title: Demonstrating formation of potentially persistent transformation products from the herbicides bromoxynil and ioxynil using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
abstract: It is shown that potentially persistent transformation products can be formed from the herbicides bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) and ioxynil (3,5-diiodo-4-hydroxybenzonitrile), and possible leaching to groundwater is discussed. A similar process to the formation of BAM (2,6-dichlorobenzamide) from the herbicide dichlobenil (2,6-dichlorobenzonitrile) can be anticipated as bromoxynil and ioxynil are analogues of dichlobenil and they are degraded by the enzymes nitrilase, nitrile hydratase and amidase. A biodegradation study using cultured Variovorax sp. DSM 11402, a species commonly found in soil, demonstrated that ioxynil and bromoxynil were fully transformed into their corresponding amides in 2-5 days. These amides were not further degraded within 18 days, and formation of other degradation products was not observed. These results are in agreement with biodegradation experiments with dichlobenil. In soil, dichlobenil is transformed into its only observed degradation product BAM, which is persistent and mobile, and has been found in 19% of 5000 samples of Danish groundwater. Variovorax sp. is known to degrade the non-halogenated analogue benzamide, suggesting that degradation of the three amides may be hindered by the halogenated substituents (meta-Br; meta-I; ortho-Cl). This hypothesis is supported by QSAR modelling of fundamental properties. Using a new optimised liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, the sorption and desorption properties of bromoxynil and ioxynil were characterised in sandy topsoil at four concentration levels. The estimated sorption coefficient K(d) was 1.4 L kg(-1) for bromoxynil and 5.4 L kg(-1) for ioxynil, indicating weak to moderate sorption to topsoil. Desorption of the herbicides showed that they were strongly and irreversible bound to the soil (K(des) > K(d)). The amount of herbicide desorbed depended on the initial concentration level. At low levels, K(des) values were higher, indicating stronger binding than at higher levels. The isocratic LC-MS/MS method developed for simultaneous detection of bromoxynil, ioxynil and their main degradation products is described. Using negative electrospray ionisation (ESI-), the detection limits were 0.4-1.0 microg L(-1), with relative standard deviations of 4-10% (n = 10) using direct injection without clean-up steps. The standard curves showed linearity in the range 5-100 microg L(-1) with r(2) > 0.992.
authors.affiliation: Geological Survey of Denmark and Greenland (GEUS), Copenhagen, Denmark.
authors.forename: Marie K K
authors.initials: MK
authors.lastname: Nielsen
authors.forename: Maria S
authors.initials: MS
authors.lastname: Holtze
authors.forename: Bo
authors.initials: B
authors.lastname: Svensmark
authors.forename: René K
authors.initials: RK
authors.lastname: Juhler
mesh_terms: D000327:Adsorption; D001673:Biodegradation, Environmental; D002853:Chromatography, Liquid / Q000379:methods; D004305:Dose-Response Relationship, Drug; D006540:Herbicides / Q000737:chemistry* / Q000378:metabolism; D007462:Iodobenzenes / Q000737:chemistry; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods; D009570:Nitriles / Q000737:chemistry*; D020560:Proteobacteria / Q000378:metabolism*; D021281:Quantitative Structure-Activity Relationship; D012988:Soil Microbiology; D013045:Species Specificity; D014881:Water Supply
chemical_list: D006540:Herbicides; D007462:Iodobenzenes; D009570:Nitriles; C008161:ioxynil; C006826:bromoxynil