【作者】 王金花；

【导师】 操继跃；

【作者基本信息】 华中农业大学 ， 基础兽医学， 2007， 硕士

【摘要】 本研究建立了高效液相色谱（HPLC）法测定盐酸多西环素在猪血浆和组织中的方法，探讨了盐酸多西环素以2.5 mg／kg体重肌内注射后在猪（n=6）体内的药物动力学和残留消除规律。色谱条件是：色谱柱为：Agilent Zorbax TC-C18；流动相：乙腈：甲醇：0.01 mol／L草酸溶液（20：10：70）；流速：0.8 ml／min；检测波长：350 nm。样品处理：称取匀质样品，加Na₂EDTA-Mcllvaine缓冲溶液提取，然后用5％高氯酸溶液去除蛋白，上清液过SPE HLB oasis离子交换柱，取60μl进样分析。盐酸多西环素注射液在猪血浆、肌肉、肝脏、肾脏、皮和脂肪等组织中的线性范围均为0.05～12.8μg／g，线性方程和相关系数分别为Y=0.0078X-0.0682（R=0.9999）（血浆）；Y=0.0042X-0.0291（R=0.9998）（肌肉）；Y=0.0064X-0.0250（R=0.9997）（肝脏）；Y=0.0060X-0.0276（R=0.9997）（肾脏）；Y=0.0028X-0.0252（R=0.9999）（皮和脂肪）。测得该方法样品回收率大于69.4％，各样品日内变异系数小于5.90％，日间变异系数小于9.60％，血浆的最低检测限（0.025μg／ml）和最低定量限（0.05μg／ml），组织的最低检测限（0.0125μg／g）和最低定量限（0.025μg／g），均满足残留检测的要求。结果表明，药物动力学模型符合有吸收一室模型，其药物动力学方程：C=3.372×(e^-0.073t-e^-1.733t)。药物动力学参数：吸收半衰期(t_1／2ka)、消除半衰期(t_{1／2 ke})分别为0.40±0.31 h、9.53±0.95 h，药时曲线下面积（AUC）为44.41±4.12μg·h／ml，最大血药浓度(C_max)为2.81±0.13μg／ml，达峰时间(T_max)为1.91±0.21 h。另外，以相同剂量肌内注射给猪（n=6），每天一次，连续给药4 d后，在不同时间测定盐酸多西环素在猪的肌肉、肝脏、肾脏、皮和脂肪中的残留量。在给药16 d后，盐酸多西环素在各组织均能检测到且残留量均低于残留限量。盐酸多西环素注射液在猪体内消除缓慢，残留期较长，建议休药期不低于16 d。更多还原

【Abstract】 High performance liquid chromatorgraphic method （HPLC） to detect Doxycyclinehydrochloride in pigs plasma and tissues were developed, pharmacokinetics and tissueresidues of Doxycycline hydrochloride were investigated, following intramuscularinjection at dose of 2.5 mg/kg body weight （b.w） in healthy pigs（n=6）.chromatographyseparation was achieved on a Agilent Zorbax TC-C18 column using a mixature ofacetonitrile, methanol and 0.01M oxalic acid（v/v 20: 10: 70） as the mobile phase,thedetection wavelength is 350 nm and mobile speed is 0.8 ml/min. The Doxycyclinehydrochloride was extracted by by McIlvaine buffer from plasma and tissues. Thesupernatant was added 5% perchlorate to remove protein,and 60μl samples were injectedin high performance liquid chromatography to analyses after clean-up procedure bySPE HLB Oasis cartridge. Their linearity are all good over the concentration range（0.05～12.8μg/g） in plasma and tissues,The calibration of doxycycline hydrochloride andCorrelation coefficient（R） in plasma, muscle, liver, kidney, skin and fat were Y=0.0078X-0.0682（R=0.9999）, Y=0.0042X-0.0291（R=0.9998）, Y=0.0064X-0.0250（R=0.9997）,Y=0.0060X-0.0276（R=0.9997）, Y=0.0028X-0.0252（R=0.9999）, respectively. Themean recoveries for all samples were all exceed 69.4%,Intra-day CV and inter-day CV areunder 5.90% and 9.60% respectively. The limits of detection （LOD） were 0.025μg/ml,0.0125μ/g in plasam, in tissues respectively. The limit of quatition （LOQ） were 0.05μg/ml, 0.025μg/g in plasam,in tissue respectively. Both can satisfy the need of theresidual detection.The kinetics of Doxycycline Hydrochloride had been fit to one compartment modelwith first order absorption after intramuscular injection at dose of 2.5 mg/kg body weight（b.w） in pigs. The main parameters were as follows: the half-lives of absorption (t_{1/2 ka})and elimination (t_{1/2 ke}) were0.400±0.312 h, and 9.530±0.956 h respectively, area underthe plasma drug concentration-time curve from 0 to∞（AUC） was 44.414±4.123μg.h/ml, the peak plasma concentration (C_max) was 2.811±0.136μg/ml, peaking at 1.910±0.213 h (T_pn)after dosing. Moreover, each healthy pig was administered DoxycyclineHydrochloride i.m. once a day at dose of 2.5 mg/kg body weight （b.w） for four days （n=6）.The tissue residues of Doxycycline Hydrochloride were determined in muscle, liver,kidney, skin and fat. Tissue residues of Doxycycline Hydrochloride was to be lower to themaximum residue limit （MRL） after 16 days. The results indicated that the elimination ofDoxycycline Hydrochloride in tissue was slow. It is proposed that withdrawal periodshould be not less than 16 days after an intramuscular injection at a dose of 2.5 mg/kgb.w.更多还原