Enantiomeric separation of quorum sensing autoinducer homoserine lactones using GC-MS and LC-MS

Abstract

Homoserine lactones (HSLs) are signaling molecules synthesized by Gram-negative bacteria in order to communicate in a process termed "quorum sensing." Until recently, only the l-stereoisomers of HSLs were thought to be produced and able to incite quorum sensing. However, recent studies have shown that select Gram-negative bacteria additionally produce non-trivial amounts of d-HSLs which may also play a role in quorum sensing. Current methods for the separation of HSL enantiomers cannot effectively separate all classes of HSLs and its enantiomers. More robust methods of separation and detection of d-HSLs are necessary. We have developed rapid and selective methods using liquid chromatography (LC) and gas chromatography (GC) coupled with mass spectrometry (MS) which can simultaneously enantiomerically separate all classes of HSLs. The advantages of these methods are in the MS compatibility as well as the ability to enantiomerically separate all classes of HSLs in a single run. The first enantiomeric separations of oxo- and hydroxy-HSLs by GC-MS, through the use of N,O-bis(trimethylsilyl)trifluoroacetamide-derivatizing reagents are discussed.