达氟沙星在史氏鲟体内药物代谢动力学比较研究

THE PHARMACOKINETIC OF DANOFLOXACIN IN HEALTHY AND DISEASED ACIPENSER SCHRENCKII INFECTED BY AEROMONAS HYDROPHILA

采用高效液相色谱法测定以10mg/kg体重剂量静脉注射和口服给药后史氏鲟血浆中达氟沙星的浓度。该法采用C18色谱柱,流动相为乙腈-水相(15∶85),荧光激发波长和发射波长分别为280nm和450nm,样品用甲醇沉淀蛋白,离心取上清液进样。达氟沙星在0.005—1.0μg/mL范围内线性关系良好,本方法的最低检测限为0.005μg/mL。健康鱼单剂量静注达氟沙星(10mg/kg),其药时数据符合无吸收的三室开放模型,方程为C=5.830-5.582t+4.162-1.157t+0.852-0.029t,主要动力学参数如下:t1/2α0.552h;t1/2β22.186h;AUC34.226mg/(L.h);V 10.922L/kg;Vb10.144L/kg;ke 10.317h.Ah感染组的V1减小至0.290L/kg,静注感染组鱼体内达氟沙星的消除没有显著的改变。健康口服组数据结果符合一级吸收二室开放模型,血药浓度和时间方程为C=1.278e-0.073t+0.177e-0.089t-1.455e-0.329t。药动学常数分别为:t1/2ka9.491h,t1/2β78.267h,Tmax6.284h,Cmax0.791mg/mL;α0.073h。但Ah感染改变达氟沙星口服给药后在史氏鲟体内的吸收、分布和消除。分布速率常数降低为0.050/h。消除减慢,消除半哀期延长为93.988h,达峰时间延长为至9.060h,峰浓度降低为0.585mg/mL。口服达氟沙星水溶液,健康及感染组史氏鲟对达氟沙星生物利用度分别为96.503%和94.435%。本实验结果表明达氟沙星在健康史氏鲟体内分布广泛、吸收较完全。感染Ah对达氟沙星在史氏鲟体内的吸收、分布及消除规律均有不同程度的影响,其中口服给药的影响更为显著。达氟沙星可用于史氏鲟感染Ah的治疗。

The pharmacokinetics of danofloxacin in healthy and Aeromonas hydrophila infected Acipenser schrenckii were investigated, so as to provide scientific basis for the application of danofloxacin in aquaculture. About 106 CFU per mL was inoculated to Amur sturgeon to induce the disease model. The 150 healthy fish and over 200 infected fish randomly divided into two groups according to different routes of administration. The Amur sturgeon are divided into 4 groups for the experiment, the healthy group for intravenous, the healthy group for oral administration, the diseased group for intravenous and the diseased group for oral ad- ministration. The dose for intravenous and oral administration were all 10mg/kg. At different times after administration the blood were collected. High-performance liquid Chromatography （HPLC） was used to determine the concentration of danofloxacin in plasma of Amur Strugeon and the MCPKP pharmacokinetic software was used to analyze the medicinal concentration-time courses. Using a C18 colume, the mobile phase consisted of Acetonitrile/tridistilled water （15：85）. The fluorescence acceleration and emission wavelenght were 280 nm and 450 nm. The protein of samples were deprived with methyl alcohol. The precipitated mixture was shaken and then centrifuged, the filtrate was evaporated to dryness with Nitrogen fumes. Within the range of 0.005- 1.0μg/mL, danofloxacin had a good linearity. The lowest detectability was 0.005μg· mL^-1 and the average recovery of danofloxacin was above 84% . The within day and day-to-day precision expressed by RSD was less than 10% at three drug levels （0. 005,0.1 and l μg/mL）. The pharmacokinetics and bioavailability of danofloxacin in healthy and diseased Amur sturgeon infected with Aeromonas hydrophila were studied. The pharmacokinetic characteristics of danofloxacin in healthy group is described by the three compartment open model by intravenous. The data is C = 5.830^-5.582t ＋ 4. 162^-1.157t ＋ 0.852^-0.029t, the main pharmacokinetic parameters of danofloxin ; distrbution half-time （t1/2α） 0.55 h ; elimination half-time （t1/2β） 22.19 h ; area under the plasma drug concentration-time curve （AUC） 34.23 mg/（L·h）, apparent volume of the central compartment （V1） 0.92L/kg, apparent volume of a drug distribution in body （Vb） is 10.14L/kg, kel is 0.32/h. The V1 of diseased sturgeon by single intravenous danofloxacin decreased to 0.29L/kg. The elimination of danofloxacin between healthy and diseased sturgeon by intravenous is same. The pharmacokinetic characteristics of danofloxacin by oral administration in healthy sturgenon is described by the two compartment open moldel with first order absorb, the data is described with C=1.278e^-0.073t＋0.177e^-0.089t-1.455e^-0.329t, the main pharmacokinetic parameters of danofloxin t1/2ka 9.49h, t1/2β 78.27h, time of reaching maximum drug concentration after a single i. m. injection of a drug （Tmax） 6.28h, maximum drug concentration after a drug （Cmax） 0.79mg/mL, slopes of distribution phrase （α） 0.07/h, The result showed that the absorption distribution and elimination of danofloxacin changed markedly. The a decreased to 0.05/h. Meanwhile the rate of elimination slowed, t1/2β increased to 93.99h, tmax prolonged to 9.06h, the Cmax decreased to 0.59mg/mL. the bioavailability of danofloxacin in healthy and diseased sturgeon is 96.50 % and 94.44 % by oral administration. The high bioavailability in healthy and diseased group showed that danofloxacin is suitable to cure the bacterial infection Amur sturgeon. Danofloxacin can be given by oral administration at dose of 10mg/kg interval 2 days to prevent infected Ah Amur sturgeon.