CN111513000B - 青蒿素在消除养殖鱼类体内磺胺类药物残留中的应用 - Google Patents
青蒿素在消除养殖鱼类体内磺胺类药物残留中的应用 Download PDFInfo
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Abstract
本发明公开了青蒿素在消除养殖鱼类体内磺胺类药物（尤其是磺胺甲噁唑）残留中的应用，一方面青蒿素可使养殖鱼类体内磺胺类药物残留浓度降低，另一方面青蒿素可使磺胺类药物在养殖鱼类体内消除半衰期和休药期缩短。青蒿素可用于加速养殖鱼类体内磺胺类化学药物残留的消除，对于保障水产品质量安全，减少人体摄入含有化学药物残留的水产品的风险具有重要意义。
Description
技术领域
本发明属于水产动物药理学技术领域，具体涉及青蒿素在加速消除养殖鱼类体内磺胺类药物(尤其是磺胺甲噁唑)残留中的应用。
背景技术
斑点叉尾鮰(Ietalurus punetaus)原产于北美洲，属鲇形目(Silurfornes)、鮰科(Ictaluridae)。斑点叉尾鮰是一种大型淡水鱼类，具有生长快、适应性广、抗病力强、肉质上乘并且价格低廉等优点，深受消费者的喜爱。从2015年起，中国已成为斑点叉尾鮰养殖第一大国。但伴随着养殖规模的扩大，集约化养殖程度的提高、品种退化、药物滥用等状况的出现，药物的使用需求也在不断增加。药物残留问题已成为阻碍我国斑点叉尾鮰产业发展的关键问题之一。
人长期摄入含有磺胺类药物残留的水产品，药物不断在人体内蓄积，当积累到一定程度后，就会对人体产生毒性作用，可引起肾损害，特别是乙酰化磺胺在尿中溶解度低，析出结晶后对肾脏损害更大。磺胺类药物也能破坏人的造血系统，引起溶血性贫血症、粒细胞缺乏、血小板减少症等。
为了减少磺胺类药物在鱼体内的残留，我们基于药物的相互作用理论为基础，采用天然产物来调控半点叉尾鮰体内化学药物的代谢，可使一些化学药物在斑点叉尾鮰体内的残留加速消除，进而缩短化学药物在斑点叉尾鮰体内残留的休药期，对于缩短用药后斑点叉尾鮰的养殖周期，减少投入，提高水产品质量，增加收入具有重要价值。
发明内容
为了快速消除养殖鱼类体内残留的磺胺类药物，本发明提供了青蒿素在加速养殖鱼类体内磺胺类药物(尤其是磺胺甲噁唑)残留消除中的应用。
为了达到上述目的，本发明采取以下技术措施：
在本发明的实施例中，斑点叉尾鮰口灌磺胺甲噁唑(口灌剂量为60mg/kg鱼体重)12h后，分别口灌蒸馏水和青蒿素(10mg/kg鱼体重)，在不同的时间点检测斑点叉尾鮰血浆和带皮肌肉中磺胺甲噁唑残留浓度。结果表明，与对照组(口灌磺胺甲噁唑+蒸馏水)相比，试验组(口灌磺胺甲噁唑+青蒿素)斑点叉尾鮰血浆中磺胺甲噁唑消除半衰期缩短10.75h，带皮肌肉中磺胺甲噁唑的半衰期的缩短1/2；试验组磺胺甲噁唑休药期较对照组缩短37.4h(约为1.5d)。
青蒿素在消除养殖鱼类体内磺胺类药物残留中的应用：养殖鱼类口灌青蒿素，或者饲喂含有青蒿素的饲料；
进一步地，所述的磺胺类药物为磺胺甲噁唑；
进一步地，青蒿素的有效剂量是10mg/kg鱼体重；
进一步地，所述的养殖鱼类包括但不限于鲶形目鱼类如斑点叉尾鮰、黄颡鱼等，鲤科鱼类如草鱼、青鱼、鲢、鳙、鲤、团头鲂等，鲈形目鱼类如罗非鱼、乌鳢、鳜鱼、鲈鱼等。
与现有技术相比，本发明具有以下优点和有益效果：
(1)青蒿素可以使磺胺类药物--磺胺甲噁唑在斑点叉尾鮰自然比例带皮肌肉的残留时间由89.76h缩短至52.36h，即休药期缩短了1.5d，同一时间点磺胺甲噁唑在斑点叉尾鮰体内的残留量也明显降低为原浓度的3/10～4/5(自然比例带皮肌肉)和1/5～9/10(血浆)。
(2)本发明可使斑点叉尾鮰中磺胺类药物(尤其是磺胺甲噁唑)的残留时间缩短和残留量降低，对于保障水产品质量安全，减少人体摄入含有磺胺类药物残留的水产品的风险具有重要意义。
附图说明
图1为对照组和试验组斑点叉尾鮰血浆中磺胺甲噁唑残留消除曲线图。
图2为对照组和试验组斑点叉尾鮰自然比例带皮肌肉中磺胺甲噁唑残留消除曲线图。
图3为对照组斑点叉尾鮰自然比例带皮肌肉中磺胺甲噁唑休药期计算结果。
图4为试验组斑点叉尾鮰自然比例带皮肌肉中磺胺甲噁唑休药期计算结果。
具体实施方式
试验鱼：斑点叉尾鮰，平均体重(180±32)g，购自武汉市白沙洲大市场，运至实验室后，放置于已清洗并用高锰酸钾消毒的水族箱(122cm×80cm×80cm)中暂养1周，试验用水为充分曝气的自来水。暂养结束后，选择无病无伤，规格均匀的个体用于试验，控制水温为24±1℃。
青蒿素(纯度≥98.0％，购自四川省维克奇生物科技有限公司)，磺胺甲噁唑(纯度≥98.0％，购自北京索莱宝科技有限公司)。
