Withdrawal Period of Oxolinic Acid in Cobia (Rachycentron canadum): Validation of an LC-MS/MS Method

Abstract

Oxolinic acid (OXO) has been routinely used in aquaculture as both prophylactic and chemotherapeutic agents, principally against systemic bacterial infections. The residue depletion of OXO was investigated in cobia following multiple (5 days) in-feed dosing (30 and 60 mg/kg body weight/day) at 25°C; we assessed OXO concentrations of liver and muscle (+skin) samples post-treatment for three weeks. The OXO residues were quantified using a validated ultra-high-performance liquid chromatographytandem mass spectrometry (UPLC-MS/MS) method and the results indicated recoveries of 86.6-95.8%, with relative standard deviations (RSD) less than 8.5% for intra- and inter-day precisions. Within the range of 10−500 ng/g, the calibration curves for all matrices presented a determination coefficient greater than 0.99. Limit of detection and quantification were estimated at 6 and 10 ng/g, respectively. In order to account for the high variability of the residue data, a safety span corresponding to 50% of depletion time could be seen as appropriate. Considering a maximum residue limit (MRL) of 50 ng/g established by the Taiwan FDA for OXO in muscle with skin in natural proportions, a withdrawal period of 15 days was calculated for the sum of 10day depletion time and 5-day safety span. These results revealed that the analytical method is developed for residue OXO and that the withdrawal period is appropriate.