A simple HPLC-MS method for the quantitative determination of the composition of bacterial medium chain-length polyhydroxyalkanoates.
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.
Grubelnik
Andreas
A
EMPA, Swiss Federal Laboratories for Materials Testing and Research, Biomaterials, St. Gallen, Switzerland. Fax: +41-71-274-77-88.
Wiesli
Luzia
L
Furrer
Patrick
P
Rentsch
Daniel
D
Hany
Roland
R
Meyer
Veronika R
VR
D000438:Alcohols / Q000737:chemistry; D017607:Aluminum Compounds / Q000032:analysis; D001704:Biopolymers / Q000737:chemistry; D002845:Chromatography / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004952:Esters / Q000737:chemistry; D005285:Fermentation; D018020:Lithium Compounds / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D008956:Models, Chemical; D054813:Polyhydroxyalkanoates / Q000032:analysis* / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013696:Temperature; D013997:Time Factors
D000438:Alcohols; D017607:Aluminum Compounds; D001704:Biopolymers; D004952:Esters; D018020:Lithium Compounds; D054813:Polyhydroxyalkanoates; D011108:Polymers; C042073:lithium aluminum hydride