CN115047193A - 一种检测污水中痕量毒品的方法 - Google Patents
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Abstract
本发明公开了一种测污水中痕量毒品的方法,包括以下步骤:过滤待检测污水除去杂质,将过滤后污水加入固相萃取柱OASIS HLB,萃取之后,向固相萃取柱OASIS HLB中加入洗脱液进行洗脱;氮吹所得洗脱液至无液体,向氮吹所得物中加入稀释液复溶;将复溶后的液体滴在试纸条上,上机测试。本发明的有益效果为:本发明所述检测污水中痕量毒品的方法采集污水样本,评估大、小区域特定人群的毒品消费量,可通过短期或长期取样检测获取吸食毒品类型的变化趋势或新型毒品种类信息。
Description
技术领域
本发明属于生物鉴定技术领域,具体涉及一种检测污水中痕量毒品的方法。
背景技术
药物滥用也称为“吸毒”或“毒品滥用”。药物滥用(drug abuse)是指与医疗目的无关地反复大量使用一些具有依赖性或称依赖性潜力的药物,用药者采用自身给药形式,导致精神依赖性和身体依赖性,造成精神混乱和一些异常行为,其严重危害社会安全和人类生命健康。
污水中毒品残留物及其代谢产物的来源主要为吸毒人员的排泄物,突击缉查时吸毒人员为毁灭证据经下水道冲走丢弃及制毒场所排放的废水等。2001年以来,在欧洲、拉丁美洲、亚洲等国家的城市污水收集系统、河流、湖泊中,甚至在某些城市自来水中,均有该类化合物被检出的报道,其浓度为ng/L水平。由于水环境中药物污染物含量极低,通常在ng/L水平,同时种类较多,常规的分析手段很难满足痕量或超痕量分析的实际需要,因此一种可精准检测污水中痕量毒品的方法是目前市场的空缺。
发明内容
本申请的主要目的在于提供一种检测污水中痕量毒品的方法。
为了实现上述目的,本发明提供如下技术方案:
一种检测污水中痕量毒品的方法,包括以下步骤:
(1)过滤待检测污水除去杂质,将过滤后污水加入固相萃取柱OASIS HLB,萃取之后,向固相萃取柱OASIS HLB中加入洗脱液进行洗脱;
(2)氮吹步骤(1)所得洗脱液至无液体,向氮吹所得物中加入稀释液复溶;
(3)将复溶后的液体滴在试纸条上,上机测试。
固相萃取柱,用于酸性、中性和碱性化合物的通用型吸附剂,具有高而稳定的回收率,即使柱床干涸,回收率也不受影响。与硅胶相比,本申请所述固相萃取柱的Oasis吸附剂的表面积增大2-3倍,容量因子大大提高,30mg Oasis HLB的吸附容量相当于100mgC18。
本发明所述检测污水中痕量毒品的方法采用抗体-抗原特异结合反应及免疫层析技术,通过免疫竞争法来检测样本中出现的吗啡(甲基苯丙胺,氯胺酮)。抗原事先固定在硝酸纤维素膜的检测区(T区),毒品特异性单克隆抗体用荧光微球进行标记后固定在玻璃纤维上。测试时样本提取出的毒品物质和膜上抗原竞争荧光标记抗体上有限的抗原结合位点。样品中吗啡(甲基苯丙胺,氯胺酮)药物的浓度不同检测区T区荧光值不同,荧光值的变化反映出吗啡(甲基苯丙胺,氯胺酮) 药物的浓度。
上述一种检测污水中痕量毒品的方法,作为一种优选的实施方案,步骤(1) 中,采用0.45um的滤膜过滤待检测污水。
上述一种检测污水中痕量毒品的方法,作为一种优选的实施方案,步骤(1) 中,洗脱采用的洗脱液为含5%氨的甲醇溶液。
上述一种检测污水中痕量毒品的方法,作为一种优选的实施方案,步骤(2) 中,氮吹的温度为60℃。
上述一种检测污水中痕量毒品的方法,作为一种优选的实施方案,步骤(2) 中,所述稀释液为pH为7.5的50mM Tris-Hcl、体积浓度为0.5-1%Triton X-100、体积浓度为0.5-1%Tween-20、质量浓度为0.5%NaN3混合溶液。
