文章摘要
杨先乐.中华绒螯蟹血淋巴内盐酸环丙沙星的反相高效液相色谱法的建立[J].水产学报,2001,25(4):
中华绒螯蟹血淋巴内盐酸环丙沙星的反相高效液相色谱法的建立
The method of RP-HPLC determination of ciprofloxacinum in hemolymph of Eriocheir sinenses
投稿时间:2014-04-15  修订日期:2014-04-15
DOI:
中文关键词: 中华绒螯蟹 盐酸环丙沙星 药物动力学 反相高效液相色谱法 血淋巴
英文关键词: Eriocheir sinensis  ciprofloxacinum  pharmacokinetics  RP-HPLC
基金项目:农业部渔业局资助项目
作者单位地址
杨先乐 上海水产大学农业部水产增养殖生态、生理重点开放实验室!上海200090 上海水产大学农业部水产增养殖生态、生理重点开放实验室!上海200090
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中文摘要:
      应用反相高效液相色谱法(RP-HPLC)建立了盐酸环丙沙星在中华绒螯蟹血淋巴内含量的测定方法.本法采用肌肉注射的给药方法,给药剂量为8.17±0.56mg·kg-1和6.25±0.85mg·kg-1.血淋巴从心区抽取.经各种抗凝方法的比较,认为等量的ACD与等量的血淋巴混合,可得到持久的抗凝效果.血淋巴样经甲醇去蛋白,3000r/min离心10min后,取上清液上HPLC.色谱柱,RP-ODS-5μl50×4.6mm;流动相,甲醇-磷酸缓冲液-四丁基溴化铵=250:750:20(v/v/v);流速,l mL·min-1;进样量,2μL;检测波长,280nm.研究表明,本法线性关系与重复性良好;雌、雄蟹平均回收率分别为100.08±3.43%和99.11±8.20%;日间精密度、平均提取精密度分别为1.19±0.92%、1.55±0.01%.用该方法对肌肉注射给药后48h内中华绒螫蟹雄蟹血淋巴中的盐酸环丙沙垡的血药浓度进行检测,揭示出由37.13±0.08μg·mL-1降至5.96±0.18μg·mL-1的代谢规律.
英文摘要:
      A method of determination of ciprof loxacinum in the hemolymph of Chinese mitten-handed crab, Eirocheir sinensis, was established by RP-HPLC. The drug was administered by intramuscular at a dose of 8. 17 ? 0. 56 and 6. 25 ? 0. 85 mg#kg - 1 to male and female crab, repectively . The hemolymph was extracted from the cardiac region. By comparison among various ant-i coagulant methods, the permanent result of ant icoagulation was obtained through ACD mixing with equal volume hemolymph. Hemolymph samples got rid of protein by methanol, centrifuged at 3000 r#min- 1 for 10minutes and then the supernatant was used to HPLC. The RP-ODS- 5 LL 50 @ 4. 6mm ID column was used, with a mobile phase of methano-l phosphate buffer-tertrabutylammonium bromide= 250B750B20 ( v/ v/ v ) ; The f low rate was 1 mL#min - 1 ; the auto-injected volume, 2 LL and the uv detection wavelength, 280nm. The study showed that the linearity and repeatability were good; the average recovery of both male and female crab were : 100. 08 ? 3. 43% and 99. 11 ? 8. 20% ; the intra-day precision, the average extracted-precision were 1. 19 ? 0. 92%, 1. 55 ? 0. 01% and 2. 31 ? 1. 03% , respectively. The law of the metabolism of blood-drug concentration was disclosed dropping from 37. 12 ? 0. 08Lg#mL- 1 to 5. 96 ? 0. 18 Lg#mL- 1, by using RP-HPLC method to determine the ciprofloxacinum in hemolymph within 48 hours in male crab after administration of the drug through intramuscular.
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