盐酸氯苯胍和恩诺沙星在异育银鲫体内药动学研究

发布时间：2018-05-04 14:11

本文选题：盐酸氯苯胍 + 恩诺沙星　；参考：《上海海洋大学》2017年硕士论文

【摘要】：近年来,异育银鲫养殖发展迅速,已成为我国主要的淡水养殖品种之一,然而大规模集约化养殖带来了严重的病害问题,尤其是细菌性疾病和寄生虫病最为严重。粘孢子虫可寄生于鲫的鳃、表皮、喉部、肠、肝等多个部位,其造成的危害已给产业带来重大的经济损失,因此需要一种具有较强杀虫驱虫效果的药物来减少病害的发生。越来越严重的细菌耐药性问题,对抗菌药的使用造成了很大麻烦,为了更好的治疗疾病和减少耐药细菌的产生,进行贴近生产的药物代谢动力学研究显得尤为重要。盐酸氯苯胍是我国国标渔药,关于盐酸氯苯胍在异育银鲫体内的药物代谢动力学未见有研究报道。针对我国鲫养殖现状,本文首先建立了盐酸氯苯胍在异育银鲫组织中残留的检测方法,又研究了通过药饵给药,盐酸氯苯胍在异育银鲫体内的药物代谢动力学研究,确定其在异育银鲫组织内的分布与消除规律,根据肌肉可食组织中的消除规律,结合相关国家和组织的最高残留限量标准,确定盐酸氯苯胍在异育银鲫体内的休药期,为鲫生产中盐酸氯苯胍的合理使用提供科学依据。1.异育银鲫组织中盐酸氯苯胍的反相高效液相色谱法测定方法的建立在比较不同流动相对盐酸氯苯胍色谱行为影响,以及不同提取方法对盐酸氯苯胍回收率影响的基础上,建立了测定异育银鲫血浆、肝胰脏、肌肉、喉、肾脏、肠、鳃、脑和胆汁等组织中盐酸氯苯胍含量的反相高效液相色谱法。方法以乙腈和0.1%甲酸水为流动相,色谱柱为Aglient Zorbax SB-C18(4.6×150mm,5μm),柱温30℃,紫外检测器波长317nm,进样量20μL,流速1.0 m L·min-1。盐酸氯苯胍在0.01~10μg·m L-1范围内线性关系良好,相关系数R2=0.9998。采用乙腈提取异育银鲫血浆、肝胰脏、肌肉、喉、肾脏、肠、鳃、脑和胆汁组织中盐酸氯苯胍,加标回收率为80.06%~102.02%,精密度为2.16%~7.41%,检测限为0.01μg·m L-1。该方法操作简单,重现性好,药峰无干扰,适用于异育银鲫生物样品中盐酸氯苯胍含量的分析。2.盐酸氯苯胍在异育银鲫体内药动学和组织分布与消除研究水温25±1℃条件下,盐酸氯苯胍以20 mg·kg-1单剂量拌饲药饵给药后在异育银鲫体内药动学,以及在肝胰脏、肌肉、喉、肾脏、肠、鳃、脑和胆汁中的分布和消除规律,为其科学安全使用提供理论支撑。结果显示,药饵给药后异育银鲫血浆中盐酸氯苯胍的药时数据符合一级吸收二室模型,血药达峰时间(Tmax)为4 h,血药浓度峰值(Cmax)为1.117 mg·L-1、药时曲线下面积(AUC0-∞)为68.39mg·L-1·h和消除半衰期(t1/2β)为56.86 h。盐酸氯苯胍在异育银鲫其他组织中分布较广,Cmax大小依次为:肠(8.53mg·kg-1)、肾脏(4.64mg·kg-1)、肝胰脏(3.27mg·kg-1)、胆汁(2.68mg·kg-1)、鳃(1.38mg·kg-1)、喉(1.17mg·kg-1)、脑(0.82mg·kg-1)、肌肉(0.43mg·kg-1);AUC0-∞大小依次:肠(156.39mg·kg-1·h)、肾脏(122.911mg·kg-1·h)、胆汁(83.772mg·kg-1·h)、肝胰脏(68.444mg·kg-1·h)、喉(53.659mg·kg-1·h)、鳃(39.142mg·kg-1·h)、脑(32.904mg·kg-1·h)、肌肉(12.448mg·kg-1·h);t1/2z大小依次为:脑(85.549h)、鳃(83.263h)、肾脏(76.541h)、喉(69.334h)、肝胰脏(67.197h)、胆汁(55.852h)、肌肉(54.506h)、肠(48.581h)。异育银鲫摄食药饵后,盐酸氯苯胍通过肠道吸收进入肝胰脏,经血液循环广泛分布于各组织器官中,最后主要从肾脏排出,这也与其较大的Vd值相印证。研究还发现,粘孢子虫感染异育银鲫的喉、鳃和脑等组织均有盐酸氯苯胍分布,为该病的治疗提供了理论依据。若以10μg·kg-1为-肌肉中最高残留限量,在本试验条件下,建议休药期不少于15 d。3.恩诺沙星临床用药药动学与药效评价本研究深入生产进行采样分析,实验池塘位于江苏省盐城市射阳盐场鲫大规模养殖区,选取发生细菌性疾病的用药池塘且按照当地用药习惯进行给药,以获得更加真实、可靠的数据,方便对实际生产中用药情况的分析和指导。采样方法上,在多次用药前后采集样品,以获得药物在动物体内的大致代谢情况和在血浆中可以达到的最高药物浓度。结果显示,异育银鲫恩诺沙星最高血药浓度为1.130μg·m L-1,环丙沙星的最高浓度为0.026μg·mL-1,恩诺沙星及其代谢产物环丙沙星在异育银鲫体内消除较慢,存留时间较长。所得的实验数据表明,池塘的用药剂量偏小,最高的药物浓度仅为1.130μg·m L-1,面对日益严重的细菌耐药问题,很难达到良好的治疗效果。模拟生产进行的30mg/kg的恩诺沙星药饵多次给药,获得的最高血药浓度为5.600μg·mL-1,已经大于多数细菌的MIC,理论上可以起到杀灭细菌并抑制耐药菌株产生,达到治疗细菌性疾病的目的。所以,在临床实际用药时,要综合考虑多种因素,病情的严重程度,科学的使用一定剂量的药物来达到好的治疗效果并减少耐药的发生。
[Abstract]:In recent years, the culture of crucian carp has been developing rapidly, and it has become one of the main varieties of freshwater aquaculture in our country. However, the large-scale intensive culture has brought serious disease problems, especially the most serious bacterial and parasitic diseases. The parasitic spore can parasitism on the gills, epidermis, larynx, intestines and liver of Carassius auratus. The harm caused by it has been given to the carp. Industry brings great economic loss, so it needs a drug with strong insecticidal and insect repellent effects to reduce the occurrence of disease. More and more serious bacterial resistance problems, and the use of anti bacteria drugs has caused great trouble. In order to better treat disease and reduce the production of drug resistant bacteria, the