Structural characterisation of humic acid-bound PAH residues in soil by 13C-CPMAS-NMR-spectroscopy: evidence of covalent bonds.
The fate of 13C-labelled phenanthrene and fluoranthene in different soil systems during biodegradation was studied. The soil humic acid fraction was isolated followed by structural characterisation using 13C-cross polarisation magic angle spinning nuclear magnetic resonance spectroscopy (13C-CPMAS-NMR). It could be demonstrated that especially the ratio between the concentrations of polycyclic aromatic hydrocarbons (PAHs) and soil humus matrix limits the usefulness of this analytical tool. Based on these results a ratio of 13C-activity(PAH)/13C-activity(soil) approximately 1.5/1.0 in the test material was suggested. The chemical transformation of a PAH and its bound residue formation in a soil system detected by changes of chemical shifts in the 13C-NMR spectrum was proven for the first time. Structural information obtained by NMR spectra were verified by alkaline hydrolysis of PAH/humus-associations and following identification of cleavage products. Ester-bound phenanthrene metabolites such as 1-hydroxy-2-naphthoic acid, ortho-phthalic acid and 3,4-dihydroxybenzoic acid were detected. Additional structural assignments indicated the presence of ether-bound phenanthrene derivatives as well. Using isotopic labelling techniques a quantitative evaluation of bound residue distribution was undertaken. Fifty to seventy percent of phenanthrene metabolites which could be related to the added 13C(1)-phenanthrene were ester bound via their carboxyl groups.
Institut für Biochemie und Lebensmittelchemie, Universität Hamburg, Germany.
Thomas
T
Käcker
Erhard T K
ET
Haupt
Christian
C
Garms
Wittko
W
Francke
Hans
H
Steinhart
D001673:Biodegradation, Environmental; D004952:Esters / Q000737:chemistry; D006812:Humic Substances / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011084:Polycyclic Aromatic Hydrocarbons / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D012989:Soil Pollutants / Q000032:analysis*
D004952:Esters; D006812:Humic Substances; D011084:Polycyclic Aromatic Hydrocarbons; D012989:Soil Pollutants