【作者】 王翔凌；

【导师】 方之平；

【作者基本信息】 华中农业大学 ， 基础兽医学， 2005， 硕士

【摘要】 本研究建立了沙拉沙星（Sarafloxacin）在鲫（Carassius auratus）血浆和组织中反相高效液相色谱检测方法,并对盐酸沙拉沙星（Sarafloxacin Hydrochloride）在鲫体内的药物动力学和残留做了研究。在21±1℃下,选用90尾雌性鲫,随机分为18组,单剂量（20mg/kg）口灌给药,在不同的时间点分别选取一组鱼自尾部静脉采血,并取肌肉、皮肤、肝胰脏、肾脏、卵巢5种组织。用甲磺酸达氟沙星（Danofloxacin mesylate）作内标,二氯甲烷提取样品中的药物,60℃恒温水浴中氮气流吹干,流动相溶解,正己烷去脂,反相高效液相色谱法测定其中的药物浓度。流动相为:乙腈:0.2mol/L Na₃PO₄缓冲液（pH7.4）（含0.5%四丁基溴化铵）=10:90（V/V）,用H₃PO₄调pH至3.0,采用ZORBAX SB-C₁₈色谱柱,紫外检测波长280nm。经测得该方法样品回收率均大于82.97%,各样品日内变异系数（intra-day CV）小于6.04%,日间变异系数（inter-day CV）小于7.11%,最低检测险（limit of detection,LOD）和最低定量限（limit of quantitation,LOQ）均满足残留检测的需要。 盐酸沙拉沙星在鲫体内的血浆药时数据符合一级吸收二室开放模型,方程为:C_血=1.28e^-0.26t+0.31e^-0.01t-1.59e^-4.46t。主要药物动力学参数为:分布半衰期(t_1/2α)、吸收半衰期(t_1/2kα)、消除半衰期(t_1/2β)分别为2.72、0.16、67.97h,药时曲线下面积（AUC）、达峰时间(T_max)、达峰浓度(C_max)分别为35.25μg·mL^-1·h、0.72h、1.31μg/mL。在组织药物动力学研究中,肌肉、皮肤、卵巢的经时过程均符合二项指数方程,理论方程分别为:C_肌=5.35(e^-0.03t-e^-0.11t、C_皮=1.69(e^{-0.01t-e-0.24t}、C_卵=3.32(e^-0.02t-e^-0.51t);肝胰脏和肾脏的经时过程均符合三项指数方程,其理论方程分别为:C_肝=37.60e^-0.23t+4.48e^-0.03t-42.08e^-0.37t、C_肾=15.11e^-0.08t+5.68e^-0.01t-20.79e^-0.22t。肌肉主要药动学参数为:t_1/2kα6.16h,消除半衰期(t_1/2ke)27.49h,T_max17.18h,C_max2.69μg/g,AUC 164.44μg·g^-1·h;皮肤主要药动学参数:t_1/2ka2.87h,t_1/2ke51.74h,T_max12.68h,C_max1.35μg/g,AUC 119.18μg·g^-1·h;卵巢主要药动学参数:t_1/2ka1.36h,t_1/2ke36.24h,T_max6.68h,C_max2.81μg/g,AUC167.16μg·g^-1·h;肝胰脏主要药动学参数:t_1/2α2.96h,t_1/2β24.37h,t_1/2ka1.90h,更多还原

