Determination of metronidazole and hydroxymetronidazole in trout by a high-performance liquid chromatographic method.
A high-performance liquid chromatographic (HPLC) method based on solid phase extraction was developed for determination of metronidazole (MNZ) and its metabolite hydroxymetronidazole (MNZ-OH) in muscle and skin tissues of rainbow trout. Tinidazole (TNZ) was used as internal standard. The compounds were extracted with acetonitrile and the extract was evaporated and redissolved in a mixture of ethyl acetate and hexane (1:2). The extract was cleaned up by solid phase extraction on a silica cartridge. The extract was analysed by reverse phase gradient HPLC on a C18 column followed by ultraviolet detection at 325 nm. The limit of detection was 2.8 micrograms/kg for both compounds in muscle. The estimated limits of detection in skin tissue were 3 micrograms/kg for MNZ and 5 micrograms/kg for MNZ-OH. The mean recoveries of MNZ in muscle calculated without use of internal standard were 93% and 81% at levels of 10 micrograms/kg and 25-100 micrograms/kg respectively. The mean recovery of MNZ-OH in muscle was 79% at a level of 10-100 micrograms/kg. The mean relative repeatability standard deviations on spiked muscle tissue were 3.3% for MNZ and 3.2% for MNZ-OH at a level of 10-100 micrograms/kg. The method was applied to trout given feed containing MNZ in an aquaculture pilot plant. Residues of MNZ and MNZ-OH were detected in muscle and skin tissues shortly after the administration period but not 3 weeks later.
Sørensen
L K
LK
Steins Laboratorium, Brørup, Denmark.
Hansen
H
H
D000818:Animals; D000890:Anti-Infective Agents / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005506:Food Contamination / Q000032:analysis; D008795:Metronidazole / Q000032:analysis* / Q000378:metabolism; D009132:Muscles / Q000737:chemistry*; D012867:Skin / Q000737:chemistry; D014337:Trout
D000890:Anti-Infective Agents; D008795:Metronidazole