Development of analytical methods for the determination of 3-hydroxy fatty acids and muramic acid as bacterial markers in airborne particles and settled dust

J Chromatogr A. 2023 Jan 11:1688:463708. doi: 10.1016/j.chroma.2022.463708. Epub 2022 Dec 9.

Abstract

Inhalation of airborne bacteria in indoor environments is known to be associated with respiratory diseases. Analytical methods for the determination of 3-hydroxy fatty acids (3-OHFAs) and muramic acid (MA) as chemical markers of gram-negative and gram-positive bacteria, respectively, were developed for airborne particle and dust samples in this study. 3-OHFAs as markers of endotoxin were released and esterified during the hydrolysis process under methanolic acid conditions, and their hydrolysates, i.e., 3-OHFA methyl esters, were cleaned up by solid-phase extraction using silica sorbent that provided more effective separation from interferents than polymeric sorbent through elution pattern. The SPE eluent was analyzed by GC-MS/MS measurement after the trimethylsilylation reaction. The recovery of the method ranged from 82.1 % to 103.2 %, with a limit of detection ranging from 0.5 to 1.1 ng/filter and good linearity (R2 > 0.991). For the analysis of MA, muramic acid methyl ester (MAME), a product formed during methanolic hydrolysis, was selected as a specific marker of peptidoglycan. It was the first proposed compound identified and confirmed with MS and MS/MS spectra using high-resolution measurement. In particular, the measurement of MAME providing 12.5 times greater sensitivity than MA with the application of the LC-MS/MS method is one of the notable findings of this study. The recovery by simple liquid extraction was 99.4 % following the removal of the hydrophobic matrix and neutralization with solvent reconstruction. The method displayed a LOD of 0.7 ng/filter and linearity (R2) of 0.997 through a simple pretreatment process. Both developed methods were applied and evaluated by determining 3-OHFAs and MA in airborne particles collected from multipurpose facilities and settled dust in the laboratory and office.

Keywords: 3-hydroxy fatty acid; Airborne dust; Bacterial marker; Muramic acid.

MeSH terms

  • Bacteria / chemistry
  • Chromatography, Liquid
  • Dust* / analysis
  • Fatty Acids / analysis
  • Muramic Acids* / analysis
  • Tandem Mass Spectrometry

Substances

  • Dust
  • Muramic Acids
  • Fatty Acids