Tris-Hcl为三羟甲基氨基甲烷盐酸盐,被广泛用作核酸和蛋白质的溶剂。
Triton X-100为聚乙二醇辛基苯基醚,是一种有机高分子化合物,为无色或几乎无色透明黏稠液体。在显微镜和组织学实验室,其稀释的溶液可作为润湿剂,协助染色,并可作为清洁剂对钻石刀进行清洁。
Tween-20是一种非离子型表面活性剂,用途为水包油乳化剂、可用作溶剂、扩散剂、稳定剂、润滑剂和抗静电剂等。Tween-20有复性抗原的作用,可提高特异性的识别能力。其分子中含有较多的亲水性基团,可与水、甲醇、乙醇、异丙醇、丙二醇、乙二醇、棉子油等混溶,是常用的非离子性去垢剂。
NaN3又名三氮化钠,是一种无机化合物,呈白色六方系晶体,无味、无臭,无吸湿性,在真空中加热不爆炸,可逐渐分解为金属钠及氮气,是高纯度金属钠的实验室制造方法之一,也是高纯度N2实验室制造方法之一。叠氮化钠为照相乳剂的一种防腐剂,可加入乳剂中,或加到中间层及保护层中,不影响乳剂照相性能,具有优良的防腐杀菌性能。
上述一种检测污水中痕量毒品的方法,作为一种优选的实施方案,步骤(3) 中,所述试纸条为毒品荧光检测试剂条。
上述一种检测污水中痕量毒品的方法,作为一种优选的实施方案,步骤(3) 中,所述上机测试为采用干式免疫分析仪检测毒品荧光检测试剂条。
本发明的有益效果为:本发明所述检测污水中痕量毒品的方法采集污水样本,评估大、小区域特定人群的毒品消费量,可通过短期或长期取样检测获取吸食毒品类型的变化趋势或新型毒品种类信息。
具体实施方式
为了使本技术领域的人员更好地理解本申请方案,下面将结合案例对本申请实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本申请一部分的实施例,而不是全部的实施例。基于本申请中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都应当属于本申请保护的范围。
实施例1
实施例1所述检测污水中痕量毒品的方法,包括以下步骤:
(1)采用0.45um的滤膜过滤待检测污水除去杂质,将过滤后污水加入固相萃取柱OASIS HLB进行萃取,待萃取完成向固相萃取柱OASIS HLB中加入洗脱液进行洗脱,所用洗脱液为含5%氨的甲醇溶液,洗脱至玻璃试管内;
(2)氮吹步骤(1)所得洗脱液至无液体,氮吹的温度为60℃,向氮吹所得物中加入稀释液复溶;所加稀释液为pH为7.5的50mM Tris-Hcl、体积浓度为0.5%Triton X-100、体积浓度为0.5%Tween-20、质量浓度为0.5%NaN3混合溶液;
(3)将复溶后的液体滴在毒品荧光检测试剂条上,采用干式免疫分析仪检测毒品荧光检测试剂条。
实施例2
实施例2所述检测污水中痕量毒品的方法,包括以下步骤:
(1)采用0.45um的滤膜过滤待检测污水除去杂质,将过滤后污水加入固相萃取柱OASIS HLB进行萃取,待萃取完成向固相萃取柱OASIS HLB中加入洗脱液进行洗脱,所用洗脱液为含5%氨的甲醇溶液,洗脱至玻璃试管内;
(2)氮吹步骤(1)所得洗脱液至无液体,氮吹的温度为60℃,向氮吹所得物中加入稀释液复溶;所加稀释液为pH为7.5的50mM Tris-Hcl、体积浓度为0.8% Triton X-100、体积浓度为0.8%Tween-20、质量浓度为0.5%NaN3混合溶液;
(3)将复溶后的液体滴在毒品荧光检测试剂条上,采用干式免疫分析仪检测毒品荧光检测试剂条。
实施例3
实施例3所述检测污水中痕量毒品的方法,包括以下步骤:
(1)采用0.45um的滤膜过滤待检测污水除去杂质,将过滤后污水加入固相萃取柱OASIS HLB进行萃取,待萃取完成向固相萃取柱OASIS HLB中加入洗脱液进行洗脱,所用洗脱液为含5%氨的甲醇溶液,洗脱至玻璃试管内;