试验分为两组，一组为口灌斑点叉尾鮰磺胺甲噁唑(口灌剂量为60mg/kg鱼体重)的对照组，对照组口灌磺胺甲噁唑12h后口灌蒸馏水；另一组斑点叉尾鮰口灌磺胺甲噁唑(60mg/kg鱼体重)后12h，再口灌青蒿素(10mg/kg鱼体重)。试验组和对照组斑点叉尾鮰分别在口灌青蒿素和蒸馏水之后2h、4h、8h、12h、1d、2d、3d、5d、7d采集斑点叉尾鮰血浆和自然比例带皮肌肉。
肌肉样品前处理：
取2g肌肉样品到15mL离心管中，加入50μL 1μg/mL的磺胺甲噁唑-d4再加入5mL乙腈，振荡30s，超声3min，再加入2g无水硫酸钠，振荡30s，8000r/min离心5min，转移上层液体到10mL离心管中，再向残渣中加入4mL乙腈，振荡30s，8000r/min离心5min，合并两次乙腈提取液于同一10mL离心管中，置50℃氮吹仪上氮吹至干，加入1mL定容溶液(甲醇：水＝20：80，体积比)溶解残渣，振荡30s，加入1mL正己烷去脂，过0.22μm滤头过滤，进HPLC-MS/MS分析。
血浆样品的处理：
取1mL血浆样品到10mL离心管中，加入50μL 1μg/mL的磺胺甲噁唑-d4，加入1mL50mM磷酸盐缓冲溶液(pH 7.0)振荡30s，再加入5mL乙酸乙酯，振荡30s，8000r/min离心5min，转移上层液体到10mL离心管中，再向残渣中加入4mL乙酸乙酯，振荡30s，8000r/min离心5min，合并两次乙酸乙酯提取液于同一10mL离心管中，置40℃氮吹仪上氮吹至干，加入1mL定容溶液(甲醇：水＝20：80，体积比)溶解残渣，振荡30s，过0.22μm滤头过滤，进HPLC-MS/MS分析。
仪器分析方法：液相色谱条件：A相为甲醇，B相为0.1％甲酸水溶液，初始条件为20％的甲醇，维持1min，A相由20％A升为40％A，5.1min由40％A变为20％A，5.1min～6.00min维持20％A。进样量为10μL。
质谱条件：喷雾电压3500V，蒸发气温度为300℃，鞘气压力位35kPa，辅助气压力为5kPa，离子传输管温度：350℃，碰撞气及压力：氩气，1.5MPa；磺胺甲噁唑母离子m/z为254，定量离子m/z为156，定性离子m/z 108；磺胺甲噁唑-d4母离子m/z为258，定量离子m/z为160。
数据处理：磺胺甲噁唑按一级动力学过程从斑点叉尾鮰体内消除，即在消除后期服从指数消除：C＝C0e-kt，C表示药物浓度，C0为残留消除对数曲线的截距(μg/kg)，k表示消除速率常数，消除半衰期T1/2＝0.693/k，休药期采用WT1.4软件计算。药物经时曲线图，采用Microsoft Excel 2013，进行计算和绘制。
结果分析
对照组和试验组斑点叉尾鮰血浆中磺胺甲噁唑消除半衰期分别为35.32h和24.57h，试验组(口灌磺胺甲噁唑+青蒿素)斑点叉尾鮰血浆中磺胺甲噁唑消除半衰期较对照组(口灌磺胺甲噁唑+蒸馏水)缩短10.75h，对照组和试验组斑点叉尾鮰带皮肌肉中磺胺甲噁唑的消除半衰期分别为32.69h和15.40h，试验组磺胺甲噁唑消除半衰期约是对照组磺胺甲噁唑消除半衰期的1/2。若以0.1μg/g为磺胺甲噁唑在水产品中的最大残留限量值(MRL)，采用欧盟WT1.4软件计算的试验组和对照组休药期分别为89.76h和52.36h，试验组休药期较对照组休药期缩短37.4h(约为1.5d)。由此看出，口灌青蒿素可以促进斑点叉尾鮰体内磺胺甲噁唑残留的消除。本实施例以斑点叉尾鮰为代表验证青蒿素对养殖鱼类体内磺胺类药物的清除作用，所述的养殖鱼类包括但不限于鲶形目鱼类如斑点叉尾鮰、黄颡鱼等，鲤科鱼类如草鱼、青鱼、鲢、鳙、鲤、团头鲂等，鲈形目鱼类如罗非鱼、乌鳢、鳜鱼、鲈鱼等。
Claims (3)
1.青蒿素在消除养殖鱼类体内磺胺类药物残留中的应用。
2.根据权利要求1所述的青蒿素在消除养殖鱼类体内磺胺类药物残留中的应用，其特征在于，所述的磺胺类药物是磺胺甲噁唑。
3.根据权利要求2所述的青蒿素在消除养殖鱼类体内磺胺类药物残留中的应用，其特征在于，青蒿素的有效剂量是10mg/kg鱼体重。
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| US4374826A (en) * | 1980-07-21 | 1983-02-22 | Pfizer Inc. | Sulfonamide compositions |
| DE3613794A1 (de) * | 1986-04-24 | 1987-10-29 | Frank Joachim Mueller | Verfahren zum nachweis von antibiotika- und sulfonamidrueckstaenden in biologischen fluessigkeiten oder nahrungsmitteln |
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