Residue depletion of melamine and cyanuric acid in catfish and rainbow trout following oral administration.

The intentional addition of triazines such as melamine to animal feeds and the lack of information about residue accumulation in food animals caused global concerns for food safety during 2007 and 2008. We report the results of a good laboratory practices (GLP) study to determine melamine and cyanuric acid residues in catfish and trout filets harvested at 1, 3, 7, 14, 28, and 42 days after a single oral dose of 20 mg/kg body weight of melamine, cyanuric acid, or melamine and cyanuric acid together. Peak melamine concentrations were 12.73 mg/kg (ppm) in catfish (mean = 9.98), 12.26 mg/kg in trout (mean = 7.89) on day 1. Within 7 days (catfish) or 14 days (trout) residues were <2.5 mg/kg, a level in foods accepted by many risk assessors worldwide to be unlikely to pose health risks to consumers. Peak cyanuric acid residues also occurred on day 1, 0.68 mg/kg in catfish (mean = 0.46), 2.59 mg/kg in trout (mean = 0.86). Cyanuric acid muscle residues were <2.5 mg/kg by day 3. The half-lives for melamine and cyanuric acid ranged between 1 and 4 days. Renal crystals formed in fish given both melamine and cyanuric acid, persisting for weeks after the single dose.

Metabolism and depletion of albendazole in the muscle tissue of channel catfish following oral treatment.

The residue depletion of albendazole (ABZ) and its metabolites was studied in channel catfish muscle tissue. Channel catfish were dosed once with 10 mg/kg ABZ via stomach tube with manual restraint. Muscle tissue samples were collected at 8, 16, 24, 48, 72, 96 and 120 h postdose. A high-performance liquid chromatographic method was used to assay ABZ and its major metabolites: ABZ sulfoxide (ABZ-SO), ABZ sulfone (ABZ-SO2) and ABZ aminosulfone (ABZ-2-NH2SO2) in the muscle tissue. The results indicate that ABZ and ABZ-SO were present in low concentrations, i.e. <15 and <10 microg/kg, respectively, at 8 h postdose in catfish muscle with and without skin. ABZ-SO2 was present at 1 microg/kg concentration levels until 48 h in muscle alone and 72 h in muscle with skin. ABZ-2-NH2SO2 was not detected at any withdrawal periods.

Determination of 14C residue in eggs of laying hens administered orally with [14C] sulfaquinoxaline.

Ten layer hens were dosed for 5 consecutive days with 6.2 mg kg(-1) [14C] sulfaquinoxaline (SQX). Eggs were collected from the hens during the 5-day dosing period and during a 10-day post-dose withdrawal period. Egg yolk and albumen were separated and assayed for total radioactive residues (TRR) using a combustion oxidizer and liquid scintillation counting techniques. Significant amounts of radioactivity were detected on the second day of dosing (greater than 24h after the initial dose) in both egg yolk and albumen. First eggs were collected about 8 h after dosing; the second-day eggs were collected during 8-h period after the second dose. Radioactive residues reached a maximum on the fifth day of dosing in albumen, whereas on the second day of withdrawal in egg yolk, the peak TRR levels in albumen were about threefold higher than in yolk. Thereafter, the TRR levels declined rapidly in albumen and were detectable up to withdrawal day 6, whereas the TRR levels in egg yolk declined more slowly and were detectable up to withdrawal day 10. High-performance liquid chromatography analysis indicated that the parent drug sulfaquinoxaline was the major component in both the egg albumen and yolk. Additionally, this work suggests that egg yolk is the appropriate matrix for monitoring SQX residues

Metabolism and residue depletion of albendazole and its metabolites in rainbow trout, tilapia and Atlantic salmon after oral administration.

Metabolic and residue depletion profiles of albendazole (ABZ) and its major metabolites in three fish species, rainbow trout, tilapia and Atlantic salmon are reported. Based on these profiles, similarities (or dissimilarities) between species will determine the potential to group fish species. ABZ at 10 mg/kg body weight was incorporated into fish food formulated in a gelatin base or in gel capsule and fed as a single dose to six fish from each species. Rainbow trout were held three each in a partitioned 600-L tank. Tilapia and Atlantic salmon were housed in separate 20-L tanks. Samples of muscle with adhering skin were collected at 8, 12, 18, 24, 48, 72, and 96 h postdose from trout kept at 12 degrees C, at 4, 8, 12, 24, 48, 72, 96, 120, and 144 h postdose from tilapia kept at 25 degrees C and at 8, 14, 24, 48, 72, and 96 h postdose from Atlantic salmon kept at 15 degrees C. The samples were homogenized in dry ice and subjected to extraction and cleanup procedures. The final extracts were analyzed for parent drug ABZ and its major metabolites, albendazole sulfoxide (ABZ-SO), albendazole sulfone (ABZ-SO2) and albendazole aminosulfone using high-performance liquid chromatography with fluorescence detection. ABZ was depleted by 24 h in trout and tilapia and by 48 h in salmon; ABZ-SO, a pharmacologically active metabolite, was depleted by 48 h in tilapia, by 72 h in rainbow trout and was present until 96 h in salmon; and low levels of ABZ-SO2 and albendazole aminosulfone, both inactive metabolites, were detectable at least till 96 h in all three fish species.

