromolecular structure of Class Ia and Class Id resinite is presented. Using a novel gas chromatographic methodology, communol (Class Ia) and ozol (Class Id) moieties within the polylabdane matrix are shown to be cross-linked with succinic acid. Samples were analyzed using pyrolysis-gas chromatography-mass spectrometry with in situ hexamethyldisilazane derivatization, using a thermal separation probe to perform the pyrolysis and sample introduction. The relatively slow rate of heating and prolonged pyrolysis of resinites using this new methodology, combined with the use of a mild derivatization reagent, allowed communol pyrolysates from Class Ia resinite and ozol pyrolysates from Class Id resinite to elute with unbroken succinyl ester cross-linkages. These results provide direct molecular evidence that the key role of succinic acid within Class Ia and Class Id resinite is to cross-link the macromolecular structure. In the Class Id resinite, the methodology also allowed the detection of succinyl ester linkages between ozol pyrolysates and dehydroabietol, thus demonstrating that nonpolymerized diterpenes contribute structurally to the macromolecular structure of Class Id resinite.