【Abstract】 This study developed a method to detect sarafloxacin in goldfish plasma and tissues using reverse phase high performance liquid chromatorgraphic method （RP-HPLC）. Pharmacokinetics and residues of sarafloxacin hydrochloride were investigated, following single oral administration at dose of 20mg/kgb · w in healthy female goldfish （Carassius auratus）. 90 female goldfish were randomly divided into 18 groups for the experiments. The plasma and five tissues （muscle, skin, hepatopancreas, kidney, ovary） were collected at different intervals after administration of sarafloxacin hydrochloride. Danofloxacin mesylate was used as inner standard and dichloromet- hane was used as extractant. The extracts were evaporated to dryness by N₂ flow at water bath of 60℃ and were dissolved in mobile phase. Then the fat of the solute was degreased was by hexane and analysed by RP-HPLC. The mobile phase was acetonitrile and 0.2mol/L phosphate buffer （pH 7.4） （including 0.5% tetrabutyl ammonium bromide） （10/90, V/V） adjusted to pH 3.0 with phosphoric acid. Sarafloxacin hydrochloride and danofloxacin mesylate were quantitated by using ZORBAX SB-C₁₈ column and ultraviolet detector was set at 280nm. The mean recoveries for all samples were all exceed 82.97%, Intra-day CV and inter-day CV are under 6.04% and 7.11% respectively. Both LOD and LOQ can meet the requirement of the residual detection.The concentration-time data of sarafloxacin hydrochloride in plasma of healthy female goldfish after single oral administration was fitted to two-compartment model with first order absorption. The pharmacokinetic equation was : C_plasma=l.28e^-0.26t +0.31e^-0.01t-1.59e^-4.46t. The main pharmacokinetic parameters for plasma were: distribution half- life (t_1/2α), absorption half-life (t_1/2ka) and elimination half-life (t_1/2β) were 2.72 h, 0.16 h, 67.97h respectively. The area under the curve （AUC）, the time to peak concentration and the peak concentration were 35.25μg · mL^-1 · h, 0.72h and1.3 l^fg/mL respectively. After single oral administration of sarafloxacin hydrochloride to healthy female goldfish, the concentration-time data of sarafloxacin hydrochloride in muscle, skin and ovary were best described by biexponential equations, while those in hepatopancreas and kidney were best described by triexponential equations. The pharmacyokinetic equations were shown as follow:n c? /_.-0.03t -0.11K. r> 1 /-o/- -O.Olt a-0.24tvr. - ^<sup>, -0.02t -0.51(xCmuSc!e= 5.35 （e -e ）; Cskin=1.69（e -e ）;COVary=3.32（e -e ）;r -rx-i {.no-0-231 a. a ziR^-003’ 40 n?p-°-37t ? r -1 ^ ii<.-oo8ij.<;C hepatopancreas =■$ I-OVC + 4.48e - 4Z.Ube , Ckidney=l^&gt;-lle +5.20.79e’°22t. The main pharmacokinetic parameters for muscle were ti/2ka 6.16h, elimination half-life（t1/2ke）27.49h, Tmax 17.18h, Cmax 2.69/ig/g, AUC 164.44fig-g’1 -h; The main pharmacokinetic parameters for skin were ty2ka2.87h, tj/2ke 51.74h, Tmax 12.68h, Cmax l-35^g/g, AUC 119.18/ig ? g"1 ? h; The main pharmacokinetic parameters for ovary were ti/2ka l-36h, t1/2ke 36.24h, Tmax 6.68h, Cmax 2.81fig/g, AUC 167.16 jug ■ g"1 ? h; The main pharmacokinetic parameters for hepatopancreas were ti/2<. 2.96h, ti^ 24.37h, ti/2ka 1.90h, Tmax 4.04h, Cmax 8.97^g/g, AUC 202.91 fig ? g1 - h. The main pharmacokinetic parameters for kidney were ti/2? 8.47h, ti/2? 61.50h, ti/2ka 3.11h, Tmax 8.74h, Cmax 9.58/ig/g, AUC 595.43^g -g’1 -h. The elimination equations for tissues were as follows: CmUscie=4.38e"a02l;CSkin=1.82e"0’011; C hepatopancreas =5.65e-003t;f ’ Q 1A -°-02t. p _i (yj _-0.02t^=-’-’^e ’, COvary—" FThe results showed that the pharmacokinetic character of sarafloxacin hydrochloride in plasma of healthy female goldfish was as follows: Sarafloxacin hydrochloride was absorbed fast and the time to peak concentration was short. The drug was slowly eliminated after a single oral aministration in healthy goldfish. The pharmacokinetic character of tissues were as follows: Sarafloxacin hydrochloride concentration were high and distributed extensively in goldfish. The penetration into tissues was strong and this drug could be effective in the treatment of whole body and deep tissues infections in goldfish. The character of residue and elimination was both residual quantity of sarafloxacin hydrochloride and elimination rate were different significantly in every tissues. The concentration of sarafloxacin hydrochloride in kidney and hepatopancreas was more higher than other tissues and skin showed the slowest depletion among five tissues, The target tissue of residue was kidney. From the formula of withdrawl period , It was suggested that the withdrawl period shouldnot be less than 14 days after single oral administration of sarafloxacin hydrochloride in healthy female goldfish.更多还原