RP-HPLC法测定中华绒螯蟹主要组织中的恩诺沙星及其代谢产物

Development of RP-HPLC for determination of enrofloxacin and its metabolite in tissues of Chinese mitten-handed crab(Eriocheir sinensis) by RP-HPLC

建立了反相高效液相色谱法(RP-HPLC)测定中华绒螯蟹肌肉、肝胰腺、精巢和卵巢组织内的恩诺沙星、环丙沙星含量。结果表明,恩诺沙星和环丙沙星在四种组织中的平均回收率分别为70.5%±12.61%,70.25%±14.73%;日内精密度分别为3.27%±0.84%,2.14%±0.68%;日间精密度分别为5.38%±2.46%,3.48%±1.16%。恩诺沙星、环丙沙星的最低检测限分别为0.02μg/g和0.004μg/g。以10 mg/kg蟹体重剂量单次肌肉注射给药后,恩诺沙星在中华绒螯蟹肌肉、肝胰腺、精巢和卵巢内的Tmax,Cmax分别为:1 h,4.323±0.56μg/g;1 h,6.042±0.72μg/g;3 h,2.381±0.43μg/g;3 h,2.101±0.29μg/g,各组织的药物消除半衰期(t1/2β)分别为:45.186 h,73.93 h,45.577 h,38.081 h。各组织中均能检测到环丙沙星,但含量均处较低水平,且代谢和消除起伏波动较大。本方法快速、灵敏、准确,适合于中华绒螯蟹组织中恩诺沙星、环丙沙星含量分析。

The methods of reverse-phase high performance liquid chromatography （RP-HPLC） were developed to determine concentration of enrofloxacin（ENR） and ciprofloxacin（CIP） in muscle, hepatopancreas, spermary, ovary of Chinese mitten-handed crab（ Eriocheir sinensis ）. The average recovery of ENR and CIP in four tissues is 70.5% ±12.61%,70.25% ± 14.73%;The intra-day precision of ENR and CIP is 3.27% ± 0.84%, 2.14% ± 0.68%, respectively; The inter-day precision is 5. 38% ± 2.46%, 3. 48% ± 1. 16%, respectively. The lowest detected limit of enrofloxacin in tissues is 0.02 μg/g, and cipmfloxacin is 0. 004 μg/g. ENR was administered to E. sinensis in a single dose（ 10 mg/kg b. w. ） following intramuscular administration. The Tmax, Cmax of ENR in four tissues are 1 h,4.323 ± 0.56μg/g; lh,6.042 ± 0.72μg/g;3 h,2.381 ± 0.43 μg/g;3 h, 2.101 ± 0.29 μg/g, respectively, and the half-life of elimination in these tissues is 45.186 h, 73.93 h, 45. 577 h, 38.081 h, respectively. The method is fast, sensitive, precise and suitable to determine the concentrations of enrofloxacin and cipmfloxacin in Chinese mitten-handed crab, Eriocheir sinensis.