Marker residue determination of tritium-labeled ivermectin in the muscle of aquacultured largemouth bass, hybrid striped bass, and yellow perch following oral treatment.

The residue depletion profiles of tritium-labeled ivermectin and its metabolites in the muscle of aquacultured largemouth bass (LMB), hybrid striped bass (HSB), and yellow perch (YP) following oral treatment are reported. Fish were administered ³H-ivermectin at the dose level of 0.1 mg/kg body weight (7-9 µCi) in a gel capsule via stomach tube. At each postdose withdrawal time, six fish of each species were sedated with buffered MS-222 and blood samples taken. Fish were then euthanized, and fillets with adhering skin (scales removed) and bile samples were collected. The muscle fillets were homogenized in dry ice to a fine powder. Aliquots of tissue, plasma, and bile were assayed for total radioactive residue (TRR). The homogenized muscle was extracted in acetonitrile or methanol followed by high-performance liquid chromatographic (HPLC) analysis to determine the presence of parent ivermectin and its potential metabolites. The highest TRR concentrations (ivermectin equivalents) of 53, 45, and 44 ng/g (ppb) were obtained on postdose day 1 for HSB, LMB, and YP, respectively. The TRR depleted most slowly in HSB to 25 ppb at day 91, followed by YP to 19 ppb at day 42 and then by LMB to 22 ppb at day 35. The total residue of ivermectin and its metabolites by HPLC analysis followed the same depletion pattern in the three species. Additionally, the depletion rate of TRR of ³H-ivermectin in the three species followed the pattern bile > plasma > muscle. The results further indicate that one of the polar metabolites of ivermectin could serve as a potential marker residue as an indication of use, rather than the parent ivermectin.

Development of a method to determine albendazole and its metabolites in the muscle tissue of yellow perch using high-performance liquid chromatography with fluorescence detection.

An HPLC method was developed for the determination of albendazole (ABZ) and its metabolites, a sulfoxide (ABZSO), a sulfone (ABZSO2), and albendazole-2-aminosulfone (ABZ-2-NH2SO2), from yellow perch muscle tissue with adhering skin. The muscle tissue samples were made alkaline with potassium carbonate and extracted with ethyl acetate, followed by a series of liquid-liquid extraction steps. After solvent evaporation, the residue was reconstituted in the initial mobile phase combination of the gradient. The mobile phase consisted of a buffer, 50 mM ammonium acetate (pH = 4.0) in 10% methanol-water, and 100% acetonitrile. The gradient was from 20% acetonitrile to 85% acetonitrile. The analytes were chromatographed on an RP Luna C18(2) column and detected by fluorescence with excitation and emission wavelengths of 290 and 330 nm, respectively. The average recoveries from fortified muscle tissue for ABZ (20-100 ppb), ABZ-SO (20-200 ppb), ABZSO2 (8-100 ppb), and ABZ-2-NH2SO2 (20-100 ppb) were 85, 95, 101, and 86%, respectively, with corresponding CV values of 9, 3, 6, and 4%, respectively. Their LOQ values were 10, 10, 1, and 10 ppb, respectively. The procedure was applied to determine ABZ and its major metabolites in the incurred muscle tissue of yellow perch obtained after orally dosing the fish with ABZ.

Residue depletion of albendazole and its metabolites in the muscle tissue of large mouth and hybrid striped bass after oral administration.

The residue depletion profiles of albendazole (ABZ) and its major metabolites: albendazole sulfoxide (ABZ-SO), albendazole sulfone (ABZ-SO(2)) and albendazole aminosulfone (ABZ-2-NH(2)SO(2)) were studied in the muscle tissues of large mouth (LMB) and hybrid striped bass (HSB). A single oral dose of 10mg/kg albendazole was given to the two fish species via intra-gastric tube. The muscle tissues with adhering skin were collected at 8, 16, 24, 48, 72, 96 and 120h post dose from both species. The samples were homogenized in dry ice and subjected to extraction and cleanup procedures. The final sample extracts were analyzed by high performance liquid chromatography. The results indicate that both ABZ and its pharmacologically active metabolite ABZ-SO were retained longer in LMB than in HSB after oral treatment. Albendazole was detectable until 8h or 6.7 degree days ( degrees D) and 48h (40 degrees D) in HSB and LMB, respectively. However, ABZ-SO was detectable up to 48h (40 degrees D) and 96h (80 degrees D) in HSB and LMB, respectively. Among the inactive metabolites, ABZ-SO(2) was present in both fish species; however, ABZ-2-NH(2)SO(2) was detected only in LMB.

Kinetics of hepatic phase I and II biotransformation reactions in eight finfish species.

