Liquid chromatography-electrospray ionization mass spectrometry for direct identification and quantification of iophenoxic acid in serum.

A liquid chromatographic-electrospray ionization mass spectrometric technique was developed for direct quantitation of iophenoxic acid (IA) in serum. IA was spiked into canine, feline, bovine, equine, and porcine sera, extracted, and quantified using negative ion monitoring following chromatographic separation on a Luna C18(2) 3 microm (100 mm x 2.1mm) reversed-phase column. The limit of detection was 25 ng/mL and the limit of quantification was 50 ng/mL. Inter- and intra-assay accuracy (86-113% and 87-115%, respectively) and precision (1.8-7.7%) were calculated. Analysis of serum collected from feral pigs, raccoons, and opossums following ingestion of IA-marked baits confirmed the appropriateness of this method for bait acceptance studies.

Matrix-immobilized organoclay for the sorption of polycyclic aromatic hydrocarbons and pentachlorophenol from groundwater.

Sorbent materials consisting of organoclay immobilized onto the surface of a solid support were evaluated for use in pentachlorophenol (PCP) and polycyclic aromatic hydrocarbon (PAH) remediation of groundwater at a creosote-contaminated Superfund site. Cetylpyridinium-exchanged low pH montmorillonite clay (CP-LPHM) was bonded to either sand (CP-LPHM/sand) or granular activated carbon (GAC) (CP-LPHM/GAC) using the free acid form of carboxymethylcellulose as an adhesive. Effluent from an oil-water separator was eluted through equal bed volumes of composite (4 g 3:2 CP-LPHM/GAC or 13 g CP-LPHM/sand), affinity-extracted, and quantitatively analyzed by GC/MS. PCP, naphthalene, fluorene, phenanthrene, pyrene, and total PAHs were initially reduced by both CP-LPHM/GAC (> or =99%, 61%, 99%, > or =99%, 97%, and 94%, respectively) and CP-LPHM/sand (90%, 70%, 94%, 95%, 93%, and 86%, respectively). Complete breakthrough of naphthalene occurred after approximately 15 h of elution through 3:2 CP-LPHM/GAC and 22 h through CP-LPHM/sand. PCP showed complete breakthrough following 18 h of elution through 3:2 CP-LPHM/GAC and 26 h through CP-LPHM/sand. However, 50% breakthrough was not attained for higher molecular weight PAHs, as fluoranthene, pyrene, benzo[a]anthracene, and chrysene continued to be greatly reduced with both 3:2 CP-LPHM/GAC (98%, 95%, 94%, and 95%, respectively) and CP-LPHM/sand (75%, 73%, 76%, and 78%, respectively) after 48 h of continuous elution. Results confirm prior studies, indicating that these organoclay-containing composites have a high capacity for contaminants found in wood preserving waste. Further, results suggest that the inclusion of CP-LPHM may be useful as part of an effective strategy for groundwater remediation of high concentrations of PCP and PAHs, in particular high molecular weight and carcinogenic PAHs.

Prevention of zearalenone-induced hyperestrogenism in prepubertal mice.

Previous methods for the control of zearalenone (ZEN)-induced hyperestrogenism in animals have proven largely ineffective. The main objective in this study was to identify an enterosorbent that decreases the dietary bioavailability, and subsequent estrogenic effects, of ZEN. Initial in vitro screenings in aqueous solution (4 microg ZEN/ml) indicated that an activated carbon (AC) was the most efficient sorbent (99%), followed by a combination of 2 parts AC plus 3 parts HEC (hectorite) (69%), cetylpyridinium-exchanged low-pH montmorillonite (CP-LPHM) clay (58%), hexadecyltrimethylammonium-exchanged low-pH montmorillonite (HDTMA-LPHM) clay (54%), and HEC alone (28%). Results from the adult hydra bioassay suggested that the addition of either AC or HEC effectively decreased the effects of ZEN on Hydra attenuata without toxicity, as was observed with the use of either CP-LPHM or HDTMA-LPHM. Based on these results, AC, HEC, and 2AC:3HEC were evaluated in prepubertal mice. At a dietary inclusion level of 0.8% (w/w), AC alone significantly protected mice against the estrogenic effects induced by 35 mg ZEN/kg feed. Inclusion of 1.2% HEC with the 0.8% AC showed no additional protection; whereas 1.2% HEC alone failed to decrease the estrogenic effects. Ground flaxseed (25% w/w) in the diet also elicited protection, but to a lesser extent. Preliminary studies suggested that three similar carbons failed to decrease ZEN bioavailability. These findings suggest that the AC used in this study may be efficacious as an enterosorbent in animals consuming ZEN-contaminated diets. However, further studies are needed to evaluate the binding specificity, as well as the safety of chronic exposure.

Tri16 is required for esterification of position C-8 during trichothecene mycotoxin production by Fusarium sporotrichioides.

We previously characterized Tri1, a gene required for hydroxylation of the C-8 position during trichothecene mycotoxin biosynthesis in Fusarium sporotrichioides NRRL 3299. Sequence analysis of the region surrounding Tri1 revealed a gene, named Tri16, which could encode an acyltransferase. Unlike the wild-type parent strain NRRL 3299, which accumulates primarily T-2 toxin along with low levels of diacetoxyscirpenol (DAS) and neosolaniol (NEO) and trace amounts of 8-propionyl-neosolaniol (P-NEO) and 8-isobutyryl-neosolaniol (B-NEO), mutants containing a disruption of Tri16 were blocked in the production of the three C-8 esterified compounds T-2 toxin, P-NEO, and B-NEO and accumulated the C-8-hydroxylated compound NEO along with secondary levels of DAS. These data indicate that Tri16 encodes an acyltransferase that catalyzes the formation of ester side groups at C-8 during trichothecene biosynthesis. We also report the presence of a Tri16 ortholog in Gibberella pulicaris R-6380 that is likely linked to a presumably inactive ortholog for Tri1.