Identification of cephapirin metabolites and degradants in bovine milk by electrospray ionization--ion trap tandem mass spectrometry.

Liquid chromatography-ion trap tandem mass spectrometry (LC-MS/MS) with electrospray ionization was used to identify cephapirin metabolites and degradants in milk from cows dosed with cephapirin. The milk was extracted according to a previously published procedure. Structures for various components were tentatively identified by their molecular weight, product ion mass spectra, and/or correspondence to standard mass spectra. These components may have occurred as metabolites or as degradants that occurred on storage or during extraction. Compounds identified in the milk included cephapirin, desacetylcephapirin, cephapirin lactone, hydrolyzed cephapirin, and a reduced cephapirin lactone that has not previously been reported. Methylcephapirin was also identified, possibly as a trace contaminant in the formulation. Analysis of incurred milk extracts showed that cephapirin and desacetylcephapirin are the major residues in milk. Desacetylcephapirin residues persisted about as long as the parent drug. The detection limit for both residues by LC-MS/MS was approximately 1 ng/mL in milk. These results have implications for microbiological methods or rapid test kits, if such methods or kits respond to cephapirin metabolites and degradants present in the milk.

Determination of trichlormethiazide in bovine milk by high-performance liquid chromatography.

A liquid chromatographic procedure was developed and validated for the quantitative determination of trichlormethiazide (TCMTZ) in bovine milk. Whole milk was defatted by initial centrifugation at 4 degrees C. The resulting skim milk was treated with lead acetate and acetonitrile, vortex mixed, and centrifuged. The acetonitrile from the supernatant was back extracted in ethyl acetate. The organic solvent mixture which contained TCMTZ was further treated with sodium tungstate, vortex mixed, and centrifuged. The top organic layer was removed and evaporated to dryness; the resulting residue was reconstituted in the mobile phase, and the final extract was analyzed by high-performance liquid chromatography (HPLC). The HPLC conditions employed included a polymer column, a mobile phase consisting of 30% acetonitrile or 30% acetonitrile-tetrahydrofuran (2:1, v/v) in a phosphate buffer (pH 3), and a UV detection at 225 nm. The average recoveries of TCMTZ from milk fortified at 7, 14, 35, 70, and 140 ppb were 88, 93, 117, 110, and 99%, respectively, with corresponding C.V. values of 7, 18, 11, 9, and 21%. The method was validated by assaying milk obtained from a cow dosed with Naquasone. TCMTZ concentration was detected only in the 8 h post dose milk samples and was determined to be 6 ppb.

Colorimetric determination of selenium in mineral premixes .

A method is described for determination of sodium selenite or sodium selenate in mineral-based premixes. It is based on the formation of intense-yellow piazselenol by Se(IV) and 3,3'-diaminobenzidine. Mineral premixes typically contain calcium carbonate as a base material and magnesium carbonate, silicon dioxide, and iron(III) oxide as minor components or additives. In this method, the premix is digested briefly in nitric acid, diluted with water, and filtered to remove any Iron(III) oxide. Ethylenediaminetetraacetic acid and HCl are added to the filtrate, which is heated to near boiling for 1 h to convert any selenate to selenite. After heating, the solution is buffered between pH 2 and 3 with NaOH and formic acid and treated with NH2OH and EDTA; any Se present forms a complex with 3,3'-diaminobenzidine at 60 degrees C. The solution is made basic with NH4OH, and the piazselenol is extracted into toluene. The absorbance of the complex in dried toluene is measured at 420 nm. The method was validated independently by 2 laboratories. Samples analyzed included calcium carbonate fortified with 100, 200, and 300 micrograms Se in the form of sodium selenite or sodium selenate, a calcium carbonate premix containing sodium selenite, a calcium carbonate premix containing sodium selenate, and a commercial premix; 5 replicates of each sample type were analyzed by each laboratory. Average recoveries ranged from 89 to 109% with coefficients of variation from 1.2 to 13.6%.