伊维菌素微乳在羊体内的药代动力学与残留研究

发布时间：2018-12-19 08:50

【摘要】：本论文对一种伊维菌素微乳新制剂在羊体内的药物动力学特性与药物残留消除规律进行了研究。研究建立了新型伊维菌素微乳在羊血浆内的药物浓度的测定方法,测定了其在羊体内的药代学数据,分析了其药动学特特性;建立了新型伊维菌素微乳制剂在靶动物羊不同组织的药物残留测定方法,获得了其在不同组织内的残留消除规律,制定了伊维菌素微乳的休药期。伊维菌素微乳的药代动力学试验,建立了高效液相色谱-串联质谱法测定血浆中药物浓度的方法,试验结果表明,前处理方法简单有效,最低定量限为0.5ng/mL,最低检测限为0.3ng/mL,标准曲线线性良好,R2为0.9995,平均回收率大于90%,日内精密度和日间精密度均小于10%,灵敏度高、干扰小、重复性好。通过皮下、肌肉、静脉注射3种途径对羊进行给药,试验结果显示,皮下给药组的AUC为1343.07±101.17h·ng/mL,达峰浓度Cmax为50.61±2.96ng/mL,达峰时间Tmax为7.14±0.80h,吸收半衰期T1/2(α)为4.46h,消除半衰期T1/2(β)为25.45h;肌肉给药组的AUC为1814.24±56.06h·ng/mL,达峰浓度Cmax为70.34±1.89ng/mL,达峰时间Tmax为3.99±0.20,吸收半衰期T1/2(α)为4.52h,消除半衰期T1/2(β)为20.08h。肌肉注射与皮下注射伊维菌素微乳的绝对生物利用度分别为91.33%和67.62%。通过对不同的给药方式进行比较发现,肌肉注射与皮下注射相比具有更高的达峰浓度,更快的达峰时间,更高的生物利用度。伊维菌素微乳药物残留消除试验,建立了以高效液相色谱-串联质谱检测羊各组织中伊维菌素残留的方法。样品使用乙腈提取,使用碱性氧化铝固相萃取小柱净化,以高效液相色谱-串联质谱进行检测。试验结果表明,伊维菌素基质匹配标准曲线在1~3000ng/mL的药物浓度范围内,药物峰面积与线性关系良好,R2均大于0.999;在颈部肌肉、肾脏、脂肪、背最长肌和肝脏五种组织的最低检测限均为0.3ng/mL,最低定量限均为0.5ng/mL,该方法的定量限低于欧盟、美国和我国对于伊维菌素残留限量的规定。样品的平均回收率大于70%,日内精密度和日间精密度均小于10%,采用高效液相色谱串联质谱法检测羊各组织中的伊维菌素残留浓度,方法简单、快速、准确、灵敏度高。结合实测浓度判断法与采用WT1.4计算软件拟合,颈部肌肉的休药期计算为14.71d,肝脏的休药期计算为18.27d,肾脏的休药期计算为17.49d,脂肪的休药期计算为18.61d,背长肌的休药期计算为12.90d。依据靶动物羊体内的消除规律研究数据,结合伊维菌素在羊各组织中的休药期结果,建议该伊维菌素微乳注射液在羊体内的休药期为19天。本研究为伊维菌素微乳的新药评价和指导临床应用奠定了基础。
[Abstract]:In this paper, the pharmacokinetic characteristics and drug residue elimination of a new Ivermectin microemulsion in sheep were studied. A new method for the determination of the concentration of ivermectin microemulsion in sheep plasma was established. The pharmacokinetic characteristics of ivermectin microemulsion in sheep were analyzed. A new method for determination of Ivermectin microemulsion in different tissues of target animal sheep was established. The law of its elimination in different tissues was obtained, and the withdrawal period of ivermectin microemulsion was established. The pharmacokinetic test of ivermectin microemulsion was carried out. A method for the determination of drug concentration in plasma by high performance liquid chromatography-tandem mass spectrometry (HPLC / MS) was established. The results showed that the pretreatment method was simple and effective, and the minimum quantitative limit was 0.5 ng / mL. The lowest detection limit was 0.3 ng 路mL ~ (-1), the linearity of the standard curve was good, R2 was 0.9995, the average recovery was more than 90, the intra-day precision and inter-day precision were less than 10, the sensitivity was high, the interference was small, and the repeatability was good. The AUC of the subcutaneous administration group was 1343.07 卤101.17 h ng/mL, peak concentration (Cmax) was 50.61 卤2.96 ng / mL, and the peak time Tmax was 7.14 卤0.80 h. Absorption half life T 1 / 2 (伪) was 4.46 h, elimination half life T 1 / 2 (尾) was 25.45 h; In the intramuscular administration group, AUC was 1814.24 卤56.06 h, Cmax was 70.34 卤1.89 ng / mL, Tmax was 3.99 卤0.20, absorption half life T 1 / 2 (伪) was 4.52 h, elimination half life T 1 / 2 (尾) was 20.08 h. The absolute bioavailability of ivermectin microemulsion by intramuscular injection and subcutaneous injection was 91.33% and 67.62%, respectively. It was found that intramuscular injection had higher peak concentration, faster peak time and higher bioavailability than subcutaneous injection. A method for the determination of ivermectin residues in sheep tissues by HPLC-tandem mass spectrometry (HPLC-MS) was established by the drug residue elimination test of ivermectin microemulsion. The samples were extracted by acetonitrile and purified by alkaline alumina solid phase extraction column. The results showed that the peak area of ivermectin matrix matching standard curve was linear in the range of 1~3000ng/mL concentration, and R2 was greater than 0.999. In neck muscle, kidney, fat, longissimus dorsi muscle and liver, the minimum detection limit is 0.3 ng / mL, and the minimum quantitative limit is 0.5 ng / mL. the quantitative limit of this method is lower than that of the European Union. Ivermectin residue limits in the United States and China. The average recovery of the sample was more than 70 and the intra-day precision and inter-day precision were less than 100.The method was simple, rapid, accurate and sensitive for the determination of Ivermectin residues in sheep tissues by HPLC tandem mass spectrometry (HPLC / MS). Combined with the method of judging the measured concentration and using WT1.4 software, the discontinuation period of neck muscle, liver, kidney, fat and fat were 14.71 days, 18.27 days, 17.49 days and 18.61 days, respectively. The discontinuation period of the dorsal longus muscle was calculated to be 12.90 days. According to the study data of the elimination rule in the sheep of target animal and the results of the discontinuation period of ivermectin in sheep tissues, it is suggested that the drug withdrawal period of Ivermectin microemulsion injection in sheep body should be 19 days. This study laid a foundation for the new drug evaluation and clinical application of ivermectin microemulsion.
【学位授予单位】：中国农业科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S859.79

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