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

J Sep Sci. 2008 Jun;31(10):1739-44. doi: 10.1002/jssc.200800033.

Abstract

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 (1)H NMR.

MeSH terms

  • Alcohols / chemistry
  • Aluminum Compounds / analysis
  • Biopolymers / chemistry
  • Chromatography / methods
  • Chromatography, High Pressure Liquid / methods*
  • Esters / chemistry
  • Fermentation
  • Lithium Compounds / analysis
  • Magnetic Resonance Spectroscopy / methods
  • Mass Spectrometry / methods*
  • Models, Chemical
  • Polyhydroxyalkanoates / analysis*
  • Polyhydroxyalkanoates / chemistry
  • Polymers / chemistry
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Temperature
  • Time Factors

Substances

  • Alcohols
  • Aluminum Compounds
  • Biopolymers
  • Esters
  • Lithium Compounds
  • Polyhydroxyalkanoates
  • Polymers
  • lithium aluminum hydride