D-alanine as a chemical marker for the determination of streptococcal cell wall levels in mammalian tissues by gas chromatography/negative ion chemical ionization mass spectrometry.
A gas chromatography/mass spectrometry method using selected ion monitoring with negative ion detection and methane chemical ionization was employed to quantitate a marker for bacterial peptidoglycan, D-alanine, in mammalian tissues. D-Alanine originating from bacterial peptidoglycan was obscured by substantial amounts of D-alanine generated by racemization from L-alanine present in tissue protein. To overcome this problem, samples were enzymatically treated and hydrolyzed in deuterated hydrochloric acid. Newly formed D-alanine derived from protein was labeled with deuterium and bacterial D-alanine remained unlabeled, enabling differentiation by the molecular weight increase. Butyl heptafluorobutyryl derivatives of the D- and L-amino acids were separated on a fused silica capillary column coated with Chirasil-val. The amounts of bacterial D-alanine found in livers of arthritic rats were consistent with previously reported levels of other carbohydrate-derived markers for bacterial peptidoglycan-polysaccharide complexes.
Ueda
K
K
Morgan
S L
SL
Fox
A
A
Gilbart
J
J
Sonesson
A
A
Larsson
L
L
Odham
G
G
D000409:Alanine / Q000032:analysis*; D000818:Animals; D002473:Cell Wall / Q000032:analysis; D002849:Chromatography, Gas; D006868:Hydrolysis; D008099:Liver / Q000032:analysis / Q000382:microbiology; D013058:Mass Spectrometry; D010457:Peptidoglycan / Q000032:analysis; D051381:Rats; D013291:Streptococcus / Q000032:analysis*
D010457:Peptidoglycan; D000409:Alanine