4.5 Article

A simple HPLC-MS method for the quantitative determination of the composition of bacterial medium chain-length polyhydroxyalkanoates

Journal

JOURNAL OF SEPARATION SCIENCE
Volume 31, Issue 10, Pages 1739-1744

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/jssc.200800033

Keywords

biopolymers HPLC-MS; medium chain-length PHA; poly((R)-3-hydroxyalkanoate); reductive depolymerization

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Bacterial poly(hydroxyalkanoates) (PHAs) vary in the composition of their monomeric units. Besides saturated side-chains, unsaturated ones can also be found. The latter leads to unwanted by-products (THF ester, secondary alcohols) during acidic cleavage of the polymer backbone in the conventional analytical assays. To prevent these problems, we developed a new method for the reductive depolymerization of medium chain-length PHAs, leading to monomeric diols that can be separated and quantified by HPLC/MS. Reduction is performed at room temperature with lithium aluminum hydride within 5-15 min. The new method is faster and simpler than the previous ones and is quantitative. The results are consistent with the ones obtained by quantitative H-1 NMR.

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