CN103969240B - 一种基于sers检测瘦肉精类物质的高通量纸质微流控芯片 - Google Patents
一种基于sers检测瘦肉精类物质的高通量纸质微流控芯片 Download PDFInfo
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Abstract
本发明属于食品安全检测领域,涉及一种基于SERS检测瘦肉精类物质的高通量纸质微流控芯片。该高通量纸质微流控芯片包含两部分:样品预处理区和SERS检测区,样品预处理区和SERS检测区都有M×N个瘦肉精测定单元,样品预处理区和SERS检测区折叠后的每个瘦肉精测定单元为一一对应,且形状一致;每个瘦肉精测定单元检测的瘦肉精类物质相同或不同;样品预处理区具有对样品过滤、分离、富集及pH调节功能;SERS检测区的功能包括免疫反应发生、SERS增强纳米粒子引入和团聚剂加入。本发明的高通量纸质微流控芯片可以同时完成多个瘦肉精样品的同时检测,实现瘦肉精类物质的即时现场检测,步骤简单快速。
Description
技术领域
本发明属于食品安全检测领域,涉及一种基于表面增强拉曼散射(SERS,surface-enhancedRamanscatting)检测瘦肉精类物质的高通量纸质微流控芯片。
背景技术
瘦肉精,属于β-受体激动剂,典型的有盐酸克伦特罗、莱克多巴胺、沙丁胺醇等。而人吃了残留有β-受体激动剂的肉类食品后,会产生肌肉颤动、呕吐、神经过敏和心跳加速等症状。因此,世界大多数国家都明令禁止使用瘦肉精作为饲料添加剂。
目前,我们将常用的瘦肉精检测手段分为两类:(1)胶体金试纸条法;(2)酶联免疫吸附法、高效液相色谱-质谱法和气相色谱-质谱法等。第一类方法假阳性率很高,第二类方法只能在中心实验室进行,不能做到即时现场检测。
纸质微流控芯片便宜、快速、能实现即时诊断,已经成为热门的研究领域之一,在发展中国家尤为如此。其制作和操作方法很简单,可用于检测临床、环境及食品安全等领域内的相关物质。可与纸制微流控芯片联用的检测手段有很多,例如电化学、比色、化学发光检测等,但鉴于庞大的实际需求,目前的这些检测方法仍显不足,开发新的检测方法迫在眉睫。
纸质微流控芯片很可能解决现有瘦肉精检测技术的诸多弊端,实现瘦肉精的即时现场高通量可靠检测,填补国内外相关研究尚处于空白。
发明内容
为了实现瘦肉精的即时现场高通量可靠检测,本发明提供了一种实际样品中瘦肉精类物质多重检测、高通量集成的纸质微流控芯片,基于此微流控芯片的瘦肉精检测快速、准确、便携。以此为切入点,可以实现人头发中生物标记物的检测。
本发明的技术方案:一种基于SERS检测瘦肉精类物质的高通量纸质微流控芯片,该高通量纸质微流控芯片包含两部分:样品预处理区和SERS检测区;样品预处理区和SERS检测区都有M×N瘦肉精测定单元,M≥1,N≥1,M与N相同或不同;样品预处理区和SERS检测区折叠后的每个瘦肉精测定单元为一一对应,且形状一致;每个瘦肉精测定单元检测的瘦肉精类物质相同或不同;样品预处理区具有对样品过滤、分离、富集及pH调节功能;SERS检测区的功能包括免疫反应发生、SERS增强纳米粒子引入和团聚剂加入等。
其中,所述高通量纸质微流控芯片的材料为定性滤纸、定量滤纸、打印纸或硝酸纤维素膜。
所述的瘦肉精测定单元在芯片上的位置、尺寸和形状可根据需要进行调节,可以根据需要改变形状(方形、菱形、三角形等)、尺寸大小及阵列范围。
本发明的高通量纸质微流控芯片可以同时完成多个瘦肉精样品的同时检测,实现瘦肉精类物质的即时现场检测,步骤简单快速。
附图说明
图1是一种用于瘦肉精检测的高通量微流控芯片平面示意图。
图2是一种用于瘦肉精检测的高通量微流控芯片折叠示意图。
图3是3种瘦肉精类物质混合后同时检测得到的SERS谱图。
图4是一种瘦肉精物质在不同pH条件下的SERS光谱图。
图5是富集不同次数后瘦肉精的SERS谱图。
图中:Ⅰ样品预处理区;ⅡSERS检测区;---代表折叠线。
具体实施方式
以下结合技术方案和附图具体说明本发明的具体实施方式。
实施例1
芯片设计如图1所示,所用材料为滤纸,采用喷蜡打印方法制作疏水性障碍,在纸上形成亲-疏水性区域。具体操作如下:实验中的三维纸芯片包括两层:Ⅰ层作为预处理区,实现过滤、分离、富集、pH调节功能;Ⅱ层作为SERS检测区。首先,我们在检测区分别加入3uL盐酸或氢氧化钠,提供瘦肉精类物质检测的pH环境(图4),然后我们对齐折叠纸芯片(图2),连通预处理区和检测区,然后从预处理区加入5uL猪毛提取液。此时,提取液中的猪毛颗粒沉积在预处理区,而样品渗透到检测区。当样品在检测区晾干后,重复上样,实现样品在检测区的富集,然后在检测区加入3uL金纳米粒子,同时进行SERS光谱的测定。值得一提的是,纸与纳米粒子的复合产生的等离子耦合可以猝灭干扰信号而增强分析物的信号。清晰地证明了随着上样次数的增加,瘦肉精的SERS信号逐渐增强。当上样次数达到累积时(图5),克伦特罗的检测限为ng/mL,可以满足实际检测要求。
实施例2
芯片设计如图1所示,所用材料为滤纸,采用喷蜡打印方法制作疏水性障碍,在纸上形成亲-疏水性区域。具体操作如下:实验中的三维纸芯片包括两层:一层作为预处理区(过滤、分离、富集、pH调节);另一层作为SERS检测区。首先,我们在检测区分别加入适量盐酸或氢氧化钠,提供瘦肉精类物质检测的pH环境,然后我们折叠纸芯片,连通预处理区和检测区,然后从预处理区加入人毛发提取液。此时,提取液中的头发颗粒沉积在过滤区,而样品渗透到检测区。当样品在检测区晾干后,重复上样,实现样品在检测区的富集,然后在检测区加入纳米粒子,同时进行SERS光谱的测定。上样次数的增加,生物标记物(人头发中的肺癌标记物)的SERS信号逐渐增强,检测限为10ng/mL。
Claims (2)
1.一种基于SERS检测瘦肉精类物质的高通量纸质微流控芯片,其特征在于,该高通量纸质微流控芯片包含两部分:样品预处理区和SERS检测区,样品预处理区和SERS检测区都有M×N个瘦肉精测定单元,沿样品预处理区和SERS检测区之间的折叠线对齐折叠微流控芯片,连通样品预处理区和SERS检测区后,样品预处理区的每个瘦肉精测定单元与SERS检测区的每个瘦肉精测定单元为一一对应,且形状一致;每个瘦肉精测定单元检测的瘦肉精类物质相同或不同;样品预处理区具有对样品过滤、分离、富集及pH调节功能;SERS检测区的功能包括免疫反应发生、SERS增强纳米粒子引入和团聚剂加入。
2.根据权利要求1所述的高通量纸质微流控芯片,其特征在于,所述高通量纸质微流控芯片的材料为定性滤纸、定量滤纸、打印纸或硝酸纤维素膜。
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