(2)氮吹步骤(1)所得洗脱液至无液体,氮吹的温度为60℃,向氮吹所得物中加入稀释液复溶;所加稀释液为pH为7.5的50mM Tris-Hcl、体积浓度为1% Triton X-100、体积浓度为1%Tween-20、质量浓度为0.5%NaN3混合溶液;
(3)将复溶后的液体滴在毒品荧光检测试剂条上,采用干式免疫分析仪检测毒品荧光检测试剂条。
对比例1
对比例1所述检测污水中痕量毒品的方法与实施例1所述的方法不同之处在于:对比例1采用甲醇作为洗脱液。
对比例2
对比例2所述检测污水中痕量毒品的方法与实施例2所述的方法不同之处在于:对比例2采用二氯甲烷和丙酮的混合液作为洗脱液,二氯甲烷和丙酮的体积比为1:4。
对比例3
对比例3所述检测污水中痕量毒品的方法与实施例3所述的方法不同之处在于:对比例3采用丙酮、甲醇和乙酸乙酯的混合液作为洗脱液,丙酮、甲醇和乙酸乙酯的体积比为:2:2:1。
本发明所述检测污水中痕量毒品的方法检测效果研究
测试方法:取城市生活污水50ml,通过0.45μm滤膜过滤除去杂质,向污水内加入20微升浓度均为100微克/毫升(吗啡、甲基苯丙胺、氯胺酮)的混合质控,将混合完成的污水加入固相萃取柱OASIS HLB浓缩净化,所用洗脱液为含5%氨的甲醇溶液(甲醇,体积比为1:4的二氯甲烷和丙酮,或体积比为:2:2:1的丙酮、甲醇和乙酸乙酯),在温度为60℃的条件下氮吹洗脱液至无液体,向氮吹所得物中加入0.5ml稀释液复溶(复溶液的浓度是2ng/ml);所加稀释液为pH为7.5 的50mM Tris-Hcl、体积浓度为1%Triton X-100、体积浓度为1%TWEEN-20、质量浓度为0.5%NaN3混合溶液,将复溶后液体滴在毒品荧光检测试剂条上,采用干式免疫分析仪检测毒品荧光检测试剂条,等待5分钟后记录测试结果。
表1
本发明所述检测污水中痕量毒品的方法采用不同洗脱液对萃取率的影响如表 2所示:
表2
从表2可以看出,采用本发明所述5%氨-甲醇作为洗脱液对痕量毒品的萃取率最高,对吗啡的萃取率达到88.00%,对甲基苯丙胺的萃取率达到83.00%。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员,在不脱离本发明方法的前提下,还可以做出若干改进和补充,这些改进和补充也应视为本发明的保护范围。
Claims (7)
1.一种检测污水中痕量毒品的方法,其特征在于,包括以下步骤:
(1)过滤待检测污水除去杂质,将过滤后污水加入固相萃取柱OASIS HLB,萃取之后,向固相萃取柱OASIS HLB中加入洗脱液进行洗脱;
(2)氮吹步骤(1)所得洗脱液至无液体,向氮吹所得物中加入稀释液复溶;
(3)将复溶后的液体滴在试纸条上,上机测试。
2.根据权利要求1所述检测污水中痕量毒品的方法,其特征在于,步骤(1)中,采用0.45um的滤膜过滤待检测污水。
3.根据权利要求1所述检测污水中痕量毒品的方法,其特征在于,步骤(1)中,洗脱采用的洗脱液为含5%氨的甲醇溶液。
4.根据权利要求1所述检测污水中痕量毒品的方法,其特征在于,步骤(2)中,氮吹的温度为60℃。
5.根据权利要求1所述检测污水中痕量毒品的方法,其特征在于,步骤(2)中,所述稀释液为pH为7.5的50mM Tris-Hcl、体积浓度为0.5-1%Triton X-100、体积浓度为0.5-1%Tween-20、质量浓度为0.5%NaN3混合溶液。
6.根据权利要求1所述检测污水中痕量毒品的方法,其特征在于,步骤(3)中,所述试纸条为毒品荧光检测试剂条。
7.根据权利要求1所述检测污水中痕量毒品的方法,其特征在于,步骤(3)中,所述上机测试为采用干式免疫分析仪检测毒品荧光检测试剂条。
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