【作者】 于晓；

【导师】 罗勇；

【作者基本信息】 大连理工大学 ， 药物化学， 2015， 硕士

【摘要】 体内外源性小分子含量测定的传统方法有高效液相色谱法、气相色谱法、高效液相色谱-质谱联用法、酶联免疫法等。这些检测方法大都前处理过程复杂,仪器庞大,检测耗时耗力。微流控芯片技术作为一种新兴的检测平台,具有耗样量小、检测速度快、可以实现高通量检测等优势。本论文以盐酸克伦特罗和冰毒为案例,结合表面增强拉曼技术,开发了两种微流控芯片小分子检测平台,展示了微流控芯片在小分子检测方面的巨大应用潜力。(1)盐酸克伦特罗(Clenbuterol Hydrochloride)是一种广泛使用的违禁饲料添加剂,它可以增加肉制品的瘦肉含量,可是会对人体产生一系列的毒副作用。通过检测猪毛中是否有盐酸克伦特罗可以判断养殖场是否存在违规操作的情况。本文基于表面增强拉曼光谱技术(SERS).磁性纳米颗粒和微流控芯片平台发展了一种新型的盐酸克伦特罗免疫测定方法,检测限可至飞克量级。具体说来,就是猪毛提取液、修饰有盐酸克伦特罗抗体的磁性纳米颗粒和修饰有克伦特罗抗原和拉曼探针(对巯基苯甲酸)的金纳米粒子在微流控芯片主通道内充分流动混合,发生竞争性免疫反应。当反应混合物流至主通道下游时,在外加磁铁的作用下,磁纳米粒子-抗体-克伦特罗抗原-金纳米粒子-拉曼探针复合物被富集并产生拉曼信号,拉曼信号越强,猪毛提取液中的盐酸克伦特罗含量越低。整个实验完成需要1小时,具有很高的特异性和灵敏度,且可应用于猪毛提取液中的盐酸克伦特罗的定量测定。(2)甲基苯丙胺(Methamphetamine),俗称冰毒,是一种具有剧烈毒性的兴奋剂药物,公安部门常用的毒品检测手段有试纸条检测、高效液相色谱检测。上述检测可能会出现假阳性结果,检测耗时较长,对仪器以及操作人员要求较高等缺点。本文基于表面增强拉曼光谱技术和玻璃微流控芯片平台,开发了一种冰毒的快速检测方法。将20μL的100ng/mL的冰毒血浆样本、增强拉曼用纳米粒子、团聚剂在芯片通道内反应,3-5分钟后即可进行拉曼检测。这种方法不仅可以用于检测血浆样本,还可以用于检测尿液、唾液样本,对其它人体中小分子的检测也具有一定的意义。同时,当装置与便携式拉曼检测仪器搭配使用时,有望成为公安部门毒品检测的新手段。更多还原

【Abstract】 The traditional methods for the determination of small molecules in the body are high performance liquid chromatography, gas chromatography, high performance liquid chromatography-mass spectrometry. But the process of the pretreatment is always complex and time-consuming, the tedious, cumbersome equipment makes analysis only possible in the central lab. Microfluidic chip technology is a new platform on analysis. It needs less sample consumption and makes the detection faster and high-throughput. This study took the clenbuterol and methamphetamine as the examples, combined Surface-enhanced Raman Spectroscopy (SERS) to develop two kinds of microfluidic platforms on small molecules detection, showed the huge potential application microfluidic chip in the field of small molecules detection.Clenbuterol hydrochloride is an illegal but widely-used feed additive, it can increase the lean meat content, but will produce a series of side effects to the human body. Detecting the clenbuterol hydrochloride in swine hair is an effective supervision of irregularities. This study developed a competitive immunoassay to determine the clenbuterol hydrochloride in the swine hair extract at femtogram level, which integrated the power of magnetic nanoparticles, microfluidics and surface-enhanced Raman spectroscopy (SERS). In the main microchannel, magnetic nanoparticles on which the clenbuterol hydrochloride monoclonal antibodies were immobilized were mixed and reacted with the swine hair extract doped with gold nanoparticles labelled with Raman reporter (4-Mercaptobenzoic acid) and clenbuterol standard. The magnetic immunocomplex produced in the reaction was then immoblized and enriched in the downstream microchannel by a magnet for Raman detection. A washing microchannel was designed to remove the interference of fluorescent species in the swine hair extract. The immunoassay was accomplished in minutes due to chaotic mixing in the microchannel. The total analytical time was reduced to about 1 hour. This method allows on-site fast analysis of clenbuterol hydrochloride in the swine hair extract with high specificity and sensitivity.Methamphetamine, commonly known as crystal meth, is a kind of stimulant drugs with severe toxicity. The common analytical technique used by the public security departments are strip test and high performance liquid chromatography (HPLC). Strip tests may appear false positive results, so the high performance liquid chromatography usually comes after the strip tests for the accurate results. We developed a fast detection of methamphetamine in plasma samples based on Surface-enhanced Raman Spectroscopy (SERS) and glass microfluidic chip. The samples, salts and nanoparticles flew into the micro channel with the help of an injection pump at the outlet. Raman signals could be detected in only 3-5 minutes. This method not only can be used to detect the plasma samples, but also for the detection of urine and saliva samples. When the device is combined with the portable Raman instruments, it is expected to become a new detection method for public security department.更多还原