pharmacokinetics of drug metabolism close to production is carried out. The study appears particularly important. Chlorobenzidine hydrochloride is a national standard fishing drug in China. The pharmacokinetics of chlorobenzidine hydrochloride in the body of crucian carp is not reported. In view of the current situation of carp culture in China, the detection method of chlorphenidine hydrochloride in the tissue of crucian carp is first established, and the drug bait, hydrochloric acid, and hydrochloric acid are also studied. The pharmacokinetics of chlorobenzidine in the crucian carp was studied, and its distribution and elimination in the tissue of the crucian carp were determined. According to the elimination rule in the edible tissues of the muscle and the standard of the highest residue limit of the related countries and organizations, the period of the taking of chlorobenzidine hydrochloride in the crucian carp was determined, and the chlorobenzene hydrochloride in the Carassius auratus production was found. The rational use of guanidine provides a scientific basis for the determination of chlorphenanidine hydrochloride in the tissue of.1. silver crucian carp by reversed phase high performance liquid chromatography (RP HPLC). On the basis of the influence of different flow relative to chlorobenzidine hydrochloride and the effect of different extraction methods on the recovery rate of chlorphenanidine hydrochloride, the determination of plasma and hepatopancreanopancreas of crucian carp was established. The reversed phase high performance liquid chromatography of chlorobenzidine hydrochloride content in tissues such as dirty, muscle, larynx, kidney, intestines, gills, brain and bile. Acetonitrile and 0.1% formate water were used as the mobile phase, the column was Aglient Zorbax SB-C18 (4.6 x 150mm, 5 mu m), the column temperature was 30, the UV detector was 317nm, the sample volume was 20 mu L, and the flow rate 1 m L min-1. hydrochloric guanidine was in 0.01~. The linear relationship between 10 g and m L-1 is good. Correlation coefficient R2=0.9998. uses acetonitrile to extract plasma, hepatopancreas, muscle, muscle, larynx, kidney, intestine, gill, brain and bile of chlorobenzidine hydrochloride, the recovery rate is 80.06%~102.02%, the precision is 2.16%~7.41%, the detection limit is 0.01 mu g. M L-1. this method is simple operation, reproducibility, peak drug peak Analysis of the content of chlorphenanidine hydrochloride in the biological samples of crucian carp (.2.), the pharmacokinetics and tissue distribution of chlorphenidine hydrochloride in the crucian carp were 25 + 1 centigrade, and the pharmacokinetics of chlorphenanidine hydrochloride in the crucian carp with 20 mg kg-1 single dose of chlorphenididine hydrochloride, and in the liver, pancreas, muscle and larynx were studied. The distribution and elimination of the kidney, intestines, gills, brain and bile provide theoretical support for the scientific and safe use of the drug. The results show that the data of chlorphenidine hydrochloride in the plasma of