Hepatic microsomes and cytosols of channel catfish (Ictalurus punctatus), rainbow trout (Oncorhynchus mykiss), Atlantic salmon (Salmo salar), red tilapia (Oreochromis sp.), largemouth bass (Micropterussalmoides), striped bass (Morone saxatilis), hybrid striped bass (M. saxatilis x M. crysops), and bluegill (Lepomis macrochuris) (n=8) were used to study the kinetics of phase I (ECOD, EROD, PROD, BROD) and phase II (UDP-glucuronosyltransferase (UDPGT)-, sulfotransferase (ST)- and glutathione-s-transferase (GST)-mediated) reactions. The best catalytic efficiency for ECOD and GST activities was performed by channel catfish, Atlantic salmon, rainbow trout and tilapia. The highest EROD catalytic efficiency was for Atlantic salmon. None of the species had either PROD or BROD activities. Rainbow trout had very similar UDPGT catalytic efficiency to tilapia, channel catfish, Atlantic salmon, largemouth bass and bluegill. Sulfotransferase conjugation had no significant differences among the species. In summary, tilapia, channel catfish, Atlantic salmon and rainbow trout had the best biotransforming capabilities; striped bass, hybrid striped bass and bluegill were low metabolizers and largemouth bass shared some capabilities with both groups.

Determination of albendazole and its metabolites in the muscle tissue of hybrid striped and largemouth bass using liquid chromatography with fluorescence detection.

A liquid chromatographic method was developed for the determination of albendazole and its metabolites albendazole sulfoxide, albendazole sulfone, and albendazole-2-aminosulfone from largemouth and hybrid striped bass muscle tissue with adhering skin. The muscle tissue samples were made alkaline with potassium carbonate and extracted with ethyl acetate. The extracts were further subjected to cleanup by using a series of liquid-liquid extractions. After solvent evaporation, the residue was reconstituted in mobile phase and chromatographed. The chromatography was carried out on a reversed-phase Luna C18 column, using acetonitrile-methanol buffer as the mobile phase. The analytes were detected by fluorescence with excitation and emission wavelengths of 290 and 330 nm, respectively. The average recoveries from the fortified muscle tissue of the 2 fish species for albendazole (25-100 ppb), albendazole sulfoxide (8.75-52.5 ppb), albendazole sulfone (1-10 ppb), and albendazole-2-aminosulfone (10-100 ppb) were 89, 82, 99, and 74%, respectively. The coefficient of variation for each compound was <20% in all cases. The procedure was applied to the determination of albendazole and its 3 metabolites in the muscle tissue of the 2 fish species after orally dosing them with albendazole.

Fish drug analysis--Phish-Pharm: a searchable database of pharmacokinetics data in fish.

Information about drug residues and pharmacokinetic parameters in aquatic species is relatively sparse. In addition, it is difficult to rapidly compare data between studies due to differences in experimental conditions, such as water temperatures and salinity. To facilitate the study of aquatic species drug metabolism, we constructed a Fish Drug/Chemical Analysis Phish-Pharm (FDA-PP) database. This database consists of more than 400 articles that include data from 90 species (64 genera) of fish. Data fields include genus, species, water temperatures, the average animal weight, sample types analyzed, drug (or chemical) name, dosage, route of administration, metabolites identified, method of analysis, protein binding, clearance, volume of distribution in a central compartment (Vc) or volume of distribution at steady-state (Vd), and drug half-lives (t((1/2))). Additional fields list the citation, authors, title, and Internet links. The document will be periodically updated, and users are invited to submit additional data. Updates will be announced in future issues of The AAPS Journal. This database will be a valuable resource to investigators of drug metabolism in aquatic species as well as government and private organizations involved in the drug approval process for aquatic species.

Determination of albendazole and its major metabolites in the muscle tissues of Atlantic salmon, tilapia, and rainbow trout by high performance liquid chromatography with fluorometric detection.

A liquid chromatographic procedure for the determination of albendazole ([5-(propylthio)-1H-benzimidazol-2yl]carbamic acid methyl ester) and its major metabolites, albendazole sulfoxide, albendazole sulfone, and albendazole-2- aminosulfone in rainbow trout, tilapia, and salmon muscle with adhering skin tissue is described. The muscle tissue samples are made alkaline with potassium carbonate and extracted with ethyl acetate. The extracts are further subjected to cleanup by utilizing a number of liquid-liquid extraction steps. After solvent evaporation, the residue is reconstituted in mobile phase and chromatographed. The chromatography is carried out on a reversed phase Luna C(18) column, using acetonitrile/methanol/buffer as a mobile phase and a fluorescence detector. The average recoveries from the fortified muscle tissue of the three fish species for albendazole (25-100 ppb), albendazole sulfoxide (15.5-62 ppb), albendazole sulfone (1-10 ppb), and albendazole-2- aminosulfone (10-100 ppb) were 94, 77, 82, and 67%, respectively. The average CV for each compound was < or =10%. The procedure was validated and then applied to the determination of albendazole and its three major metabolites in the muscle tissue of the three fish species obtained after orally dosing with albendazole.