Carassius auratus after drug delivery are in accordance with the first order absorption two compartment model, the peak time of blood drug (Tmax) is 4 h, the peak value of blood drug concentration (Cmax) is 1.117 mg. L-1, under the curve of the drug time The product (AUC0- infinity) is 68.39mg / L-1 / h and the elimination half life (t1/2 beta) is 56.86 h. chlorobenzene guanidine in the other tissues of the crucian carp. The Cmax size is the following: the intestine (8.53mg. Kg-1), the kidney (4.64mg. Kg-1), the hepatopancreas (3.27mg.), the gills, the larynx, the brain and the muscles. Kg-1 (156.39mg. Kg-1. H), kidney (122.911mg. Kg-1. H), bile (83.772mg kg-1 / h), hepatopancreas (68.444mg. H), larynx (H), muscles (veins), gills, kidneys, larynx (69.33) 4h), hepatopancreas (67.197h), bile (55.852h), muscle (54.506h), and intestine (48.581h). Chlorphenanidine hydrochloride is absorbed into the hepatopancreas through intestinal absorption through intestinal absorption, and is widely distributed in the tissues and organs through the blood circulation. Finally, it is mainly discharged from the kidney, which is also evidenced by its larger Vd value. The distribution of chlorphenanidine hydrochloride in the larynx, gills and brain tissues of the Carassius auratus has provided a theoretical basis for the treatment of the disease. If 10 g kg-1 is the highest residual limit in the muscle, it is suggested that the drug pharmacokinetics and efficacy of enrofloxacin are not less than 15 d.3.. The pond is located in the large aquaculture area of the Sheyang salt carp in Yancheng City, Jiangsu province. The drug ponds of the bacterial diseases are selected and given according to the local drug use, so as to obtain more real and reliable data and facilitate the analysis and guidance of the use of drugs in the actual production. The highest concentration of the blood was 1.130 u g. M L-1, the highest concentration of ciprofloxacin was 0.026 G. ML-1, and enrofloxacin and its metabolite ciprofloxacin were slow to be eliminated in the crucian carp and the retention time was more than that in the Carassius auratus. The experimental data showed that the dosage of the drug in the pond was small and the highest drug concentration was only 1.130 G. M L-1. In the face of the increasingly serious bacterial resistance problem, it was difficult to achieve a good therapeutic effect. The 30mg/kg enrofloxacin bait for the simulated production was given many times, the highest concentration of blood drug was 5.600 mu g. ML-1, which was more than a lot. The MIC of several bacteria, in theory, can kill bacteria and inhibit the production of resistant strains to achieve the purpose of treating bacterial diseases. Therefore, in clinical practice, we should consider a variety of factors, the severity of the disease, the scientific use of a certain dose of drugs to achieve good therapeutic effects and reduce the occurrence of drug resistance.

【学位授予单位】：上海海洋大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S943

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